Contents
Pages
Contents……………………………………………..……………………………......….1
Executive Committee and Meeting Organizers………………….……………….......2
Welcome……………………………………………..………..……………….…..……..3
General Information…………………………………………....…………………......…4
Events Summary………………………………….………….…………………………..5
INRC 2019 Awardees…………………………….……….……………………...……..7
Program………….………………………………………….…………………………….9
Symposia Abstracts…………………………………………………………………….17
Poster Abstracts…………………………………………………………………...……42
Exhibitor and Sponsor Information……………………………………………..……133
Floor Plan……………………………………………..………….…………….….…...134
Participants………………………………………………………………..…………...135

1

Executive Committee and Meeting Organizers
INRC Officers
Larry Toll (President, USA)Fred Nyberg (Past-President, Sweden)
Lee Yuan Liu-Chen (Treasurer, USA)
Amynah Pradhan (Information Officer, USA)

INRC Executive Committee
Ream Al-Hasani (USA)Meritxell Canals (Australia)
Lucia Hipolito (Spain)
Emily Jutkiewicz (USA)
Eamon Kelly (UK)
Minoru Narita (Japan)
Stefan Schulz (Germany)
Shogo Tokuyama (Japan)
James Zadina (USA)INRC 2019 Local Organizing Committee
Eduardo Butelman
Lakshmi Devi
Sandra Comer
Lloyd Fricker
Mary Jeanne Kreek
Brian Reed
Larry Toll2

Welcome
50th International Narcotics Research Conference(INRC)

INRC 2019 Program Committee
Lakshmi Devi
Mary Jeanne Kreek
Laura Bohn
Eduardo Butelman
Meritxell Canals
Sandra Comer
Lloyd Fricker
Lucia Hipolito
Susan Ingram
Emily Jutkiewicz
Minoru Narita
Susruta Majumdar
Brian Reed
Vivianne Tawfik
Larry Toll

3

General Information
INRC Registration Desk and Message Center will be located at the Grand Hyatt
New York, Lobby level (109 East, 42nd Street at Grand Central Terminal)
The registration desk hours are:Sunday, July 7, 20194:00 PM – 9:00 PM

Monday, July 8, 20198:00 AM – 6:00 PM

Tuesday, July 9, 20198:30 AM – 6:00 PM

Wednesday, July 10, 20198:30 AM – 5:30 PM

Thursday, July 11, 20198:30 AM – 5:00 PM

Pick up your badge and program book at the INRC Registration Desk at the Manhattan Ballroom
Foyer of the Lobby Level of Grand Hyatt New York (109 East 42nd Street at Grand Central Terminal).

Scientific Presentations will be in the Manhattan Ball Room (see floor plan on page 134).
Poster Sessions will be at the Museum Space (see floor plan on page 134).
The list of poster titles and authors are on pages 42-51
Poster Session 1. Monday July 8th, 2019
Posters may be set up between 12:30 – 2:00 PM and should be removed by 7:00 PM. Presenterswill be at their posters from 4:30 – 6:00 PM on Monday.
Poster Session 2. Tuesday July 9th, 2019
Posters may be set up before 9:00 AM and should be removed by 7:00 PM. Presenters will be attheir posters from 4:30 – 6:00 PM on Tuesday.
Poster Session 3. Wednesday July 10th, 2019
Posters may be set up before 9:00 AM and should be removed by 3:00 PM. Presenters will be attheir posters from 12:30 – 2:00 PM on Wednesday.

Lunches will be served at the Gallery on Lex (see floor plan on page 134) from Monday throughWednesday and open to all delegates.

Banquet will be at the Empire Ballroom III; the pre-Banquet reception will be at the Empire
Ballroom Foyer (4th floor of Grand Hyatt New York).

4

Events Summary
Sunday July 7, 2019
Welcome Reception - 7:00 PM – 9:00 PM
Monday July 8, 2019
Welcome Address - 8:45 AM – 9:00 AM
Plenary Lecture - 9:00 AM – 10:00 AM
Mary Jeanne Kreek, Rockefeller University, NY, USA
Symposium 1 - 10:00 AM – 12:40 PM
Opioid Receptor Structure/Ultra Structure/Single Molecules
Lunch - 12:30 PM – 2:00 PM
Symposium 2 - 2:00 PM – 4:00 PM
Biased Signaling
Data Blitz 1 - 4:00 PM – 4:30 PM
Poster Session 1 - 4:30 PM – 6:00 PM
Tuesday July 9, 2019
Plenary Session - 9:00 AM – 10:00 AM
George Koob, National Institute on Alcohol Abuse and Alcoholism, NIH, USA
Nora Volkow, National Institute on Drug Abuse, NIH, USA
Symposium 3 - 10:00 AM – 12:40 PM
Albert Herz Memorial Symposium: Targeting Kappa Opioid Receptors
Lunch - 12:30 PM – 2:00 PM
Executive Committee meeting - 12:30 PM – 2:00 PM (by invitation only)
Symposium 4 - 2:00 PM – 4:00 PM
Gavril Pasternak Memorial Symposium: Chemical Biology of Diverse Opioids
Data Blitz 2 - 4:00 PM – 4:30 PM
Poster Session 2 - 4:30 PM – 6:00 PM

5

Wednesday July 10, 2019
Founders’ Lecture - 9:00 AM – 10:00 AM
Solomon H. Snyder, Johns Hopkins University, MD, USA
Symposium 5 - 10:00 AM – 12:30 PM
Circuits in Neuropathic Pain
Poster session - 12:30 PM – 2:00 PM
Lunch - 12:30 PM – 2:00 PM
INRC Founders’ Round Table Discussion - 2:00 PM – 3:00 PM
Career Development Session: Grant Writing Workshop - 3:30 PM – 5:00 PM

Thursday July 11, 2019Young Investigator Award Lecture - 9:00 AM – 10:00 AM
Emily Jutkiewicz, University of Michigan, MI, USA
Symposium 6 - 10:00 AM – 12:30 PM
Endogenous Opioid System: Progress and New Concepts
Lunch - 12:30 PM – 2:00 PM (On your own)
Symposium 7 - 2:00 PM – 4:00 PM
Atypical Opiates
Business Meeting: 4:00 PM – 5:00 PM
All attendees are encouraged to attend and elect new members of the executive
committee. Conference updates, future meeting locations and other businesswill be discussed.
Reception: 6:00 PM – 7:00 PM
Banquet and Dance: 7:00 PM – 10:00 PM
Banquet dinner ticket is in your registration packet.

6

INRC 2019 AWARDEE
Founders’ Lecture – Professor Solomon Snyder
Dr. Solomon H. Snyder is the Distinguished Service
Professor of Neuroscience, Pharmacology and Psychiatryat the Johns Hopkins University School of Medicine. He is
the founder of the Solomon H. Snyder Department of
Neuroscience and served as its director from 1980 to 2006.
Born in Washington, D.C. on December 26, 1938,
Dr. Snyder received his undergraduate and medical
training at Georgetown University (MD in 1962); Research
Associate training with Julius Axelrod at the NIH (19631965); and psychiatric training at the Johns Hopkins
Hospital (1965-1968). In 1966 he joined the faculty of theJohns Hopkins University School of Medicine (AssistantProfessor Pharmacology, 1966-1968; Associate Professor Pharmacology/Psychiatry (19681970); Professor (1970)). In 1980 he established the Department of Neuroscience andserved as Director (1980-2006). He is presently Distinguished Service Professor of
Neuroscience, Pharmacology and Psychiatry.
Dr. Snyder is the recipient of numerous professional honors, including the Albert
Lasker Award for Basic Biomedical Research (1978), the National Medal of Science (2005),
the Albany Medical Prize (2007), the Warren Alpert Prize, Harvard University Prize (2014);
Honorary Doctor of Science degrees from Northwestern University (1981), Georgetown
University (1986), Ben Gurion University (1990), Albany Medical College (1998), Technion
University of Israel (2002), Mount Sinai Medical School (2004), University of Maryland(2006), Charles University, Prague (2009), Johns Hopkins University (2018); the Wolf
Foundation Prize in Medicine (1983), the Dickson Prize of the University of Pittsburgh(1983), the Bower Award of the Franklin Institute (1991), the Bristol-Myers Squibb Award forDistinguished Achievement in Neuroscience Research (1996), the Gerard Prize of theSociety for Neuroscience (2000) and the Salk Institute Science Award (2016). He is amember of the United States National Academy of Sciences and a Fellow of the American
Academy of Arts and Sciences and the American Philosophical Society. He is the author of
more than 1000 journal articles and several books including Uses of Marijuana (1971),
Madness and the Brain (1974), The Troubled Mind (1976), Drugs and the Brain (1986), andBrainstorming (1989).
Many advances in molecular neuroscience have stemmed from Dr. Snyder's
identification of receptors for neurotransmitters and drugs, and elucidation of the actions of
psychotropic agents, making him one of the most highly cited biologists in the world. His
1973 Science publication describing the opiate receptor was a major advance in the field.

7

INRC 2019 AWARDEE
Young Investigator Award – Dr. Emily Jutkiewicz
Dr. Emily Jutkiewicz is currently an Associate Professor
with tenure in the Department of Pharmacology at the
University of Michigan Medical School. She attended Tufts
University in Medford, Massachusetts for her undergraduate
studies and graduated cum laude with a B.S. inbiopsychology. She worked for a couple of years as a
research technician at McLean Hospital before starting
graduate school in the Department of Pharmacology at the
University of Michigan in the laboratory of Dr. James H
Woods. She received her PhD in 2004 with a thesis entitled“Antidepressant effects of the nonpeptidic delta-opioid
receptor agonist SNC80.” She completed postdoctoral fellowships at the University ofMichigan in the mass spectrometry core facility and in the laboratories of Drs. Margaret
Gnegy and Jim Woods. In 2009, she became a research investigator in the laboratory of Dr.
John Traynor and then became a tenure-track faculty in 2013. Dr. Jutkiewicz’s research has
focused on the behavioral effects of opioids and other drugs of abuse, such as nicotine andamphetamine. She identified that delta opioid receptor agonists produce antidepressant-like
effects in animal models and determined that these effects were independent of their seizure
activity. Her current research aims to elucidate further the role of opioid systems, in particular
the delta opioid receptor, in disease states. More specifically, her laboratory investigates thein vivo effects of delta opioid and multifunctional opioid receptor ligands and the cellularmechanisms underlying the in vivo effects of delta opioid agonists. Her lab is also interested
in identifying novel treatments for addiction.
Thus far, Dr. Jutkiewicz has had a productive career. She has published 58 peerreviewed journal articles and edited a volume of Handbook of Experimental Pharmacology
on Delta Opioid Receptor Pharmacology and Therapeutic Applications (to which many INRC
members contributed). In addition, she is a co-applicant on two patents and currently serveson the Editorial Boards for the journals Psychopharmacology, and Experimental and ClinicalPsychopharmacology. She has also received grant awards from NIDA, NIMH, and the
PhRMA Foundation. Dr. Jutkiewicz has been awarded several honors, including CPDD Early
Career Investigator Award, Basic Sciences Teaching Award in Pharmacology, and theJacoby Lectureship at Ferris State University College of Pharmacy. In addition, her trainees
have won a number of awards within the Department of Pharmacology, at the annual
meeting of the American Society of Pharmacology and Experimental Therapeutics (ASPET),
and at the Behavior, Biology, and Chemistry (BBC) Conference. She has attended INRC
meetings consistently since 2009 and currently serves on the INRC Executive Committee.

8Program
Sunday, July 7, 20194:00 PM – 9:00 PM
Registration

Manhattan Ball Room Foyer

7:00 PM – 9:00 PM
Welcome Reception

Manhattan Ball Room Foyer and Museum Space

Monday, July 8, 20198:45 AM – 9:00 AM
Welcome Address
9:00 AM – 10:00 AM
Plenary Lecture
Mary Jeanne Kreek, Rockefeller University, USA
50 Years of INRC: 1969 to 2019 - 55 Years of our Rockefeller University Researchand 50 to 60 Years of Opioid Research

10:00 AM – 12:40 PM
Symposium 1 - Opioid Receptor Structure/Ultra Structure/Single
Molecules
Chairs: Meritxell Canals, University of Nottingham, UK
Mark von Zastrow, University of California at San Francisco, USA
10:00 AM – 10:30 AM - Damien Jullié, University of California at San Francisco, USA
A discrete presynaptic vesicle cycle for neuromodulator receptors

910:30 AM – 11:00 AM - Vsevolod Katritch, University of Southern California, USA
High resolution 3D structures are now available for opioid receptors
11:00 AM – 11:30 AM - Coffee Break
11:30 AM – 11:50 AM - Seksiri Arttamangkul, Oregon Health & Science University, USA
Visualizing endogenous opioid receptors in living neurons using ligand-directedchemistry
11:50 AM – 12:10 PM - Marta Filizola, Icahn School of Medicine at Mount Sinai, USA
Understanding µ-opioid receptor activation and allosteric modulation via molecular
dynamics simulations for the design of safer analgesics
12:10 PM – 12:30 PM - Aashish Manglik, University of California at San Francisco, USA
Traversing chemical space to understand opioid function
12:30 PM – 12:40 PM - Hot Topic Presentation - Alexander Gillis, Univ. of Sydney, Australia
Low intrinsic G-protein efficacy, not reduced ß-arrestin 2 signaling, as a mechanism
for improved opioid analgesics

12:30 PM – 2:00 PM
Lunch
2:00 PM – 4:00 PM
Symposium 2 - Biased Signaling
Chairs: Laura Bohn, Scripps Institute, USA
Chris Bailey, University of Bath, UK
2:00 PM – 2:20 PM - Laura Bohn, Scripps Institute, USA
Differential adaptations to chronic biased opioid administration in different neuronal
populations
2:20 PM – 2:40 PM - Andrea Kliewer, Jena University, Germany
Phosphorylation-deficient G-protein-biased µ-opioid receptors improve analgesia anddiminish tolerance but worsen opioid side effects
2:40 PM – 3:00 PM - Lee-Yuan Liu-Chen, Temple University, USA
Phosphoproteomic approach for agonist-specific signaling in mouse brains: mTOR
pathway is involved in kappa opioid aversion
3:00 PM – 3:20 PM - Venetia Zachariou, Mount Sinai School of Medicine, USA
A “complex” role of RGS proteins in responses to opioid analgesics
3:20 PM – 3:40 PM - Michelle Mazei-Robison, Michigan State University, USA
Molecular and cellular adaptations of VTA dopamine neurons induced by morphine10

3:40 PM – 4:00 PM - Yavin Shaham, National Institute on Drug Abuse, USA
Effect of chronic buprenorphine and the biased mu-opioid receptor agonist TRV130
on relapse to oxycodone seeking in a rat model of opioid maintenance therapy

4:00 PM – 4:30 PM
Data Blitz
Organizers: Meritxell Canals, University of Nottingham, UK
Lucia Hipolito, University of Valencia, Spain

4:30 PM – 6:00 PM
Poster Session 1 (Poster number M1 – M52)

Tuesday, July 9th, 20199:00 AM – 10:00 AM
Plenary Session
George Koob, National Institute on Alcohol Abuse and Alcoholism, NIH, USA
Alcohol and drug addiction: The gain the brain is in the pain
Nora Volkow, National Institute on Drug Abuse, NIH, USA
Scientific priorities to address the opioid crisis

10:00 AM – 12:40 PM
Symposium 3 - Albert Herz Memorial Symposium: Targeting Kappa
Opioid Receptors
Chairs: Eduardo Butelman, Rockefeller University, USA
Thomas Prisinzano, University of Kentucky, USA
10:00 AM – 10:15 AM - Fred Nyberg, Uppsala University, Sweden
Tribute in memory of Albert Herz
10:15 AM – 10:40 AM - Michael Bruchas, University of Washington, USA
Dissecting nucleus accumbens opioid circuits in reward seeking1110:40 AM – 11:00 AM - Sara Jones, Wake Forest University, USA
Kappa opioid receptors drive dopamine deficiency and dysphoria during withdrawal
from alcohol, cocaine and heroin
11:00 AM – 11:30 AM - Coffee Break
11:30 AM – 11:50 AM - Brian Reed, Rockefeller University, USA
In vivo effects of kappa agonists with differential in vitro efficacy
11:50 AM – 12:10 PM - Thomas Prisinzano, University of Kentucky, USA
Design and synthesis of natural product based kappa opioid receptor ligands
12:10 PM – 12:30 PM - Pierre-Eric Lutz, McGill University, Canada
Epigenetic programming of the kappa opioid receptor by child abuse: a post-mortem
brain study
12:30 PM – 12:40 PM - Hot Topic Presentation - Andrea Bedini, University of Bologna, Italy
Functional selectivity and analgesic effects of CL39, a novel kappa opioid receptor(KOR) selective agonist

12:30 PM – 2:00 PM
Lunch
2:00 PM – 4:00 PM
Symposium 4 - Gavril Pasternak Memorial Symposium: Chemical
Biology of Diverse Opioids
Chairs: Susruta Majumdar, Washington University School of Medicine, USA
Nurulain Zaveri, Astraea Therapeutics, LLC, USA
2:00 PM – 2:15 PM - Ying-Xian Pan, Memorial Sloan Kettering, USA
Tribute to Gavril Pasternak
2:15 PM – 2:40 PM - Susruta Majumdar, Washington University School of Medicine, USA
Structure based design and synthesis of diverse opioids
2:40 PM – 3:05 PM - Nurulain Zaveri, Astraea Therapeutics, LLC, USA
Bifunctional NOP/MOP agonists as a new approach for non-addicting analgesics andtreatments for opioid use disorders
3:05 PM – 3:25 PM - Ream Al-Hasani, Washington University School of Medicine, USA
Determining the neurochemical profile of endogenous opioids during withdrawal
3:25 PM – 3:40 PM - Saheem Zaidi, The Bridge Institute, USC Dornsife, USA
Designing bitopic opioids: Activity modulation in targeting sodium binding pocket ofthe opioid receptors
12

3:40 PM – 4:00 PM - James Zadina, Tulane University School of Medicine, USA
A mu opioid with substantially reduced side-effects shortens recovery from chronic
pain relative to morphine

4:00 PM – 4:30 PM
Data Blitz
Organizers: Meritxell Canals, University of Nottingham, UK
Lucia Hipolito, University of Valencia, Spain

4:30 PM – 6:00 PM
Poster Session 2 (Poster number T1 – T52)

Wednesday, July 10th, 20199:00 AM – 10:00 AM
Founders’ Lecture
Solomon H. Snyder, Johns Hopkins University, MD, USA
Messenger molecules of the mind

10:00 AM – 12:30 PM
Symposium 5 - Circuits in Neuropathic Pain
Chairs: Vivianne Tawfik, Stanford University, USA
Minoru Narita, Hoshi University, Japan
10:00 AM – 10:20 AM - Vivianne Tawfik, Stanford University, USA
Glial cells in opioid tolerance and chronic pain
10:20 AM – 10:40 AM - Minoru Narita, Hoshi University, Japan
Influence of pain-related neurocircuits on tumor immune response
10:40 AM – 11:00 AM - Gregory Corder, University of Pennsylvania, USA
Pain & the amygdala
11:00 AM – 11:30 AM - Coffee Break13

11:30 AM – 11:50 AM - Ru-Rong Ji, Duke University Medical Center, USA
Distinct roles of b-arrestin-2 in regulating opioid analgesia, opioid hyperalgesia, andneuropathic pain
11:50 AM – 12:10 PM - Isaac Chiu, Harvard Medical School, USA
Pain, neuro-immune interactions and bacterial infections
12:10 PM – 12:30 PM - Anna Taylor, University of Alberta, Canada
The gut microbiome contributes to neuroinflammation, pain and affect during opioid
withdrawal

12:30 PM – 2:00 PM
Lunch
Poster Session 3 (Poster number W1 – W52)
2:00 PM – 3:00 PM
INRC Founders’ Round Table Discussion
Moderators: Charles Chavkin, University of Washington, USA
Larry Toll, Florida Atlantic University, USA

3:00 PM – 5:00 PM
Career Development Session: Grant Writing Workshop
Organizers: Susan Ingram, Oregon Health & Sciences University, USA
Roger Sorensen, National Institute on Drug Abuse, USA

Thursday, July 11th, 20199:00 AM – 10:00 AM
Young Investigator Award Lecture
Emily Jutkiewicz, University of Michigan, USA
Deciphering the function of delta-opioid receptors

1410:00 AM – 12:30 PM
Symposium 6 - Endogenous Opioid System: Progress and New
Concepts
Chair: Emily Jutkiewicz
10:00 AM – 10:20 AM - Elena Bagley, University of Sydney, Australia
Drug dependence dials down the effects of endogenous opioids
10:20 AM – 10:40 AM - Catherine Cahill, University of California Los Angeles, USA
Kappa opioid modulation of negative affect and pain aversion associated with ongoing
chronic pain
10:40 AM – 11:00 AM - Hervé Poras, Pharmaleads SA, France
DENKI (Dual ENKephalinase Inhibitors), a new class of painkillers devoid of abuse potential
and of opiate side-effects
11:00 AM – 11:30 AM - Coffee Break
11:30 AM – 11:50 AM - Leslie Sombers, North Carolina State University, USA
Concurrent voltammetric detection of catecholamine and met-enkephalin dynamics in rat
striatum and adrenal tissue
11:50 AM – 12:10 PM - David Hsu, Stony Brook University, USA
Endogenous opioid regulation of emotions in the social environment
12:10 PM – 12:20 PM – Hot Topic Presentation – Hiroshi Ueda, Kyoto University, Japan
Chronic pain status and loss of morphine analgesia have independent brain
mechanisms in mice with fibromyalgia-like generalized pain
12:20 PM – 12:30 PM – Hot Topic Presentation – Kabirullah Lutfy, Western University of
Health Sciences, USA
The role of endogenous opioids in amphetamine-induced locomotor sensitization

12:30 PM – 2:00 PM
Lunch (on your own)
2:00 PM – 4:00 PM
Symposium 7 - Atypical Opiates
Chairs: Sandra Comer, Columbia University, USA
Ellen Walker, Temple University, USA
2:00 PM – 2:20 PM - Gretchen Snyder, Intra-cellular Therapies, USA
A new approach to the treatment of opioid use disorder15

2:20 PM – 2:40 PM - Jonathan Javitch, Columbia University, USA
The atypical antidepressant tianeptine is an efficacious mu opioid receptor agonist
2:40 PM – 3:00 PM - Joseph Stauffer, Nektar Therapeutics, USA
A novel pegylated opioid in development for the treatment of chronic pain
3:00 PM – 3:20 PM - Jay McLaughlin, University of Florida, USA
Evaluation of kratom and mitragynine as safer treatments for pain and opioid
dependence
3:20 PM – 3:40 PM - Ellen Walker, Temple University, USA
Preclinical assessment of novel psychoactive substances: Opioids
3:40 PM – 4:00 PM - S. Stevens Negus, Virginia Commonwealth University, USA
Learning from failure: Kappa agonists as negative controls in analgesic drug
development

4:00 PM – 5:00 PM
Business Meeting

Manhattan Ballroom

6:00 PM – 7:00 PM
Reception

Empire Ballroom Foyer

7:00 PM – 10:00 PM
BanquetEmpire Ballroom III

16

Symposium Abstracts
Monday July 8, 201910:00 AM – 12:40 PM
Symposium 1 - Opioid Receptor Structure/Ultra Structure/Single Molecules
Chairs: Meritxell Canals, University of Nottingham, UK
Mark von Zastrow, University of California at San Francisco, USA
Presenter – Jullié, Damien
A discrete presynaptic vesicle cycle for neuromodulator receptors
Damien Jullié1,2, Miriam Stoeber1,2, Eric Hosy3,4, Jean-Baptiste Sibarita3,4, Thomas M. Bartol5, Seksiri
Arttamangkul6, John T. Williams6, Terrence J. Sejnowski5, Mark von Zastrow1,2*
1
Department of Cellular and Molecular Pharmacology and 2Department of Psychiatry, University ofCalifornia at San Francisco, USA, 3Interdisciplinary Institute for Neuroscience, UMR, Centre National dela Recherche Scientifique, Bordeaux, France, 4Interdisciplinary Institute for Neuroscience, University of
Bordeaux, Bordeaux, France, 5Howard Hughes Medical Institute, Salk Institute for Biological Studies, La
Jolla, USA, 6The Vollum Institute, Oregon Health and Science University, Oregon, USA
A major function of GPCRs is to inhibit neurotransmitter release from presynaptic terminals but little is
known about the cell biology of GPCRs at the presynapse. Presynaptic inhibition can achieve andmaintain high ligand sensitivity despite requiring receptors to mediate allosteric coupling to effectors
locally at terminals or active zones therein, supporting the previously established paradigm of GPCR
immobilization or pre-coupling at terminals. We identify a fundamentally different cell biology bydemonstrating that opioid receptors are laterally mobile throughout the axonal plasma membrane andinternalize after ligand-induced activation specifically at terminals. We delineate a discrete regulated
endocytic cycle that operates at the presynapse, separate from the synaptic vesicle cycle, assuring rapid
clearance of activated receptors from terminals. We show that, when such local regulation occurs, GPCR
diffusion provides a robust, alternate strategy for local presynaptic ligand detection with performance thatis inherently superior to receptor immobilization or pre-coupling.
Presenter – Katritch, Vsevolod
High resolution 3D structures are now available for opioid receptors
Vsevolod Katritch1, Saheem A. Zaidi1, Nilkanth Patel1, Tao Che2, Xi-Ping Huang2,3, Bryan L. Roth2,3,4,
Susruta Majumdar5
1
Departments of Biological Sciences and Chemistry, Bridge Institute, University of Southern California,
Los Angeles, California, USA, 2Department of Pharmacology, 3Division of Chemical Biology andMedicinal Chemistry, 4National Institute of Mental Health Psychoactive Drug Screening Program,
University of North Carolina, Chapel Hill, USA, 5Center for Clinical Pharmacology, St. Louis College of
Pharmacy Washington University School of Medicine St. Louis, USA.
High resolution 3D structures are now available for opioid receptors in both active and inactive-states.
Combined with molecular modeling, this structural information provides the key insight into molecularinteractions of small molecule and peptide agonists and antagonists, while establishing a platform forunderstanding functional mechanisms of the receptors, including allosteric modulation and biased
signaling. Moreover, it presents a unique opportunity for in silico discovery of new lead chemotypes andnew probes with desired subtype selectivity and functional profiles. One of the recent examples of such
structure-guided design involves screening for derivatives with dual MOR/KOR G-protein biased profile,where the resulting compounds show effective analgesia with reduced side effects and tolerance in micemodels. Other structure-based designs include bitopic ligands that combine orthosteric moieties withextensions targeting conserved sodium allosteric pocket deep in the 7TM bundle of the receptor,
covalently bound selective antagonists, and photoswitchable ligands that allow control of the receptor
activation by light. Supported by P01 DA035764 and R21/R33 DA038858.
17

Presenter – Arttamangkul, Seksiri
Visualizing endogenous opioid receptors in living neurons using ligand-directed chemistry
Seksiri Arttamangkul1, Andrew Plazek1, Emily Platt1, Haihong Jin1, William Birdsong1, Thomas
Murray2, Kenner Rice3, David Farrens1, John Williams1
1
Oregon Health Science University, USA, 2 Creighton University, USA, 3 National Institute on Alcohol
Abuse and Alcoholism, NIH, USA
Identifying single neurons that have functional opioid receptors for cellular, synaptic and system
actions of opioids is a key to advance the understanding of the opioid system. This study used achemical approach termed “traceless affinity labeling” to develop a probe to fluorescently label opioid
receptors. Naltrexamine, an opioid antagonist, was linked to a reactive acylimidazole and fluorescentdye. Naltrexamine-guided the binding such that the acylimidazole could react with the receptor thus
attaching the fluorescent dye to the receptor and at the same time free naltrexamine. The reaction
was selective and labeled opioid receptors in multiple areas from mouse and rat brains. Thismethod allows visualization of living opioid-sensitive neurons in heterogeneous populations of thebrain without loss of function. The ability to locate endogenously expressed receptors in living tissues
will aide considerably in establishing the distribution and physiological role of opioid receptors in the
CNS of wild type animals.
Presenter – Filizola, Marta
Understanding µ-opioid receptor activation and allosteric modulation via molecular dynamics
simulations for the design of safer analgesics
Marta Filizola, PhD1
1
Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York,USA
Understanding the molecular mechanisms underlying the diverse µ-opioid receptor (MOR) signaling
pathways at the atomistic level is essential for discovering analgesics with reduced side effects.
Although recent high-resolution experimental structures of MOR provide important insights into thisunderstanding, they are limited to a few conformational states stabilized by specific ligands boundat the orthosteric biding site on the receptor. To provide an understanding of the MOR allostericmodulation by metal ions, positive allosteric modulators, and balanced or functionally selective MOR
ligands, we use enhanced molecular dynamics simulations combined with Markov state models,
information theory, and/or machine learning analyses. I will provide an overview of recent studies wehave carried out in the lab to investigate how to fine tune opioid receptor signaling towards thedesired therapeutic pathways and away from those regulating adverse effects, with the ultimate goalof using this information to design improved therapeutics.

18

Presenter – Manglik, Aashish
Traversing chemical space to understand opioid function
Aashish Manglik1
1
Departments of Pharmaceutical Chemistry and Anesthesia, University of California, USA
Despite two centuries of medicinal chemistry since the isolation of morphine from the opium poppy,
the ideal opioid analgesic devoid of key liabilities like addiction and respiratory depression remains
elusive. Recently determined structures of opioid receptors have provided atomistic insights into themolecular mechanisms that drive opioid receptor activation. Though highly informative, thesebeautiful structures conceal the remarkable conformational plasticity that drives opioid receptor
signaling complexity. We are developing new tools to interrogate the atomic basis of opioid receptor
regulation and conformational plasticity, with an eye towards a complete description of the physicalbasis of ligand-specific opioid receptor signaling. Our hope is to combine this insight with new virtual
screens employing more chemical diversity than has ever been tested at these important receptors.
Traversing such chemical space may enable the identification of completely new opioidergic
molecules with unprecedented biological effects.
Hot Topic Presenter – Gillis, Alexander
Low intrinsic G-protein efficacy, not reduced ß-arrestin 2 signaling, as a mechanism for
improved opioid analgesics
Alexander Gillis1, Claudia Alamein1, Preeti Manandhar2, Arisbel Gondin3, Andrea Kliewer4, Stefan
Schulz4, Meritxell Canals3,5, Mac Christie1
1
University of Sydney, Australia, 2Macquarie University, Australia, 3Monash University, Australia,
4Friedrich Schiller University, Jena, Germany, 5University of Nottingham, UKß-arrestin 2 dependent signaling via the µ-opioid receptor (MOPr) has been proposed to mediate
opioid analgesic side effects. We failed to observe an effect of either ß-arrestin2 knock-out or 11S/TA phosphomutant MOPr knock-in on morphine respiratory depression in mice, challenging this‘arrestin hypothesis’. We re-examined relative opioid efficacy using irreversible antagonism in amembrane-potential dye assay of G-protein activated potassium channels. Buprenorphine andfentanyl have extremely low and extremely high efficacy respectively. Putatively G-protein biased
MOPr agonists SR17018, PZM21 and oliceridine (TRV130) that do not strongly induce MOPr-ßarrestin2 interactions were all observed to have low intrinsic efficacy compared to morphine, itself apartial agonist relative to fentanyl.
Selected opioids were profiled in a hot-plate assay of anti-nociception and two respiratory side-effectassays in C57BL/6 mice. Contrary to published results, both SR17018 and PZM21 induced
significant respiratory depression in a whole-body plethysmography assay. SR17018, PZM21 andoliceridine were found to have therapeutic windows slightly superior to morphine. Buprenorphine wasobserved to have an extremely wide therapeutic window with a ‘ceiling’ effect on respiratory
depression, whereas fentanyl had a narrow window. Fentanyl caused substantial blood oxygen
desaturation compared to morphine as assessed using conscious pulse oximetry, whereas
oliceridine had minimal effect at equi-active antinociceptive doses. Therapeutic window between
anti-nociception and respiratory depression for the agonists studied was robustly predicted bydifferences in intrinsic G-protein efficacy. Low intrinsic efficacy is a plausible alternative mechanism
for the improved pre-clinical profile of putatively G-protein biased MOPr agonists that aligns withexisting opioid analgesics.

19

Monday July 8, 20192:00 PM – 4:00 PM
Symposium 2 - Biased Signaling
Chairs: Laura Bohn, Scripps Institute, USA
Chris Bailey, University of Bath, UK
Presenter – Bohn, Laura
Differential adaptations to chronic biased opioid administration in different neuronal
populations
Laura M. Bohn, PhD1
1
The Scripps Research Institute, Jupiter, USA.
It has been proposed that “biased agonists” at opioid receptors may serve as a means to refine
opioid therapeutics in a manner to eliminate certain side effects. Opioid receptors are expressed
throughout the body, and throughout the brain. In the brain, opioid receptors reside in different
neuronal populations and their signaling and regulation may differ. The purpose of this session is toprovide understanding as to how these drug targets may be functioning in vivo. If we understandhow the receptor functions in its endogenous environment and how such signaling relates tophysiological function, then we can begin to understand what signaling “bias” we wish to refine. The
selection of speakers will focus on how we are understanding diverse opioid receptor function in
vivo.
Presenter – Kliewer, A.
Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia anddiminish tolerance but worsen opioid side effects
A. Kliewer1, F. Schmiedel1, S. Sianati2, A. Bailey3, J. T. Bateman4, E. S. Levitt4, J. T. Williams5, M. J.
Christie2 and S. Schulz1*
1
Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich-Schiller-University,
Germany, 2Discipline of Pharmacology, School of Medical Sciences, University of Sydney, NSW,
Australia, 3Institute of Medical and Biomedical Education, St George’s University of London,
London , UK, 4Department of Pharmacology and Therapeutics, University of Florida, Gainesville,
USA, 5The Vollum Institute, Oregon Health and Science University, Portland, USA.
Opioid analgesics are powerful pain relievers; however, over time, pain control diminishes as
analgesic tolerance develops. The molecular mechanisms initiating tolerance have remainedunresolved to date. We have previously shown that desensitization of the μ-opioid receptor andinteraction with β-arrestins is controlled by carboxyl-terminal phosphorylation. Here we createdknockin mice with a series of serine- and threonine-to-alanine mutations that render the receptor
increasingly unable to recruit β-arrestins. Desensitization is inhibited in locus coeruleus neurons ofmutant mice. Opioid-induced analgesia is strongly enhanced and analgesic tolerance is greatly
diminished. Surprisingly, respiratory depression, constipation, and opioid withdrawal signs areunchanged or exacerbated, indicating that β-arrestin recruitment does not contribute to the severityof opioid side effects and, hence, predicting that G-protein-biased µ-agonists are still likely to elicit
severe adverse effects. In conclusion, our findings identify carboxyl-terminal multisite
phosphorylation as key step that drives acute μ-opioid receptor desensitization and long-term
tolerance.

20

Presenter – Liu-Chen, Lee-Yuan
Phosphoproteomic approach for agonist-specific signaling in mouse brains: mTOR pathway is
involved in kappa opioid aversion
Lee-Yuan Liu-Chen1, Jeffrey J. Liu2#, Yi-Ting Chiu1#, Kelly M. DiMattio1#, Chongguang Chen1#, Peng
Huang1, Taylor A. Gentile1, John W. Muschamp1, Alan Cowan1, and Matthias Mann2#
These authors contributed equally 1Temple University Lewis Katz School of Medicine, Philadelphia,
USA, 2Max Planck Institute of Biochemistry, Martinsried, Germany
Kappa opioid receptor (KOR) agonists produce analgesic and anti-pruritic effects, but their clinicalapplication was limited by dysphoria and hallucinations. Nalfurafine, a clinically used KOR agonist, does
not cause dysphoria or hallucinations at therapeutic doses in humans. We found that in CD-1 mice
nalfurafine produced analgesic and anti-scratch effects dose-dependently, like the prototypic KOR
agonist U50,488H. In contrast, unlike U50,488H, nalfurafine caused no aversion, anhedonia, sedation or
motor incoordination at the effective analgesia and anti-scratch doses. Thus, we established a mousemodel that recapitulated important aspects of the clinical observations. We then employed aphosphoproteomics approach to investigate mechanisms underlying differential KOR-mediated effects.
A large-scale mass spectrometry (MS)-based analysis on brains revealed that nalfurafine perturbed
phosphoproteomes differently from U50,488H in a brain-region specific manner after 30-minutetreatment. In particular, U50,488H and nalfurafine imparted phosphorylation changes to proteins found indifferent cellular components or signaling pathways in different brain regions. Notably, we observed that
U50,488H, but not nalfurafine, activated the mammalian target of rapamycin (mTOR) pathway in the
striatum and cortex. Inhibition of the mTOR pathway by rapamycin abolished U50,488H-induced
aversion, without affecting analgesic, anti-scratch, and sedative effects and motor incoordination. The
results indicate that the mTOR pathway is involved in KOR agonist-induced aversion, but not othereffects. This is among the first demonstrations that phosphoproteomics can be applied to agonist-specificsignaling of G protein-coupled receptors (GPCRs) in mouse brains to unravel pharmacologicallyimportant pathways. Furthermore, this is one of the first two reports that the mTOR pathway mediates
aversion caused by KOR activation. Supported by R01 DA041359 from NIDA.
Presenter – Zachariou, Venetia
A “complex” role of RGS proteins in responses to opioid analgesics
Venetia Zachariou1
1
Nash Family Department of Neuroscience, Icahn School of Medicine at Mount Sinai, USA
Regulators of G protein signaling (RGS) proteins are multifunctional GPCR signaling modulators, withdynamic roles in physiology and disease. RGS proteins control functional responses of GPCRs by
multiple mechanisms: they may bind to activated G alpha subunits to accelerate their GTPase activity or
to prevent activation of their effectors; they may also bind directly to GPCRs, act as kinetic scaffolds andthey may control transcriptional and epigenetic processes. We have demonstrated that RGS proteins
exhibit brain region and agonist-specific roles in behavioral responses to opioid analgesics. The striatal
RGS9-2 modulates the rewarding and analgesic efficacy of opioids in an agonist-dependent manner.
RGS4 is a smaller RGS protein distributed throughout the pain matrix which does not affect the analgesic
efficacy of morphine, but it acts as a positive modulator of fentanyl, oxycodone and methadone. Our latest
studies reveal that RGSz1, a small RGS protein that is upregulated in the mouse periaqueductal gray
(PAG) upon repeated exposure to opioid analgesics, modulates in the opposite manner analgesia versus
reward. Specifically, genetic inactivation of RGSz1 enhances the analgesic efficacy of synthetic opioids
and delays the development of tolerance, while decreasing sensitivity to the rewarding actions ofmorphine. Using RNA Sequencing, bioinformatic analysis and biochemical assays we identified a novel
mechanism by which RGSz1 in the mouse PAG promotes the development of analgesic tolerance.
RGSz1 competes with Axin2 (Wnt\beta-catenin pathway component and member of the RGS family) for
binding to activated Gaz subunits. Multiprotein complexes containing Axin2 control the nuclear
translocation and transcriptional activity of beta-catenin, which in turn promotes the expression of genesthat oppose the development of analgesic tolerance. Several RGS proteins are expressed throughout
the nociceptive pathway, suggesting that in addition to analgesia they may also modulate the processingof nociceptive information.
21

Presenter – Mazei-Robinson, Michelle
Molecular and cellular adaptations of VTA dopamine neurons induced by morphine
Sarah Cooper1, Amber Garrison1, Marie Doyle1, Qiwen Hu3, Elizabeth Heller3, Michelle Mazei-Robison1,2.
1
Neuroscience Program, Michigan State University, East Lansing, USA, 2Department of Physiology,
Michigan State University, East Lansing, USA, 3Department of Pharmacology, University ofPennsylvania, USA
Despite the prevalent use and abuse of opioid drugs, much remains unknown about the neuroadaptations
that occur with chronic use. We previously determined that opiate exposure decreases the soma size of
dopamine (DA) neurons in the ventral tegmental area (VTA), an effect that appears to be drug-classspecific, suggesting that distinct molecular changes drive opioid-induced plasticity in the VTA.
Consistently, in an RNA sequencing (RNAseq) screen, we identified different patterns of gene expression
changes in the VTA following morphine vs. cocaine treatment. However, this study was limited tohomogenization of the entire VTA, potentially obscuring changes that occur specifically in VTA DA
neurons. We sought to address this knowledge gap through the use of using Translating Ribosome
Affinity Purification (TRAP), completing RNAseq analysis on VTA samples from sham- and morphinetreated DATEGFP-L10a mice. We first confirmed that our samples were enriched for DA transcripts anddepleted of GABA, glutamatergic and glial transcripts. Excitingly, while we identified a similar number ofdifferentially expressed genes in the input and IP fractions, there was little overlap between the two
samples, suggesting that morphine-induced changes in gene expression specific to VTA DA neurons are
not evident in whole VTA analyses. We have now validated by RT-PCR a number of novel targets
including genes involved in transcriptional regulation, cytoskeletal remodeling, neuronal activity, andneuropeptide signaling. We are currently interrogating the functional impact of a subset of these novel
gene targets, as well as defining their expression pattern within the VTA.
Presenter – Shaham, Yavin
Effect of chronic buprenorphine and the biased mu-opioid receptor agonist TRV130 on relapse tooxycodone seeking in a rat model of opioid maintenance therapy
Jennifer M. Bossert1, Eugene Kyatkin1, Hannah Korah1, Jennifer K. Hoots1, Ida Fredriksson1, Bruce E.
Blough2, and Yavin Shaham1
1
Behavioral Neuroscience Branch, IRP/NIDA/NIH, Baltimore, USA, 2Center for Drug Discovery, Research
Triangle Institute, Research Triangle Park, USA
High relapse rate is a major obstacle in addressing the current opioid crisis. Opioid agonist maintenancetherapy (buprenorphine, methadone) is an effective treatment for opioid addiction, but does not prevent
relapse in all opioid users. We modified the context-induced reinstatement model in rats trained to selfadminister the prescription opioid oxycodone and compared the efficacy of buprenorphine with that of thenovel opioid mu opioid receptor (MOR) biased agonist TRV130. We trained rats to self-administer
oxycodone (6-h/d, 14 d) in context A; infusions were paired with a discrete tone-light cue. We thenimplanted Alzet osmotic pumps containing vehicle or 3, 6, or 9 mg/kg/d of buprenorphine or TRV130 and
performed three relapse-related tests: (1) responding for oxycodone-paired discrete cues under extinction
conditions in a non-drug context (context B), (2) context-induced reinstatement of oxycodone seeking incontext A, and (3) reacquisition of oxycodone self-administration in context A. We also assessed theeffect of chronic TRV130 delivery on acute oxycodone-induced decreases in oxygen levels in nucleus
accumbens.
Chronic buprenorphine delivery significantly decreased cue-induced oxycodone seeking in context B
and reacquisition of oxycodone self-administration in context A. Chronic delivery of TRV130 significantly
decreased relapse-related behaviors on all three measures. Chronic TRV130 delivery also prevented
acute oxycodone-induced brain hypoxia. Chronic delivery of TRV130 decreased oxycodone seeking on
multiple measures of relapse and prevented acute opioid-induced brain hypoxia. We propose that biased
MORs agonists, currently developed for pain treatment, should also be considered as opioid agonist
maintenance treatment for opioid addiction.

22

Tuesday July 9, 201910:00 AM – 12:40 PM
Symposium 3 - Albert Herz Memorial Symposium: Targeting Kappa Opioid Receptors
Chairs: Eduardo Butelman, Rockefeller University, USA
Tom Prisinzano, University of Kansas, USA
Presenter – Nyberg, Fred
Tribute in Memory of Albert HerzFred Nyberg1
1
Department of Pharmaceutical Biosciences, Uppsala University, Sweden
With sadness we learned that one of our well-known pioneer INRC members professor Albert Herz
passed away on November 9, 2018. He reached an age of 97 and an essential part of his scientificlife was given to INRC, where he belonged to the core of highly qualified and successful scientists
contributing to our organization. Albert Herz was born in Sonthofen, Bavaria in Germany on June 5,
1921. He studied medicine at the Ludwig-Maximilians-Universität in Munich, where he obtained his
doctorate in 1948. After continuous work in pathology and internal medicine he was qualified as auniversity professor in pharmacology and toxicology. Albert Herz moved to the Max Planck Institute(MPI) of Psychiatry in 1962, where he ten years later was appointed as Director of the
Neuropharmacological Department at MPI. A great part of his career was centered on theinvestigation of processes underlying the action of morphine and other opiates on brain function. His
work clarified an essential part of mechanisms involved in drug addiction and in pain relief andaddiction therapy as well as in relation to the addictive potential of painkillers. His work did create adeeper understanding of the complexities of addiction. Dr. Herz worked as professor at MPI, where
he became an Emeritus Scientific Member of the institute in 1989. Albert Herz has meant a lot for
the INRC organization. He was among the group of scientists who initiated INRC in 1969 he acted
as responsible chairperson for the INRC at several annual meetings and from 1988 to 1990 Dr. Herz
served as president for the INRC. We hereby express our deep sadness but also our thankfulness
to Albert Herz, thanking him for all his outstanding contribution to the INRC. The memory of his highquality in science and nice personality will remain.

23

Presenter – Bruchas, Michael
Dissecting nucleus accumbens opioid circuits in reward seeking
Christian E. Pedersen1,2,3,4, Daniel C. Castro2,3,4, Michael R. Bruchas1,2,3,4
1
Department of Bioengineering, 2Department of Pharmacology, 3Department of Anesthesiology, and4Center for the Neurobiology of Addiction, Pain and Emotion, University of Washington, Seattle, USA
Kappa and Mu-opioid receptors are highly enriched at both pre and post-synaptic locations within the
nucleus accumbens. Numerous studies have been linked these receptors and peptides within this region
to both naturalistic reward, motivation, and aversive behavior. However, how neuronal ensembles of
opioid peptide-containing medium spiny neurons (MSNs), encode these behaviors remains unknown. In
vivo electrophysiology experiments have previously revealed that neural activity of medium spiny neurons(MSNs) in the medial shell of nucleus accumbens correlates to the palatability of reward. The use of credriver lines and optogenetics has recently enabled the cell-type specific dissection of medial shell MSN
circuits in modulating reward behavior. It has been found that genetically distinct MSN cell types can have
opposite effects on valence and reinforcement, suggesting that individual cell types have diverse function.
Advances in in vivo 2-photon calcium imaging through endoscopic lenses allows for the measurement ofneural activity in deep brain structures with single cell resolution. Here we leverage this approach toobserve ensemble neural activity of functionally diverse Dynorphinergic and Enkephalinergic MSNs
across multiple days while animals consume rewards of varied value and learn associations between
rewards and reward predictive cues. Specifically, we imaged the calcium dynamics of dozens ofdynorphin and enkephalin MSNs simultaneously while animals consumed 4 different concentrations of
sucrose reward and tracked changes in single cell activity throughout 7 days of Pavlovian conditioning
where animals learned auditory cues predicted subsequent reward delivery. Furthermore, using
retrograde methods, and dual-site fiber photometry approaches we examined how specific MSN outputs
to the ventral tegmental area (VTA), lateral hypothalamus (LH) or ventral palladum (VP) respond undervarious reward conditions, to determine how specific NAc projects encode reward value. Understanding
the complex nature of endogenous opioid-containing neurons could help to better dissect mechanism forkappa and mu-opioid dependent behaviors, as well as aberrant behavior as a result of depression,anxiety, or addiction behavior. Supported by R37 DA033396 from NIDA.
Presenter – Jones, Sara
Kappa opioid receptors drive dopamine deficiency and dysphoria during withdrawal from alcohol,
cocaine and heroin
Sara R. Jones1, Paige Estave1, Brianna George1, Anushree Karkhanis1, Cody Siciliano2, Katie Holleran1
1
Wake Forest University School of Medicine, Winston Salem, USA, 2Massachusetts Institute ofTechnology, Cambridge, USA
Chronic exposure to alcohol and other drugs of abuse results in a reduction in overall dopamine (DA)neurotransmission in the ventral striatum, contributing to negative affective states during withdrawal. Oneof the mechanisms driving hypodopaminergia may be drug-induced hyperactivity of brain stress systems,
including the dynorphin/kappa opioid receptor (KOR) system. KORs are present on dopamine axon
terminals in the nucleus accumbens, and when activated by dynorphin or an exogenous agonist, theyrobustly inhibit dopamine release. We found that KORs on dopamine terminals are supersensitive toagonist activation after chronic alcohol exposure in multiple species, including mice, rats and monkeys.
More recently, in rats we have documented increased activity of KORs after 2-4 weeks of cocaine selfadministration and withdrawal under several different schedules of reinforcement. Finally, heroin selfadministration also increased KOR activity. All of these abused substances acutely elevate dopamine,
but after repeated intensive exposure, compensatory changes reduce basal levels of dopamine, andthese reductions can be reversed with KOR antagonists. In addition to reducing dopamine release andbasal levels, KOR activation also influences dopamine transporter function through direct protein-protein
interactions. We have explored the reciprocal interactions of KORs and dopamine transporters and foundthat there is extensive cross-talk between the two proteins. Thus the dynorphin/KOR system acts on atleast two important nodes of dopamine neurotransmission regulation, contributing to withdrawal-relatedanhedonia and dysphoria. We therefore suggest that KOR antagonists may be useful as therapeutic
agents to reduce negative affect during acute and extended withdrawal from multiple drugs of abuse.
24

Presenter – Reed, Brian
In vivo effects of kappa agonists with differential in vitro efficacy
Brian Reed1, Amelia Dunn1, Eduardo Butelman1, Mary Jeanne Kreek1
1
Laboratory of the Biology of Addictive Diseases, Rockefeller University, USA
Addictions to cocaine and methamphetamine continue to be major public health problems. No
pharmacotherapeutic interventions have been approved to help treat these addictions, in spite ofintense efforts. Although cocaine’s primary neurochemical mediators are members of thecatecholamine system, studies from our laboratory and others have implicated the endogenous
opioid system, with persistent changes in the levels of mRNA and proteins associated with thissystem following chronic cocaine administration. In particular, the counter modulatory dynorphin
peptide/kappa opioid receptor subsystem has been shown to be dysregulated in both human andanimal models, and activation of this system in turn has been demonstrated to attenuate the effectsof cocaine, both behaviorally and neurochemically. Thus we are working to develop novel
therapeutics which selectively target the kappa opioid receptor as partial agonists or biased agonists.
Working with a team of medicinal chemists, we have screened hundreds of compounds, using invitro cell binding and signaling assays, allowing us to identify several novel kappa opioid receptor
agonists as having reduced efficacy (partial agonists) and/or differential signaling (biased agonists),
indicating pyrrolidine- and arylacetamide-substituted pyrano[3,4]-pyrazine fused-ring scaffold canserved as a backbone for development of novel kappa ligands. In rodent models, using several
compounds of differential ligand induced signaling bias, we have identified that signaling via the
kappa opioid receptor beta arrestin signaling pathways is responsible for the sedative properties of
kappa signaling, indicating that, for kappa agonists, G-protein signaling bias is a desirable property
to reduce the side effect of sedation.
Presenter – Prisinzano, Thomas
Design and synthesis of natural product based kappa opioid receptor ligands
Thomas E. Prisinzano1
1
Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, USA
CNS active natural products have provided valuable, sometimes serendipitous, scientific andmedicinal discoveries in the past that have furthered our understanding of the mind and body. One
such example is the neoclerodane diterpene salvinorin A which has emerged as a valuable tool for
gaining additional insight into the pharmacology of κ-opioid receptors (KORs). However, thediscovery and optimization of meaningful transformations leading to useful semisynthetic and fully
synthetic analogues of salvinorin A has been limited, making continued structure-activity relationship(SAR) studies a major challenge. Using information gathered from previously described SAR studies,
as well as our own ongoing semisynthetic work on salvinorin A, a modular total synthesis has beendeveloped capable of producing natural product-based ligands that interact with KORs. The strategy
has permitted the deliberate simplification and introduction of functionality about the target molecule
to provide access to molecular features on salvinorin A otherwise unattainable by semisynthesis.
Using this approach, a potent pseudo-neoclerodane ligand has been realized.

25

Presenter – Lutz, Pierre-Eric
Epigenetic programming of the kappa opioid receptor by child abuse: a post-mortem brain study
Pierre-Eric Lutz1, Jeffrey A. Gross1, Sabine K. Dhir2, Gilles Maussion1, Jennie Yang1, Alexandre
Bramoulle1, Michael J. Meaney2, and Gustavo Turecki1
1
McGill Group for Suicide Studies, Douglas Mental Health University Institute, McGill University, Verdun,
Quebec, Canada, 2Douglas Mental Health University Institute, McGill University, Verdun, Quebec,
Canada.
Experiences of abuse and neglect during childhood are major predictors of the emergence of depressive
and suicidal behaviors throughout life. The underlying biological mechanisms, however, remain poorly
understood. Here, we focused on the opioid system as a potential brain substrate mediating these effects.
Postmortem samples from three brain structures regulating social bonds and emotions were analyzed.
Groups were constituted of depressed individuals who died by suicide, with or without a history of severe
child abuse, and of psychiatrically healthy controls. Expression of opioid peptides and receptors wasmeasured using qPCR. DNA methylation, a major epigenetic mark, was investigated using targetedbisulfite sequencing and characterized at functional level using in vitro reporter assays. Finally, oxidative
bisulfite sequencing was used to differentiate methylation and hydroxymethylation of DNA. Results
indicated that a history of child abuse specifically associated in the anterior insula with a downregulationof the kappa opioid receptor, as well as decreased DNA methylation in the second intron of thecorresponding gene. In vitro assays further showed that this intron functions as a genomic enhancer
where glucocorticoid receptor binding regulates kappa expression, unraveling a new mechanism
mediating the well-established interactions between opioids and stress. Finally, results showed that childabuse is associated in the kappa intron with a selective reduction in levels of DNA hydroxymethylation,
likely mediating the observed downregulation of the receptor. Altogether, our findings uncover new facetsof Kappa physiology, whereby this receptor may be epigenetically regulated by stressful experiences, inparticular as a function of early social life.
Hot Topic Presenter – Bedini, Andrea
Functional selectivity and analgesic effects of CL39, a novel kappa opioid receptor (KOR)
selective agonist
Andrea Bedini1, Lorenzo di Cesare Mannelli2, Laura Micheli2, Rossella De Marco3, Luca Gentilucci3, Carla
Ghelardini2, Santi Spampinato1
1
Department of Pharmacy and Biotechnology - University of Bologna, Italy, 2Department of Neuroscience,
Psychology, Drug and Children Health, University of Florence, Italy, 3Department of Chemistry G.
Ciamician, University of Bologna, Italy
Kappa opioid receptor (KOR) agonists determine antinociception by G protein signaling; conversely, they
trigger relevant side effects through arrestin 3-dependent p38MAPK activation. Thus, raising significant
interest in developing G protein biased KOR agonists as innovative analgesics. Recently, we identified
CL39 (H-Tyr-Amo-Trp-PheNH2), an endomorphin-1 analogue with high affinity, selectivity and partial
agonist activity at KOR; here we investigate its functional selectivity and analgesic effects as comparedto U50,488. CL39 weakly recruited arrestin 3 at KOR, as assessed via arrestin complementation assay
in U2OS cells and by means of a BRET assay in HEK-293 cells; calculated bias factor for U50,488 and
CL39 were equal to 1 and 65, respectively. CL39 inhibited adenylyl cyclase and activated early (5-15min),G protein-dependent component of ERK1/2 phosphorylation in HEK-hKOR and U87-MG cells, asassessed by ELISA and western blot, but did not trigger in the same cells late (60min), arrestin-dependent
component of ERK1/2 or p38MAPK phosphorylation. On the contrary, U50,488 activated all the above
mentioned signaling pathways. Moreover, U50,488, but not CL39, significantly increased U87-MG cellproliferation in an arrestin 3-, p38MAPK-dependent fashion, as assessed by [3H]-thymidine incorporation.
Experiments on normal human astrocytes are ongoing and will be presented at the conference. In vivo,
both CL39 and U50,488 (0-30 mg/kg; 0-60 min; i.p.) induced a significant, KOR-mediated, dosedependent antinociception in warm-water tail-withdrawal test, displaying the ligands comparable ED50.
CL39-mediated analgesia in a model of oxaliplatin-induced neuropathic pain is currently underinvestigation. CL39 is a novel, G protein biased KOR agonist displaying analgesic effects in vivo.

26

Tuesday July 9, 20192:00 PM – 4:00 PM
Symposium 4 - Gavril Pasternak Memorial Symposium: Chemical Biology of Diverse Opioids
Chairs: Susruta Majumdar, Washington University School of Medicine, USA
Nurulain Zaveri, Astraea Therapeutics, LLC, USA
Presenter – Pan, Ying-Xian
Tribute to Gavril Pasternak
Ying-Xian Pan1
1
Memorial Sloan Kettering, USA
It is with profound sadness and heavy hearts that we learned the passing of Dr. Gavril Pasternak, adistinguished physician/scientist and long-time INRC member, who dedicated his entire career to opioid
research and pain management. In 2015, Gav was honored with the Founder’s lecture by INRC torecognize his contribution. Gav’s lifelong passion in studying opioids began when he joined the laboratoryof Dr. Sol Snyder for his PhD work, which led to the seminal work of identifying opioid receptors using
radioligand binding assays and contributing to identification of endogenous opioid peptides. He was thefirst to show that sodium ions stabilized the receptor in an antagonist configuration, a finding that is now
recognized as a generalized GPCR effect with a sodium binding site identified in several GPCR crystal
structures. In 1979, Gav joined the Department of Neurology at Memorial Sloan-Kettering Cancer Center,
where he defined several mu opioid receptor subtypes using a combination of classic pharmacologicalapproaches and novel antagonists. This work led to the main rationale and effort for identifying an arrayof alternatively spliced variants of mu opioid receptor genes decades later, providing distinct targets fordeveloping novel analgesics with fewer side effects. Gav published over 430 scientific papers, five books
and a dozen of patents and received numerous awards. In addition to being a great mentor for many
young people worked in his lab, Gav was also known for his passion for lacrosse and for founding NYC’s
first youth lacrosse program.

27

Presenter – Majumdar, Susruta
Structure based design and synthesis of diverse opioids
Susruta Majumdar1, Gavril W Pasternak2, Rajendra Uprety2, Tao Che3, Saheem Zaidi4, Steven
Grinnell5, John Pintar6, Ying-Xian Pan2, Jay P McLaughlin7, Jonathon Javitch5, Vsevolod Katritch4
and Bryan Roth3
1
Center for Clinical Pharmacology, St. Louis College of Pharmacy and Washington University Schoolof Medicine, St. Louis, USA., 2Department of Neurology and Molecular Pharmacology, Memorial
Sloan Kettering Cancer Center, USA, 3Department of Pharmacology, University of North Carolina,
Chappell Hill, USA, 4Bridge Research Institute, University of Southern California, Los Angeles,
5
Department of Psychiatry, Columbia University, Piscataway, USA, 6Department of Neuroscience
and Cell Biology, Rutgers Robert Wood Johnson Medical School, Piscataway, USA, 7Department ofPharmacodynamics, University of Florida, Gainesville, USA.
Utilizing the crystalized structure of the active state kappa opioid structure with MP1104, we now
have rationally designed compounds which are G-protein biased at both mu as well as kappa opioid
receptor. Our leads target a region residing between transmembrane 5 (TM5) and extracellular loop
(ECL2). We propose this region allosterically controls arrestin recruitment at both mu and kappa
opioid receptors.
Several chemical probes designed using structure based approaches were synthesized inboth the morphinan as well as fentanyl templates were tested to probe arrestin recruitment throughTM5-ECL2 region. The model was validated using mutational analysis of key residues in this region.
The lead compounds, MP1207 and 1208 are analgesics devoid of respiratory depression andconditional place preference and/or aversion at the highest dose tested suggestive of a possibility ofachieving opioid functional selectivity using dual activation of mu and kappa receptors while notrecruiting βarrestin-2. Supported by NIDA (R01DA046487 and R21DA045884) and through the Peer
Reviewed Medical Research Program (W81XWH-17-1-0256).
Presenter – Zaveri, Nurulain
Bifunctional NOP/MOP agonists as a new approach for non-addicting analgesics andtreatments for opioid use disorders
Nurulain Zaveri1
1
Astraea Therapeutics, LLC, USA
Agonists for the nociceptin opioid receptor (NOP) have been known to modulate the pharmacologyof mu opioid (MOP) agonists in pain and drug reward. We show that bifunctional agonists targeting
the NOP and MOP receptor show enhanced analgesia without opioid adverse effects such as
respiratory depression, tolerance and dependence. An appropriate balance of NOP agonist efficacy
to MOP agonist efficacy is important to obtain a non-addicting analgesic profile. Such bifunctional
NOP/MOP agonists can also decrease the reinforcing effects of prescription opioids, providing an
innovative, dual action for the opioid crisis, for those experiencing pain and addiction to prescription
opioids. In this talk, we will report on the design and identification of NOP/MOP bifunctional agonists,
using structure-based drug design and medicinal chemistry optimization, to modulate the NOP and
MOP binding affinity and balance of functional efficacy. The in vivo pharmacology of suchbifunctional NOP/MOP agonists in rodents and nonhuman primates will be discussed.

28

Presenter – Al-Hasani, Ream
Determining the neurochemical profile of endogenous opioids during withdrawal
Ream Al-Hasani1
1
Washington University School of Medicine, USA
The US Department of Health states that 42,249 people died from overdosing on opioids in 2016, 2.1
million people had an opioid use disorder and the economic cost of this epidemic was $504 billion and is
still rising. The reasons for this are multifaceted and my research focuses on understanding the circuitry
and endogenous peptide profile during drug withdrawal, with an emphasis on the kappa opioid system.
The goals of my research are two-fold: 1) to better understand the role of dynorphin during drug
withdrawal and 2) to develop innovative, novel approaches to allow in vivo detection of endogenously
release opioid peptides. The ability to better understand the role of the dynorphin/kappa opioid system inmodulating negative affect, such as anxiety and depression, associated with drug withdrawal will be
critical in reducing the incidence of relapse and overdose. This will allow us to consider alternative more
affective long-term treatments for addiction. To do this we use mouse models to directly measure in vivochanges in dynorphin levels in the nucleus accumbens region of the brain, a key region in the modulationof reward/addiction behavior, during withdrawal from opioids such as morphine and fentanyl. We use
microdialysis paired with liquid chromatography/mass spectrometry to allows sub-fmol range detection of
peptides. In addition, we are also developing micro-immunoelectrodes to allow more spatiotemporally
controlled detection of opioid peptide release in vivo. This technology allows us to pair detection of peptidechanges on a second to minute timescale during negative affective/anxiety-like behaviors characteristicof withdrawal.
Presenter – Zaidi, S.A.
Designing bitopic opioids: Activity modulation in targeting sodium binding pocket of the opioid
receptors
Zaidi, S. A.1, Che, T.2, Kormos, C.3, Faouzi, A.4, Majumdar, S.4, Carroll, F. I.3, Roth, B.2, Katritch, V.1
1
Department of Biological Sciences, Bridge Institute, University of Southern California, Los Angeles, USA,2
Department of Pharmacology, University of North Carolina at Chapel Hill School of Medicine, Chapel
Hill, USA, 3Center for Organic and Medicinal Chemistry, Research Triangle Institute, Research Triangle
Park, USA, 4Center for Clinical Pharmacology, St Louis College of Pharmacy and Washington University
School of Medicine, St Louis, USA
Design and development of functionally biased opioid receptor ligands, able to selectively recruit Gprotein over β-arrestin, offers a very promising approach towards development of limited liability
analgesics. Engagement and stabilization of the specific sub-pockets within the receptor by ligands can
induce such functional response. The “sodium binding” pocket, an allosteric site present two helical turns
below the orthosteric ligand binding region, could be a very effectual receptor sub-pocket to induce suchfunctional profile. The sodium binding pocket consists of 16 highly conserved amino acid residues in thecenter of the transmembrane bundle of class A GPCRs, and undergoes major conformational changes
upon activation, as elucidated by several active state GPCR crystal structures. These changes lead tocollapse of the sodium pocket, controlled by outward movement of helix 6 and inward movement of helix
7 of GPCR. The coupling of the conformational changes between the sodium pocket and the functionally
relevant helices highlights the pocket as a compelling target for modulation of agonist action andfunctionality. We hypothesized that displacement of sodium and stabilization of the pocket in a specificconformational state by bitopic ligands targeting the pocket could lead to distinct functional profiles,
including biased G-protein recruitment profile of the ligands. Two bitopic analogs for nalfurafine scaffold
were designed via ligand docking and ligand-receptor optimization protocols using the conformationallyoptimized active/inactive state kappa opioid receptor crystal structures. Pharmacological evaluation of
the synthesized ligands revealed them to be indeed G-protein biased in kappa opioid receptor, thus
validating our hypothesis. Moreover, the highly conserved constitution of the sodium binding pocket
grants a generic receptor sub-pocket to be targeted to control functional profile in other class A GPCRs.
Supported by P01 DA035764 and R21/R33 DA038858 from NIDA.

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Presenter – Zadina, JamesA mu opioid with substantially reduced side-effects shortens recovery from chronic pain relative
to morphine
James Zadina1
1
Neuroscience Laboratory, V.A. Medical Center, Tulane University, New Orleans, USA
In addition to well-known negative side effects of currently used opioids, including abuse liability andtolerance, recent studies have shown a lesser-known effect; chronic exposure can paradoxicallyexacerbate and prolong pain hypersensitivity. It has been proposed that injury causes pro-inflammatory
CNS signaling which is exacerbated by morphine, causing more intense and longer-lasting pain. Knownas the “two hit” hypothesis, this exacerbation can occur with either injury-then-drug or vice-versa, and cancontribute to the transition from acute to chronic pain. This transition can also occur through “latentsensitization” (LS), pathological pain following injury or inflammation that is “masked” by changes in opioid
receptor function during apparent recovery and “unmasked” by treatment with an opioid inverse agonist
like naltrexone, or by stress. We have developed an analog (ZH853) of endomorphin with a reduction orabsence of several major side effects, including abuse liability, respiratory depression, tolerance, andimpairment of motor function. In addition, while morphine produces pro-inflammatory glial responses,
ZH853 does not. Acute administration of ZH853 produces longer antinociception than morphine inneuropathic, inflammatory, postoperative and visceral pain. Chronic administration of morphine before orafter inflammatory (CFA) or postoperative (paw incision) pain prolonged and intensified hypersensitivity.
By contrast, ZH853 unexpectedly reduced the total time and intensity of hypersensitivity. In addition,
latent sensitization occurred after vehicle and morphine treatment, but was blocked by ZH853. These
results show two mechanisms of reduced transition to chronic pain after treatment with ZH853, andtogether with its low side-effect profile, indicate that ZH853 could transform pain management.

30

Wednesday July 10, 201910:00 AM – 12:30 PM
Symposium 5 - Circuits in Neuropathic Pain
Chairs: Vivianne Tawfik, Stanford University, USA
Minoru Narita, Hoshi University, Japan
Presenter – Tawfik, Vivianne
Glial cells in opioid tolerance and chronic pain
Vivianne Tawfik, MD, PhD1
1
Department of Anesthesiology, Perioperative & Pain Medicine, Stanford University School of
Medicine, Stanford, USA
Chronic pain is a common and often debilitating condition that affects 100 million Americans. A better
understanding of pain’s molecular mechanisms is necessary for developing safe and effective
therapeutics, especially in the context of an ongoing Opioid Epidemic. Chronic opioid use can resultin analgesic tolerance and paradoxical opioid-induced hyperalgesia (OIH). Tolerance and OIH are
primary drivers of diminished pain control and dose escalation; however, their underlying
mechanisms remain unclear. Studies have suggested that glial cells, and microglia in particular, areessential contributors to chronic pain, opioid tolerance and OIH. We will discuss the case for indirect
involvement of microglial cells in opioid tolerance and OIH as histological and transcriptomic analysisof microglia failed to identify expression of the µ opioid receptor (MOR) in these cells. In contrast,
using spatiotemporal microglial ablation strategies we identified a key direct contribution of microglia
to the transition from acute to chronic pain. Overall, our data highlight the multidimensional
importance of glial cells in pain and opioid circuits.
Presenter – Narita, Minoru
Influence of pain-related neurocircuits on tumor immune response
Minoru Narita, PhD1
1
Department of Pharmacology, Hoshi University School of Pharmacy and Pharmaceutical Sciences,
Tokyo, Japan
Pain is a natural alert system that protects the body from potential tissue injury. However, emerging
evidence has shown that chronic pain does not serve this purpose; it only provides a persistentsensation of pain and a negative experience. Interestingly, a growing body of evidence suggests that
intractable pain is accompanied by a decreased survival rate in cancer patients and chronic stressful
conditions suppresses the systemic immune response, whereas pleasant conditions can enhance
the immune response to pathogens on an empirical basis. These facts indicate that both chronic
pain and stressful conditions affect central-peripheral immune interaction. Recently, our study has
revealed that 1. either pain or systemic inflammation could be a priming factor involved in tumor
progression, 2. a stressful condition involving activation of the hypothalamic-pituitary-adrenal axis
related to an increase in peripherally released cortisol may aggravate tumor progression, and 3. apleasant condition involving activation of the central opioid-dopamine circuits associated with thebrain-reward network may affect tumor progression. These results suggest that crosstalk between
pain/stress-related central neurocircuits and peripheral immune systems may be crucial for tumor
progression. In this symposium, we will present topics related to the complex mechanisms that
underlie pain and the tumor immune response.

31

Presenter – Corder, Gregory
Pain & the amygdala
Gregory Corder, PhD1
1
Departments of Psychiatry & Neuroscience, University of Pennsylvania, Philadelphia, USA
The same neural circuits that drive adaptive behavior can also be maladaptive. Those involved in
the detection, processing and transformation of nociception into pain affective-motivational
information are likely the most evolutionarily crucial functions of the nervous system. Thoughevidence for nociceptive-specific circuits in the peripheral and spinal nervous systems has existed
for decades, it remains unclear how pain affective circuits are embedded in the brain and how are
they modulated during neuropathic pain. Here, we will discuss our recently reported discovery of anamygdalar neural ensemble, at single neuron resolution, that selectively encodes theunpleasantness of all types of acute pain, regardless of the peripheral sensory mechanisms, andthat attributes negative affect to chronic neuropathic pain. While this work provides a critical point ofentry into the affective circuits of pain, it is not clear how these circuits are controlled, and to whichbrain regions they signal. To determine other circuit elements of this pathway, we are identifying thebrain-wide, input-out neural architecture using activity-dependent viral circuit tracing. We find that
amygdalar nociceptive neurons send long-range projections that preferentially innervate specificsubregions of the nucleus accumbens. This suggests that transmitted pain valence informationtransmitted could influence motivational and hedonic behavior, e.g., depression co-morbid withchronic-pain. Constructing a brain-wide global map of this pain affect pathway is our first step toward
identifying a series of candidate brain regions for therapeutic targeting of chronic pain.
Presenter – Ji, Ru-Rong
Distinct roles of β-arrestin-2 in regulating opioid analgesia, opioid hyperalgesia, andneuropathic pain
Ru-Rong Ji, PhD1
1
Center for Translational Pain Medicine, Departments of Anesthesiology & Neurology, Duke
University Medical Center, Durham, USA
β-arrestin 2 (Arrb2) is scaffold protein and plays an important role in regulating GPCR signaling andacute opioid analgesia. However, the role of Arrb2 in regulating opioid-induced hyperalgesia andneuropathic pain is unclear. It is well-established that activation of spinal cord NMDA receptor(NMDAR) is essential for the development of opioid-induced hyperalgesia and neuropathic pain.
Here we demonstrate an active role of Arrb2 in regulating spinal cord NMDAR function and theduration of pain. Intrathecal DAMGO induced early-phase analgesia, which is potentiated andprolonged in Arrb2 knockout (KO) mice. Notably, intrathecal DAMGO also induced late-phasemechanical allodynia after 24 hours, and this pain hypersensitivity is mediated by NMDAR andprolonged in Arrb2 KO mice. Spinal administration of NMDA induces GluN2B-dependent mechanicalallodynia, which is prolonged in Arrb2-KO mice. Loss of Arrb2 also increases the duration of
inflammatory pain and neuropathic pain. Strikingly, loss of Arrb2 results in a profound increase in
GluN2B-mediated NMDA currents in spinal lamina IIo neurons. Finally, spinal over-expression of
Arrb2 using lentivirus can rescue chronic neuropathic pain after nerve injury. Our data show that βarrestin 2 (Arrb2) not only regulates mu opioid receptor to suppress opioid analgesia but also
regulates NMDAR to suppress opioid-induced hyperalgesia and neuropathic pain. Thus, spinal
Arrb2 is critical for the transition from acute pain to chronic pain. While Arrb2 inhibitor can enhance
acute opioid analgesia via opioid receptors, it may also promote opioid-induced hyperalgesia andchronic pain via NMDARs. Supported by NIH R01 grants NS87988 and DE17794.

32

Presenter – Chiu, Isaac
Pain, neuro-immune interactions and bacterial infections
Isaac Chiu, PhD1
1
Harvard Medical School, Department of Immunology, Boston, USA
Pain is a hallmark of many bacterial infections. Nociceptor neurons densely innervate barrier tissues
that are exposed to bacterial pathogens. We find that nociceptors detect bacteria to produce pain
during infection, and signal to the immune system to regulate bacterial killing. Therefore, the nervous
system participates in host defense against pathogens, and blocking neuronal signaling hasconsequences on infection. Understanding the role of pain in host defense could inform the treatmentof pain and infection.
Presenter – Taylor, Anna
The gut microbiome contributes to neuroinflammation, pain and affect during opioid
withdrawal
Anna Taylor, PhD1
1
Department of Pharmacology, University of Alberta, Edmonton, Alberta, Canada
Opioid withdrawal is associated with an unpleasant constellation of symptoms including hyperalgesia(increased pain) and dysphoria that contribute to addictive behaviors leading to relapse. One of the
major advancements in this area of research has been the discovery that inflammation within thecentral nervous system contributes to many of the unwanted side effects of chronic opioid use.
Recently, our group has begun to investigate the contribution of the resident bacteria of the gut (i.e.
gut microbiota) to the full expression of opioid dependent behaviors including neuroinflammation.
Male C57Bl/6J mice were treated with escalating doses of morphine sulfate for four days. Tailwithdrawal and conditioned place preference were used to assess changes in evoked pain
thresholds and impaired reward, respectively. Fecal samples were processed for 16s rDNA
sequencing. Manipulation of the gut microbiome was used to assess the causal relationship between
the gut microbiome and opioid withdrawal.
Morphine withdrawal was associated with microglial activation in the brain and spinal cord,
hyperalgesia, and negative affect. Withdrawal also produced a significant shift in the gut microbiome
when compared to saline treated controls. Recolonization of a drug-naïve mouse with a ‘morphinewithdrawn’ gut microbiome produced microglial activation, hyperalgesia, and negative affect.
Blocking gut motility during opioid withdrawal with a peripherally restricted opioid agonist was
sufficient to reverse these withdrawal phenotypes. This study suggests that changes in the gut
microbiome are causally related to the opioid withdrawal behaviors, and strategies that mitigate gut
effects during opioid exposure may improve treatment outcomes during withdrawal.

33

Thursday July 11, 20199:00 AM – 10:00 AM
Young Investigator Award Lecture
Emily Jutkiewicz, University of Michigan, USA
Deciphering the function of delta-opioid receptors
10:00 AM - 12:40 PM
Symposium 6 - Endogenous Opioid System: Progress and New Concepts
Chair: Emily Jutkiewicz
Presenter – Bagley, Elena
Drug dependence dials down the effects of endogenous opioids
Elena Bagley1
1
The University of Sydney School of Medical Sciences, Australia
Endogenously released opioid peptides regulate the development of drug addiction, including aversive
responses to drug withdrawal, reward processing and associative learning. Opioid peptide signaling isterminated by peptidases, which cleave active peptides into inactive fragments. During opioid withdrawal,
peptidase activity increases in brain regions important for drug addiction, such as the ventral tegmental
area and the periaqueductal gray, suggesting a dynamic role for peptidases in the regulation of peptide
signaling. Whether this translates to reduced endogenous opioid actions is unknown. We have recently
shown that endogenous opioids strongly regulate the intercalated cells (ITCs) of the amygdala, a brain
region implicated in drug seeking and relapse behaviors. Here, we investigated whether increased
peptidase activity during withdrawal limits endogenous opioid effects on amygdala neural circuits. Using
patch-clamp electrophysiology of ITCs in brain slices isolated from Sprague-Dawley rats, we found thatthe activity of one peptidase, thiorphan, was markedly increased during withdrawal from chronic morphine
treatment. The increased peptidase activity results from elevated protein kinase activity during withdrawal
and reduced the actions of both exogenous and endogenous enkephalin at the δ-opioid receptor (DOR)
and exogenous nociception on basolateral amygdala synaptic outputs to the ITC. This effect was not due
to changes in receptor activity because, while the µ receptor showed homologous desensitization after
chronic opioid treatment, DORs remained unaffected. These data indicate that greater peptidase activity
during morphine withdrawal dials down the cellular actions of endogenous opioids on amygdala neuralcircuits which could contribute to the development of drug seeking and relapse behaviors.
Presenter – Cahill, Catherine
Kappa opioid modulation of negative affect & pain aversion associated with ongoing chronic pain
Catherine Cahill, PhD1
Shirley and Stefan Hatos Center for Neuropharmacology, UCLA, USA
Chronic pain is second only to bipolar disorder as the major cause of suicide among all medical illnesses.
Co-occurring psychopathology in chronic pain patients significantly impacts pain perception (heightened
pain intensity), increases pain-related disability, decreases response to treatment and increases risk ofprescription opioid misuse. In humans, kappa opioid receptor (KOR) activation causes anxiety,discomfort, agitation, depression and dysphoria. Considering that the circuitry involved in pain processing
and affective/motivational systems overlaps extensively, we asked whether KOR contributes to theaversive nature of chronic pain. In a rodent model of chronic neuropathic pain, we show that theendogenous tone of the KOR system within mesolimbic dopaminergic circuitry is robustly increased.
Importantly, we show that KOR blockade or elimination of KOR in midbrain dopamine neurons alleviatesa tonic-aversive component of chronic neuropathic and inflammatory pain in male, but not female mice.
KOR blockade also alleviates depressive and anxiogenic effects associated with neuropathic pain but
this effect is not sex-dependent, suggesting a diversion of mechanisms between affective dimensions ofchronic pain and the on-going tonic-aversive states. Our results strongly support the use of KOR
antagonists as therapeutic adjuvants to alleviate the emotional, tonic-aversive component of chronic pain.
34

Presenter – Poras, Hervé
DENKI (Dual ENKephalinase Inhibitors), a new class of painkillers devoid of abuse potential
and of opiate side-effects
Hervé Poras1, Gwendolyn E Burgess2, Bryan F Sears2, Tanja Ouimet1, Michel Wurm1, Marie-Claude
Fournié-Zaluski1, Bernard P Roques1, Emily M Jutkiewicz2
1
Pharmaleads SA, Paris, France, 2 Department of Pharmacology, University of Michigan, Medical
School, Ann Arbor, USA
Pharmaleads develops two Dual Enkephalinase Inhibitors (DENKI®), PL37 and PL265, active on
different types of pain models and devoid of opiate side effects. Enkephalins are penta-peptides
produced, e.g. during extreme physical workout or during long lasting painful stimuli, in the central
nervous system and in peripheral tissues. Their release elicits a “physiological” analgesia, providing
the body with its own pain modulation system. Once released, enkephalins are rapidly inactivatedby two membrane-bound exo-metalloproteases, neprilysin (NEP) and aminopeptidase N (APN).
DENKI® specifically protect enkephalins from NEP and APN in those areas where pain is perceived,transmitted and integrated. Unlike exogenous opiates, naturally secreted enkephalins do not induce
rewarding effects, reinforcing effects, withdrawal symptoms or dependence. In animal models of
pain, DENKI® induce antinociceptive responses similar to those of morphine. They remain active
after chronic treatment with morphine, indicating that DENKI®-induced analgesia persists inmorphine-tolerant mice. DENKI® do not induce constipation in animals or humans.
PL37 and PL265 do not produce morphine-like discriminative stimulus effects. They fail to maintainresponding or maintain significantly less responding in rats trained to self-administer remifentanil andmorphine. In addition, they induce delta-opioid receptor-mediated antidepressant-like effects in theforced swim test and do not induce convulsions. PL37 and PL265 appear to be devoid of abuseliability even at high doses and after repeated administration and could represent a new class of
painkillers in all types of severe pain as an affordable solution to the opioid crisis.
and of opiate side-effects
Presenter – Sombers, Leslie
Concurrent voltammetric detection of catecholamine and met-enkephalin dynamics in rat
striatum and adrenal tissue
Dr. Leslie A. Sombers1
1
W.M. Keck Center for Behavioral Biology, North Carolina State University, USA
Opioid peptides modulate a wide range of biological processes in both the peripheral and central
nervous system. However, a precise understanding of the functional significance of specific opioid
peptides is lacking, largely due to analytical challenges associated with monitoring theseendogenous species in situ. Our group has developed a multiple-scan-rate waveform to enable realtime voltammetric detection of M-ENK dynamics in tissue. The waveform parameters have beensystematically evaluated to increase sensitivity to M-ENK, specifically. This talk will demonstratethat the optimized methodology can be used to reveal co-fluctuations of putatively identified M-ENK
and catecholamine in both striatum and adrenal tissue with sub-second temporal resolution. Finally,
data will be presented to demonstrate that chemically selective electrode coatings can beincorporated to further enhance selectivity against larger peptidergic species. Overall, this work
describes a suite of electroanalytical tools that, when combined, offer the sensitivity, selectivity, andspatial resolution required to monitor peptidergic concentration fluctuations in situ on a time scalecommensurate with that of neuronal firing.

35

Presenter – Hsu, David
Endogenous opioid regulation of emotions in the social environment
David T. Hsu, PhD1
1
Stony Brook University, Health Sciences Center, Stony Brook, USA
Humans depend on others for survival and emotional well-being. Social rejection – when one is notwanted or liked – is a direct threat to this need. Several psychiatric disorders stem from exaggerated
responses to rejection, including major depressive, social anxiety, borderline personality, andsubstance/alcohol use disorders, yet the neural regulation of rejection is poorly understood. In thistalk, I will present laboratory models of social rejection and how we can study neural responses tosocial rejection. I will present our work using functional magnetic resonance imaging (fMRI) andpositron emission tomography (PET) to examine the endogenous opioid response to rejection, andshow that this response is reduced in depressed individuals, suggesting an inability to decreasesocial “pain.” I will also present work suggesting that the experience of rejection can be manipulatedby pharmacological and nonpharmacological methods traditionally used to reduce physicalpain. These represent new approaches to treating psychiatric disorders stemming from exaggerated
responses to rejection.
Hot Topic Presenter – Ueda, Hiroshi
Chronic pain status and loss of morphine analgesia have independent brain mechanisms inmice with fibromyalgia-like generalized pain
Hiroshi Ueda1,2, Hiroyuki Neyama2,3
1
Kyoto University, Japan, 2Nagasaki University, Japan, 3Riken Kobe, Japan
Fibromyalgia (FM) is an unmet medical need in terms of poor understanding of pathogenesis. We
attempted to study the molecular mechanisms underlying the lack of morphine analgesia, which isone of FM-specific pharmacotherapeutic features, by using the intermittent cold stress (ICS)-induced
generalized pain disease model in mice, which show fibromyalgia-like pathophysiological andpharmacotherapeutic features. First, we confirmed the lack of morphine analgesia in ICS-treatedmice by measuring mechanical somatic and muscular hyperalgesia and electrical stimulationinduced nociceptive responses. Next, we obtained the recovery of morphine analgesia in NMDA
receptor subunit NR2A-deficient (KO) mice, which blocks the morphine analgesic tolerance, possibly
through a blockade of anti-opioid counterbalance mechanisms. These results suggest that the lackof morphine analgesia in ICS mice is presumably mediated by μ-opioid receptor (MOPr)
desensitization/tolerance through a repeated stimulation by stress-induced endogenous opioid
release. The involvement of NR2A was supported by the successful recovery of morphine analgesia
by microinjection of NR2A siRNA into the PAG, but not VTA of ICS mice. In accordance, thepretreatment with dextromethorphan with NMDAr antagonist activity recovered morphine analgesia,
and single administration of methadone with NMDAr antagonist and MOPr agonist activities showed
potent analgesia. The ICS-induced hyperalgesia and lack of morphine analgesia remain unchangedin MOPr- and LPA1-KO mice, respectively. These results suggest that both mechanisms underlying
chronic pain and lack of morphine analgesia are independent to each other. Supported by JSPS
KAKENHI Grants JP17H01586 (H.U.) and JP26253077 (H.U.).

36

Hot Topic Presenter – Lutfy, Kabirullah
The role of endogenous opioids in amphetamine-induced locomotor sensitization
Kabirullah Lutfy1
1Western University of Health Sciences, Pomona, USA
The endogenous opioid system has been implicated in amphetamine-induced hyperlocomotion andlocomotor sensitization. However, the role of each opioid peptide in these actions is unknown. Using micelacking mu opioid receptors (MOPr) and their wild-type controls, we determined the role of MOPr in theseactions of amphetamine. Mice of both genotypes were brought to the laboratory, habituated to thelocomotor activity chambers for 1 h, injected with saline or amphetamine (3 mg/kg) and motor activity wasrecorded at 15 min bin for 1 h. Mice were treated with their respective treatments for three days and thenleft untreated until day 8 when they were tested for locomotor sensitization. On this day, mice were treatedas described above except a lower dose of amphetamine (1 mg/kg) was used. Our results showed that
locomotor sensitization but not acute motor stimulatory action of amphetamine was reduced in micelacking MOPr compared to their wild-type controls. We then assessed which opioid peptide acts on thisreceptor to alter the action of amphetamine. Mice lacking either beta-endorphin or enkephalin and theirrespective wild-type controls were treated and tested for amphetamine-induced hyperlocomotion andsensitization as described above. Mice lacking beta-endorphin showed reduced locomotor stimulation to
the lower dose of amphetamine but exhibited a comparable locomotor sensitization to their wild-type
controls. In contrast, mice lacking enkephalin showed comparable hyperlocomotion but reduced
locomotor sensitization. These results suggest that enkephalin is possibly the endogenous ligand thatacts on MOPr to alter amphetamine-induced locomotor sensitization.

37

Thursday July 11, 20192:00 PM – 4:00 PM
Symposium 7 - Atypical Opiates
Chairs: Sandra Comer , Columbia University, USA
Ellen Walker, Temple University, USA
Presenter – Snyder, Gretchen
A new approach to the treatment of opioid use disorder
Gretchen L. Snyder1, Kimberly E. Vanover1, Peng Li1, Stephanie Cruz1, Wei Yao1, Terry Martin1, Lisa R
Gerak2, David R Maguire2, Charles P France2, Nigel Calcutt3, and Robert E Davis1
1
Intra-Cellular Therapies, Inc. Alexandria Center for Life Science, New York, USA, 2University or Texas
Health Science Center at San Antonio, Texas, USA, 3University of California, San Diego, USA
Medications are needed to treat opioid use disorder (OUD) by easing the somatic symptoms of drug
withdrawal and mitigating the dysphoria and psychiatric comorbidities that drive opioid use/abuse. Here,
we describe ITI-333, a novel molecule combining high affinity binding (Ki < 50 nM) to receptors (i.e., mu
opioid (MOP), serotonin 5-HT2A, and dopamine D1) that individually are known to treat substance use
disorders and psychiatric comorbidities. Data from cell-based and in vivo assays show that ITI-333
functions as a biased MOP receptor ligand with partial agonist activity, acting as an antagonist to block
effects of high doses of morphine in both pain and motor activity models, while acting alone as a potent
analgesic against acute and neuropathic pain in rodents. ITI-333 (0.01-3 mg/kg, SC), produces naloxonesensitive analgesia in the tail flick mouse (TF) assay, while attenuating morphine-induced analgesia. ITI333 blocks morphine hyperactivity without significant effects on spontaneous locomotor activity. In drug
abuse assays, ITI-333 dose-dependently suppresses the somatic signs of opioid withdrawal precipitatedby naloxone injection in oxycodone-dependent mice and blocks heroin cue-induced reinstatement
responding in rats (0.3- 3mg/kg, SC). Chronic treatment with ITI-333 (0.3 or 3mg/kg, SC) does not result
in tolerance or physical dependence in rats; acute doses of ITI-333 do not induce GI or pulmonary side
effects. Finally, ITI-333 is not self-administered by heroin-maintained rats (0.003-0.01mg/kg/inj) or rhesus
monkeys (0.01-0.1mg/kg/inj). In summary, evidence supports a role for ITI-333 in mitigating opioid
withdrawal and supports its potential as a treatment for OUD, associated comorbid dysphoric symptoms,
and pain.

38

Presenter – Javitch, Jonathan
The atypical antidepressant tianeptine is an efficacious mu opioid receptor agonist
Andrew Kruegel1, Dalibor Sames1, Rene Hen2,3, Jonathan A. Javitch2,31
Department of Chemistry, Columbia University, New York, USA, 2Department of Pharmacology,
Columbia University Vagelos College of Physicians & Surgeons, New York, USA, 3New York State
Psychiatric Institute, New York, USA
Tianeptine is an atypical antidepressant that has been used clinically in Europe, Asia and SouthAmerica for decades. We recently discovered that tianeptine acts as a selective and efficacious
agonist of the mu opioid receptor (MOR), signaling in a manner analogous to enkephalins andendorphins, the endogenous opioid peptides. Using both genetic knockout and pharmacologicalinhibition in mouse models for the study of depression, we have demonstrated that MOR is the direct
molecular target by which tianeptine exerts its antidepressant effects, a unique mechanism among
existing antidepressants. Importantly, at clinical doses, tianeptine does not produce euphoria; in fact,
at 6 times the normal dose, patients reported no euphoria. Moreover, in mice, after 10 days of twice
daily administration at equianalgesic doses, morphine produces dramatic tolerance and profoundphysical withdrawal in response to naloxone treatment, whereas tianeptine leads to no tolerance andno naloxone-induced withdrawal. Furthermore, unlike treatment of pain by traditional opioid agonists,
prolonged occupancy of the receptor is not required for the antidepressant effect. Instead, much likethe hypothesized pulsatile and time-limited action of endogenous opioids in response to social
rejection and acceptance, tianeptine-induced MOR agonism sets in place a signaling cascade that
leads to lasting physical changes in dendritic structure in key brain circuits. Interestingly, PET studies
have shown that endogenous opioids act at MOR in response to social rejection and that this
response is dramatically blunted in depressed individuals, raising the possibility that tianeptine acts
postynaptically to remedy this deficit in endogenous opioid signaling.
Presenter – Stauffer, Jonathan
A novel pegylated opioid in development for the treatment of chronic pain
Jonathan Joseph Stauffer1
1
Nektar Therapeutics and Inheris, Chicago, USA
Opioids are effective agents for the relief of moderate-severe pain. However, their clinical utility isoften limited by adverse effects, particularly those involving the central nervous system (CNS).
Sedation, cognitive impairment, and respiratory depression often limit the doses that patients cantolerate. In addition, opioid abuse may arise from CNS effects experienced as pleasurable by someusers. The issues of misuse, abuse, and diversion associated with opioids constitute a significant
public health crisis. Although sustained-release formulations and other abuse-deterrent products
designed to make access to the active opioid ingredient more difficult have been approved by the
FDA, the safety mechanisms built into these drug products can be subverted and/or are not optimal.
NKTR-181, an investigational new molecular entity (NME) opioid analgesic, is a selective mu-opioid
receptor agonist providing effective chronic pain relief with a lower potential for abuse. Although rateof movement across the blood brain barrier (BBB) is not the sole determinant of abuse potential, it
is understood that rapid entry into the central nervous system (CNS) is an important factor in theoverall attractiveness of a drug as a target for abuse. Therefore, it is our hypothesis that drugs with
inherently slow CNS uptake could have lower abuse potential and improved safety. The combinationof several physicochemical properties, inherent to the molecular structure of NKTR-181, reduces its
intrinsic membrane permeability and is a core factor in the slow CNS uptake of this drug candidate.
The clinical effects of PEGylation of the morphinan pharmacophore in NKTR-181 will be presented.

39

Presenter – McLaughlin, Jay
Evaluation of kratom and mitragynine as safer treatments for pain and opioid dependence
Jay P. McLaughlin,1 Lisa Wilson,1 Shainnel O. Eans,1 Heather M. Stacy,1 Rajendra Uprety,2 Dalibor
Sames,3 Gavril Pasternak,2 Susruta Majumdar 2,4
1
Department of Pharmacodynamics, University of Florida, Gainesville, USA, 2Molecular
Pharmacology and Chemistry Program and Dept. of Neurology, Memorial Sloan Kettering Cancer
Center; New York, USA, 3Department of Chemistry, Columbia University, USA, 4Center for ClinicalPharmacology, St Louis College of Pharmacy and Washington University School of Medicine, St
Louis, USA
Although facing DEA Schedule I designation, users claim Kratom and mitragynine ameliorate opioid
withdrawal and produce analgesia without liabilities. We evaluated Kratom and mitragynine for
analgesia, as a treatment for opioid withdrawal, and for direct liabilities. C57BL/6J and knockout(KO) mice for each opioid receptor were administered graded intracerebroventricular, subcutaneous(s.c) or oral (p.o.) doses (n=8/dose) of morphine, Kratom alkaloid or mitragynine and tested for
antinociception in the 55oC warm-water tail-withdrawal test. Additional C57BL/6J mice so treated
were evaluated for liabilities of acute antinociceptive tolerance, respiratory depression andambulations, opioid withdrawal signs or conditioned place preference (CPP). Subcutaneousadministration of Kratom alkaloid or mitragynine produced antinociception, albeit 10.8- and 17.1-foldless potent, respectively, than morphine (ED50=3.91 (2.92-5.17) mg/kg) that was abolished in muopioid receptor KO mice. Both samples were more potent after oral administration, and ameliorated
precipitated withdrawal in morphine-dependent animals. However, while producing lesspsycholocomotor activity and respiratory depression than morphine, both samples produced
significant conditioned-place preference that was abolished in MOR KO mice. In conclusion, while
retaining some liabilities, kratom and mitragynine produced significant MOR-mediatedantinociception and eased physical withdrawal in morphine-dependent animals, suggesting someclinical value. Supported by NIDA (DA06214), McManus Charitable Trust, and Mayday Foundation(to GWP), DA 034106 and DA 045884 (to SM), NCI CA008748 (to MSKCC) and the University of
Florida (to JPM).
Presenter – Walker, Ellen
Preclinical assessment of novel psychoactive substances: Opioids
Ellen Walker1
1
Department of Pharmaceutical Sciences, Temple University, USA
The emergence of a large number of fentanyl analogs and novel opioid-like substances on the illicit
market have contributed to considerable increases in unintentional overdoses in the past decade. In
addition, new compounds are appearing on the global market every few months. However, there isa lack of preclinical data about the basic pharmacology of these novel synthetic opioids until relatively
recently. Investigators are currently examining these fentanyl analogs and novel opioid-like
substances alone and in combination with naltrexone and are comparing them to reference opioids
fentanyl and morphine using preclinical abuse liability assessment models such as locomotor
assays, antinociceptive assays, and drug discrimination in rodents. Potency comparisons of anumber of the fentanyl analogs and novel opioid-like substances reveal similar efficacies to
traditional opioids and an overall trend of intermediate potencies between fentanyl and morphinewith some notable exceptions. To date, all fentanyl analogs and novel opioid-like substances that
produce effects similar to morphine or fentanyl are sensitive to naltrexone antagonism although there
may be potency differences for naltrexone among the different analogs related to the structure of thenovel opioid substance. These data contribute to the growing catalog of in vitro and in vivo results
required to prioritize scheduling decisions and understand the impact of the novel synthetic opioidson the current opioid epidemic. Supported by DJD-15- HQ-P- 0740 and DJD-17-HQ-P-0646

40

Presenter – Negus, S.
Learning from failure: Kappa agonists as negative controls in analgesic drug development
S. Stevens Negus1
1Department of Pharmacology and Toxicology, Virginia Commonwealth University, USA
The crisis in opioid addiction and overdose deaths has stimulated interest in development of safeand effective alternatives to mu opioid receptor (MOR) agonist analgesics for treatment of pain.
Preclinical behavioral procedures will likely play a key role in testing candidate analgesics andprioritizing their advancement to studies in humans. However, conventional approaches havehistorically generated a large number of “false positive” outcomes for drugs that appeared promisingin preclinical studies but then failed in translation to the clinic due either to insufficient analgesic
efficacy or to dose-limiting side effects. Centrally acting kappa opioid receptor (caKOR) agonists
illustrate this type of failed translation. This talk will review evidence from preclinical studies that led
to initial enthusiasm for caKOR agonists and from clinical experience to show their limited utility. The
talk will then consider how these caKOR agonists can be used as active negative controls to refine
preclinical testing strategies and improve preclinical-to-clinical translation in analgesic drug
development. Specifically, novel procedures of pain-related behavioral depression have beendeveloped that clearly differentiate between the effects of MOR and caKOR agonists. These novel
procedures validated with MOR agonists as positive controls and caKOR agonists as active negative
controls can then be used to provide novel insights on the analgesic potential of atypical opioids orother classes of candidate analgesics. More generally, progress in analgesic drug development will
benefit from a perspective that views failed translation as an opportunity to improve on our existingprocedures rather than to indict them.

41Poster Abstracts
MONDAY POSTERS
Poster Title

ID

First nameLast nameM1

Wen-Yu

Hsu

M2

Beth

Wiese

M3

Kevin

Beier

M4

Lindsay

Lueptow

M5

Abdelfattah Faouzi

Design of MP135, a novel mu-delta heteromer
selective Gi-protein biased agonist

M6

Eriko

NakataNovel delta-opioid receptor agonist NC-2800 has a
low risk of addiction

M7

SophiaFang

Increased Abuse of Illicit Fentanyls are Associatedwith Opioid-Related Overdose Deaths in the U.S.

M8

Aya

Matsui

MOR in direct-pathway medium spiny neurons
contribute to the preference for alcohol drinking

M9AmynahPradhan

The critical role of central delta opioid receptors inmodels of migraine and opioid-induced hyperalgesia

M10

David

ReinerRole of the orbitofrontal cortex in relapse to fentanyl
seeking after choice-induced voluntary abstinence

M11

Amelia

Dunn

M12

James

Ryan

M13

Susruta

Majumdar

M14

OliviaLevine

M15

Kylie

McPherson

M16

Nora

McCallM17

Gwendolyn

Burgess

Association between Opioids use disorder andFractures
The Role of Cannabinoid Receptor Mediation of
Opioid Induced Respiratory Depression in the
Prebötzinger Complex
Identification of neuronal ensembles that promote
opiate addiction
Chronic pain alters the intravenous selfadministration profile of remifentanil in mice

Correlation between Beta-Arrestin-2 signaling at the
kappa opioid receptor and rotarod sedation
Interrogating the Role of Brain-Derived Neurotrophic
Factor and the Ventral Hippocampus in Mediating
Fear Generalization
Oxidative metabolism as a modulator of kratom’s
opioid analgesic activity
Acute estradiol modulation of a discrete limbic circuit
driving binge alcohol drinking
Diverse cell types within the vlPAG exhibit opioidinduced GIRK currents.
Mapping the affective component of pain: amygdalar
nociceptive neurons project to the nucleus
accumbens
Evaluation of the Abuse Potential of AAH8, a Mixed
Efficacy Opioid Ligand

42

M18

Amie

Severino

M19

Erin

BobeckM20

Brady

AtwoodM21

Ram

Kandasamy

M22

Brian

Cox

M23

Rob

Hill

M24

WilliamBirdsong

M25

Sam

Groom

M26

Gary

Matyas

M28

Lucia

Hipólito

M29

Kyle

Windisch

M30

YolandaCamposJurado

M31

Akihiko

Ozawa

M32

ChihiroNozaki

M33

ArianaBrice-Tutt

M34

Kabirullah

Lutfy

M35

Abhijeet

KapoorKappa opioid receptors on dopaminergic projections
from the ventral tegmental area mediate distinct
aversion behaviors in chronic pain.
Differential intracellular signaling within theperiaqueductal gray contributes to differences in
opioid tolerance
Mu opioid receptors on vGlut2-expressing glutamate
neurons regulate aversion to oxycodone
Positive allosteric modulators of the mu-opioid
receptor produce pain relief without disruptive side
effects
Gene expression changes in the ventral tegmental
area (VTA) and lateral habenula (LHb) of rats after asingle morphine treatment.
Fentanyl depression of respiration: differential
reversal by antagonists and reduced cross tolerance
to morphine.
Acute desensitization and long-term tolerance toopioids are differentiated by multiple phosphorylation
motifs in the C-terminus of the mu-opioid receptor‘Compound 1’: a G-protein-biased MOPr agonist thatinduces MOPr desensitization apparently through
GRK
A Stable Heroin Hapten (6-AmHap) Vaccine
Formulation Slated for a Phase 1 Clinical Trial
Induces Long Duration Antibody Titers that Block theAntinociceptive Effects of Heroin and other Opioids
Ethanol intra-VTA induces CPP through the activationof local MORs and increases NR2A expression in the
NAc.
Investigating the Effect of Nalmefene on Extended
Access Cocaine Self-Administration in Male Mice
Inflammatory pain dysregulates MORs in the VTA:
effect on ethanol induced dopamine release in NAc
and ethanol relapse
Neuroanatomical analysis of chronic paindependently activated neurons in the brain and spinal
cord
CB2 receptor deletion on monocytes enhances
neuropathic pain development in mice
Mixed Opioid Receptor Agonism and Antagonism
Demonstrated by a Novel Cyclic Tetrapeptide
Prevents Drug- and Stress-Induced Reinstatement ofMorphine- and Cocaine- Seeking Behavior.
The role of endogenous PACAP in morphine-induced
state-dependent CPP and reinstatement
The Unique Conformational Ensemble and Active
Conformation of µ-Opioid Receptor Induced by
Methadone43Microglial P2X4 Receptors Contribute to Morphine
MorganTolerance Mediated by the Periaqueductal Gray in
the Rat
Glycosylation of Opioid Neuropeptides to Produce
Polt
Brain-Penetrant Drugs
The Mu-Delta Opioid Receptor Heterodimer Acts as a
Negative Feedback Brake to Reduce Opioid AntiStreicher
Nociception by Repression of CaMKII and Src
Signaling
The contribution of cocaine- and amphetamineKirály
regulated transcript (CART) peptide fragments tospinal nociceptive transmission in rats
In vivo identification of Rab10 as a regulator of the δDegrandmaison
opioid receptor (DOPr) trafficking
Inhibition of HDAC6 activity alleviates symptoms of
Saklothchronic pain
Mu opioid receptor expression and function in mouse
SéguélaACC versus S1 cortex

M36

Michael

M37

Robin

M38

John

M39

Kornél

M40

Jade

M41

Farhana

M42

Philippe

M43

Mariana

Lemos Duarte

Opiate-mediated protein kinase C signaling examinedusing novel antibodies that recognize active protein
kinase C and phosphorylated mu opioid receptors.

M44

Gavin

Bart

Altered Methadone Pharmacokinetics: impact ofantiretrovirals and an independent effect of HIV

M45

Cassandra

Retzlaff

Investigating the Role of Striatal Neuropeptide Y
Expressing Interneurons in Opioid Action

M46

LoubenDorval

M47

Grace

Rossi

M48

Brian

Knapp

M49

Mikko

Airavaara

M50

Lovisa

Holm

M51

Devon

Collins

M52

Evan

Fullerton

Fibroblast Growth Factor 21 (FGF21) Attenuates
Morphine Preference in Mice
The pharmacological and molecular characterizationof mu opioid receptor splice variants in the rat using
gene targeting models generated by Easi-CRISPR
G-Protein subunits alpha and gamma influencefunctional activation profiles of opioids at the mu
opioid receptor
Alcohol drinking and craving in female mice housed
in automated cages
TRV130 modulates reward in a beta-arrestindependent manner
Oxycodone conditioned place preference, locomotor
activity, and self-administration in male and female
Oprm1 A112G mice
Impact of Sex and Advanced Age on Opioid
Analgesia in the Rat Periaqueductal Gray

44

TUESDAY POSTERS
ID

First nameLast nameT1

MihályBaloghT2

AndreaKliewerT3

Gerta

Cami-Kobeci

T4

Yan

Zhou

T5

Hervé

Poras

T6

Orna

LevranT7

Yu-JunWang

T8

Saheem

Zaidi

T9

Bryan

McElroy

T10

Matthew

Hearing

T12

Zachariah

Bertels

T13

Jessica

Anand

T14

Wallace

Chan

T15

Jacob

Beierle

T16

Leela

Afrose

T17

Chase

Webb

Poster Title
High efficacy opioids compared to morphine produce
promising analgesia in advanced diabetic neuropathic
pain in rats
Phosphorylation-deficient G-protein-biased μ-opioid
receptors improve analgesia and diminish tolerance
but worsen opioid side effects
Naltrexone esters and close analogs as potential new
analgesics with reduced side effect profile
Combination of clinically utilized kappa opioid receptor
agonist nalfurafine with low-dose naltrexone reduces
excessive alcohol drinking in male and female mice
DENKI (Dual ENKephalinase Inhibitors), a new classof painkillers devoid of abuse potential and of opiate
side-effects
Combinations of polymorphisms in circadian rhythm
genes are associated with cocaine addiction in African
Americans
Discovery of a highly selective and potent kappa
opioid receptor agonist with reduced CNS side effects
Designing bitopic opioids: Activity modulation intargeting sodium binding pocket of the opioid receptors
Impact of short acting synthetic kappa-opioid
antagonists LY2444296 and LY2795050 on selfgrooming and anhedonic-like behaviors in male miceCell-type and region-specific nucleus accumbens
AMPAR plasticity associated with opioid reward, drugseeking, and withdrawal

T11
Differential Effects of Mu and Delta Opioid Receptor
Agonists in Models of Migraine
Structure Activity Relationships of Illicit Fentanyl
Analogs
Docking-Based Virtual Screen for Bifunctional μ-Opioid
Agonist/δ-Opioid Antagonist Compounds
Mouse inbred strain survey of oxycodone addiction
traits in an opioid multi-stage addiction assessment
paradigm
Interaction of a Newly Deorphanized G ProteinCoupled Receptor, GPR171, in Chronic Morphine
Treatment
Computational Discovery of Novel µ Receptor
Agonists
45

T18

David

MaguireT19

Lawrence

Kuo

T20

Max

McDermott

T21

Amy

Alder

T22

AndreaCippitelli

T23

TeresaMilner

T24

Ivone

Gomes

T25

Matthew

Randesi

T26

SeshatMack

T27

AmandaFakira

T28

Ying

He

T29

Li

Zhang

T30

Chaim
(Chagi)

Pick

T31

Ying-Xian

Pan

T32

Sofia

Zelleroth

T33

Rossella

De Marco

T34

Lisa

WilsonT35

CamronBryant

The long-acting mu opioid receptor selective
antagonist methocinnamox (MCAM) prevents andreverses abuse- and overdose-related effects of
opioids
Novel Biased Mu Opioid Agonists for the Treatment of
Pain
Exploring GPR171 as a novel pain therapeutic with
reduced abuse liability
Preclinical Evaluation of the Novel Mu Opioid Receptor
Agonists Kurkinorin and Kurkinol
PPL-103: A kappa partial agonist approach that
potently reduces compulsive cocaine taking andprime-induced cocaine seeking
Sex and chronic stress influence the distribution ofdelta opioid receptors within the rat hippocampal CA1
region following oxycodone conditioned place
preference
The intricacies of opioid receptor signaling byendogenous opioid peptides.
Sex differences in opioid, plasticity and stress-related
genes in rat caudate putamen following oxycodone
conditioned place preference
Characterization of small molecule ligands targeting
GPR83, a newly deorphanized striatal receptor that
could modulate opiate reward
Investigating the novel PEN-GPR83 receptor system
in morphine reward and analgesia.
Long term opioid use is required for sickle cell disease
after curative hematopoietic stem cell transplantation
Morphine tolerance is attenuated in germ-free miceand reversed by probiotics implicating the role of gut
microbiome
Mirtazapine and Trazodone: A possible opioid
involvement in their use (at low dose) for sleep?
Biased signaling through Oprm1 full-length 7TM exon
7-associated variants: Insights into in vivo mu opioid
actions
Anabolic Androgenic Steroids Affect Weight Gain,
behavior and Brain Gene Expression in Male Rats
Tryptophan-Containing Non-Cationizable Opioid
Peptides Derived from CJ-15,208: from dual
KOR/MOR affinity profile to selective interactions for
MOR, DOR, or KOR
Kratom Tea as a therapeutic agent for opioid
dependence
Gene mapping and validation of a functional intronic
variant in Gabra2 underlying reduced Gabra2
expression, increased naloxone aversion andincreased methamphetamine stimulant sensitivity
46

T36

Kabirullah

Lutfy

T37

Naoko

Kuzumaki

T38

Indrajeet

SharmaT39

Mark

Nilges

T40

FrederickTejadaT41

Jean K.

Rivera Irizarry

T42

Hiroshi

Ueda

T43

AlexisBaileyT44

Masahiro

Shibasaki

T45

Myurajan

Rubaharan

T46

Ariel

Ben-Ezra

T47

Alexander

Gillis

T48

Lih-Chu

Chiou

T49

Kate

Brown

T50

Brian

Trieu

T51

AnnikaThorsell

T52

Ariel

Amgott-Kwan

The role of endogenous PACAP and dynorphin innicotine-induced conditioned place aversion
Changes in the expression of opioids and opioid
receptors in the sensory neurons of fibromyalgia andParkinson’s disease patients using disease-specific
iPS cells
Structure–Activity Relationships on Collybolide:
Discovery of a Potent Kappa-Opioid Agonist with
Enhanced Metabolic Stability
Effects of Cannabinoid and Opioid Combinations on
the Learning and Performance of Response
Sequences in Rhesus Macaques
Opioid Awareness Among UMES Pharmacy Students
Anterior Paraventricular Thalamus to Nucleus
Accumbens Projection is Involved in Reward-seeking
Behaviors
Chronic pain status and loss of morphine analgesia
have independent brain mechanisms in mice withfibromyalgia-like generalized pain
Effect of A1 vs A2 Milk Casein Exposure at an Early
Developmental Age on the Endogenous Opioid
System of the Rat Brain
Identification of cells in the ventral tegmental area
activated by systemic treatment with opioids andmethamphetamine
The impact of age, sex, and chronic pain on microgliarelated neuroinflammation in the rat Periaqueductal
Gray
The Effect of Kappa Opioid Receptor MediatedNalfurafine on Cocaine Self-administration in MiceLow intrinsic G-protein efficacy, not reduced ß-arrestin
2 signaling, as a mechanism for improved opioid
analgesics
Acupuncture relieved neuropathic pain in opioidtolerant mice via orexin-initiated & endocannabinoidmediated disinhibition in the periaqueductal grayComparison of cortisol serum levels in stressminimized normal volunteers and drug-free former
cocaine dependent volunteers
Modulation of accumbal synaptic transmission by theenkephalin heptapeptide MERF (Tyr-Gly-Gly-Phe-MetArg-Phe)
The OPRM1-A118G modulates reward sensitivity to
opiates as well as the biased MOR agonist TRV130
Lack of withdrawal aversion and drug-induced
reinstatement of extinguished morphine conditioned
place preference by endomorphin analog ZH853.

47

WEDNESDAY POSTERS
ID

First name

Poster Title

Last name

W1

Einat

Peles

W2

CarinaChen

W3

Daisuke

YamadaW4

Jing-Gen

Liu

W5

Sarah

Simmons

W6

Matthew

Hearing

W7

NicoleArguelles

W8

Vishakh

Iyer

W9

Sweta

Adhikary

W10

Bryan

Sears

W11

KelseyKochan

W12

Courtney

Bouchet

W13

Keith

Olson

W14

Michael

Emery

Polymorphisms in three stress-related
genes are associated with reduced cocaine
abuse and longer retention in methadone
maintenance treatment (MMT)
Age trajectory of exposure to cannabis andalcohol, in volunteers diagnosed with opioid
dependence, opioid and cocaine
dependence, or cocaine dependence
KNT-127, a selective agonist for deltareceptors, modulates excitatory synaptictransmission in the medial prefrontal cortex
in mice
Activation of k opioid receptor-induced
mGluR2/3 reduction in prelimbic cortex
contributes to acute stress-induced social
avoidance
Endogenous Kappa Opioid Systemregulation of discrete populations of Lateral
Habenula neurons.
Sex-Dependent Opioid-Induced Plasticity inprefrontal-accumbens circuits: Implications
for Cognitive Control and Relapse
Sex/Cycle differences in CNS oxycodone
metabolism and resulting analgesia: A
potential role for brain CYP2D
Negative allosteric modulation of CB1
cannabinoid receptor signaling suppresses
opioid-mediated reward and dopamine
effluxReceptor, Cellular, and Synaptic
adaptations after chronic morphine
treatment
In Vivo Evaluation of AMB67 and AAH8,
Mixed Efficacy MOR Agonist/DOR
Antagonist Ligands: An Alternative
Approach for Pain Management
Effects of a G-protein βγ subunit inhibitor ondelta opioid receptor (DOR) mediatedbehaviors
Presynaptic Opioid and Cannabinoid
Receptor Signaling Within the Descending
Pain Modulatory Pathway
Naltrexone C7-Esters with Improved
Nociceptin Agonism as Potential Analgesics
with Reduced Rewarding Effects
Differential behavioral responses tomorphine in rats selectively bred for highversus low response to novelty
48

W15

Kristyn

Borrelli

W16

ShahinSaberi

W17

Alexandra

Conibear

W18

Yuki

MoriyaW19

DamienJullie

W20

SaadetInan

W21

Saadet

Inan

W22

Achla

Gupta

W23

Akiyoshi

SaitohW24

WakakoFujitaW25

PreetiManandhar

W26

Anushree

Karkhanis

W27

Nillu

Ghosh

W28

Felix

Effah

W29

Richardvan Rijn

W30

Suda

Yukari

W31

Frida

Stam

Neonatal morphine administration inoutbred CFW mice induces behavioral signsof withdrawal and delayed development
Small molecule inhibitors of PSD95–nNOS
protein–protein interactions suppress
opioid-induced reward in rats
A G protein-biased δ opioid receptor agonist
with improved therapeutic profile
Opioid system is involved in modulating thedevelopment of behavioral sensitization to
METH
A discrete presynaptic vesicle cycle forneuromodulator receptors
Nalbuphine, a kappa opioid receptor agonist
and mu opioid receptor antagonist,
suppresses scratching in mice induced byHIV-1 TAT and also by chloroquine, an antimalarial agent
Nalbuphine suppresses deoxycholic acidinduced scratching in male, Swiss Webster
mice
A role for an endocytic peptide processing
enzyme in modulating opioid receptor
trafficking and function
KNT-127, but not SNC80, facilitates
extinction learning of contextual fear
memory via delta opioid receptor in miceRole of an endogenous chaperone protein,
RTP4, on MOPr-DOPr Heterodimer
upregulation.
Does Pregabalin or Gabapentin modulate µopioid receptor signaling?
Dysregulation of KOR-DAT interaction in theNAc mediates enhanced cocaine seeking
following adolescent social isolation in rats
Morphine induced gastric pathology isprevented by proton pump inhibitor
Effect of Prolonged Milk-Casein Exposure
During Early Development on Mood and mOpioid Receptor Density (MOPr) of
Conventional and Germ-Free Rats: An
Investigation of Diet x Gut Microbiota-Brain
Axis Interaction
Structural analysis of the G protein-biased
delta opioid peptide rubiscolin to probe thereceptor’s ‘biased’ binding pocket
Parkinson’s disease-related overexpressionof COMT in nigrostriatal dopaminergic
neurons inhibits μ-opioidergic systems
Commonly Used Prescription Opioids
Induce Membrane Damage and49

Mitochondrial Dysfunction in the NeuronalLike NG108-15 Cells
W32

AndreaBedini

W33

Yong

Zhang

W34

Jenny

He

W35

Kabirullah

Lutfy

W36

Katarzyna

TargowskaDuda

W37

Mark

Presker

W38

Fabio

BelliaW39

Julia

Brynildsen

W40

Cynthia WeiSheng

Lee

W41

Mariana

Rae

W42

Kerri

Pryce

W44

Neil

Varshneya

W45

Bernard P.

Roques

W46

Emily

Leff

W47

MiriamBarnett

W48

Katherine

Martucci

Functional Selectivity and Analgesic Effectsof CL39, a Novel Kappa Opioid Receptor(KOR) Selective Agonist
Effects of nalfurafine on oxycodone induced conditioned place preference andlocomotor activity in adolescent male andfemale C57BL/6J mice
Mechanisms of presynaptic suppression of
GABA release by opioid receptors in the
hippocampus
The role of endogenous opioids inamphetamine-induced locomotor
sensitization
NOP receptors system in migraine
Morphine and cocaine cues differentiallycontrol natural reward motivated behavior:role of locus coeruleus.
Alcohol consumption during pregnancy
induces altered transcriptional regulation inmothers and offspring partially counteracted
by environment enrichment
Functional network connectivity in opiatenaïve and opiate-dependent states
Cellular Mechanisms of Metabolic
Adaptations Mediated by μ-Opioid andNociceptin Receptors
Effects of Environmental Enrichment on
Alcohol and Social Reward and Modulationof The Oxytocinergic System
Oxycodone-induced gene expression
adaptations in the brain reward center in amurine model of neuropathic pain
The effects of N-phenyl-fluorinatedbutyrylfentanyl regioisomers in mice
Exaltation of natural opioids enkephalinsfunctions by inhibiting their synaptic
peptidases inactivation in painful anddepressive conditions. Extension to
migraine
GRK2/3 dependent desensitization of the
mu-opioid receptor is eliminated following
chronic morphine treatment
Differential signaling patterns of the kappa
opioid receptor through specific Gα and Gγ
subunits

Examining the Effects of Opioid Use on
Neural Responses to Monetary Incentives in
Chronic Pain
50

W49

CarleeToddesW50

Jennifer

Whistler

W51

Kelly

Standifer

W52

YusukeHamadaThe social brain and mu: Modulation of social
behavior by mu opioid receptors in the brain
reward network
Balanced opioids provide excellent
analgesia and reduced side effects
Sex differences in Nociceptin/Orphanin FQ
Peptide (NOP) Receptor knockout rat
response to traumatic stress: Tumor
Necrosis Factorα (TNFα) andprepronociceptin (PNOC) mRNA
Possible involvement of μ-opioid receptorpositive neurons located in the PVN in thecontrol of the tumor-bearing state

Poster abstracts on pages 52 to 128 are in alphabetical order based on presenter’s last name.

51

Presenter – Adhikary, Sweta
Abstract ID – W9
Receptor, cellular, and synaptic adaptations after chronic morphine treatment
Sweta Adhikary1, Stefan Schulz2, John T. William1, William T. Birdsong3
1
OHSU/ Vollum Institute, USA, 2Institute of Pharmacology and Toxicology, Jena University Hospital,
Germany,3University of Michigan Medical School, Department of Pharmacology, Ann Arbor, USA
Opioids produce both pain relief and reward through agonist activity at the µ-opioid receptor (MOR).
Long-term use results in the development tolerance whereby increased dosage of drug is required
to maintain therapeutic efficacy. MOR phosphorylation is a key molecular step that mediates
tolerance after chronic opioid treatment in the somato-dendritic compartment of neurons, butmechanisms underlying tolerance in the presynaptic compartment are unknown. Phosphorylationdeficient mice with alanine mutations of 10 serine (S) and threonine (T) residues in the C-terminusof MOR (10S/T-A) were used to examine the role of phosphorylation in mediating tolerance in thepresynaptic compartment. Glutamate projections from the medial thalamus to the dorsal striatum
were isolated using an optogenetic approach. AAV2-CsChR was injected into the medial thalamus
and optically evoked thalamo-striatal EPSCs (oEPSCs) were recorded in medium spiny neurons(MSNs) in striatal brain slices. Chronic opioid treatment decreased morphine-induced inhibition of
oEPSCs in WT mice but this decrease in inhibition was less in 10S/T-A mice, suggesting a role forphosphorylation in mediating receptor adaptations to chronic treatment. There was also greater tonic
inhibition of thalamo-striatal synapses by adenosine in 10S/T-A mice compared to WT, suggesting
increased cellular and synaptic adaptations in 10S/T-A mice. Additionally, adenosine tone in the
striatum was evaluated using electrical stimulation of striatal afferents, and optogenetic stimulationof cortico-striatal afferents in both naïve and chronically treated animals. Increased adenosine tone
developed in approximately 50 percent of MSNs, suggesting a role of postsynaptic MORs in
modulating adenosine in the striatum in a cell-type specific fashion.
Presenter – Afrose, Leela
Abstract ID – T16
Interaction of a newly deorphanized G protein-coupled receptor, GPR171, in chronicmorphine treatment
Leela Afrose1, Max McDermott1, Erin N. Bobeck1
1
Department of Biology, Utah State University, Logan, USA
Opioid analgesics, such as morphine, remain a common pain therapeutic despite having unwantedside effects with repeated use. Therefore, the need for novel therapeutics is crucial. Recently our lab
has found the newly deorphanized G protein-coupled receptor, GPR171 as a regulator of morphine
antinociception. The purpose of this study was to evaluate GPR171 in morphine induced tolerance
and withdrawal. GPR171 was found to be highly expressed within the Periaqueductal Gray area
which is known to be involved in opioid tolerance. To test the role of GPR171 in morphine tolerance
and withdrawal mice were injected twice daily for 4 days with GPR171 agonist or antagonist or DMSO
10 min prior to injections of morphine or saline. On day 5, cumulative quarter log doses of morphine
were administered and mice were tested on the hotplate and tail flick tests. We observed that micetreated with the GPR171 antagonist had no effect on morphine tolerance and it is expected that theagonist will reduce tolerance given its enhanced morphine antinociception during acute treatment.
Mice were treated similarly on days 1-4 to investigate this receptor’s role in morphine withdrawal. On
day 5, mice were injected with their designated drug combination followed by naloxone-precipitatedwithdrawal where naloxone was administered two hours later and withdrawal behaviors wereobserved via video recording. Those mice pretreated with the antagonist, showed a decrease inwithdrawal symptoms. Taken together this data reveals the functional interaction of the mu opioid
receptor and GPR171 during repeated morphine treatment.

52

Presenter – Airavaara, Mikko
Alcohol drinking and craving in female mice housed in automated cages
Maryna Koskela1, Vootele Voikar1, Mikko Airavaara1
1
Neuroscience Center, University of Helsinki, Finland

Abstract ID – M49

We aimed to establish an alcohol craving model in group-housed mice in automated cages withminimal human interference. For this purpose, alcohol drinking training was paired with light cues toestablish conditioning. Alcohol was available in three corners, and one corner was allocated to water.
Water in the water corner was available with opened doors all the time during training, whereas nosepoke was needed to open the door for alcohol drinking. Mice were housed in a group of eight. Ethanol
concentration started from 4%, and on every two days following, the concentration of ethanol wasincreased by 1% until 12% ethanol was reached. Then the mice continued the training for ten days
with access to 12% ethanol. The extinction test was carried out without alcohol but in the presenceof light cues and empty bottles. We measured the number of visits, nose-pokes, and licks. We foundthat the number of visits and nose-pokes were much higher in alcohol group as compared to thewater group in conditioned stimuli (CS)+ corner, and no difference is observed in CS- corner during
training. Also, during training, the alcohol dose was steadily increasing. In the extinction tests, wefound that number of nose-pokes in the CS+ corner was increased in alcohol group on bothwithdrawal days 1 and 10 whereas no difference was observed in the number of visits between thegroups. We conclude that we have established the model to study alcohol drinking behavior andcraving in female mice.
Presenter – Alder, Amy
Abstract ID – T21
Preclinical evaluation of the novel mu opioid receptor agonists Kurkinorin and Kurkinol
Amy F Alder1, Nirajmohan Shivaperumal1, Rachael S Crowley2, Samuel Williamson2, Thomas E
Prisinzano2, Bronwyn Kivell1
1
Victoria University of Wellington, Wellington, New Zealand, 2The University of Kansas, Lawrence,
Kansas, USA
Chronic pain (CP) is a complex and difficult to treat condition that affects 1 in 10 Americans. It is aleading cause for accessing healthcare, costing $635 billion USD annually. Currently, first line
therapeutics include opioids such as morphine and fentanyl that target the mu opioid receptor (MOR).
These compounds are limited by side effects induced by the recruitment of β-arrestin, including
tolerance, respiratory depression and addiction. Generating compounds with reduced β-arrestin
recruitment provides an avenue for the development of non-addictive opioids for CP. Here we show
the novel MOR agonists Kurkinorin and Kurkinol have reduced abuse liability and induce potent
analgesic effects in multiple rodent models of anti-nociception. In the dose-response tail flick assay
Kurkinorin (ED50 = 5 mg/kg) was equipotent with Morphine (ED50 = 5.3 mg/kg), while Kurkinol had
increased potency (ED50 = 2.35 mg/kg). Both show a similar duration of action to morphine in tailflick and hotplate assays. In the Paclitaxel-induced neuropathic pain model, Kurkinol was potent at
reducing mechanical (ED50 = 1.5 mg/kg) and cold allodynia (ED50 = 1.74 mg/kg) while Kurkinorin(mechanical ED50 = 9.2 mg/kg; cold ED50 = 14.82 mg/kg) was equipotent with morphine(mechanical ED50 = 9.2 mg/kg; cold ED50 = 9.03 mg/kg). Evaluation of the side effect Profiles
showed both have reduced abuse liability in conditioned place preference, while Kurkinorin also
showed reduced respiratory depression in unrestrained whole body plethysmography. Overall
Kurkinorin and Kurkinol both show promise as potent non-addictive analgesics for the treatment of
acute thermal and neuropathic pain.

53

Presenter – Amgott-Kwan, Ariel
Abstract ID – T52
Lack of withdrawal aversion and drug-induced reinstatement of extinguished morphine
conditioned place preference by endomorphin analog ZH853.
Ariel Amgott-Kwan1, James Zadina1,2,3,4
1
Tulane Brain Institute, USA, 2Department of Medicine Tulane University, USA, 3Department ofPharmacology Tulane University, USA, 4SE Louisiana Veterans HCS, New Orleans, USA
Opioids provide potent analgesia. However, their long-term use is limited by side effects including
abuse liability, tolerance, and drug dependence. We previously demonstrated the reduced abuseliability of endomorphin analog ZH853, indicated by a lack of conditioned place preference (CPP) or
self-administration. Here we further examine the addiction potential of ZH853 through theassessment of conditioned place aversion (CPA), a sensitive index of the affective-motivational
component of morphine withdrawal, and reinstatement of morphine seeking. CPA measures theaversive state associated with naloxone-precipitated withdrawal. Naloxone produced an aversive
state in morphine-treated male and female rats but not in those treated with equi-antinociceptive
doses of ZH853. We also examined reinstatement of morphine CPP elicited by priming injections of
intravenous morphine or ZH853 in male rats. Following extinction of morphine CPP, rats werechallenged with a priming dose of morphine or an equi-antinociceptive dose of ZH853 and tested forreinstatement of drug-seeking behavior. Morphine, but not ZH853, reinstated morphine CPP. The
ability of ZH853 to provide antinociceptive effects while not contributing to relapse of previous drugseeking would support its potential use as a safer pain treatment for patients previously addicted to
opioids, and as a potential novel opioid maintenance therapy for opioid use disorder. These findings
further support the reduced potential of ZH853 to be abused by demonstrating that ZH853 shows
less negatively reinforcing properties and relapse vulnerability as compared to morphine.
Presenter – Anand, Jessica
Abstract ID – T13
Structure activity relationships of illicit fentanyl analogs
Jessica P. Anand1,2, Carmelita M. Naccarato1, Jack J. Twarozynski1, Sherif Hassanien3, Jonathon
Bassman3, Donna Iula3, John R. Traynor1,2
1
University of Michigan, Ann Arbor, USA, 2Edward F. Domino Research Center, Ann Arbor, USA,
3
Cayman Chemical, Ann Arbor, USA
The number of drug seizures and overdose deaths involving fentanyl and its analogs has risen
steeply since 2016 and as a result emergency scheduling for fentanyl analogs was put into place inlate 2017. Fentanyl is a highly potent and efficacious mu opioid receptor (MOR) agonist withsignificant abuse potential. However, little is known about the novel fentanyl analogs (fentalogs)
flooding the illicit drug market. It is presumed that these novel fentalogs are also potent, highly
efficacious MOR agonists and they are driving the increase in opioid overdoses, but pharmacological
data on these compounds are scarce. We synthesized a library of fentalogs organized aroundcompounds commonly found in drug search and seizure samples. We determined their affinity for
and agonist activity at MOR in vitro using competitive ligand binding and GTPγS binding assays. We
found that structure greatly alters binding affinity and agonist activity; in this library, affinity at MOR
ranged from 0.4 nM to almost 400 nM – fentanyl displays 1.6 nM affinity for MOR. Based on the datacollected from this library, we have been able to construct structure-activity relationships which may
allow us to predict which fentalogs are potent agonists and are likely to carry significant abuseliability.
Supported by DA 048129 and resources provided by the Edward F. Domino Research Center.

54

Presenter – Arguelles, Nicole
Abstract ID – W7
Sex/cycle differences in CNS oxycodone metabolism and resulting analgesia: A potential rolefor brain CYP2D
Nicole Arguelles1,2, Fariba Baghai Wadji2, Bin Zhao1,2, Sharon Miksys1,2, Rachel Tyndale1,2,3
1Campbell Family Mental Health Research Institute, CAMH, Canada, 2Department of Pharmacology& Toxicology, University of Toronto, Canada,3Department of Psychiatry, University of Toronto,
CanadaSex differences are recognized in opioid analgesia and dependence, in rodents and humans. Numerous
opioids are CYP2D enzyme substrates and variation in CYP2D alters opioid response. We havepreviously shown that inhibiting brain CYP2D in male rats increased oxycodone analgesia and brain, notplasma, levels. Here we explored sex/cycle differences in oxycodone-induced analgesia, drug levels, andbrain CYP2D. Male and female (in diestrus and estrus) rats were administered oxycodone orally (7.5mg/kg). Propranolol (20 µg), a CYP2D-specific mechanism-based inhibitor, or vehicle was delivered into
cerebral ventricles 24 hours before oxycodone testing. Oxycodone-induced analgesia was measured
using tail-flick latency and brain drug and metabolites levels were measured in vivo by microdialysis.
Analgesia and brain drug levels were analyzed using AUCs(0-120min) by one-way or two-way ANOVA, with
post-hoc analyses. Females were each tested in diestrus and estrus phases with evident cycle effects;
analgesia was higher (p<0.001) during diestrus than during estrus. Between rats, females in diestrus had
1.6-times higher analgesia (p<0.0001) and 1.8-times higher brain oxycodone levels (p<0.01) than females
in estrus, and 2.0-times higher analgesia (p<0.0001) and 3.2-times higher brain oxycodone levels(p<0.001) than males. Analgesia correlated with brain oxycodone (R2=0.67, p<0.0001) but not
oxymorphone levels. Propranolol (vs vehicle) increased oxycodone analgesia and brain levels in males(1.8-fold, p<0.001), females in estrus (1.4-fold, p<0.001), and females in diestrus (1.1-fold). Plasma
oxycodone levels were unchanged. These findings demonstrate that both sex and estrous cycle impact
oxycodone analgesia and brain levels, likely through differences in brain CYP2D oxycodone metabolism.

Presenter – Atwood, Brady
Abstract ID – M20
Mu opioid receptors on vGlut2-expressing glutamate neurons regulate aversion tooxycodone
Megan Kube1, Brandon Fritz1, Braulio Munoz1, David Haggerty1, Kaitlin Jones1, Fuqin Yin1, Brady
Atwood1,2
1Indiana University, USA, 2Stark Neurosciences Research Institute, Indianapolis, USA
Prescription opioid drugs such as oxycodone produce reward largely through mu opioid receptors(MORs). We previously demonstrated that oxycodone alters the ability of MORs to modulate
corticostriatal glutamatergic transmission. We also found that MORs regulate thalamostriatal
glutamatergic transmission. Using conditional MOR knockout mice (MORflox-vGlut2cre), we now
demonstrate the localization of these MORs as being in vGlut2-expressing thalamic neurons. Whereas
our previous work found that MORs are expressed on small populations of corticostriatal glutamatergic
neurons, indicating a likely more behavioral restricted role of these receptors, MORs are more broadly
expressed in vGlut2 neurons in the brain. Therefore, we decided to explore the role of MORs on vGlut2
neurons in oxycodone-related behaviors. Adult male and female MORflox-vGlut2cre and wild-type
littermate controls underwent behavioral tests including oxycodone-induced conditioned place preference(CPP), home cage 2-bottle choice drinking, and locomotion testing. Whereas wild-type mice showed
CPP to oxycodone, MORflox-vGlut2cre mice displayed conditioned place aversion. MORflox-vGlut2cre
mice also did not show oxycodone-induced increases in locomotor behavior. In 2-bottle choice oxycodone
drinking, MORflox-vGlut2cre mice consumed less and showed less preference for oxycodone compared
to controls. These data reveal for the first time that MORs on vGlut2-expressing neurons play a significant
role in opioid reward circuitry, specifically modulating aversion to oxycodone. Current work is determining
what other opioid-related behaviors are mediated by MORs on this class of glutamatergic neurons andidentifying the specific brain regions in which these neurons reside that mediate these behaviors.

55

Presenter – Bailey, Alexis
Abstract ID – T43
Effect of A1 vs A2 milk casein exposure at an early developmental age on the endogenous
opioid system of the rat brain
Aya Osman1, Elizabeth Fagbodun2, Simone Zuffa3, Ian Kitchen1, Jonathan Swann3, Alexis Bailey2
1
School of Biosciences and Medicine, Faculty of Health and Medical Sciences, University of Surrey,
Guildford, Surrey, UK, 2Pharmacology Section, St George's University of London, London, UK,
3
Imperial College London, South Kensington Campus, London, UK
Nutrition can affect early brain development leading to changes in behaviour. We showed that prolonged
milk casein exposure post-weaning in rats affects the central opioid system and results in a depressivelike phenotype. Casein metabolite beta-casomorphin-7 (BCM-7) is biologically active. It acts on mu opioid
receptors (MOPr) and is implicated in neurodevelopmental deficits. Bovine milk has two major types ofcasein; the A1 type which yields BCM-7 and the A2 type which does not. We assessed the effect of
prolonged exposure of rats to A1 casein milk or A2 milk during early post-weaning age, on mood, thecentral opioid receptor system and frontal cortex metabolome. Rats were given either A1 or A2 milk, orno milk from post-natal day (PND) 21 until PND26. Depressive-like behaviour was assessed with the useof the forced swim test and brains removed for autoradiographic assessment of MOPr. NMR
metabolomic analysis was also performed in the frontal cortex. Exposure to both milks induced adepressive-like phenotype. However, only exposure to A1 milk caseins led to increases in MOPr binding
in the somatosensory and motor cortex and hippocampus. No alterations were observed in rats exposed
to A2 milk. Twenty seven metabolites were raised in the frontal cortex of rats fed an A1 but not A2 milk
diet including GABA, glutamate and glutamine. The findings suggest that ingestion of BCM-7 at an early
developmental age modulates the central opioid receptor system and induces broad changes in brain
metabolomic profile which may impact on behavior.
Presenter – Balogh, Mihály
Abstract ID – T1
High efficacy opioids compared to morphine produce promising analgesia in advanced diabetic
neuropathic pain in rats
Mihály Balogh1, Ferenc Zádor2, Zoltán S. Zádori1, Shaqura Mohammed3, Kornél Király1, Amir
Mohammadzadeh1, Bence Varga1, Dávid Karádi1, Bernadette Lázár1, Shaaban A. Mousa3, Sándor
Hosztafi4, Pál Riba1, Sándor Benyhe2, Michael Schäfer3, Susanna Fürst1, Mahmoud Al-Khrasani1
1
Department of Pharmacology and Pharmacotherapy, Faculty of Medicine, Semmelweis University,
Budapest, Hungary, 2Institute of Biochemistry, Biological Research Center of the Hungarian Academy of
Sciences, Szeged, Hungary, 3Department of Anesthesiology and Intensive Care Medicine, Charité
University Berlin, Campus Virchow Klinikum and Campus Charite Mitte, Berlin, Germany, 4Department ofPharmaceutical Chemistry, Semmelweis University, Budapest, Hungary
In this study we (i) Assessed the antiallodynic effect of 14-O-methylmorphine-6-O-sulfate (14-OMeM6SU), a novel opioid of high efficacy, in rats with DNP, after systemic administration, (ii) Investigatedthe peripheral component in the antinociceptive actions, and (iii) Analyzed receptor functions at CNSlevels. Fentanyl and morphine were used as reference drugs. Diabetic polyneuropathy was induced by
ip. streptozocin (STZ) injection in male Wistar rats. Mechanical allodynia was assessed by dynamic
plantar aesthesiometer prior to and weekly after STZ injection for 12 weeks. Experiments were performedat the 9th week. The role of peripheral opioid receptors in antinociception was investigated by the coadministration of naloxon methiodide (NAL-M, opioid antagonist with limited CNS penetration). For in vitro
analysis G-protein functioning and immunohistological assays were performed. 14-O-MeM6SU andfentanyl displayed similar antinociception in diabetic and non-diabetic rats. However, a 7-times
impairment in morphine antinociception was measured. NAL-M failed to alter the antiallodynic effect oftest compounds, indicating the contribution of central opioid receptors. Significant impairment in G-protein
coupling was measured for morphine, yet no change in case of 14-O-MeM6SU or fentanyl at spinal levelof diabetic- and non-diabetic animals. Furthermore, reduction in MOR immunoreactivity of nerve terminals
was detected in the spinal cord and dorsal root ganglia of diabetic rats. The reduction of spinal opioid
receptors concomitant with reduced analgesic effect of morphine may be circumvented by prescribing
high efficacy opioids, which provide superior analgesia over morphine.

56

Presenter – Barnett, Miriam
Abstract ID – W47
Differential signaling patterns of the kappa opioid receptor through specific Gα and Gγ subunits
Miriam E. Barnett1, Brian I. Knapp1, Jean M. Bidlack1
1
University of Rochester Medical Center, Rochester, USA
The kappa opioid receptor (KOR) receives and converts extracellular cues into precise intracellularsignaling events by coupling to heterotrimeric G proteins, Gα•βγ. It is not understood how the couplingwith the different members of the Gαi class, including Gαi1, Gαi2, Gαi3, GαoA, GαoB and Gαz, affects
KOR signaling. Additionally, the Gγ subunits, Gγ1 and Gγ2, have different dissociation rates and may
affect KOR signaling. Thus, the milieu of G proteins within a particular cell type may greatly influence thepharmacological properties of opioid ligands. Using a bioluminescent resonance energy transfer (BRET)
sensor, the effect of various Gα and Gγ subunits on KOR signaling was studied. In this novel system,changes in opioid-induced interactions of Venus-tagged Gβγ with its effector G protein-coupled receptor
kinase 3 tagged with Renilla luciferase were characterized after a 30-sec and 50-min incubation. We
observed that various agonists, partial agonists, and antagonists activate the KOR to different extents
depending on which Gα protein was coupled. For example, dynorphin A (1-17) was a full agonist
regardless of which Gα or Gγ protein was coupled. In contrast, partial agonists, such as the
benzomorphans, cyclazocine and (-)pentazocine, and the morphinan nalmefene, activated the KOR to
the greatest extent when signaling through Gαz. These findings suggest that the composition of the Gα
and γ subunits coupled to the KOR influences the pharmacological properties of opioids.
Supported by the Margo Cleveland Fund and John R. Murlin Fund.
Presenter – Bart, Gavin
Abstract ID – M44
Altered methadone pharmacokinetics: impact of antiretrovirals and an independent effect of HIV
Gavin Bart1,2,3, Le Minh Giang4, Huu Anh Nguyen4, Linnae Biard3, Carnayla Johnson3, Richard C.
Brundage5
1Hennepin Healthcare, USA, 2University of Minnesota School of Medicine, USA, 3Hennepin Healthcare
Research Institute, USA, 4Center for Training and Research on Substance Use-HIV, Hanoi Medical
University, Vietnam, 5University of Minnesota College of Pharmacy, USA
Drug-drug interactions between methadone and antiretrovirals such as efavirenz increase methadone
clearance and potentially destabilize patients, which may lead to medication non-adherence andincreased risk behavior. We used population pharmacokinetics to evaluate methadone-efavirenz
interactions in a real-world methadone-maintained population. Plasma levels of R- and S-methadone
were measured approximately 4 and 24 hours after daily methadone dosing in 308 methadonemaintained patients. A linear one-compartment population pharmacokinetic model with first-orderconditional estimation with interaction (FOCE-I) was used to evaluate methadone CL/F, V/F, and F. Thesecan be separated in a population analyses because increases in CL will decrease AUC and shorten halflife, whereas decreases in F will decrease AUC but not change half-life. The influence of covariates HIV
status, efavirenz, gender, weight, BMI, age, and methadone dose on parameter estimates was evaluatedusing stepwise covariate modeling. Of 308 patients (male=292), 155 were HIV-negative and 153 HIVpositive. Mean (SD) methadone dose was 65 (35) mg in the HIV- group and 135 (70) mg in the HIV+
group. Base model CL and Vd for R methadone were 6.29 L/h and 379 L, respectively. Efavirenz
increased R-methadone CL by 79% and HIV reduced F by 29%. Efavirenz increased S-methadone CL
by 113% and HIV reduced F by 34%. This is the first real-world clinical study showing efavirenz increases
methadone clearance. It is the first to identify an effect of HIV on methadone oral bioavailability in a realworld clinical population. These results may allow for future therapeutic drug monitoring in methadone
patients.

57

Presenter – Bedini, Andrea
Abstract ID – W32* Hot Topics Selection, Symposium 3, Tuesday, July 9 AM
Functional selectivity and analgesic effects of CL39, a novel kappa opioid receptor (KOR)
selective agonist
Andrea Bedini1, Lorenzo di Cesare Mannelli2, Laura Micheli2, Rossella De Marco3, Luca Gentilucci3, Carla
Ghelardini2, Santi Spampinato1
1
Department of Pharmacy and Biotechnology - University of Bologna, Italy, 2Department of Neuroscience,
Psychology, Drug and Children Health - University of Florence, Italy, 3Department of Chemistry G.
Ciamician - University of Bologna, Italy
Kappa opioid receptor (KOR) agonists determine antinociception by G protein signaling; conversely, they
trigger relevant side effects through arrestin 3-dependent p38MAPK activation. Thus, raising significant
interest in developing G protein biased KOR agonists as innovative analgesics. Recently, we identified
CL39 (H-Tyr-Amo-Trp-PheNH2), an endomorphin-1 analogue with high affinity, selectivity and partial
agonist activity at KOR; here we investigate its functional selectivity and analgesic effects as comparedto U50,488. CL39 weakly recruited arrestin 3 at KOR, as assessed via arrestin complementation assay
in U2OS cells and by means of a BRET assay in HEK-293 cells; calculated bias factor for U50,488 and
CL39 were equal to 1 and 65, respectively. CL39 inhibited adenylyl cyclase and activated early (5-15min),G protein-dependent component of ERK1/2 phosphorylation in HEK-hKOR and U87-MG cells, asassessed by ELISA and western blot, but did not trigger in the same cells late (60min), arrestin-dependent
component of ERK1/2 or p38MAPK phosphorylation. On the contrary, U50,488 activated all the above
mentioned signaling pathways. Moreover, U50,488, but not CL39, significantly increased U87-MG cellproliferation in an arrestin 3-, p38MAPK-dependent fashion, as assessed by [3H]-thymidine incorporation.
Experiments on normal human astrocytes are ongoing and will be presented at the conference. In vivo,
both CL39 and U50,488 (0-30 mg/kg; 0-60 min; i.p.) induced a significant, KOR-mediated, dosedependent antinociception in warm-water tail-withdrawal test, displaying the ligands comparable ED50.
CL39-mediated analgesia in a model of oxaliplatin-induced neuropathic pain is currently underinvestigation. CL39 is a novel, G protein biased KOR agonist displaying analgesic effects in vivo.
Presenter – Beier, Kevin
Abstract ID – M3
Identification of neuronal ensembles that promote opiate addiction
Kevin Beier1,2, Christina Kim3, Paul Hoerbelt1, Lin Wai Hung1, Boris Heifets1,4, Kathy DeLoach2,7, Timothy
Mosca2, Sophie Neuner1, Karl Deisseroth5,6,7, Liqun Luo2,7, Robert Malenka1
1
Nancy Pritzker Laboratory, Stanford University School of Medicine, Stanford, USA, 2Department of
Biology, 4Department of Anesthesiology, Perioperative and Pain Medicine, 5Department of
Bioengineering, 6Department of Psychiatry and Behavioral Sciences, Stanford University School of
Medicine, USA, 7Howard Hughes Medical Institute, Stanford University, Stanford, USA
Identification of neural circuit changes contributing to behavioral plasticity has routinely been conductedon candidates that were selected based on past results. We recently developed an unbiased method for
identifying experience-triggered circuit-level changes in neuronal ensembles. Using rabies virus
monosynaptic tracing we mapped cocaine-induced global input changes onto ventral tegmental area
neurons, and identified a novel drug-induced enhancement of activity in parvalbumin-positive globus
pallidus neurons. We then demonstrated using inhibitory manipulations that these GPe neurons play avital role in two early forms of both opioid and psychostimulant-triggered behavioral plasticity, locomotor
sensitization and conditioned place preference. This behavioral inhibition was affected, at least in part,by GPe-mediated local disinhibition of dopamine neuron activity. This modification in GPe neuron activity
likely represents the initial adaptation in a cascade of changes that drive drug addiction. To follow up on
this finding, we are now using a suite of unbiased technologies, including the rabies virus activity screen
as well as automated analyses of brain-wide activity in cleared tissue, and are examining different points
during the progression to addiction to identify how activity in neural ensembles is modulated during
progressive opiate administration and relapse. Our ultimate goal is to elucidate how each of thesechanges contributes to the development of opiate addiction specifically, and to motivated behavior moregenerally.

58

Presenter – Beierle, Jacob
Abstract ID – T15
Mouse inbred strain survey of oxycodone addiction traits in an opioid multi-stage addiction
assessment paradigm
Jacob A Beierle1,2,3, Juila Scotellaro1,4, Emily Yao1, Julia C Kelliher1, Richard K Babbs1, Ming Zheng5,
Gray Peltz5
1
Laboratory of Addiction Genetics, Department of Pharmacology and Experimental Therapeutics andPsychiatry, 2T32 Biomolecular Pharmacology Training Program, 3Transformative Training Program in
Addiction Science, Boston University School of Medicine, USA, 4Undergraduate Research Opportunity
Program (UROP), Boston University, USA, 5Department of Anesthesiology, Perioperative, and Pain
Medicine, Stanford University School of Medicine, USA
Opioid use disorder (OUD) is a heritable addictive disorder with a largely unknown genetic basis. Murine
forward genetic studies of addiction traits can identify novel genetic factors and biological pathways
relevant to humans, potentially leading to improved therapeutics. We are conducting a large strain survey
in a panel of inbred mouse strains for oxycodone (OXY)-induced behaviors using our multi-stage addiction
assessment protocol (MSAAP). Our long-term goal is to conduct high resolution haplotype association
mapping of the genetic basis of opioid addiction traits. These traits include acute locomotor response,
locomotor sensitization, conditioned place preference (CPP), extinction, antinociceptive tolerance,
spontaneous anxiety-like withdrawal, and several protracted withdrawal phenotypes. We have thus far
tested 20 inbred strains (A/J, ALR/LtJ, AKR/J, BALB/cJ, BALB/cByJ, BPH/2J, BTBR T+ Itpr3tf/J, BUB/BnJ,
C3H/HeJ, C57BL/6J, CBA/J, DBA/1J, DBA/2J, FVB/J, LG/J, MRL/MpJ, NOD/ShiLtJ, NZL/LtJ, SB/LeJ,
SM/J). The large phenotypic variance in these behaviors across strains, and high narrow-sense
heritability estimates indicate their suitability for haplotype association mapping. Notably, we identified
behavioral differences between BALB/cJ and BALB/cByJ substrains in state-dependent CPP, elevatedplus maze, weight loss, and open field behaviors. These substrain differences are particularly interesting,as their reduced genetic complexity facilitates rapid identification of novel genetic factors underlying
behavioral addiction traits. Complete testing of 30 classical inbred strains will lead to the identification ofhigh resolution genetic loci, candidate genes, and functional variants to be confirmed via gene editing.

Presenter – Bellia, Fabio
Abstract ID – W38
Alcohol consumption during pregnancy induces altered transcriptional regulation in mothers
and offspring partially counteracted by environment enrichment
Bellia F1, Wille-Bille A2, Pucci M1, Miranda-Morales RS2, Pautassi RM2,3, D'Addario C1,4
1
Faculty of Bioscience, University of Teramo, Italy, 2Instituto de Investigación Médica M. y M.
Ferreyra (INIMEC–CONICET-Universidad Nacional de Córdoba), Córdoba, Argentina, 33Facultadde Psicología, Universidad Nacional de Córdoba, Córdoba, Argentina, 4Karolinska Institutet,
Department of Clinical Neuroscience, Center for Molecular Medicine, Stockholm, Sweden
Alcohol consumption by mothers during pregnancy may lead to mental or physical issues for the newborn,as well as be potentially dangerous to themselves after delivery, possibly increasing the risk of alcohol
use due to heightened stress. The different phenotypes occurring in both mothers and offspring might
involve the epigenetic regulation of genes transcription. Using an animal model of prenatal ethanol
exposure, we here studied in mothers postpartum and in their offspring the effects of brain transcriptional
regulation of target genes and how environmental enrichment might modulate possible alterations. The
dams were intragastrically administered once daily with a 16.8% ethanol solution or vehicle. After delivery
litters were divided into two groups and starting from PD14 in both infants and adolescents were evaluatedthe exploratory activity, risk-taking, and anxiety-like behavior. Mothers and offspring were sacrificed after
the behavioral tests by decapitation, brain dissected, nucleic acids extracted for genes expression studies
and genes promoter DNA methylation/hydroxymethylation analysis. Our findings so far show selective
altered expression for BDNF and prodynorphin genes in the VTA of adolescent rats prenatally exposed
to alcohol and of dams exposed to alcohol during pregnancy, whereas environmental enrichment partiallyreverted these changes at least in adolescent offspring. Moreover, altered methylation at specific CpG
sites at both gene promoters was observed consistently with genes transcription’s changes. These data,
even if preliminary, might be promising in order to understand the protective role of the environmental
switch on the effects evoked by alcohol also suggesting molecular mechanisms accounting for it.
59

Presenter – Ben-Ezra, Ariel
Abstract ID – T46
The effect of kappa opioid receptor mediated nalfurafine on cocaine self-administration in
mice
Ariel Ben-Ezra1, Amelia D. Dunn1, Jose Erazo1, Brian Reed1, Mary Jeanne Kreek1
1
Laboratory of the Biology of Addictive Diseases, The Rockefeller University, USA
The kappa opioid receptor (KOR) is a target for the modulation of mood and reward, specifically thetreatment of psychostimulant addiction. Most KOR agonists with CNS activity have aversive and/or
sedative effects. However, nalfurafine is a potent KOR agonist clinically utilized in Japan for pruritus
with few and moderate side effects. The aim of the current study was to determine whether
nalfurafine alters the profile of acquisition of cocaine self-administration behavior. Additionally, theshort-acting KOR antagonist LY2444296 was used to test KOR specificity. Following intrajugularcannulation surgery, adult, male C57BL6 mice (n=36) were given access to 0.5mg/kg/infusion
cocaine infusions in response to FR1 nosepokes during daily 2-hour sessions with immediately priorinjection of saline (group 1) or 10 µg/kg nalfurafine (group 2). Increased intake of cocaine wasobserved in the nafurafine treated group. On days 7 and 8, both groups were treated with saline, andgroup 2 decreased cocaine intake to be commensurate with group 1. Upon reintroduction of
nalfurafine to group 2, cocaine intake was escalated approximately 50%. In a separate experiment,
the nalfurafine-induced increase in cocaine intake was replicated, and was blocked by pretreatment
with subcutaneous 3mg/kg LY2444296 15 minutes before each self-administration session.
Nalfurafine (10 µg/kg) increases intake of cocaine in a mouse self-administration model. Additional
neurochemical studies are needed to determine the mechanism underlying this effect. Further
exploration into KOR modulation of cocaine’s biological effects is warranted.
Supported by the Miriam and Sheldon Adelson Medical Research Foundation and Robertson
Therapeutic Discovery Fund.
Presenter – Bertels, Zachariah
Abstract ID – T12
Differential effects of mu and delta opioid receptor agonists in models of migraine
Zachariah Bertels1, Kendra Siegersma1, Alycia Tipton1, Serapio M. Baca2, Richard van Rijn3,
Amynah Pradhan1
1University of Illinois at Chicago, Chicago, USA, 2University of Colorado at Denver, Aurora, USA,
3
Purdue University, West Lafayette, USA
Mu opioid receptor agonists are still commonly prescribed for migraine despite potential adverse
effects including medication overuse headache/opioid induced hyperalgesia (OIH). Comparatively,delta opioid receptor agonists reduced correlates of migraine allodynia and aura, while showing
limited OIH. Our aim was to develop mouse models reflecting exacerbation of migraine symptoms
by mu activation, and contrasting it to anti-migraine effects of the non-internalizing/non-convulsantdelta agonist, KNT-127. An established model of OIH was used to examine the effect of morphine
on a physiological correlate of migraine aura, cortical spreading depression (CSD). Escalating dosesof morphine or vehicle was given twice daily for 4 days. On day 5, CSD was induced by administrationof KCl onto the dura. Morphine treatment significantly increased CSD events relative to controls. To
model exacerbation of headache-associated pain by mu agonists we investigated the effect ofchronic morphine in a nitroglycerin (NTG) model of migraine. Chronic periorbital allodynia was notobserved following treatment with chronic morphine (20 mg/kg, SC) or low dose of the humanmigraine trigger NTG (0.1 mg/kg IP). However, combination of both treatments resulted in sustained
and severe allodynia. Conversely, the delta agonist, KNT-127 (5 mg/kg IP) inhibited CSD events,
and blocked NTG-induced chronic allodynia. Our results show that we can model the exacerbationof migraine-related symptoms by mu agonists, and confirms previous findings that delta agonists
have anti-migraine properties. This study highlights differential effect of mu and delta opioid receptor
agonists on migraine-associated symptoms, and further supports development of delta agonists fortreating headache disorders.
60Presenter – Birdsong, William
Abstract ID – M24
Acute desensitization and long-term tolerance to opioids are differentiated by multiple
phosphorylation motifs in the C-terminus of the mu-opioid receptor
Emily Leff1, Seksiri Arttamangkul1, Omar Koita1, William Birdsong2, John Williams1
1
Oregon Health & Science University, Portland, USA, 2University of Michigan, Ann Arbor, USA
Prolonged exposure to opioids results in acute desensitization of signaling and long-term tolerance
at the cellular and behavioral levels. The processes of desensitization and tolerance are thought tobe related, in that desensitization may lead to the development of tolerance. Both desensitizationand tolerance are mediated by phosphorylation of the C-terminus of the mu-opioid receptor (MOR)
and previous work demonstrated that eliminating 11 potential phosphorylation sites in the MOR Cterminus abolished both desensitization and tolerance measured at the cellular level. Here wesystematically expressed GFP-tagged MOR mutants with individual serine and threonine
phosphorylation clusters in the MOR C-terminus mutated to alanine via viral delivery into the locus
coeruleus of MOR knockout rats to assess acute desensitization and the development of cellular
tolerance. Using two measures of cellular tolerance to morphine and two measures of desensitization
to the endogenous agonist [Met5] -enkephalin, the results demonstrate that multiple phosphorylation
motifs are involved in mediating both desensitization and tolerance and that these two events arepotentially separable, with mutations to different phosphorylation motifs affecting each process
somewhat uniquely. Thus, while all 11 potential phosphorylation sites appear to contribute to acutedesensitization and the development of tolerance, differing patterns of phosphorylation may mediate
unique aspects of desensitization and tolerance.
Presenter – Bobeck, Erin
Abstract ID – M19
Differential intracellular signaling within the periaqueductal gray contributes to differencesin opioid tolerance
Akila Ram1, Taylor Edwards1, Erin Bobeck1
1
Utah State University, Logon, USA
Opioids remain commonly used analgesics despite their very high propensity for tolerance
development. Antinociceptive tolerance to a variety of opioids, including morphine and fentanyl, hasbeen shown to be mediated by the midbrain ventrolateral periaqueductal gray (vlPAG). However,recently we have found that cross-tolerance does not develop between different opioids whenmicroinjected into the vlPAG. This lack of cross tolerance is likely caused by differences inintracellular signaling profiles despite primary antinociceptive properties being mediated by the mu
opioid receptor. We have found that extracellular signal-regulated kinase 1/2 (ERK1/2) is one ofthese downstream kinases that is differently regulated in vlPAG, such that acute DAMGO andfentanyl activate ERK1/2 morphine does not. However, in the current study using
immunohistochemistry we find that after chronic morphine there is an increase activation of ERK1/2
that is localized to endosomes within vlPAG. In addition, we evaluated activation of protein kinase A(PKA) and protein kinase C (PKC) and both were found to be increased by acute and chronicmorphine at equal levels compared to saline treated animals. Interestingly, there is significant
reduction in nuclear transport of PKA after chronic morphine. Taken together, this data indicates that
acute and chronic treatment of opioids produce distinct signaling profiles within the vlPAG which may
contribute to differences in tolerance development for these different agonists.

61

Presenter – Borrelli, Kristyn
Abstract ID – W15
Neonatal morphine administration in outbred CFW mice induces behavioral signs ofwithdrawal and delayed development
Kristyn N. Borrelli1, Emily J. Yao1, William W. Yen2, Qiu T. Ruan1, Julia C. Kelliher1, Melanie M.
Chen1, Richard K. Babbs1, Jacob A. Beierle1, Elisha M. Wachman3,4, Alberto Cruz-Martin2, Camron
D. Bryant1
1
Laboratory of Addiction Genetics, Department of Pharmacology and Experimental Therapeutics andPsychiatry, Boston University School of Medicine, Boston, USA, 2Department of Biology, Boston
University, Boston, USA, 3Department of Pediatrics, Boston Medical Center, Boston, USA, 4Grayken
Center for Addiction Medicine, Boston Medical Center, Boston, USA
Neonatal Opioid Withdrawal Syndrome (NOWS)- is a condition manifested by withdrawal symptoms
in newborns born to mothers exposed to opioids during pregnancy. Behavioral symptoms include
irritability, poor feeding and sleep, and inconsolability. Amidst the current opioid epidemic in the U.S.,
NOWS incidence has risen to alarming rates. NOWS is driven by hyperactivity of the central andautonomic nervous systems. Treatment includes non-pharmacologic care and replacement therapy
with morphine, methadone, or buprenorphine to alleviate withdrawal symptoms. Both the short- andlong-term effects of perinatal opioid exposure on neurodevelopment are largely unknown and could
take decades to establish through human studies. To model aspects of NOWS, we treated neonatal
pups from outbred CFW mice with twice daily injections of morphine sulfate (15 mg/kg, s.c.) frompostnatal day (P) 1-14 (the approximate third trimester and early postnatal period in humans).
Morphine-treated pups showed increased sensitivity to thermal nociception and an increase inultrasonic vocalizations (USVs) at P14. Furthermore, female P7 mice showed earlier evidence forincreased USVs, which could indicate sex differences in mu opioid receptor system development orfunction. Unbiased classification of USV syllables further supported a developmental delay,
characterized by an increase in syllable similarity between P7 saline mice and P14 morphineexposed mice. We also observed an increase in latency to self-right, suggesting deficits in motor
development. We are currently conducting transcriptome analysis via mRNA-sequencing of
brainstem, a region critical for opioid withdrawal, which will likely provide new insight to molecular
adaptations driving NOWS-like phenotypes.
Presenter – Bouchet, Courtney
Abstract ID – W12
Presynaptic opioid and cannabinoid receptor signaling within the descending pain modulatory
pathway
Courtney A. Bouchet1, Minghua Li1, Susan Ingram1
1
Oregon Health & Science University, Portland, USA
Opioids are the “gold standard” treatments for pain, yet they lack long-term efficacy and are highly
addictive. Cannabinoids have been proposed as a reasonable alternative to opioids and their legalization
has dramatically increased access for patients. However, there are substantial gaps in our understandingof their cellular and molecular actions in brain circuits. In this study, we compare the presynaptic actionsof the mu opioid receptor (MOR) and cannabinoid receptor subtype 1 (CB1) within the descending pain
modulatory pathway, specifically the ventrolateral periaqueductal gray (vlPAG) and the rostral
ventromedial medial (RVM) in male and female Sprague Dawley rats. Activation of MOR and CB1
receptors inhibit GABA release in both the vlPAG and RVM. Interestingly, CB1 receptor inhibition is
reduced in both vlPAG and RVM slices from rats pretreated with Complete Freund’s Adjuvant (CFA) for
5-7 days to induce persistent inflammation. Additional experiments will test whether MOR inhibition of
GABA release is also altered following CFA treatment. Finally, the signaling pathways used by MOR and
CB1 receptors will be analyzed to further understand the adaptations in presynaptic terminals induced bypersistent inflammation. Adaptations in presynaptic GPCR function and signaling are an understudied
area of pain, especially considering the strong expression of both opioid and cannabinoid receptors inpresynaptic terminals. Understanding differences between the functions of MOR and CB1 receptors will
provide new avenues for development of pain therapies.
62Presenter – Brice-Tuitt, Ariana
Abstract ID – M33
Mixed opioid receptor agonism and antagonism demonstrated by a novel cyclic tetrapeptide
prevents drug- and stress-induced reinstatement of morphine- and cocaine- seeking behavior.
Ariana C. Brice-Tutt1, Lisa L. Wilson1, Shainnel O. Eans1, Heather M. Stacy1, Chloe A. Simons1, Grant
Simpson2, Jeremy S. Coleman2, Michael J. Ferracane2, Jane V. Aldrich2, Jay P. McLaughlin1
1
Department of Pharmacodynamics, 2Department of Medicinal Chemistry, College of Pharmacy,
University of Florida, Gainesville, USA
There is an urgent need for new, more effective treatments for both substance abuse and pain. The
macrocyclic tetrapeptide natural product CJ-15,208 (cyclo[Phe-D-Pro-Phe-Trp]) is a multifunctional muopioid receptor (MOR) and kappa-opioid receptor (KOR) agonist and KOR antagonist that produces
antinociception and prevents stress-induced reinstatement of extinguished cocaine-conditioned place
preference (CPP). We hypothesized that a mixed MOR/KOR agonist/KOR antagonist analog of CJ15,208, JVA4001, would produce potent antinociception with fewer liabilities while preventing both drugand stress-induced reinstatement of CPP. Opioid receptor agonist and antagonist activity of JVA4001
was assessed in mice using the 55ºC warm-water tail-withdrawal assay. Liabilities of respiration,
spontaneous ambulation, and direct rewarding or aversive properties were assessed. Finally, the abilityof JVA4001 to block drug- and stress- induced reinstatement of extinguished morphine or cocaine
conditioned place preference (CPP) was determined. JVA4001 demonstrated dose-dependent, shortlasting antinociception upon oral and peripheral administration, mediated by MOR and (to a lesser extent)
KOR. The cyclic tetrapeptide demonstrated multifunctional MOR and KOR agonism and potent dosedependent KOR antagonism lasting up to 4 h. JVA4001 displayed reduced liabilities compared tomorphine, and pretreatment with JVA4001 blocked both stress- and drug-induced reinstatement of both
extinguished morphine and cocaine place-preference responses in a time-dependent manner. Overall,these data suggest that JVA4001 is a promising lead compound for the treatment of pain with reduced
liabilities as well as the prevention of both drug- and stress-induced relapse in abstinent morphine- or
cocaine-seeking subjects.
Supported by W81XWH-15-1-0452 and W81XWH-15-1-0464 from DoD CDMRP
Presenter – Brown, Kate
Abstract ID – T49
Comparison of cortisol serum levels in stress-minimized normal volunteers and drug-free former
cocaine dependent volunteers
Kate Garland Brown1, Brian Reed1, Jose Erazo1, Michelle Morochnik1, Ariel Ben-Ezra1, Mary Jeanne
Kreek11
Rockefeller University, USA
The hypothalamic-pituitary-adrenal (HPA) axis plays an important role in maintaining homeostasis when
an individual experiences stress and has been shown to become disrupted following long-term exposure
to drugs of abuse. This study examined serum cortisol levels between normal volunteers (NV), (n=10)
and drug-free former cocaine dependent (DFFCD) volunteers, (last use of cocaine >6 months, n=12). To
assess differences in stress responsivity we conducted a study which involved administration ofadrenocorticotropic hormone (ACTH), 750mcg intravenous (IV), and metyrapone, 2.25gm by mouth, on
two separate study days, compared to baseline, in which 0.9% normal saline IV was administered on thefirst study day. Both groups were studied concomitantly over the past 2 years. The objective of the study
was to measure serum cortisol levels in response to ACTH and metyrapone; time points in which blood
was drawn were: baseline/ACTH days: t= -10, 0, 10, 20, 30, 40, 50, 60, 75, 90, 120, 240, 360 min.;metyrapone– t= -10, 0, 60, 120, 240, 360, 480 min. Areas under the curve (AUCs) were estimated using
the trapezoid method, from 0-360 min following injection. ANOVAs, diagnostic condition x treatment, were
performed. While there was a main effect of treatment both in ACTH and metyrapone, there was no main
effect of diagnostic condition, and no interaction of diagnostic condition x treatment. Both NV and DFFCD
volunteers show equivalent endocrine stress responses, as determined by serum cortisol levels.
Supported by the Adelson Medical Research Foundation (AMRF) and Clinical and Translational ScienceAward (CTSA)

63

Presenter – Bryant, Camron
Abstract ID – T35
Gene mapping and validation of a functional intronic variant in Gabra2 underlying reduced Gabra2
expression, increased naloxone aversion and increased methamphetamine stimulant sensitivity
Lisa R. Goldberg1, Eric R. Reed2, Jiayi Wu3, Cory Parks4, Christine E. Watkins4, Morgan Dickerson4, Sufiya
Khanam4, Emily J. Yao1, Julia C. Kelliher1, Stacey L. Kirkpatrick1, Melanie M. Chen1, Gregg E. Homanics5,
Robert W. Williams4, Megan K. Mulligan4, Camron D. Bryant1
1
Department of Pharmacology and Experimental Therapeutics and Psychiatry, Boston University School of
Medicine, USA, 2Ph.D. Program in Bioinformatics, 3Graduate Program in Genetics and Genomics, Boston
University, USA, 4Department of Genetics, Genomics, and Informatics, University of Tennessee Health
Science Center, USA, 5Departments of Anesthesiology, Neurobiology, and Pharmacology and Chemical
Biology, University of Pittsburgh, USA
Substance use disorders including opioid and psychostimulant use disorders, have a genetic component thatremain mostly unknown. Genetic correlation among SUDs indicate an overlapping, shared genetic basis. Here,we conducted a genome-wide quantitative trait locus (QTL) analysis of behavioral sensitivity to the aversive
properties of the opioid receptor antagonist naloxone and the locomotor stimulant properties of thepsychostimulant methamphetamine. To facilitate gene identification, we employed a Reduced Complexity
Cross (RCC) between C57BL/6 mouse substrains. We mapped a single, pleiotropic genome-wide significant
QTL on medial chromosome 5 that influenced naloxone-induced freezing and methamphetamine-induced
locomotor activity (LOD=3.9-4.6; peak: 35 cM; C.I. = 27-48 cM) and striatal transcript levels of Gabra2 (adj.p.=
7.2e-27). Gabra2 codes for the α2 subunit of the GABA-A receptor. The peak marker (rs29547790; 70.93 Mb)
is located near a hypomorphic intronic single nucleotide deletion in Gabra2 that is derived from C57BL/6J andreduces Gabra2 expression (Mulligan MK et al., 2019; Frontiers in Genetics). CRISPR/Cas9-mediatedcorrection of the mutant indel with a single nucleotide insertion of the wild-type C57BL/6N allele restored
Gabra2 RNA and protein expression, methamphetamine-induced locomotor activity, and naloxone freezing
and aversion toward a wild-type C57BL/6N level, thus identifying the quantitative trait nucleotide. Our studies
identify a functional, pleiotropic effect of reduced Gabra2 expression on naloxone aversion and the stimulant
response to methamphetamine and further support GABA-A receptor function in opioid and psychostimulant
use disorders.

Presenter – Brynildsen, Julia
Abstract ID – W39
Functional network connectivity in opiate-naïve and opiate-dependent states
Julia K. Brynildsen1, Carmen Weidler2, Danielle S. Bassett3, Julie A. Blendy1
1
Department of Systems Pharmacology and Translational Therapeutics, Perelman School of Medicine,
University of Pennsylvania, Philadelphia, USA; 2Department of Psychiatry, Psychotherapy andPsychosomatics, Faculty of Medicine, RWTH Aachen University, Germany; 3Department of Bioengineering,
University of Pennsylvania, Philadelphia, USA
A major challenge in addiction research is to determine why addictive behaviors endure or can be reinstatedlong after drug use is discontinued. The persistence of addiction throughout the lifetime of an individualsuggests that chronic drug exposure results in long-term adaptations in the brain. In the interest of identifying
network-level biomarkers for opioid dependence, we analyzed changes in functional connectivity between
brain regions at three time points: before chronic opiate exposure, immediately following chronic opiate
exposure when dependence has been established, and following prolonged abstinence, at a time that may be
associated with risk for relapse. In order to characterize changes in neuronal activity induced by chronic opiate
exposure, we compared Fos expression following acute morphine exposure in morphine-naïve and morphinedependent C57BL/6J mice. Mice were transcardially perfused 90 min after an injection of 10 mg/kg morphine,
and Fos expression was quantified in 20 brain regions implicated in opiate reward and dependence. Statedependent differences in Fos reactivity were observed in regions of the isocortex, striatum, and thalamus.
Interregional correlations in Fos expression were used to construct functional network models under eachstate. Finally, modularity analysis was used to identify community structure within each network, and therelative inter-community connectivity of each brain region was compared by computing its participation
coefficient. This approach identifies between-region functional associations that may be vulnerable to theeffects of chronic opiate exposure. Future studies will examine genetic influences on functional brain networks
during opiate dependence and following protracted withdrawal.

64

Presenter – Burgess, Gwendolyn
Abstract ID – M17
Evaluation of the abuse potential of AAH8, a mixed efficacy opioid ligand
Gwendolyn E. Burgess1, Ashleigh L. Matthews11, Allison G. Ladetto11, Katarzyna Sobczyk-Kojiro21,
Henry I. Mosberg21, Emily M. Jutkiewicz1.1
1
University of Michigan, USA
Mixed efficacy opioid ligands have emerged as a potential alternative to the mu opioid receptor(MOR) agonists used clinically. AAH8 is a high affinity MOR agonist/DOR antagonist with 5-foldselectivity for MOR over DOR in vitro. AAH8 produces MOR-mediated antinociception in the mousewarm water tail withdrawal assay with limited rewarding effects in the conditioned place preference
test. The purpose of this work was to further examine the abuse-related effects of AAH8. In rats
trained to self-administer 0.3 mg/kg/infusion morphine on a fixed ratio 1 (FR1) schedule ofreinforcement, AAH8 was substituted for morphine at multiple doses (0.03, 0.1, 0.3, or 1 mg/kg/inf.).
AAH8 maintained significantly less responding than morphine. The interoceptive effects of AAH8
were evaluated in rats trained to discriminate injections of either morphine or the prototypical DOR
agonist SNC80 from saline in a two-key discrimination paradigm. In rats trained to discriminate
morphine, AAH8 did not produce morphine-appropriate responding. In rats trained to discriminate
SNC80, AAH8 did not generalize to SNC80; however, pretreatment with AAH8 to small doses of
SNC80 potentiated SNC80-like responding. To further evaluate the DOR agonist properties of AAH8,
we investigated the ability of AAH8 to potentiate pentylenetetrazol (PTZ)-induced convulsions in
mice. Like SNC80, AAH8 was able to shift the PTZ curve to the left. Collectively, these data andprevious studies demonstrate that AAH8 is a MOR agonist/partial DOR agonist with antinociceptive
effects and minimal abuse potential.
Supported by DA003910 (Mosberg) and DA041565 (Jutkiewicz).
Presenter – Cami-Kobeci, Gerta
Abstract ID – T3
Naltrexone esters and close analogs as potential new analgesics with reduced side effectprofileGerta Cami-Kobeci1, Keith Olson2,3, John R. Traynor2,3, Stephen M. Husbands1
1
Department of Pharmacy and Pharmacology, University of Bath, Bath, UK, 2Department ofPharmacology, University of Michigan, Ann Arbor, USA, 3Edward F. Domino Research Center,
University of Michigan, Ann Arbor, USA
Structure-activity relationship studies have suggested that the region of space occupied by the tertbutyl group in buprenorphine is key to good NOP receptor activity.[1] We discovered that this region
can also be accessed by certain acyl and alkyl groups attached to the C14-oxygen of the MOP
antagonist ligand naltrexone (naltrexone itself has no affinity for NOP receptors). In the light of thisevidence we designed a series of C14-oxygen naltrexone esters. Within this series BU10038 showed
fivefold higher affinity to NOP and two fold higher to MOP compared to buprenorphine, with thedesired increase in efficacy at NOP in the [35S]GTPgammaS binding assays, combined with low
efficacy partial agonism at MOP receptors.[2] Although BU10038 has a promising analgesic profile,
we were interested in studying C6-analogs due to the known naltrexone metabolism to 6β-naltrexol,
as well the opportunity to further investigate the structure activity relationships (SAR) within thisseries. To this end we designed esters based on C6-desoxynaltrexone, naltrexol and nalmefene.
Using this strategy we have identified BU14003 as of particular interest as it showed the highest
affinity at NOP receptors from within this series (2 orders of magnitude better than buprenorphine)and similar affinity at MOP compared to BU10038. In the [35S]GTPgammaS binding assays thedesired high potency and increase in efficacy at NOP was observed coupled with low efficacy partial
agonism at MOP receptors. BU14003 therefore represents a potential analgesic ligand, with low side
effect profile.

65

Presenter – Campos-Jurado, Yolanda
Abstract ID – M30
Inflammatory pain dysregulates MORs in the VTA: effect on ethanol induced dopamine
release in NAc and ethanol relapse
Yolanda Campos-Jurado1, Jesus D. Lorente1, Javier Cuitavi1, Jose L. Gonzalez-Romero1, RaquelMonton-Molina1, Teodoro Zornoza1, Luis Granero1, Lucia Hipolito1
1
University of Valencia, Spain
Epidemiologic research has evidenced a relationship between pain and addiction. Indeed, recent
clinical studies have shown that pain increases the vulnerability to alcohol relapse. Previous data inrats revealed that inflammatory pain desensitizes mu opioid receptors (MORs) in the ventral
tegmental area (VTA) and increases intake of high doses of heroine. In the present study, first we
further evaluated VTA MORs function under inflammatory pain condition, showing that thesereceptors are dysregulated depending on the projection area. Thereafter, we evaluated the effect ofinflammatory pain on accumbal increase of dopamine release elicited by ethanol. This microdialysis
study showed that the presence of inflammatory pain blunted the increase of extracellular dopaminelevels in the Nucleus Accumbens (NAc) induced by ethanol. Later on, we evaluated the effect ofinflammatory pain on alcohol relapse by using two different non-operant paradigms. In the 4-bottle
choice paradigm our results showed that inflammatory pain does not affect to the magnitude ofrelapse. Interestingly, we used an intermittent 2-bottle choice paradigm in male and female rats andthe results indicated that, after an abstinence period, the increase in alcohol drinking was only
notorious in female rats in the presence of pain. Finally, the analysis by western-blot of the brainsfrom this last behavioral approach revealed that the expression of MORs and KORs in VTA, NAc
and Prefrontal Cortex is altered only in female pain-treated animals. All together our results uncover
the need to understand how inflammatory pain affects the vulnerability to suffer alcohol addiction.
Presenter – Chan, Wallace
Abstract ID – T14
Docking-based virtual screen for bifunctional μ-opioid agonist/δ-opioid antagonist
compounds
Wallace Chan1, Keith Olson1, Yang Zhang1, John Traynor1
1
University of Michigan, Ann Arbor, USA
Activation of the μ-opioid receptor while inhibiting the δ-opioid receptor has been demonstrated inanimal models to elicit analgesia with reduced tolerance and dependence. In the present study, adocking-based virtual screen was carried out for both the μ- and δ-opioid receptors to find bifunctional
ligands that act as a μ-opioid agonist/δ-opioid antagonist. Retrospective virtual screens were
performed against μ- and δ-opioid GPCR-Bench datasets consisting of between 100 – 600 active
compounds and 15 decoys per active compound. Docked poses were rescored with the scoring
functions, DSX (J. Chem. Info. Model., 2011) and X-Score (J. Comput.-Aided Mol. Des., 2002).
Overall, rescoring with XScore (AUC: μ=0.75 / δ=0.85) performed better than that with DSX (AUC:
μ=0.74 / δ=0.73) and was thus used as the protocol for the study. Furthermore, active compounds
were found at greater than 10 times over random for each receptor. Subsequently, prospective virtual
screens were run against a targeted subset of ~1.3M compounds from ZINC database. Seven highscoring compounds for both receptors were chosen for experimental validation; three compounds
were found to have better than 10 μM affinity for the δ-opioid receptor, while one compound had
moderate affinity (Ki=11 μM) for the μ-opioid receptor. Further work is needed to further explore
chemical space for the optimization of both binding and bifunctionality of the most promising
compounds. These preliminary results highlight the ability of the current virtual screening pipeline inpredicting the binding of compounds for the μ- and δ-opioid receptors.

66

Presenter – Chen, Carina
Abstract ID – W2
Age trajectory of exposure to cannabis and alcohol, in volunteers diagnosed with opioid
dependence, opioid and cocaine dependence, or cocaine dependence
Carina Chen1, Kate Brown1, Rachel Conybeare1, Eduardo Butelman1, Mary Jeanne Kreek1
1
Rockefeller University, New York, USA
Persons with opioid and cocaine use disorders often have a history of prior exposure to cannabis
and alcohol. This study examined the ages of onset of heaviest use of cannabis and alcohol acrossdifferent clinical groups, based on DSM-IV diagnoses: 1. opioid dependence but not cocaine
dependence (OD), 2. cocaine dependence but not opioid dependence (CD), 3. opioid and cocaine
dependence (OD+CD), 4. any other drug diagnoses (Any other Dx), and 5. healthy controls. Adult
volunteers (n=1,413) were sequentially ascertained from a metropolitan area in an observational
study. Study protocol was approved by the Rockefeller University Hospital IRB. Kreek-McHughSchluger-Kellogg (KMSK) scales measured maximal drug exposure and age-related data. Heavy
exposure to cannabis and alcohol was detected in groups 1-4. The age of onset of heaviest use of
cannabis and alcohol preceded those for either heroin or cocaine, in volunteers with OD, CD or
OD+CD. The ages of onset of heaviest use of cannabis and alcohol differed significantly across thefive clinical groups (Log-Rank test p<0.0001 and p=0.0005, respectively). Thus, the age of onset ofheaviest use of cannabis in the OD+CD group was the earliest among all the groups (median age=15
years). A multivariate Cox regression showed that belonging to the OD, CD and OD+CD groups was
associated with earlier onset of heaviest use of cannabis, compared to the reference "Any other Dx"group (alcohol was similarly analyzed). Overall, volunteers diagnosed with OD, CD or OD+CD havea pattern of early and heavy exposure to both cannabis and alcohol.
Presenter – Chiou, Lih-Chu
Abstract ID – T48
Acupuncture relieved neuropathic pain in opioid-tolerant mice via orexin-initiated & endocannabinoid-mediated disinhibition in the periaqueductal gray
Lih-Chu Chiou1,2,3, Ming Tatt Lee1,21
Graduate Institute of Brain and Mind Sciences, College of Medicine, National Taiwan University, Taipei,
Taiwan, 2Department of Pharmacology, College of Medicine, National Taiwan University, Taipei, Taiwan,
3
Graduate Institute of Acupuncture Sciences, China Medical University, Taichung, Taiwan
Recently, we revealed a non-opioid analgesic mechanism induced by electroacupuncture at the PC6(Neiguan) acupoint (EA-PC6), which overlays the median nerve, in mice.1 Here, we further explore thetherapeutic potential of EA-PC6 in opioid-tolerant subjects with chronic pain using a mouse neuropathic
pain model induced by chronic sciatic nerve constriction injury (CCI). On days 8-14, CCI-mice were daily
treated with morphine (10 ml/kg, i.p.), EA-PC6 (2 Hz, 2 mA, 0.15 ms) or morphine+EA-PC6. Repeated
EA-PC6 significantly suppressed mechanical allodynia in CCI-mice, and this anti-allodynic effect did not
develop tolerance, unlike repeated morphine treatments. Furthermore, morphine tolerance was reversed
by EA-PC6 on day 14 and prevented by daily EA-PC6 in a manner by i.p. blockade of OX1 or CB1
receptors. In a more clinically-relevant status, analgesia tolerance induced by escalating doses ofmorphine (10 mg/kg on days 8-14, 20 mg/kg on days 15-21, and 50 mg/kg on days 22-28) was also
reversed by single dose (on day 28) or repeated doses (days 22-28, daily) of EA-PC6. These results
suggest that repeated EA-PC6 induces an analgesic effect without tolerance, and can either prevent or
attenuate opioid tolerance via a disinhibition mechanism mediated by OX1R-initiated & endocannabinoidCB1R-mediated signaling in the periaqueductal gray.1 Thus, EA-PC6 is a potential novel pain
management strategy in patients suffered with opioid tolerance with escalating doses of opioids, such as
terminal cancer patients.
1
Chen YH et al. (2018) Median nerve stimulation induces analgesia via orexin-initiated endocannabinoid
disinhibition in the periaqueductal gray.
Proc Natl Acad Sci USA 115:E10720-E10729.

67

Presenter – Cippitelli, Andrea
Abstract ID – T22
PPL-103: A kappa partial agonist approach that potently reduces compulsive cocaine taking andprime-induced cocaine seeking
Andrea Cippitelli1, Megan Barnes1, John A. Lawson2, Lawrence Toll1
1
Department of Biomedical Science, Charles E. Schmidt College of Medicine, Florida Atlantic University,
Boca Raton, Florida, USA, 2Phoenix PharmaLabs Inc., Woods Cross, Utah, USA
Pharmacological treatments for cocaine addiction are urgently needed. Phoenix PharmaLabs (PPL) has
developed a novel family of opioid receptor ligands, including PPL-103. This compound displays a unique
combination of high binding affinities and partial agonist activities at mu, kappa, and delta opioid
receptors, with highest efficacy at kappa, and lowest at mu. Since the kappa receptor system isupregulated under conditions of chronic cocaine use and upon cocaine withdrawal, we hypothesized thatthe partial kappa agonist, PPL-103, can be effective as a pharmacotherapy for the treatment of cocaine
addiction. Accordingly, we determined whether systemic injections of PPL-103 (0.0, 0.1, 0.3, 1.0 mg/kg)
could reduce operant cocaine self-administration and reinstatement in rats. Results indicate that the
kappa partial agonist reduced the number of cocaine infusions in short (1-hour) as well as long (6-hour)access sessions. The effect was more potent in rats that escalated cocaine self-administration, a behavior
that mimics compulsive patterns of human cocaine use. Self-administration under a progressive ratioschedule was also reduced in both short and long access groups, whereas food self-administration was
not altered at doses that decreased cocaine infusions. Furthermore, PPL-103 was more effective inblocking cocaine prime-induced reinstatement than conditioned reinstatement of cocaine seeking. Finally,
PPL-103 not only prevented induction of cocaine seeking when tested under extinction conditions, butrather decreased seeking below the level of extinction at all doses examined. PPL-103 displays apromising profile as a possible pharmacotherapy for treatment of cocaine addiction.

Presenter – Collins, Devon
Abstract ID – M51
Oxycodone conditioned place preference, locomotor activity, and self-administration in maleand female Oprm1 A112G mice
Devon Collins1, Yong Zhang1, Mary Jeanne Kreek1
1
The Rockefeller University, New York, USA
Mu-opioid receptors (MORs) mediate the rewarding properties of oxycodone and other prescription
opioid medications, which have played a central role in the current opioid epidemic. A common single
nucleotide polymorphism (SNP) in the human mu-opioid receptor (MOR) gene (OPRM1), A118G,
has been demonstrated to alter MOR function in healthy humans, and is associated with opioid
addiction. Mice bearing a functionally analogous SNP, A112G, in the mouse MOR gene weregenerated, characterized, and studied (Mague et al., 2009). Male and female mice homozygous forthe 112G allele (GG) show genotype-dependent reductions in morphine-stimulated locomotor
activity and genotype-dependent, sex-specific reductions in morphine-induced conditioned place
preference versus homozygous 112A (AA) mice (Mague et al., 2009). Female mice bearing one or
two copies of the 112G allele show reduced buprenorphine-stimulated locomotor activity (Browne etal., 2017). GG mice of both sexes self-administer more heroin compared to AA littermates (Zhang etal., 2015). We examined oxycodone conditioned place preference (CPP), locomotor activation, andself-administration behavior (SA) [extended-access (4-hr) sessions] in male and female AA and GG
mice. We found that oxycodone (1 and 3 mg/kg) produced CPP in AA and GG mice of both sexes.
Oxycodone (1 and 3 mg/kg) produced a significant increase in locomotor activity during conditioning
sessions in AA mice only; there was no such increase in GG mice. Both male and female GG miceself-administered significantly more oxycodone (0.25 mg/kg unit dose) compared to their wild-type
AA littermates.

68Presenter – Conibear, Alexandra
Abstract ID – W17
A G protein-biased δ opioid receptor agonist with improved therapeutic profile
Alexandra Conibear1, Junaid Asghar3, David Kendall2, Bengt von Mentzer2, Eamonn Kelly1
1
University of Bristol, UK, 2PharmNovo AB, Sweden, 3Gomal University, Pakistan
The δ opioid receptor is a promising therapeutic target for the treatment of chronic pain states andemotional disorders. Unlike µ opioid receptor agonists, δ opioid receptor agonists have low abuseliability. However some δ opioid receptor agonists have pro-convulsant properties and tolerance tothe therapeutic effects can develop. As such, interest is now growing in the development of biased
agonists as a potential means to improve the therapeutic profile of δ opioid receptor agonists. In
collaboration with PharmNovo AB we have characterized the signalling and trafficking of a selectionof novel δ opioid receptor ligands. We find that some of the ligands display bias toward G protein
activation over arrestin recruitment. Molecular dynamics (MD) simulations of the δ opioid receptor incomplex with either the prototypical agonist SNC80, or one of the G protein-biased ligands hasrevealed distinct ligand residue interactions that may in part, underlie the structural basis of theobserved bias. Further one of the biased ligands, PN6047, is effective in rodent models of chronic
pain, shows no apparent analgesic tolerance after prolonged treatment and has no effect on
chemically-induced seizures. Thus, δ opioid receptor ligands with limited arrestin signalling may betherapeutically beneficial in the treatment of chronic pain states.
Presenter – Cox, Brian
Abstract ID – M22
Gene expression changes in the ventral tegmental area (VTA) and lateral habenula (LHb) ofrats after a single morphine treatment.
Ryan Shepard1, Shawn Gouty1, Abby Sachar1, Fereshteh S Nugent &1, Brian M Cox1
1
Department of Pharmacology & Molecular Therapeutics, Uniformed Services University, Bethesda,
USA
Dopamine (DA) released from VTA neurons plays a critical role in reward; drugs of abuse candysregulate DA release. The firing rate of VTA DA neurons is reduced by LHb projections activating
GABA interneurons in the VTA. We previously reported that 24 hr after a single morphine dose,
mEPSC frequency in VTA DA neurons was increased, with a subsequent reduction in mIPSC
frequency and amplitude. We also showed reduced acetylation of histone H3 at K9 (Ac-H3K9); an
effect reversible by the Class I HDAC inhibitor CI-994 up to 24 hrs later (Authement et al, 2016).
Following morphine, activation of CB1 receptors by WIN 55212-2 did not further reduce mIPSC
frequency (which was already reduced by the morphine treatment), suggesting that morphineincreases endogenous cannabinoids (eCBs) in VTA, inhibiting GABA release. We now show that asingle morphine treatment (10 mg/kg IP, 24 hr) also increases the expression of A-kinase anchoring
protein (AKAP150) in VTA DA neurons (with no change in AKAP150 expression in the substantia
nigra). Additionally, we show increased AKAP150 abundance and decreased levels of Ac-H3KP in
glutamatergic neurons of the LHb. We also observed reduced levels of the eCB-degrading enzyme
MAGL in LHb after morphine, suggesting morphine elevates levels of eCBs in LHb. These results
indicate that a single morphine dose exerts parallel effects on gene expression and signaling
pathways in the VTA and critical regulatory neurons in LHb. These actions may play mechanisticroles in the adaptive responses to morphine in the mediation of reward.

69

Presenter – De Marco, Rosella
Abstract ID – T33
Tryptophan-containing non-cationizable opioid peptides derived from CJ-15,208: from dual
KOR/MOR affinity profile to selective interactions for MOR, DOR, or KOR
Rossella De Marco1, Lorenzo Cavina1, Edoardo Pirazzoli1, Andrea Bedini1, Santi Spampinato1, Luca
Gentilucci1
1
University of Bologna, Bologna, Italy
The Tryptophan-Containing Non-Cationizable Opioid Peptides (in short TryCoNCOPs) are a new familyof peptides deprived of the classic pharmacophores of the opioid peptides, which can still bind to andactivate the opioids receptor (OR). The TryCoNCOPs show atypical interactions with the ORs, being themain pharmacophoric motif the dipeptide Trp&Phe, and in particular the interaction between indole-NH
and Asp (3:32) of the ORs appears fundamental for agonist activity. We started from the naturally
occurring cyclotetrapeptide CJ15,208, c[Phe1-D-Pro2-Phe3-Trp4], that showed a dual KOR/MOR affinity(Ki = 47 and 260 nM, respectively). The residues at the positions 1 and 2 have been replaced with a-, bor g-amino acids. The different compounds showed nM affinity and complete selectivity for either MOR,
DOR, or KOR, and variable functional activity (cAMP test). The peptide EDO43 showed full agonist
activity in vitro, and MOR-dependent antinociceptive effect in vivo in a mouse model of visceral pain uponsystemic administration, consistent with the cyclic nature. Conformational and Docking analyses
supported that the significant preference of each distinct peptide for the ORs seems to be correlated to
specific secondary structures. In conclusion, starting from CJ15,208 we addressed the mixed selectivity
profile and we gained high specificity for the ORs. In perspective, these atypical peptides might be utilized
for stabilizing receptor conformations not explored previously, displaying biased signaling and hence,unlike the classic opioid drugs, have more favorable in vivo profiles, reduced side effects and novel
therapeutic applications.
Presenter – Degrandmaison, Jade
Abstract ID – M40
In vivo identification of Rab10 as a regulator of the δ-opioid receptor (DOPr) trafficking
Jade Degrandmaison1,2,4,6, Khaled Abdallah2,4, Véronique Blais2,4,6, Samuel Génier1,4, Marie-Pier
Lalumière1,4, Francis Bergeron2,4,6, Catherine M. Cahill5, Jim Boulter5, Christine L. Lavoie2,4, Jean-Luc
Parent1,4, Louis Gendron2,3,4,7
1
Department of Medicine, 2Department of Pharmacology-Physiology, 3Department of Anesthesiology,
4
Faculty of Medicine and Health Sciences, Université de Sherbrooke, Canada, 5Department of Psychiatry
and Biobehavioral Sciences, Hatos Center for Neuropharmacology, David Geffen School of Medicine,
University of California Los Angeles, USA, 6Quebec Network of Junior Pain Investigators, 7Quebec Pain
Research Network, Canada
Despite their limited antinociceptive effects in acute pain models, delta-opioid receptor (DOPr)-selective
agonists have demonstrated analgesic efficacy in several chronic pain models, including neuropathic,
inflammatory, diabetic and cancer-related pain, while exhibiting milder undesired effects than the clinicallyavailable opioids. Interestingly, our research group and others have previously observed a correlation
between the translocation of DOPr from the intracellular compartments to the plasma membrane and theincrease of DOPr agonist-mediated analgesic effects. Therefore, to investigate the molecularmechanisms regulating DOPr trafficking in vivo, we generated a genetically engineered knock-in mouse
expressing a FLAG-tagged version of DOPr and characterized its expression, distribution, activation,
locomotor and antinociceptive properties. Then, using the rostral part of the brain of these mice, wedeveloped an approach to identify endogenous DOPr interactors. Mass spectrometry analyses performed
on immunoprecipitated FLAG-DOPr from brains of knock-in mice confirmed previously reported DOPrinteracting partners (e.g. CB1, Calnexin, TFRC, CDK5, SEC31A and VAPA) and classic GPCRs
interactors such as β-arrestin-1 and heterotrimeric G proteins. Among the newly identified interactors, thesmall GTPase Rab10 was selected to validate our approach. In HEK293 cells, we confirmed theinteraction between Rab10 and the human DOPr and showed that they co-localize in the trans-Golgi
network. Furthermore, we observed that upregulation or downregulation of Rab10 regulates the trafficking
of DOPr, therefore modulating its cell-surface expression. Besides reporting the first interaction between
Rab10 and a GPCR, this work provides a novel approach to identify and study endogenous mechanisms
involved in the trafficking of DOPr and presumably other GPCRs.
70

Presenter – Dorval, Louben
Fibroblast growth factor 21 (FGF21) attenuates morphine preference in mice
Louben Dorval1, Brian I. Knapp1, Jean M. Bidlack1
1
University of Rochester, New York, USA

Abstract ID – M46

Fibroblast growth factors (FGFs) are a family of growth factors involved in angiogenesis and multiple
endocrine signaling pathways. Unlike most FGFs, FGF21, has low affinity for heparan sulfate
glycosaminoglycans and is released into the bloodstream. FGF21 is expressed in liver, brown adipose
tissue, glia, and neurons. Previous studies showed that FGF21 decreased alcohol and sweet preferencein mice (Talukdar et al, 2016). Furthermore, the effects of FGF21 on taste preference correlated with adecrease in dopamine levels in the nucleus accumbens. Therefore, FGF21 may also modulate the effects
of morphine and other opioids in the reward pathway. This current study explored the effects of FGF21
on morphine preference in mice using FGF21-Tg (expressing a higher level of FGF21) and wild-type
littermate mice. FGF21-Tg mice expressed a 2,400-fold increase in FGF21 protein levels in serum
compared to wild-type mice. Transgenic and wild-type mice were injected with morphine or saline andtheir condition place preference was determined over a 3-day period. All mice showed similar initial
preference, but after the conditioning phase, male transgenic mice showed a 43% lower preference for
10 mg/kg morphine compared to wild-type male littermates. Female FGF21-Tg mice showed a 45% lower
preference compared to wild-type female littermates at 10 mg/kg morphine. Moreover, female transgenic
mice showed a 95% lower preference for 3 mg/kg morphine compared to wild-type female littermates.
These results demonstrate that FGF21 attenuates morphine preference in mice.
Supported by NIDA grant DA044766

Presenter – Dunn, Amelia
Abstract ID – M11
Correlation between beta-arrestin-2 signaling at the kappa opioid receptor and rotarod
sedation
Amelia Dunn1, Brian Reed1, Mary Jeanne Kreek1
1
Rockefeller University, New York, USA
The kappa opioid receptor (KOR) is an important drug target for diseases of mood and reward, suchas addiction and depression. KOR agonists have been shown to block drug-seeking in animalmodels, however they also cause significant side effects such as sedation and aversion. In order topreserve the therapeutic effects of KOR agonists while avoiding negative side effects, many groups
are investigating biased agonists that selectively activate only certain pathways downstream of thereceptor. We compared the signaling properties of 21 KOR agonists, including the endogenous
peptide Dynorphin A (1-17), clinically-used morphinan compounds, and 2 novel compounds
synthesized for our research. We found that most compounds tested were biased for G-protein
signaling over β-Arrestin-2 recruitment compared to the reference agonist U69,593. A subset ofthese compounds was tested for KOR-mediated sedation using the rotarod assay. We found asignificant correlation between maximum effect on the rotarod and maximum efficacy in the βArrestin-2 recruitment assay, but not the G-protein signaling assay. This suggests that β-Arrestin-2
signaling in this in vitro system can model in vivo sedation behavior, and that a G-protein biased
KOR agonist may not cause the sedative side effects associated with full, unbiased KOR activation.
Supported by the Robertson Therapeutic Development Fund and the Dr. Miriam and Sheldon G.
Adelson Medical Research Foundation.

71

Presenter – Effah, Felix
Abstract ID – W28
Effect of prolonged milk-casein exposure during early development on mood and µ-opioid
receptor density (MOPr) of conventional and germ-free rats: An investigation of diet x gut
microbiota-brain axis interaction
Felix Effah1, Fatima Joly2, Sylvie Rabot2, Imran Simreekheea1, Arundeep Sohal1, Ethan Wong1,
Claire Le Poupon3, Aurélien Raynaud3, Ian Kitchen4, Aya Osman4, Vincent Bombail3, Alexis Bailey1
1
Pharmacology Section, St George's University of London, London, UK, 2Micalis, INRA,
AgroParisTech, Université Paris Saclay, Jouy en Josas, France, 3NBO, INRA, Université Paris
Saclay, 78350, Jouy, France, 4School of Biosciences and Medicine, Faculty of Health and Medical
Sciences, University of Surrey, Guildford, Surrey, UK
There is evidence to suggest that prolonged exposure to milk caseins, the most abundant protein
component of milk, during early development could have detrimental effects on brain and behaviour.
We previously demonstrated that prolonged exposure to casein-rich but not casein-free milk in rats
at early post-weaning age induces a depression-like phenotype and region-specific alterations in
MOPr density in the brain. There is evidence suggesting that gut microbiota may be involved inmediating this effect via the gut-brain axis. To assess the involvement of gut microbiota on the effectof milk casein on mood, we analyzed depressive-like behaviour of conventional and germ free (GF)
rats exposed to casein-rich or casein-free milk from post-natal day 21 to 26, with the forced swim
test. Quantitative receptor autoradiographic binding of MOPr was carried out in the brains of thoserats to assess the involvement of gut microbiota. While no change in depressive-like behavior was
detected between conventional and GF rats exposed to casein-rich milk, a marked reduction ofdepressive-like behaviour was observed in GF rats exposed to casein-free milk. An overall
upregulation of MOPr was detected in the olfactory orbital cortex of GF animals, irrespective of theircasein exposure. Conversely, casein-rich milk exposure induced an upregulation of MOPr in the
motor cortex but a downregulation in the nucleus accumbens, irrespective of microbiota presence.
These findings unveil a key role of microbiota in mediating diet x gut-brain axis interactions which
impact on behavior.
Presenter – Emery, Michael
Abstract ID – W14
Differential behavioral responses to morphine in rats selectively bred for high versus low
response to novelty
M.A. Emery1, C. Aydin, E.K. Herda-Bauer, P.M. Maras, A. Parsegian1, A.V. Stefanov1, A. Tang1, S.
J. Watson1,2, H. Akil1,2
1
Molecular and Behavioral Neuroscience Institute, University of Michigan, USA, 2Department of
Psychiatry, University of Michigan, Ann Arbour, USA
Our lab uses a selective breeding strategy based on degree of locomotor response in a novel
environment to generate and maintain two lines of rats with distinct behavioral and neurochemical
phenotypes. Several studies using these lines have demonstrated that selectively-bred highresponders (bHRs) represent a phenotype that is vulnerable to addiction-like behavior with regard
to psychostimulants, whereas bred low-responders (bLRs) are resilient to psychostimulant addiction.
However, despite previously demonstrated strain-differences in the neurobiology of the endogenous
opioid system, bHR/bLR differences in the response to morphine have not yet been characterized.
Here, we characterize behavioral responses of bHR and bLR rats to morphine in a variety ofexperimental paradigms, including locomotor sensitization, conditioned place preference, and rateof analgesic tolerance development. In addition, we expand the characterization of basal differencesin the expression patterns of opioid receptors between these lines. Results reveal strain differencesin the behavioral response to morphine. Further, basal differences in the biology of the endogenous
opioid system may, at least in part, mediate these differential behavioral responses. These findings
provide a foundation to use this model to probe the genetic and neurobiological antecedents
underlying resilience versus vulnerability to opioid addiction.
72

Presenter – Fakira, Amanda
Abstract ID – T27
Investigating the novel PEN-GPR83 receptor system in morphine reward and analgesia.
Amanda K. Fakira1,2, Lindsay M. Lueptow11,2, Nikita A. Trimbrake1, Lakshmi A. Devi1,2
1
1Department of Pharmacological Sciences, 2Fishberg Department of Neuroscience, and Friedman
Brain Institute, Icahn School of Medicine at Mount Sinai, New York, USA, 2Fishberg Department of
Neuroscience, and Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York,USA
Opioid-mediated effects in the reward system and descending-pain pathway result in both pain relief
and addiction. The neuropeptide PEN receptor, GPR83, exhibits high expression in the nucleus
accumbens, periaqueductal gray and spinal cord of the human and rodent brain suggesting a rolefor GPR83 in reward and pain processing. Despite this, the functional role GPR83 in reward andpain behaviors in not fully understood. Using GPR83/GFP BAC transgenic mice we examined cell
type expression and find that GPR83 in cholinergic interneurons in the nucleus accumbens anddopamine neurons in the ventral tegmental area. We also find that morphine reward-learning
increases GPR83 expression in the nucleus accumbens. Using shRNA mediated GPR83 knockdown
we examined a role for this receptor in morphine reward. We find that local decrease in receptor
expression leads to attenuation of morphine reward-learning. These results suggest an exciting
possibility that small molecule ligands targeting GPR83 have the potential to blunt the rewarding
effects of opioids. In a related set of studies, we examined the potential of targeting GPR83 toenhance morphine-induced pain relief. Preliminary examination of GPR83 agonist on morphineanalgesia demonstrate a possible blunting of morphine analgesia suggesting that modulation of
GPR83 with small molecule ligands may have a functional impact on morphine analgesia. These
results highlight the possibility that GPR83 may be a therapeutic target for the treatment opioid
dependence and addiction.
Supported in part by NIH grants (DA008863 and NS026880).
Presenter – Fang, Sophia
Abstract ID – M7
Increased abuse of illicit fentanyls are associated with opioid-related overdose deaths in the
U.S.
Sophia Fang1
1
Winston Churchill High School, Potomac, USA
Illicit fentanyls are uncontrolled opioid-like substances with opioid receptor agonist’s function andproduce serious adverse health risks in the U.S. The aim of this study is to review the current dataof national illicit fentanyl use, the opioid overdose death data at both national and state level, andtheir possible correlation. The three collected cohort data include (1) numbers of illicit fentanyls
seized by the National Forensic Laboratory Information System (NFLIS), which systematicallycollects results from forensic laboratories across the nation; (2) numbers of individual deaths due toillicit fentanyl abuse from (2) The Centers for Disease Control and Prevention (CDC) Morbidity andMortality Weekly Report, and (3) The Overdose Data and Reports published by the State of
Maryland Department of Health's Behavioral Health Administration (BHA). The results demonstrated
a substantial increase in the number of illicit fentanyl abuse nationally from 2010 to 2017. A
correlation study shows that the increased illicit fentanyl number is positively correlated with theincrease in overdose death rates in the US nationally as well as in the state of Maryland. The study
results also showed that the increased number of illicit fentanyls are positively correlated withincreased deaths due to opioid overdose and a further legal and regulatory implementation may berequired to reduce the non-control access of illicit fentanyls and subsequent public risk due to thefentanyl abuse.

73

Presenter – Faouzi, Abdelfattah
Abstract ID – M5
Design of MP135, a novel mu-delta heteromer selective Gi-protein biased agonist
Abdelfattah Faouzi1,2, Rajendra Uprety3, Ivone Gomes4, Nicolas Massaly5, Gavril W Pasternak3, Jose
Moron-Concepcion5, Lakshmi A. Devi4, Susruta Majumdar1,2
1
Washington University School of Medicine, USA 2Center for Clinical Pharmacology, St Louis College of
Pharmacy, USA, 3Department of Neurology, Memorial Sloan Kettering Cancer Center, USA 4Departmentof Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, USA, 5Department ofAnesthesiology, Washington University School of Medicine, St. Louis, USA
Conventional opioid analgesics have been known for decades to induce several adverse reactions(ADRs) such as respiratory depression and addiction. To prevent this or at least, decrease these sideeffects, a keen interest has been developed towards the model of “biased-ligand”. This concept relies on
the ability of a ligand to bind to the opioid receptor (OR) and selectively activate the Gi-protein or the βarrestin intracellular signaling route. The Gi-activation is responsible for the analgesic effects while the βarrestin pathway is usually associated with the development of multiple ADRs. On the other hand, it has
been shown that upon activation, the opioid receptors form physiologically relevant dimers which arenecessary for maturation, trafficking and downstream signaling. A previous study identified CYM51010
as a small molecule probe that selectively activates μOR-δOR heteromers and exhibits antinociception
similar to morphine with reduced antinociceptive tolerance (Gomes et al, PNAS 2013). However,additional studies emphasized that this molecule was totally β arrestin-2 biased and hence our goal here
was to synthesize a Gi-biased agonist which could prevent ADRs while sustaining a strong analgesic
potency. We now report carfentanyl amide based opioid templates as novel Gi-protein biased modulatorsof μOR-δOR heteromers with a greater selectivity over the homomeric μOR and δOR (when comparedto that of CYM51010). Pharmacological characterization of our lead MP135 suggests the possibility forthe development of a unique type (heteromer-biased) of drug that is more potent and without the severeside effects associated with conventional clinical opioids.

Presenter – Fujita, Wakato
Abstract ID – W24
Role of an endogenous chaperone protein, RTP4, on MOPr-DOPr heterodimer upregulation.
Wakako Fujita1, Mini Yokote2, Ivone Gomes3, Achla Gupta3, Hiroshi Ueda4, Lakshmi A. Devi3
1
Department of Frontier Life Sciences, and 2Department of Pharmacology and Therapeutic Innovation,
Nagasaki University Graduate School of Biomedical Sciences, Nagasaki, Japan, 3Department ofPharmacological Sciences, Icahn School of Medicine at Mount Sinai, USA, 4Department of Molecular
Pharmacology, Graduate School of Pharmaceutical Sciences, Kyoto University, Japan
Mu (MOPr) and delta (DOPr) opioid receptors associate to form heterodimers (MOPr-DOPr); theseheterodimers exhibit unique intracellular signaling. Previous studies have shown that long-term MOPr
activation by chronic morphine treatment leads to the upregulation of MOPr-DOPr and development of
antinociceptive tolerance. However, little is known about the intracellular mechanisms underlying theregulation of MOPr-DOPr levels by long-term MOPr activation. Receptor transporter protein 4 (RTP4),
one of the endogenous G-protein coupled receptor chaperone molecules, has been shown to protect
MOPr-DOPr from proteasomal degradation and to facilitate MOPr-DOPr trafficking to the plasma
membrane, suggesting an important role for RTP4 in modulating MOPr-DOPr levels at the cell membrane.
In this study, we investigated the intracellular regulation of RTP4 and of MOPr-DOPr by activating MOPr.
Treatment of Neuro2a cells expressing MOPr and DOPr with DAMGO for 24 h led to Gi activation and asignificant increase in RTP4 mRNA levels. This increase was reversed by pretreatment with pertussis
toxin or actinomycin D, suggesting the involvement of transcriptional pathways under Gi activation.
Furthermore, DAMGO induced a significant increase in MOPr-DOPr expression at the cell surface in a
RTP4 siRNA reversible manner. In addition, proximity ligation assay clearly revealed the significant
increase in interaction between RTP4 and MOPr-DOPr following DAMGO stimulation. These results
suggest that MOPr stimulation leads to increases in rtp4gene transcription and in RTP4 protein, whichinteracts with MOPr-DOPr to increase MOPr-DOPr at the cell surface.

74

Presenter – Fullerton, Evan
Abstract ID – M52
Impact of sex and advanced age on opioid analgesia in the rat periaqueductal gray
Evan Fullerton1, Myurajan Rubaharan1, Mary C. Karom1, Anne Murphy1
1
Georgia State University, Atlanta, USA
Approximately 50 million adults in the United States experience chronic pain on a daily basis, withan increased prevalence observed in individuals aged ≥65. Opioid-based medications are the mostcommon treatment used for the management of chronic pain, however there is a dearth of
information regarding the impact of advanced age on opioid signaling, specifically in the midbrain
periaqueductal gray (PAG), a key neural site for opiate action. Using a model of persistent
inflammatory pain, our studies show that morphine efficacy is significantly attenuated in aged maleand female rats. The present studies test the hypothesis that reduced efficacy of opioids observedin the aged population is due to changes in mu opioidergic expression and binding properties in the
PAG. Using immunohistochemistry and autoradiography, we report decreases in both μ opioid
receptor (MOR) expression and [3H]-DAMGO binding in the ventrolateral PAG (vlPAG) in aged (1620 months) vs. adult (2-3 months) rats. Furthermore, our behavioral studies following an intraplantar
injection of Complete Freund’s show that the antihyperalgesic effect of morphine is significantly
attenuated in aged male rats compared to adults, while no shift in morphine ED50 values wasobserved in females. These analyses indicate that age and sex impact MOR expression and binding
in the PAG. Current studies are underway assessing the impact of age and sex on MOR binding
affinity using ligand binding assays. Collectively, the data provide novel insight into the impact of age
and sex on the molecular mechanisms mediating opioid analgesia.
Presenter – Ghosh, Nillu
Morphine induced gastric pathology is prevented by proton pump inhibitor
Nillu Ghosh1, Li Zhang1, Kousik Kesh1, Sundaram Ramakrishnan1, Sabita Roy1
1
Department of Surgery, University of Miami, USA

Abstract ID – W27

Opioids are the most extensively used analgesics for pain management. However chronic use of opioids
has been associated with significant comorbidities. Gastric ulcer is a multifaceted process including acidsecretion, inflammation and extracellular matrix degradation. Morphine has been demonstrated toaugment gastric mucosal injury, although the mechanism is unclear. In the current study, a mice model
was used to investigate the mechanism by which morphine induces gastric inflammation. We also
evaluated the protective role of proton pump inhibitor (omeprazole) on morphine induced gastricinflammation. Mice stomach was histologically examined to evaluate gastric damage. Morphine treatedmice showed increase gastric inflammation and apoptosis. Significant up-regulation of inflammatory
cytokines and matrix degrading enzyme MMP9 were detected in morphine treated mice. These effects
of morphine were inhibited by the opioid antagonist naloxone. Interestingly, we found significant increases
in the basal level of acid secretion upon morphine treatment. The rate of gastric emptying was alsodelayed in morphine treated mice. Level of IL-6 in mice gastric tissue increased in morphine treated mice.
The delay in gastric emptying was alleviated in IL-6 knockout mice. Mice treated with omeprazole before
morphine showed significant reduction in gastric inflammation. In conclusion, morphine induced gastricinflammation is due long-term acid retention in the stomach which induced inflammatory cytokines
including IL6. Omeprazole prevented morphine induced acid secretion and reversed morphine induced
delay in gastric emptying and thus gastric damage. Hence, our study provides a novel approach of using
proton pump inhibitor along with morphine to reduce morphine associated gastric comorbidity

75

Presenter – Gillis, Alexander *Abstract ID – T47* Hot Topics Selection, Symposium 1, Monday, July 8, AM
Low intrinsic G-protein efficacy, not reduced ß-arrestin 2 signaling, as a mechanism for improved
opioid analgesics
Alexander Gillis1, Claudia Alamein1, Preeti Manandhar2, Arisbel Gondin3, Andrea Kliewer4, Stefan
Schulz4, Meritxell Canals3,5, Mac Christie1
1
University of Sydney, Australia, 2Macquarie University, Australia, 3Monash University, Australia,
4Friedrich Schiller University, Jena, Germany, 5University of Nottingham, UKß-arrestin 2 dependent signaling via the µ-opioid receptor (MOPr) has been proposed to mediate opioid
analgesic side effects. We failed to observe an effect of either ß-arrestin2 knock-out or 11S/T-A
phosphomutant MOPr knock-in on morphine respiratory depression in mice, challenging this ‘arrestin
hypothesis’. We re-examined relative opioid efficacy using irreverisible antagonism in a membranepotential dye assay of G-protein activated potassium channels. Buprenorphine and fentanyl haveextremely low and extremely high efficacy respectively. Putatively G-protein biased MOPr agonists
SR17018, PZM21 and oliceridine (TRV130) that do not strongly induce MOPr-ß-arrestin2
interactions were all observed to have low intrinsic efficacy compared to morphine, itself a partial agonist
relative to fentanyl. Selected opioids were profiled in a hot-plate assay of anti-nociception and two
respiratory side-effect assays in C57BL/6 mice. Contrary to published results, both SR17018 and PZM21
induced significant respiratory depression in a whole-body plethysmography assay. SR17018, PZM21
and oliceridine were found to have therapeutic windows slightly superior to morphine. Buprenorphine wasobserved to have an extremely wide therapeutic window with a ‘ceiling’ effect on respiratory depression,
whereas fentanyl had a narrow window. Fentanyl caused substantial blood oxygen desaturationcompared to morphine as assessed using conscious pulse oximetry, whereas oliceridine had minimal
effect at equi-active antinocieptive doses. Therapeutic window between anti-nociception and respiratory
depression for the agonists studied was robustly predicted by differences in intrinsic G-protein
efficacy. Low intrinsic efficacy is a plausible alternative mechanism for the improved pre-clinical profile ofputatively G-protein biased MOPr agonists that aligns with existing opioid analgesics.

Presenter – Gomes, Ivone
Abstract ID – T24
The intricacies of opioid receptor signaling by endogenous opioid peptides.
Ivone Gomes1, Salvador Sierra1, Lindsey Lueptow1, Achla Gupta1, Elyssa Margolis2, Lakshmi A.
Devi1
1
Icahn School of Medicine at Mount Sinai, New York, USA, 2University of California at San Francisco,
USA
Opiates such as morphine and fentanyl are the most widely used analgesics for the treatment of
pain; however, prolonged treatment with these drugs leads to the development of tolerance andopiate addiction. Recent reports on the ‘Opioid Epidemic” have led to a great interest in a deeper
understanding of fundamental mechanisms activating opioid receptors. It is generally thought thatthe three types of opioid receptors (mu, delta, kappa), are activated by peptides from three different
precursors - respectively proopioimelanocortin, proenkephalin and prodynorphin. Post-translational
processing of these precursors leads to the generation of ~20 peptides with opioid receptor activity
and this has led to a long-standing question regarding the significance of the repertoire of
endogenous opioid peptides. Here, we address this question by evaluating ligand binding andsignaling properties of the peptides at all the three receptors. We show that all of the peptides areable to activate the three opioid receptor types, many of the peptides exhibit agonist-directed
receptor signaling (functional selectivity) and some of them exhibit agonist-directed receptor
trafficking. We also show that dynorphins and shorter forms of beta-endorphin activate all three
receptor types with high potency and are able to elicit mu receptor signaling in midbrain dopamine
neurons ex vivo. Collectively, this information leads to a better understanding of the opioid system
which is a prerequisite for the development of drugs for the treatment of pain and addiction.
This work was supported in part by NIH grants (DA008863 and NS026880).

76

Presenter – Groom, Sam
Abstract ID – M25‘Compound 1’: a G-protein-biased MOPr agonist that induces MOPr desensitization
apparently through GRKSam Groom1, Alexandra Conibear2, Alex Disney1, Stephen Husbands1, Yangmei Li3, Lawrence Toll4,
Graeme Henderson2, Eamonn Kelly2, Chris Bailey1
1
Department of Pharmacy & Pharmacology, University of Bath, Bath, UK, 2School of Physiology,
Pharmacology & Neuroscience, University of Bristol, Bristol, UK, 3Department of Drug Discovery andBiomedical Sciences, University of South Carolina, Columbia, USA, 4Florida Atlantic University, Boca
Raton, USA
In this study we investigated MOPr desensitization induced by G-protein-biased MOPr agonists
using brain slice electrophysiology. Biased agonists studied were PZM21 (Manglik et al, 2016) and‘Compound 1’ (Tyr-c[D-Lys-Phe-Tyr-Gly]) (Li et al, 2016), which in HEK293 cells has equivalent
efficacy to morphine for G-protein activation, but induces less phosphorylation of Ser375 on MOPrand arrestin-3 recruitment. Whole-cell voltage-clamp recordings were made from rat locus coeruleus(LC) neurons. MOPrs couple to GIRK channels, with the amplitude of K+ currents providing ameasure of MOPr activation. Opioids (30µM) were perfused for 10 minutes and reductions in opioidevoked currents used to assess MOPr desensitization. Kinase modulators were pre-incubated for
20 minutes prior to agonist perfusion. While PZM21 induced less MOPr desensitization thanmorphine, surprisingly, Compound 1 induced robust MOPr desensitization of greater magnitude thanmorphine. The extent of Compound 1-induced MOPr desensitization was unaffected by activation(PMA, 1µM) or inhibition (GF109203X, 1µM) of PKC but was significantly reduced by inhibition of
GRK (Compound 101, 30µM). Despite Compound 1 inducing minimal Ser375 phosphorylation andarrestin recruitment, it induced robust MOPr desensitization. Unexpectedly, our data suggests thatthe mechanisms of Compound 1-induced MOPr desensitization are GRK-dependent. Investigations
are ongoing to further detail the nature of the receptor desensitization induced by Compound 1 andits implications for the functional consequences of biased signalling at MOPr.
Manglik A et al (2016) Nature 537:185-190; Li Y et al (2016) J Med Chem 59:1239-45
Presenter – Gupta, Achla
Abstract ID – W22
A role for an endocytic peptide processing enzyme in modulating opioid receptor trafficking
and function
Achla Gupta1, Ivone Gomes1, Salvador Sierra1,2, Lakshmi A. Devi1
1
Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York,USA, 2Dept. of Physiology and Biophysics, Virginia Commonwealth University, Richmond, USA
We previously showed that following activation by endogenous opioids mu and delta receptors
undergo differential internalization and recycling; the rate and extent of recycling but notinternalization is regulated by the endocytic peptide converting enzyme, ECE2. In this study wefocused on kappa opioid receptors (KOR) and examined the ability of endogenous opioid peptides
released by post-translational processing of prodynorphin and proenkephalin to induce receptor
internalization/recycling and how ECE2 modulates these processes. We find that longer opioid
peptides induce fast and robust internalization/recycling; co-expression of ECE2 affects the rate andextent of internalization/recycling by only a subset of these peptides. Proximity-based ligation assay
shows that KOR and ECE2 are in close proximity to facilitate co-internalization. We also find thatthe kinetics of ECE2 internalization/recycling is comparable to that of KOR and that blockade of KOR
recycling leads to retention of ECE2 in an intracellular compartment. Moreover, we find that inhibitionof ECE2 leads to significant attenuation of receptor resensitization. Taken with the differential
expression of ECE2 in the brain (relatively high expression in midbrain & hypothalamus and low
expression in the striatum & hippocampus), these results support a role for ECE2 in differentiallymodulating opioid receptor function.
This work was supported in part by NIH grants (DA008863 and NS026880).
77

Presenter – Hamada, Yusuke
Abstract ID – W52
Possible involvement of μ-opioid receptor-positive neurons located in the PVN in the controlof the tumor-bearing state
Yusuke Hamada1, Sara Yoshida1, Michiko Narita1, Hideki Tamura2, Hiroyuki Tezuka3, Akihiro
Yamanaka4, Naoko Kuzumaki1,2, Minoru Narita1,21
Dept. Pharmacol., Hoshi Univ., Tokyo, Japan, 2L-StaR, Hoshi Univ., Tokyo, Japan, 3Dept. Cellular
Function Analysis, Fujita Health Univ., Nagoya, Japan, 4Dept. Neurosci.II, RIEM, Nagoya Univ.,
Nagoya, Japan
Recent studies have shown that either chronic stressful conditions or depression could suppress thesystemic immune response. In contrast, emerging evidence from the field of reward research has
shown that positive stimulation, such as pleasant sensations, could enhance the immune response.
Therefore, we hypothesized that crosstalk between changes in the central function and the peripheral
immune regulation could be crucial for tumor progression. On the other hand, the neural regulationin the hypothalamic paraventricular nucleus (PVN) is known to have the ability to change the innate
endocrine and immune functions. Notably, it has been reported that µ-opioid receptor is highly
expressed in the PVN. However, its physiological function remains unclear. In this study, weinvestigated the role of the hypothalamic μ-opioidergic system in tumor progression. To investigate
the changes in the activity of PVN neurons under the tumor-bearing condition, we used cFosCreERT2::EGFP mice which can capture activated neurons during a time windows defined by 4hydroxytamoxifen administration using targeted recombination in active populations method. Welabeled the cFos-positive neurons by the 4-hydroxytamoxifen administration 7 days after the tumor
implantation. Interestingly, we found that µ-opioid receptor-expressing neurons in the PVN weresignificantly activated under the tumor-bearing state. Under these conditions, we demonstrated thatthe continuous microinjection of morphine into the PVN significantly suppressed tumor growth.
These findings suggest that the activation of μ-opioidergic system in the PVN could directly regulate
tumor progression via the stress-related feed-forward circuit.
Presenter – He, Ying
Abstract ID – T28
Long term opioid use is required for sickle cell disease after curative hematopoietic stem celltransplantation
Ying, He1,3, Zaijie Jim Wang2
1
University of Illinois at Chicago, Chicago, USA, 2University of Illinois at Chicago, Chicago, USA,
3
University of Maryland, Baltimore, USA
Pain is the hallmark feature of sickle cell disease (SCD), which is characterized as chronic baseline
pain with intermittent episodes of acute pain crises throughout the life long course of the disease.
Hematopoietic stem cell transplantation (HSCT) offers a cure of this genetic blood disorder. While
sickle cell phenotype is corrected in patients after HSCT, it remains unknown about pain transition
and resolution in SCD following HSCT. Taking advantage of a humanized mouse model of SCD, wecharacterized pain behaviors in SCD mice before and after HSCT. The evoked hypersensitivities tomechanical and thermal stimuli were measured by the von Frey test, and the Hargreaves test,
respectively. The conditioned place preference (CPP) paradigm was applied to reveal spontaneousongoing pain in mice by a single trial conditioning (clonidine: 1 µg in 5 µL of saline, i.t.). Both
spontaneous ongoing pain and evoked sensory hypersensitivity were observed in three-month old
SCD mice. Receiving non-myeloablative allogeneic transplantation with stem cells from non-sickle
control littermates, the recipient SCD mice established quick and stable engraftment within two
weeks. After a four-week recovery period post HSCT, pain behaviors were tested once every week
for 8 weeks. We found robust spontaneous ongoing pain, mechanical allodynia and thermal
hyperalgesia remained to exist in the recipient SCD mice. In agreement with the clinical reports,
these data demonstrated the presence of chronic pain in SCD mice even after receiving successful
curative HSCT, suggesting that long term opioid usage remains to be required in post-transplant
SCD patients.
78

Presenter –He, X. Jenny
Abstract ID – W34
Mechanisms of presynaptic suppression of GABA release by opioid receptors in the
hippocampus
X. Jenny He1, Matthew Banghart1
1
Division of Biological Sciences, Section on Neurobiology, UCSD, La Jolla, USA
Opioid receptors are prominent in interneurons of the hippocampus where they modulate GABAergic
inhibition onto pyramidal cells. Mu opioid receptor (MOR) activation is known to suppress GABA
release from parvalbumin (PV) basket cells in the CA1 region, which positions opioids to affect spiketiming, oscillations, and synchronization of neuronal populations in the hippocampus. Although thedelta opioid receptor (DOR) is also prominent in the hippocampus, its functional role in distinct
interneuron classes has not been established, nor have the molecular mechanisms that underlie
MOR and DOR-mediated suppression of synaptic transmission. Using a combination ofelectrophysiology and optogenetics, we discovered that both MOR and DOR strongly inhibit GABA
release from PV+ basket cells in a mutually-occlusive manner. Although both receptors appeared toutilize a presynaptic mechanism, they were differentially prone to desensitization. We further
investigated the molecular mechanisms of opioid receptor-mediated suppression using two-photon
calcium imaging and pharmacology and found evidence for both calcium-dependent and calciumindependent modulation. Our results suggest a previously unappreciated role for DOR in modulating
perisomatic inhibition in the hippocampus and reveal the molecular mechanisms by which both MOR
and DOR regulate GABA release from PV basket cells in CA1.
Presenter – Hearing, Matthew
Abstract ID – T10
Cell-type and region-specific nucleus accumbens AMPAR plasticity associated with opioid
reward, drug-seeking, and withdrawal
Aric Madayag1, Devan Gomez1, Eden Anderson1, Anna Ingebretson2, Mark Thomas2, Matthew Hearing1
1
Marquette University, Milwakee, USA, 2University of Minnesota, USA
Despite evidence that morphine-related pathologies reflect adaptations in NAc glutamate signaling,
substantial gaps in basic information remain. Using whole-cell electrophysiology to examine AMPARmediated synaptic transmission at (NAc) medium spiny neurons (MSNs) containing the dopamine
receptor type 1 or 2 (D1, D2-MSNs), we examined region-, time-, and cell-specific adaptations associatedwith abstinence from non-contingent vs self-administered opioids and spontaneous vs. precipitatedwithdrawal. We find that protracted withdrawal from non-contingent and self-administered opioids
produces overlapping and distinct changes in pre/postsynaptic excitatory drive at D1- and D2-MSNs in
the NAc shell (NAcSh) that may reflect adaptations unique to infralimbic cortex inputs. In the NAcCore,
unlike previous findings with non-contingent morphine, self-administration produced increases anddecreases in transmission at D1- and D2-MSNs, respectively. Similar to our findings with cocaine, reexposure to morphine produced a transient reduction in synaptic strength. However, unlike cocaine,
blocking morphine-induced AMPAR endocytosis augmented, rather than inhibited, reinstatement of place
preference, suggesting that similar forms of plasticity and post-drug experience may have distinct
behavioral consequences across drug classes. In contrast to plasticity associated with acute, repeated,or drug re-exposure, we find that spontaneous withdrawal following a dependence-inducing regimen ofmorphine aligned with enhanced excitatory drive selectively at D2-MSNs in the NAcSh. Preliminary data
indicate that somatic symptoms associated with this withdrawal can be mitigated by systemic injection of
a novel G protein-gated inwardly-rectifying K+ channel agonist, ML-297. Accordingly, future studies will
determine if similar plasticity occurs with precipitated withdrawal and explore the impact of this drug onplasticity.

79Presenter – Hearing, Matthew
Abstract ID – W6
Sex-dependent opioid-induced plasticity in prefrontal-accumbens circuits: Implications forcognitive control and relapse
Eden Anderson1, Annabel Engelhardt1, Matthew Hearing1
1
Marquette University, Milwakee, USA
Even when taken as prescribed, opioid use carries significant risk for misuse, dependence, andaddiction. These risks are influenced by biological variables such as sex, with woman tending to escalate
drug use and shift towards uncontrollable drug-taking on a more rapid time scale, and deficits in cognitive
control, whether intrinsic or arising from early or prolonged drug use. Dysfunction of medial prefrontal
cortex-nucleus accumbens circuits (mPFC-NAc) have long been identified as a key substrate forpathological strengthening of addiction behavior, as they govern motivational processes, including
relapse to drug-seeking, as well as flexible and inhibitory behavior. Unlike psychostimulants, theanatomical selectivity and underlying mechanisms promoting dysfunction of these circuits is poorly
characterized. Using a mouse model of self-administration, we show that pyramidal neurons (PN) in the
prelimbic cortex (PrLC) exhibit a hypoactivation state but increased firing capacity following 5-6 weeks
of drug exposure and that this hypofrontality emerges after only 2 weeks in females, is predominantly
expressed in PN projecting to the NAc core, and aligns with time- and sex-dependent changes in thebalance of excitatory:inhibitory synaptic drive. Hypofrontality aligned with a rescuable impairment incognitive flexibility (operant attentional set-shift model) and perhaps escalation of intake and relapse
behavior, with preliminary findings indicating that hypofrontality of PrLC-Core PN is paralleled by anunderlying strengthening at PrLC-Core spiny neuron synapses. Data suggest that opioids may produce
distinct psychopathologies through discrete adaptations in the PrLC-Core circuits, and that intrinsic sex
differences in this circuit influence opioid plasticity and risk for developing addiction.

Presenter – Hill, Rob
Abstract ID – M23
Fentanyl depression of respiration: differential reversal by antagonists and reduced crosstolerance to morphine.
Rob Hill1,2, William.L.Dewey 3, Eamonn Kelly1, Graeme Henderson1
1
University of Bristol, UK, 2University of Nottingham, UK, 3Virginia Commonwealth University, USA
In this study we compared the ability of naloxone and diprenorphine to reverse fentanyl depressionof respiration and examined whether fentanyl could break through morphine-induced tolerance inmice. Respiration was measured by whole body plethysmography in male mice (CD-1) breathing
95% air/5% CO2. Prolonged morphine pre-treatment was by priming injections (3x100 mg/kg i.p.
12hr apart) followed by implantation of an osmotic minipump (s.c.) that delivered 45 mg/kg/d for 6d.
Administration of diprenorphine (30 or 90 μg/kg i.p.) produced dose-dependent equivalent reversalof fentanyl and morphine respiratory depression. In contrast, naloxone (0.3 or 1 mg/kg i.p.) produced
greater reversal of morphine respiratory depression than that induced by fentanyl. An acutechallenge dose of morphine (10 mg/kg i.p.) induced 40% depression of respiration in control micebut following prolonged morphine pre-treatment the morphine challenge did not depress respiration(i.e. mice were tolerant). In contrast the same morphine pre-treatment only partially reduced the
response to an equipotent dose of fentanyl (0.15 mg/kg i.p.) from 40% to 20% depression. Withhigher equipotent challenge doses of morphine (90 mg/kg i.p.) and fentanyl (1.35 mg/kg i.p.) the
response to fentanyl was not reduced, being 60% depression in control and morphine pre-treatedmice, whereas that to morphine was reduced from 60% to 25%. These data demonstrate that
diprenorphine is better at antagonizing fentanyl than naloxone and that prolonged morphine
treatment induces less cross-tolerance to fentanyl.

80Presenter – Hipolito, Lucia
Abstract ID – M28
Ethanol intra-VTA induces CPP through the activation of local MORs and increases NR2A
expression in the NAc
Yolanda Campos-Jurado1, Lucia Martí-Prats1, Jose Moron-Concepcion2, Ana Polache1, Luís
Granero1, Lucia Hipolito1
1
University of Valencia, Valencia, Spain, 2Washington University, St. Louis, USA
The neurobiological mechanisms underlying ethanol motivational properties are still not fully
understood. Drug-associated environmental stimuli can trigger ethanol relapse and promote ethanol
consumption and NMDA receptors have an important role in these learn-context associations. In thepresent study we evaluated, for the first time, the ability of ethanol directly administered into the VTA
to induce conditioned place preference or aversion (CPP or CPA), focusing on the role of the MORs
and followed by the analysis of changes in the expression of NMDA receptor and its different subunits
in the implicated brain areas. First, we analyzed the effect of 0, 35, 70, 150 and 300 nmol of ethanol
intra-VTA in the development of CPP or CPA. Furthermore, the involvement of local MORs in theacquisition of CPP was studied by their selective blockade with β-Funaltrexamine. Finally, we used
the western blot and In Situ Hybridization techniques to analyze changes after CPP expression in
the levels of NMDA receptor subunits in the Nucleus Accumbens (NAc) and Hippocampus. Results
showed, for the first time, that ethanol intra-VTA induces either CPP or CPA in a dose-dependent
manner. Moreover, the blockade of VTA MOR impairs the acquisition of ethanol-induced CPP. The
In Situ Hybridization results showed that, in the NAc, mRNA levels of NR2A but not NMDAR1 arehigher after ethanol CPP. These novel results set the base to further analyze the mechanism bywhich ethanol motivational properties are associated with learned environmental cues.
Presenter – Holm, Lovisa
TRV130 modulates reward in a beta-arrestin dependent manner
Lovisa Holm1, Annika Thorsell1, Markus Heilig1
1
CSAN, IKE, Linkoping University, Sweden

Abstract ID – M50

The mu-opioid receptor (MOR) is a G-protein-coupled receptor signalling through both G-proteins aswell as β-arrestins, and ligands for the receptor may be biased towards activation of either the Gprotein or β-arrestin-mediated intracellular pathways. TRV130 is a G-protein pathway biased agonistat the MOR that produces a potent antinociceptive effect in mice without inducing severe constipation
or respiratory dysfunction. Here, we examined reward-related behavior for TRV130 in a classic
conditioned place preference (CPP) paradigm using beta-arrestin 2 knockout mice (wt, het, and ko).
In addition, antinociceptive effects were measured using hot plate and tail flick, and compared to theeffects of morphine. The CPP model used was a two-chamber set-up (Med Associates Inc. ST
Albans, VT, USA). Following pre-test, conditioning was done in daily 30 minute sessions alternating
TRV130 and vehicle for 8 days (4 TRV130 (1.0 mg/kg) and 4 vehicle sessions). The test-session
was preceded by a vehicle injection and run for 30 minutes. TRV130 (1.0 mg/kg) induced CPP in wt
BARR-mice (CPP-score 68.3 +/- 1.9%), while both the het BARR and the ko BARR showed blunted
CPP (score 62.2 +/- 2.2 and 58.7 +/- 1.2, respectively). Antinociception was similar in all genotypes
indicating a beta-arrestin independent mechanism. Data presented here indicate that TRV130
induces reward-related behavior via a beta-arrestin dependent mechanism.

81

Presenter – Hsu, Wen-Yu
Abstract ID – M1
Association between opioids use disorder and fractures
Wen-Yu Hsu1,2, Cheng-Li Lin3, Chia-Hung Kao2,41
Department of Psychiatry, Changhua Christian Hospital, Changhua, Taiwan, 2Graduate Institute ofClinical Medical Science, China Medical University, Taichung, Taiwan, 3Management Office for
Health Data, China Medical University Hospital, Taichung, Taiwan, 4School of Medicine, College ofMedicine, China Medical University, Taichung, Taiwan
Patients with opioid use disorder (OUD) have increased risk of several comorbidities, such as
hepatitis C virus and human immunodeficiency virus infections, cardiac toxicity, pulmonary
disorders, and sleep apnea, importantly, with bone fractures. We hypothesize that the incidences offracture are relatively higher among patients with OUD in comparison with the general population.
The aim of this study is to investigate whether fractures are more prevalent among OUD patients in
Taiwan. For this population-based cohort study, claims data from 2000 to 2011 were obtained fromthe National Health Insurance Research Database (NHIRD) in Taiwan. We used the dataset ofLongitudinal Health Insurance Database 2000, which contains the longitudinally linked data of
1,000,000 enrollees randomly sampled from the NHIRD. We compared the incidence of fracture
between patients with OUD and non-OUD cohorts. OUD patients were 4.13 times more likely tosuffer fractures than non-OUD patients (incidence rate per 1000 person-years [IR]: 23.0 vs. 5.47,
adjusted HR: 3.74, 95% CI: 3.27–4.29). Compared with the general population, the risk of fracture
was higher among the patients with OUD. Risk of fracture was higher in male, elderly, diabetes
mellitus, alcohol related illness, or osteoporosis. The cumulative incidences of fracture of OUD andnon-OUD patients significantly differed. The patients in OUD group had higher adjusted hazard ratio
for fracture than that in the non- OUD group in this study. Further studies for the mechanism areneeded and further regular examination for osteoporosis in this population should be suggested.
Presenter – Inan, Saadet
Abstract ID – W20
Nalbuphine suppresses deoxycholic acid-induced scratching in male, Swiss Webster mice
Alan Cowan1, Nae J Dun1, Saadet Inan1
1
Temple University, Philadelphia, USA
Cholestatic itch is an ongoing challenge in medicine that is often refractory to available therapeutics.
In consolidating the concept of nalbuphine as a global antipruritic, we have tested this compoundagainst bile acid (deoxycholic acid, DCA)-induced compulsive body scratching in mice. DCA (0.3-10
mg/kg, s.c.) evoked dose-dependent scratching with hindpaws. Bile acids are believed to cause therepetitive behavior by activating MRGPRX4 receptors in this species (Meidiong et al., PNAS 2019,
In press). Nalbuphine (10 mg/kg, s.c.) was administered 30 min before a submaximal dose of DCA(3 mg/kg, s.c.) and the number of scratching bouts was counted for 30 min. Pretreatment with
nalbuphine reduced the number of DCA-induced bouts from 94 ± 17 (SEM) to 24 ±7 (P<0.01).
Nalbuphine thus joins naloxone, naltrexone and nalmefene as a promising medication for cholestaticitch. In conclusion, we call attention to nalbuphine as a pan-antipruritic drug.
Supported by CSAR Grant #DA013429

82

Presenter – Inan, Saadet
Abstract ID – W21
Nalbuphine, a kappa opioid receptor agonist and mu opioid receptor antagonist, suppresses
scratching in mice induced by HIV-1 TAT and also by chloroquine, an anti-malarial agent
Saadet Inan1, Nae J Dun1, Alan Cowan1
1
Temple University, Philadelphia, USA
Chronic itch is one of the most disturbing symptoms encountered not only in some skin diseases but
also in systemic, neuropathic and psychological disorders. There is no FDA approved medication for
the treatment of chronic itch in the USA. Nalfurafine, a kappa opioid receptor (KOR) agonist, hasbeen approved for the treatment of uremic pruritus in Japan and our previous work on this drug has
been instrumental in developing the concept that the KOR is fundamental in the anti-itch process.
Nalbuphine is an attractive commercial alternative to nalfurafine since it is (unlike nalfurafine)unscheduled and antagonizes scratching in mice precipitated by phoenixin-3. We now report thenovel finding that HIV-1 TAT (0.1-1 mg/kg, s.c.) causes excessive body scratching with the hindpaws
in a mouse standard itch assay (Kuraishi et al.,1995). Promisingly, nalbuphine (0.3-10 mg/kg, s.c.,at -20 min) attenuated this scratching. Additional positive results were obtained with nalbuphine (110 mg/kg, s.c., at -20 min) against chloroquine (a submaximal dose, 10 mg/kg, s.c.), an anti-malarial
drug that effects scratching in certain black Africans. All in all, nalbuphine, which looked like anincremental drug for pruritus, may turn into a transformational one.
Supported by CSAR Grant # DA013429 and the gift of HIV-1 TAT from Phoenix Pharmaceuticals
Presenter – Iyer, Vishakh
Abstract ID – W8
Negative allosteric modulation of CB1 cannabinoid receptor signaling suppresses opioidmediated reward and dopamine efflux
Vishakh Iyer1,2, Claudia Rangel-Barajas1,2, Jonathon D. Crystal1,2, Abhijit Kulkarni3, Ganesh A.
Thakur3, George V. Rebec1,2, Andrea G. Hohmann1,2,4
1
Program in Neuroscience, Indiana University, Bloomington, USA, 2Department of Psychological andBrain Sciences, Indiana University, Bloomington, USA, 3Department of Pharmaceutical Sciences,
Northeastern University, Boston, USA, 4Gill Center of Biomolecular Science, Indiana University,
Bloomington, USA
In recent years, the pervasiveness of the opioid crisis has underscored the urgent need to identify
novel compounds that circumvent opioid abuse liability while sparing their therapeutic efficacy.
Opioid reward and reinforcement require crosstalk between cannabinoid type 1 receptors (CB1R)
and mu-opioid receptors (MOR). While CB1R modulate the motivational properties of opioids via
dopaminergic mechanisms and are well-positioned to control critical aspects of opioid abuse, theclinical use of CB1R agonists and antagonists have been limited by adverse side effects. Allostericmodulation represents an indirect strategy to modulate endocannabinoid signaling by targeting a site
distinct from that which binds agonists like THC or the natural ligands (endocannabinoids) of CB1R.
Positive and negative allosteric modulators (PAM and NAM) increase or decrease the affinity and/or
efficacy of the orthosteric agonists, respectively and may produce an optimal and circumscribed
spectrum of physiological effects compared to the indiscriminate effects of agonists or antagonists.
The present study used GAT358, a CB1-NAM to test the hypothesis that negative allostericmodulation of CB1R would block opioid reward by decreasing dopaminergic transmission, while
bypassing the unwanted side effects of direct CB1R blockade. In vivo voltammetry findings suggestthat GAT358, a CB1-NAM can attenuate morphine-induced increases in dopamine signaling in the
nucleus accumbens shell, a key component of the mesocorticolimbic reward pathway. GAT358 also
blocked morphine mediated conditioned place preference without producing reward or aversion
when administered alone. Our results support the therapeutic potential of CB1-NAM and highlighttheir role in fine-tuning undesirable on target-effects of MOR signaling.

83Presenter – Jullie, Damien
Abstract ID – W19
A discrete presynaptic vesicle cycle for neuromodulator receptors
Damien Jullie1,2, Miriam stoeber1,2, Jean-Baptiste Sibarita3,4, Thomas M Bartol5, Seksiri
Arttamangkul6, Terrence J Sejnowski5, Eric Hosy3,4, Mark von Zastrow1,2
1
Department of Psychiatry, 2Department of Cellular and Molecular Pharmacology, University ofCalifornia, San Francisco School of Medicine, San Francisco, USA, 3Interdisciplinary Institute for
Neuroscience, UMR 5297, Centre National de la Recherche Scientifique, Bordeaux, France,
4Interdisciplinary Institute for Neuroscience, University of Bordeaux, Bordeaux, France, 5Howard
Hughes Medical Institute, Salk Institute for Biological Studies, La Jolla, USA, 6The Vollum Institute,Oregon Health and Science University, Portland, USA
A major function of GPCRs is to inhibit neurotransmitter release from presynaptic terminals. Thisrequires ligand-activated receptors to couple locally to effectors at terminals. The current
understanding of how this is achieved is based on receptor immobilization on the presynaptic
surface. Here we show that opioid peptide receptors, GPCRs which mediate highly sensitive
presynaptic inhibition, are instead dynamic in axons. Opioid receptors diffuse rapidly throughout theaxon surface and internalize after ligand-induced activation specifically at presynaptic terminals. We
delineate a parallel regulated endocytic cycle for GPCRs at the presynapse, separate from thesynaptic vesicle cycle, which clears activated receptors from the presynaptic surface and mediates
local reinsertion to maintain the diffusible surface pool. We propose an alternate strategy to achieve
local control of presynaptic effectors that, opposite to being based on receptor immobilization, isbased on lateral mobility of receptors and leverages the inherently allosteric nature of GPCR effector coupling.
Presenter – Kandasamy, Ram
Abstract ID – M21
Positive allosteric modulators of the mu-opioid receptor produce pain relief without disruptiveside effects
Ram Kandasamy1, John R. Traynor1
1
Department of Pharmacology and Edward F. Domino Research Center, University of Michigan Medical
School, Ann Arbor, USA
Opioids produce analgesia by acting at the orthosteric site on the mu-opioid receptor (MOR); however,
GPCR function can also be controlled by compounds acting at a separate (allosteric) site. One compound,
BMS-986122, binds to the allosteric site on the to enhance the affinity/potency of MOR agonists in vitro.
We hypothesized that BMS-986122 produces antinociception by enhancing endogenous and exogenous
opioid activity in vivo without disruptive side effects. Male and female mice were administered BMS986122 or vehicle (10 mg/kg, i.p.). Antinociception was measured using the hot plate, tail withdrawal, andvon Frey tests. Side effects were assessed using tests of pain-depressed nesting, constipation, andantinociceptive tolerance. Thirty min pretreatment of BMS-986122 enhanced antinociception producedby methadone (10 mg/kg) on the hot plate test. BMS-986122 alone produced antinociception on the tailwithdrawal test for 2 hours. This effect was reversible by naloxone pretreatment (10 mg/kg) suggestingthat the antinociception is caused by enhancement of endogenous opioid activity. BMS-986122 also
produced naloxone-reversible antinociception against inflammatory pain induced by intraplantar
injections of carrageenan and Complete Freund’s Adjuvant (CFA). BMS-986122 produced
antinociception against CFA-induced pain after multiple administrations, indicating a lack of tolerance
development, and prevented visceral pain-induced depression of nesting. BMS-986122 did not produce
constipation compared to morphine (10 mg/kg). Our data suggest that positive allosteric modulators of
MOR produce antinociception against acute and persistent pain by enhancing exogenous andendogenous opioids. This antinociception is produced without the development of antinociceptive
tolerance, constipation, or behavioral disruption.
Supported by R37 DA039997 and T32 DA007268.

84

Presenter – Kapoor, Abhijeet
Abstract ID – M35
The unique conformational ensemble and active conformation of µ-opioid receptor induced bymethadone
Abhijeet Kapoor1, Davide Provasi1, Marta Filizola1
1
Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, USA
Although not the most lethal opioid drug, and mainly used to curb opioid addiction and reduce cravings,
methadone still caused ~3,400 overdose deaths in 2016. The different properties of methadone comparedto other opioids (e.g., lower cross-tolerance in animal models and higher relative potency during opioid
rotation in patients) have been attributed to its ability to generate a unique conformational ensemble ofthe µ-opioid receptor (MOR). While recent estimates from a novel biophysical assay suggested that Lmethadone stabilizes distinct MOR active conformations from those induced by loperamide and the Gprotein-biased agonist PZM21, the effect of L-methadone on the conformational sampling and activation
kinetics of MOR remains largely uncharacterized at the atomic level. Here, we investigate
computationally the conformational ensemble of MOR bound to L-methadone and compare it to thereceptor conformational sampling induced by morphine and the G protein-biased ligand TRV-130. First,
we used a recently validated strategy that combines induced-fit docking and metadynamics simulations
to predict the binding pose of L-methadone at MOR in an explicit lipid-water environment. We then carriedout ~0.3 ms of enhanced molecular dynamics (MD) simulations of the methadone-bound MOR complex.
The resulting methadone-induced MOR dynamics was analyzed using Markov State Models andcompared with the results of previously published ~0.5 ms high-throughput MD simulations of MOR boundto morphine or TRV-130. The analysis confirms that methadone induces a distinct conformationallandscape and kinetics of MOR activation compared to morphine and TRV-130 and reveals the atomic
details of a unique active conformation of MOR.
Presenter – Karkhanis, Anushree
Abstract ID – W26
Dysregulation of KOR-DAT interaction in the NAc mediates enhanced cocaine seeking following
adolescent social isolation in rats
Anushree N. Karkhanis1,2, Paige M. Estave2, Steven E. Albertson2, Jeff L. Weiner2, Sara R. Jones2
1
Binghamton University, Binghamton, USA, 2Wake Forest School of Medicine, Winston-Salem, USA
Chronic stress during adolescence augments cocaine craving and withdrawal symptoms, potentiallydriving cocaine addiction in humans. Similarly, in rodents, adolescent social isolation (aSI) results inelevated cocaine seeking in adulthood compared to group housed rats (aGH). Previous studies haveshown that Kappa opioid receptors (KORs) and dopamine transporter (DATs) exhibit hyperfunction
following aSI. Both proteins are involved in substance use disorder. Here we investigated whether theinteraction between these two proteins drives the potentiated cocaine seeking in aSI rats. Following 6weeks of group (aGH; 4rats/cage) or isolated (aSI; 1rat/cage) housing, rats were given access to levers
to self-administer cocaine intravenously. A progressive ratio schedule showed enhanced cocaine seeking
in aSI rats (final ratio: aGH-77; aSI-268; 2.25mg/kg/infusion), which was reduced by KOR inhibition.
Dopamine response to acute cocaine (15mg/kg) was enhanced in in aSI rats, which was reversed by
KOR blockade, possibly indicating that the reinforcing value of cocaine is greater for aSI rats. Ex vivo
fast-scan-cyclic-voltammetry shows that acute KOR activation results in cocaine tolerance and cocaine
self-administration augments KOR response to the agonist only in aGH rats. This relationship is alteredby exposure to chronic stress. Interestingly, selectively overexpressing KORs in dopamine neurons inadulthood does not result in similar changes in cocaine potency. These data suggest that chronic stress
exposure, specifically during adolescence, dysregulates the functional interaction between KORs and
DATs, which in turn affects cocaine self-administration. These findings implicate that early-life stress may
result in a chronic KOR-dependent hypodopaminergic state that may perpetuate future cocaine seeking.

85

Presenter – Király, Kornél
Abstract ID – M39
The contribution of cocaine- and amphetamine-regulated transcript (CART) peptide fragments tospinal nociceptive transmission in rats
Kornél Király1, Zita Puskár2, Márk Kozsurek2, Erika Lukácsi2, Klára Gyimesi2, Csaba Fekete3, Balázs
Gereben3, Petra Mohácsik3, Zsuzsa Helyes4, Kata Bölcskei4, Valéria Tékus4, Edina Szűcs5, Sándor
Benyhe5, Timea Imre6, Pál Szabó6
1
Department of Pharmacology and Pharmacotherapy and 2Department of Anatomy, Histology andEmbryology, Semmelweis University, Budapest, Hungary, 3Department of Endocrine Neurobiology,
Institute of Experimental Medicine of the Hungarian Academy of Sciences, Budapest, Hungary,
4
Department of Pharmacology and Pharmacotherapy, Centre of Neuroscience, University of Pécs, Pécs,
Hungary, 5Institute of Biochemistry, Biological Research Centre and 6MS Metabolomics Laboratory,
Research Center for Natural Sciences of the Hungarian Academy of Sciences, Budapest, Hungary
Long CART peptides (55-102 and 62-102) containing C-terminal globular region are abundant in many
brain regions and in spinal cord. Although the existence of CART in the superficial dorsal horn suggests
its involvement in pain transmission, the exact role of the peptides in these processes have not been
determined. The C-terminal portion is important in the putative receptorial actions and most of thebiological effects. Short variants CART(55-76 and 62-76) are endogenously unproven and only exiguous
data are available about their activity. The N-terminal amino acids of CART(55-x) peptides are Ile-Pro-Ile(IPI), the sequence of the known DPP4 inhibitor. IPI shows MOR- mediated antihyperalgesic action in
inflammatory pain models after intrathecal (it) administration in rats. Herein we studied the effect of CART
peptides in tail flick test, carrageenan induced inflammatory hyperalgesia and PSL induced neuropathy.
Long variants caused rough ascending tremor after it. administration. IPI containing CART(55-76)
resulted in MOR mediated antihyperalgesia in inflammatory state, weak non-opioid action in neuropathic
model and no effect in tail-flick test. CART(62-76) showed KOR-mediated antihyperalgesic effect incarrageenan model, however, it was ten-fold less potent. N-terminal amino acids of CART(62-76) are
Tyr-Gly suggesting opioid receptor binding, yet on GTPgS binding assay it was undetectable. CART(5576) did not inhibit DPP4 activity in vitro, but after it. administration we measured increased IPI level inspinal cord extract by mass spectrometry. In conclusion, both short CART peptides show opioid-mediatedantihyperalgesic action in inflammatory pain with distinct mechanisms.

Presenter – Kliewer, Andrea
Abstract ID – T2
Phosphorylation-deficient G-protein-biased μ-opioid receptors improve analgesia anddiminish tolerance but worsen opioid side effects
A. Kliewer1, F. Schmiedel1, S. Sianati2, A. Bailey3, J. T. Bateman4, E. S. Levitt4, J. T. Williams5, M. J.
Christie2, S. Schulz1
1Institute of Pharmacology and Toxicology, Jena University Hospital, Friedrich-Schiller-University,
Germany, 2Discipline of Pharmacology, School of Medical Sciences, University of Sydney, Australia,
3
Institute of Medical and Biomedical Education, St George’s University of London, UK, 4Department ofPharmacology and Therapeutics, University of Florida, Gainesville, USA, 5The Vollum Institute, Oregon
Health and Science University, Portland, USA
Opioid analgesics are powerful pain relievers; however, over time, pain control diminishes as analgesic
tolerance develops. The molecular mechanisms initiating tolerance have remained unresolved to date.
We have previously shown that desensitization of the μ-opioid receptor and interaction with β-arrestins iscontrolled by carboxyl-terminal phosphorylation. Here we created knockin mice with a series of serineand threonine-to-alanine mutations that render the receptor increasingly unable to recruit β-arrestins.
Desensitization is inhibited in locus coeruleus neurons of mutant mice. Opioid-induced analgesia isstrongly enhanced, and analgesic tolerance is greatly diminished. Surprisingly, respiratory depression,
constipation, and opioid withdrawal signs are unchanged or exacerbated, indicating that β-arrestin
recruitment does not contribute to the severity of opioid side effects and, hence, predicting that G-proteinbiased µ-agonists are still likely to elicit severe adverse effects. In conclusion, our findings identify
carboxyl-terminal multisite phosphorylation as key step that drives acute μ-opioid receptor desensitizationand long-term tolerance.

86Presenter –Knapp, Brian
Abstract ID – M48
G-Protein subunits alpha and gamma influence functional activation profiles of opioids at the
mu opioid receptor
Brian I. Knapp1, Miriam E. Barnett1, Jean M. Bidlack1
1
University of Rochester Medical Center, Rochester, USA
Opioid receptors couple to the Gi family of G proteins. Heterotrimeric Gi proteins are composed ofvarious types of alpha, beta, and gamma subunits, which may affect the functional activity ofopioids. We investigated the effects of several alpha subunits (Gαi1, Gαi2, Gαi3, Gαz, GαoA, GαoB)
and two gamma subunits (Gγ1 and Gγ2) on the activity profiles of known mu opioid receptor ligands
in transfected HEK 293T cells using bioluminescence resonance energy transfer (BRET). For
example, DAMGO, methadone, and etorphine were full agonists at the mu opioid receptor when
signaling through G proteins composed of Gαz and Gγ2. Morphine and buprenorphine were partial
agonists like the benzomorphans (-) pentazocine, ethylketocyclazocine, and cyclazocine. Naloxone,
naltrexone, and samidorphan did not stimulate signaling. The agonist efficacy profile for the mu
opioids was dependent on the composition of the heterotrimeric G protein indicating that receptor
conformational changes induced by various mu opioids differentially activate G proteins. Thus, thefunctional response of an opioid may depend on the distribution and expression of various G protein
subunits and their association with the mu opioid receptor.
Presenter – Kochan, Kelsey
Abstract ID – W11
Effects of a G-protein βγ subunit inhibitor on delta opioid receptor (DOR) mediated behaviors
Kelsey Kochan1, Emily Jutkiewicz1, John Traynor1
1
University of Michigan, Ann Arbor, USA
Delta opioid receptor (DOR) agonists, such as SNC80, are known to induce convulsions, hindering
their development as analgesics or antidepressants. Determining the signaling pathways that giverise to DOR-mediated behaviors, specifically convulsions, may lead to the development of safer
therapeutics. Therefore, to better understand the mechanisms contributing to these behaviors, therole of Gβγ subunits in SNC80-induced antinociception, antidepressant-like effects, and convulsions
was explored. We evaluated these behaviors in C57BL6 mice in the presence or absence of gallein,a G-protein βγ subunit inhibitor. Antinociceptive effects were evaluated using the acetic acid writhing
assay (AAWA), antidepressant-like effects were examined using the tail suspension test (TST), andconvulsions were observed for 60 min and scored according to the Racine scale. In each of theseassays, an approximate 3-fold leftward shift in the SNC80 (s.c.) dose response curve was observed
following the administration of 30 mg/kg gallein (i.p.), indicating a potentiation of the effects of
SNC80. For example, in the TST, the dose of SNC80 that produces the maximal reduction in
immobility during the 6 min experimental test period is 3.2 mg/kg without gallein and 1 mg/kg withgallein. Together, these data suggest that inhibiting G-protein βγ subunits enhances the potency of
DOR agonists for antinociception, antidepressant-like effects, and convulsions, suggesting thesebehaviors are all mediated through Gα subunit signaling. Further understanding of thesemechanisms would be useful in the potential development of DOR therapeutics.
Supported in part by PhRMA Foundation Research Starter Grant (Jutkiewicz)

87

Presenter – Kuo, Lawrence
Abstract ID – T19
Novel biased mu opioid agonists for the treatment of pain
James E. Barrett1, Shariff Bayoumy2, Saadet Inan1, Lawrence C. Kuo2, Brett A. Tounge2
1
Center for Substance Abuse Research, Lewis Katz School of Medicine, Temple University,
Philadelphia, USA, 2Mebias Discovery, Philadelphia, USA
Although mu-opioid receptor agonists are efficacious analgesics, abuse potential and profoundrespiratory depression represent significant liabilities. Addressing these problems with twotechnologies – native GPCR purification and solution dynamic protein nuclear magnetic resonance– Mebias Discovery(http://mebiasdiscovery.com) has identified a series of ‘biased agonists’ of thehuman mu-opioid receptor that are efficacious in rodent models. Among them, two compounds(MEB-1166 and MEB-1170) are currently in pre-clinical development studies with a plan to file an
IND in 2020.3 MEB-1166 and MEB-1170 are potent analgesics with drastically diminished on-target
adverse effects of respiratory depression, constipation, and sedation. In male and female SpragueDawley rats, MEB-1166 and MEB-1170 are antinociceptive in the tail flick and formalin assays. Both
MEB-1166 and MEB-1170 have a longer duration of action than morphine and Oliceridine (TRV130). In accelerating rotarod studies, the Mebias compounds show no sedation. When tested incharcoal transit in the proximal small bowel of rats, these compounds are indistinguishable fromvehicle. Both display no respiratory depression at high doses as reflected by pO2, pCO2, pH, minutevolume, tidal volume, and respiratory rate. Importantly, in the Conditioned Place Preference model
to assess addiction potential, the Mebias‘biased’ agonists induce no reward or liking behavior. These
data demonstrate that MEB-1166 and MEB-1170 have validated the in vivo translation of Mebias’
unique platform to identify ‘biased’ mu opioid agonists with significantly improved preclinical profiles
to tackle limitations and challenges that face presently marketed opioid analgesics.
Presenter – Kuzumaki, Naoko
Abstract ID – T37
Changes in the expression of opioids and opioid receptors in the sensory neurons offibromyalgia and Parkinson’s disease patients using disease-specific iPS cells
Naoko Kuzumaki1,2, Reiko Kagawa1, Yuri Ikeda1, Yukari Suda2, Chizuru Iwasawa1, Takahiro Kato3,
Nobutaka Hattori4, Hideyuki Okano5, Minoru Narita1,21
Dept. Pharmacol., Hoshi Univ., Tokyo, Japan, 2L-StaR, Hoshi Univ., Tokyo, Japan, 3Dept.
Neuropsychiatry, Grad. Sch. Med. Sci., Kyushu Univ., Fukuoka, Japan, 4Dept. Neurol., Juntendo
Univ. Sch. Med., Tokyo, Japan, 5Dept. Physiol., Keio Univ. Sch. Med. Tokyo, Japan
Chronic pain is associated with motor restriction, depression, anxiety and reduced quality of life.
Since the mechanisms of chronic pain are different and complex under each disease, it should beneeded to find an effective treatment. In the present study, to investigate the mechanisms of sensory
neuropathy induced by fibromyalgia or Parkinson’s disease (PD), we applied for the stem cell
technology using induced pluripotent stem cells (iPSCs). We differentiated iPSCs derived fromfibromyalgia and PD patients into sensory neurons following previous methods. We found that the
iPSC-derived sensory neurons expressed dorsal root ganglion neuron markers, BRN3A and ISL1,
and nociceptive neuron markers, TRPV1, NTRK1 and vGLUT2. Under these conditions, weanalyzed changes in the gene expression of endogenous opioids and opioid receptors infibromyalgia and PD patient-specific sensory neurons derived from iPSCs. As a result, changes inlevels of opioid receptors and those ligands were found in the fibromyalgia and PD-derived sensory
neurons compared to healthy control. These findings indicate that alterations of opioidergic functions
in fibromyalgia and PD-derived sensory neurons may, at least in part, reflect the pathophysiology of
sensory neuropathy in fibromyalgia and PD. We propose here that these pain researches using thestem cell technology with disease-specific iPSCs may lead to the development of disease-specifictreatments for sustained pain.

88Presenter – Lee, Cynthia Wei-Sheng
Abstract ID – W40
Cellular mechanisms of metabolic adaptations mediated by μ-opioid and nociceptin
receptors
Tzu-Yu Lin1, Cynthia Wei-Sheng Lee1,2,3
1
Center for Drug Abuse and Addiction, China Medical University Hospital, Taichung 40447, Taiwan,
2
Graduate Institute of Biomedical Sciences, China Medical University, Taichung 40402, Taiwan,
3
Brain Disease Research Center, China Medical University Hospital, Taichung 40447, Taiwan
We aimed to explore the metabolic adaptations induced by opioids through coexpressed μ-opioid(MOP) and nociceptin/orphanin FQ (NOP) receptors in human embryonic kidney (HEK) 293 cells.
Transcriptome analysis in MOP+NOP-expressing cells treated with morphine, methadone, or
buprenorphine showed that several pathways involved in metabolic reprogramming weresignificantly affected. The lactic acid contents, ATP levels, and glucose consumption were measured
and normalized to the total protein concentrations. Expression levels of aminomethyltransferase andglycine amidinotransferase were determined by immunoblotting. Metabolic changes elicited by MOPNOP receptors after opioid treatment warrant further investigation.
Presenter – Leff, Emily
Abstract ID – W46
GRK2/3 dependent desensitization of the mu-opioid receptor is eliminated following chronicmorphine treatment
Emily Leff1, John Williams2
1
Oregon Health & Science University, Portland, USA, 2Oregon Health & Science University, Portland,
USA
Work done in the locus coeruleus (LC) has shown that acute desensitization of the mu-opioid
receptor (MOR) is a critical step leading to long-term tolerance to opioids and that phosphorylationof MOR is necessary for acute desensitization. Blocking phosphorylation of all 11 sites on the Cterminal tail of MOR blocks acute desensitization in naïve and morphine treated animals. However,
the endogenous mechanisms and kinases involved, and how they are regulated during chronic
treatment, are unknown. Therefore, using a combination of whole-cell electrophysiology andpharmacological inhibition of select kinase activity, the kinases involved in acute MOR
desensitization both before and after chronic morphine treatment are investigated. The GRK2/3
inhibitor, Compound 101, was found to significantly block acute desensitization of MOR in LC
neurons. Interestingly, Compound 101 did not block acute desensitization in morphine treatedanimals. In addition, while the JNK inhibitor, SP600125, did not block acute desensitization in naïve
animals, it significantly blocked acute desensitization in chronically treated animals. These results
indicate that kinase regulation of MOR shifts following chronic morphine treatment.

89

Presenter – Lemos Duarte, Mariana
Abstract ID – M43
Opiate-mediated protein kinase C signaling examined using novel antibodies that recognize
active protein kinase C and phosphorylated mu opioid receptors.
Mariana Lemos Duarte1, Nikita Trimbake1, Xiaomin Fan2, Catherine Woods2, Akila Ram3, Joshua
de Tenorio1, Gregory Zilberg1, Erin Bobeck3, Deborah Schechtman4, Daniel Wacker1, Lakshmi
Devi1
1
Icahn School of Medicine at Mount Sinai, USA, 2AvantGen, Inc, USA, 3Utah State University, USA,4
São Paulo University, Brasil
Recent alarming reports of the “Opioid Epidemic” have led investigators to focus efforts on
reexamining the fundamental mechanisms of action of opiates. Activation of opioid receptors leads
to initiation of signaling cascades that result in the activation of several kinases. Among them, protein
kinase C (PKC) has been proposed to play a crucial role in opioid receptor desensitization. To directly
examine the dynamics of PKC activation in an endogenous setting, we developed a conformationallyspecific antibody that detects active PKC. Using a high-throughput microscopy approach we
examined the temporal dynamics of PKC activation upon activation of mu opioid receptor (MOR) by
opiates in neuronal cells and in mice. We find that, while morphine treatment leads to rapid andtransient PKC activation, fentanyl treatment leads to a sustained activation in cells. A similar patternof activation is also seen in mice exposed to acute versus chronic morphine treatment. Next, wesought to directly investigate MOR phosphorylation using phospho-specific antibodies against
Ser363 and Ser375/Thr376in the MOR C-terminus. We find that the kinetics of MOR phosphorylation atthese sites parallels the kinetics of PKC activation and that PKCα is involved in this process. Finally,using purified human MOR reconstituted into nanodiscs (consisting by a phospholipid bilayer andmembrane scaffold protein) we carried out in vitro phosphorylation with purified PKCα and finddifferential, ligand-dependent dynamics of MOR phosphorylation at these sites. Together, we herein
generate novel tools to study PKC signaling and use them to provide insights into the dynamics of
PKC-mediated MOR phosphorylation in response to different opiates.
Presenter – Levine, Olivia
Abstract ID – M14
Acute estradiol modulation of a discrete limbic circuit driving binge alcohol drinking
Olivia Levine1,2, John Miller1, Mary Jane Skelly2, Kristen Pleil2
1
Weill Cornell Medicine, Graduate Program in Neuroscience, 2Weill Cornell Medicine, Department ofPharmacology, USA
Alcohol use disorder (AUD) is highly comorbid with anxiety disorders, both of which are debilitating
diseases that impose a tremendous social and economic burden on our society. Women are more
likely to develop both of these diseases and telescope to addiction faster upon first use compared tomen. To date, the precise neural underpinnings by which this occurs remain unknown. Females
across mammalian species binge drink more than males and the human and rodent literature
indicates a role for endogenous estrogen (E2) in the regulation of alcohol intake. Binge drinking is a
primary risk factor for these diseases, and neurons expressing the stress neuropeptide corticotropin
releasing factor (CRF) in the bed nucleus of the stria terminalis (BNST) regulate binge drinking andanxiety in mice. Preliminary data show marked differences in the synaptic and neuronal function of
BNST CRF neurons that indicate this population is more active and sensitive to synaptic inputs infemales than males. For example, females receive a more robust monosynaptic excitatory input fromthe paraventricular nucleus of the thalamus (PVT), a region that also regulates anxiety and we founddrives binge drinking. Here, I test the hypothesis that E2 plays a positive modulatory role inglutamatergic transmission at PVT-BNST CRF synapses to increase the activity of BNSTCRF neurons and drive binge drinking behavior. Because endogenous E2 levels are basally higher
in females, especially during peak estrogen states, they are at higher risk for the negative
consequences of binge drinking.

90

Presenter – Levran, Orna
Abstract ID – T6
Combinations of polymorphisms in circadian rhythm genes are associated with cocaine
addiction in african americans
Orna Levran1, Matthew Randesi1, Jurg Ott2, Mary Jeanne Kreek1
1
The Rockefeller University, New York, USA, 2Laboratory of Statistical Genetics, The Rockefeller
University, New York, USA
Dysregulation of the circadian rhythm is implicated in drug addiction. Association analysis of cocaine
addiction (CD) was performed in a sample of African Americans (cases; n=202, controls; n=153).
The sample was genotyped with the Smokescreen array by the NIDA Genetics Consortium. Ancestry
was verified by principal/MDS component analysis. Relatives were excluded based on PI_HAT >0.2. A set of 1000 SNPs from 11 circadian genes was selected and pruned based on quality, MAF,
and LD. A total of 648 tag SNPs and indels were analyzed. Several nominally significant associations
were detected. Conditional analysis of SNPs combinations among SNPs with nominal signals
revealed an association of two SNP pairs with CD (rs7080411/rs13145618 andrs35336050/rs35383884) (p <9 x10-5). rs7080411 is located upstream of the neuropeptide FF
Receptor 1 gene (NPFFR1) that is involved in regulation of the HPA axis. SNP rs35336050 is in an
open chromatin region downstream of the casein kinase 1 epsilon (CSNK1E). CSNK1E is involved
in circadian rhythm regulation by phosphorylating clock proteins. The indel rs35383884 isdownstream of the NPY receptor gene (NPY5R) and upstream of NPY1R. rs13145618 is locatedupstream of the main transcript of NPY2R and in an intron of transcript variant and was associatedwith gene expression in the pituitary and the hippocampus. Neuropeptide Y has a critical role incircadian rhythm stress response, cognition, learning, and memory. The study extends our previous
studies of CD in African Americans and provides evidence for a combined effect of SNPs in circadianrhythm genes on cocaine addiction.
Presenter – Liu, Jing-Gen
Abstract ID – W4
Activation of κ opioid receptor-induced mGluR2/3 reduction in prelimbic cortex contributes
to acute stress-induced social avoidance
Liu-Bin Guo1,2, Yu-Jun Wang1, Jing-Gen Liu1
1
Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of
Sciences, Shanghai, China, 2China Pharmaceutical University, Nanjing, China
Increasing evidence indicates that k opioid receptors and dynorphin are involving in transducing theeffects of stress, but the underlying mechanism is unclear. In this study, we found that mice exposed
to social defeat stress (SDS) markedly decreased social interaction with increased dynorphin
expression level in the prelimbic cortex, but not in the infralimbic cortex and caudate putamen.
Systemic and intra-prelimbic cortex administration of norBNI, a k opioid receptor antagonist,significantly inhibited SDS-induced social avoidance. Stress can regulate glutamate homeostasis.
We found that the metabotropic glutamate receptor 2/3 (mGluR2/3) level was reduced in the
prelimbic cortex after SDS. SDS-induced social avoidance could be suppressed or facilitated byintra-prelimbic cortex infusion of mGluR2/3 agonist or antagonist. More importantly, we found that
SDS-induced decreased of mGluR 2/3 was blocked by norBNI intra-prelimbic cortex infusion. Thus,
our preliminary results suggested that k-opioid receptor-regulated social avoidance associated with
SDS was through mGluR 2/3.

91

Presenter – Lueptow, Lindsay
Abstract ID – M4
Chronic pain alters the intravenous self-administration profile of remifentanil in mice
Lindsay M Lueptow1,2, Shadi Hanukaai1,2, Leeza Shashkova1,2, Christopher J Evans1,2, Catherine M
Cahill1,2
1
Hatos Center for Neuropharmacology, 2Department of Psychiatry and Biobehavioral Sciences,
University of California, Los Angeles, USA
Chronic pain affects a significant portion of the population, and effective pain management has
proven challenging. While opioid therapeutics are potent analgesics, their use is limited by highabuse liability. Currently, both preclinical and clinical data offer conflicting views on whether chronic
pain might impart resilience or vulnerability to opioid misuse and transition to an opioid use disorder.
Remifentanil is a potent, short-acting μ-agonist; properties that enhance abuse liability. Our current
study investigates the impact of chronic pain on the intravenous self-administration (IVSA) profile ofremifentanil, in male and female C57Bl/6J mice. Using a chronic constriction injury (CCI) model ofneuropathic pain, seven days after pain onset, mice began IVSA of remifentanil (50 ug/kg/infusion)under a FR1 schedule. After stable responding (~7-10 days), mice were transferred to a FR3
schedule. While we found no differences in the total number of reinforcers earned between groups
during FR1 IVSA, we found that within the first hour of a session, CCI mice achieved significantly
more reinforcers than naïve mice. However, during FR3 schedule, this effect disappeared, and CCI
mice reduced the number of reinforcers obtained as compared to the FR1 schedule, whereas thepain naïve mice earned similar reinforcers on both schedules. While it is unclear what caused thisswitch in IVSA responding between schedules in CCI mice, it is likely due to changing balance
between the analgesic properties and hedonic value of remifentanil. Our ongoing studies seek toparse the potential impact of time and negative affective state on this IVSA profile.
Presenter – Kabirullah. Lutfy
Abstract ID – W35* Hot Topics Selection, Symposium 6, Thursday, July 11 AM
The role of endogenous opioids in amphetamine-induced locomotor sensitization
Kabirullah Lutfy1
1Western University of Health Sciences, Pomona, USA
The endogenous opioid system has been implicated in amphetamine-induced hyperlocomotion andlocomotor sensitization. However, the role of each opioid peptide in these actions is unknown. Using
mice lacking mu opioid receptors (MOPr) and their wild-type controls, we determined the role of
MOPr in these actions of amphetamine. Mice of both genotypes were brought to the laboratory,
habituated to the locomotor activity chambers for 1 h, injected with saline or amphetamine (3 mg/kg)
and motor activity was recorded at 15 min bin for 1 h. Mice were treated with their respective
treatments for three days and then left untreated until day 8 when they were tested for locomotor
sensitization. On this day, mice were treated as described above except a lower dose ofamphetamine (1 mg/kg) was used. Our results showed that locomotor sensitization, but not acutemotor stimulatory action of amphetamine was reduced in mice lacking MOPr compared to their wildtype controls. We then assessed which opioid peptide acts on this receptor to alter the action ofamphetamine. Mice lacking either beta-endorphin or enkephalin and their respective wild-type
controls were treated and tested for amphetamine-induced hyperlocomotion and sensitization as
described above. Mice lacking beta-endorphin showed reduced locomotor stimulation to the lower
dose of amphetamine but exhibited a comparable locomotor sensitization to their wild-type controls.
In contrast, mice lacking enkephalin showed comparable hyperlocomotion but reduced locomotor
sensitization. These results suggest that enkephalin is possibly the endogenous ligand that acts on
MOPr to alter amphetamine-induced locomotor sensitization.

92

Presenter - Kabirullah. Lutfy
Abstract ID – T36
The role of endogenous PACAP and dynorphin in nicotine-induced conditioned place
aversion
Kabirullah Lutfy1
1Western University of Health Sciences, Pomona, USA
Nicotine addiction is a major public health and socioeconomic issue. While the rewarding action of
nicotine promotes its continued use, its aversive effect deters its use and abuse. Thus, drugs or
endogenous regulators of either rewarding or aversive effects of nicotine can have potential as
pharmacotherapy of nicotine dependence. In the present study, we assessed the role of endogenous
pituitary adenylyl cyclase activating polypeptide (PACAP) as well as dynorphin in the aversive effectof nicotine. Mice lacking either peptide and their wild-type controls were tested for baseline place
preference in the place conditioning paradigm on day 1, and then received conditioning withsaline/nicotine (1 mg/kg) or saline/nicotine twice daily for three days and then tested for postconditioning place preference on day 5. Mice received additional conditioning on the following week,
as described for the first week, and tested for place preference in a drug-free state again. Micelacking PACAP and their wild-type controls did not show conditioned place preference(CPP) or aversion (CPA) on day 5. On the other hand, mice lacking dynorphin exhibited a significant
CPP response on day 5. Wild-type mice of either genotype exhibited a significant CPA following
further conditioning during the second week. This aversive response was not observed in micelacking PACAP or dynorphin, showing that endogenous dynorphin may cancel the rewarding actionof nicotine at the early stages of drug administration and endogenous PACAP at later stages of drug
administration. Therefore, targeting these two systems may be a viable approach to treat nicotine
addiction.
Presenter - Kabirullah. Lutfy
Abstract ID – M34
The role of endogenous PACAP in morphine-induced state-dependent CPP and reinstatement
Kabirullah Lutfy1
1
Department of Pharmaceutical Sciences, College of Pharmacy, Western University of Health
Sciences, Pomona, USA
We have earlier showed that morphine-induced conditioned place preference (CPP) following single
but not repeated conditioning with morphine was reduced in the absence of pituitary adenylyl
cyclase-activating polypeptide (PACAP). In the present study, we determined if morphine-induced
state-dependent CPP or the reinstatement of morphine CPP would be altered in mice lacking PACAP
compared to their wild-type controls. Male mice of each genotype were tested for baseline place
preference on day 1, then received conditioning with saline/morphine (5 mg/kg) or morphine/saline
for 8 days (4 conditioning with each treatment) and then tested for the post-conditioning place
preference on day 10. Mice were then challenged with morphine (5 mg/kg) on day 17 and tested
again for morphine-induced state-dependent CPP. Our results showed that while mice of either
genotype did not show any significant CPP, wild-type mice exhibited a robust state-dependent CPP
but this response was reduced in mice lacking PACAP. This prompted us to examine if PACAP would
be involved in the reinstatement of morphine CPP. Another group of mice lacking PACAP and theirwild-type controls were tested as described above except a higher dose of morphine (10 mg/kg) wasused on the conditioning days. Mice were then exposed to extinction training in which mice wereconditioned with saline/saline instead of saline/morphine and tested for the extinction of CPP
followed by a test for morphine-induced reinstatement. Mice of both genotypes showed a robust but
comparable CPP. However, the reinstatement of morphine CPP was reduced in mice lacking PACAP
compared to wild-type controls.

93

Presenter – Mack, Seshat
Abstract ID – T26
Characterization of small molecule ligands targeting GPR83, a newly deorphanized striatal
receptor that could modulate opiate reward
Seshat Mack1, Ivone Gomes1, Amanda Fakira1, Roberto Sanchez1, Lakshmi Devi1
1
Dept. of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, USA
The worsening opioid crisis demonstrates an unmet need for novel therapeutics to treat opioid abuse.
The mesolimbic dopamine system, which includes the nucleus accumbens, plays a central role inmodulating reward for drugs of abuse. Characterizing understudied receptor systems in the nucleus
accumbens would be a key step towards identifying novel therapeutic targets for the treatment of
substance abuse disorders. GPR83 is an understudied receptor with robust expression in regions
within the reward pathway (including the nucleus accumbens) in both mouse and human. To
characterize GPR83, a potential therapeutic target for opioid abuse disorders, we initiated studies todeorphanize the receptor and identified the neuropeptide PEN (one of the most abundantneuropeptides) as its endogenous ligand. Identification and characterization of small molecule
ligands would serve as critical tools to explore the role of GPR83 in vivo. With this in mind, we
generated a homology model of GPR83 and virtually docked a small molecule library of 6 million
compounds. The top 50 predicted hits were screened in a cell-based assay leading to theidentification of two agonists and one antagonist with high selectivity. Here, we present data fromstudies characterizing the molecular pharmacological properties of these compounds. We alsopresent data on the effect of small-molecule GPR83 ligands on conditioned place preference tomorphine in mice. Taken together, these studies highlight a potential role for this novel
neuropeptide-receptor system in modulating reward behaviors, including drug-related reward
behaviors.
This work was supported in part by NIH grant DA008863 (to LAD and SM).
Presenter – Maguire, David
Abstract ID – T18
The long-acting mu opioid receptor selective antagonist methocinnamox (MCAM) preventsand reverses abuse- and overdose-related effects of opioids
David R. Maguire1, Lisa R. Gerak1, Martin A Javors1, James H. Woods1, Stephen M. Husbands2,
Alex Disney2, Charles P. France1
1
University of Texas Health Science Center at San Antonio, San Antonio, Texas, USA, 2University ofBath, Bath, UK
Opioid abuse and overdose remain a serious public health challenge despite the availability ofeffective medications. Naltrexone is the only opioid receptor antagonist currently available to treat
opioid abuse and naloxone is the only antagonist available to reverse opioid-induced respiratory
depression. Although naltrexone is effective in some patients and naloxone reverses opioid-induced
respiratory depression, a short duration of action and surmountability of antagonism limit theeffectiveness of both medications. These studies evaluated methocinnamox (MCAM), a long-acting,mu opioid receptor selective antagonist, for its ability to block abuse-related and respiratorydepressant effects of opioid agonists in rhesus monkeys. Naltrexone and MCAM decreased selfadministration of heroin and fentanyl but not cocaine and decreased choice of the opioid remifentanil
in a food versus drug choice procedure. A single injection of MCAM was effective for a week orlonger, whereas effects of naltrexone lasted less than 24 hr. MCAM and naloxone reversed heroinand fentanyl-induced respiratory depression. MCAM protected against agonist-induced respiratory
depression for up to a week after a single injection whereas effects of naloxone lasted less than 2
hr. A dose of MCAM that was effective for at least a week reached maximum blood concentration(11 ng/ml) 30 min after injection but was below the limit of detection (1 ng/ml) 24 hr later. MCAM had
no effect of heart rate, blood pressure, body temperature, or activity. These data indicate that MCAM
could be a safe, effective, and long-acting treatment for opioid abuse and overdose that could haveseveral advantages over currently available treatments.

94

Presenter – Majumdar, Susruta
Abstract ID – M13
Oxidative metabolism as a modulator of kratom’s opioid analgesic activity
Rajendra Uprety1, Andrew Kreugel2, SO Eans3, John Pintar4, Ying-Xian Pan1, Gavril W Pasternak1,
Michael Cameron5, Jay P McLaughlin3, Dalibor Sames2, Susruta Majumdar6
1
Department of Neurology and Molecular Pharmacology, Memorial Sloan Kettering Cancer Center,
USA, 2Department of Chemistry, Columbia University, USA, 3Department of Pharmacodynamics,
University of Florida, Gainesville, USA, 4Department of Neuroscience and Cell Biology, Rutgers
Robert Wood Johnson Medical School, Piscataway, USA, 5Department of Molecular Therapeutics,
Scripps Research Institute, Jupiter, USA, 6Center for Clinical Pharmacology, St Louis College of
Pharmacy and Washington University School of Medicine, St Louis, USA
Development of safe and abuse free opioid analgesics for human pain medication is a longstanding
scientific problem. Various opioids including morphine and fentanyl analogs that target mu-opioid
receptor (MOR) are ideal clinical drugs for treating moderate to severe pain. However, such MOR
agonists produce problematic side effects. Alternatively, various herbal medicine including plant
Mitragyna speciosa (Kratom) has been used as a substitute for opioids for treatment of variety of
diseases like pain, alcoholism and depression. Kratom is increasingly also becoming a recreational
drug of abuse in the United States. Here, we present a comprehensive pharmacological evaluation
on Kratom based products including kratom extract, the major alkaloid mitragynine and its key
metabolites in mice. We show that the analgesic actions of kratom is mediated by G-protein biased
agonist mitragynine and its first-pass potent metabolic products. The oxidative metabolites mediate
their biological actions through MOR and show attenuated respiratory depression while still retaining
the reinforcing behavior seen with classical opioids. Chemistry, metabolism, pharmacokinetics andpharmacology of kratom and its alkaloids will be discussed.
Supported by NIDA (R01DA046487 and R21DA045884) and through the Peer Reviewed Medical
Research Program (W81XWH-17-1-0256).
Presenter – Manandhar, Preeti
Abstract ID – W25
Does Pregabalin or Gabapentin modulate µ-opioid receptor signalling?
Preeti Manandhar1, Bridin Murnion1, Mark Connor1, Natasha Grimsey2, Marina Santiago11Macquarie University, 2University of Auckland, New ZealandPregabalin and gabapentin improve neuropathic pain symptoms but there are emerging concerns
regarding their misuse potential, which is more pronounced among patients with substance use
disorder, particularly involving opioids. Co-ingestion of gabapentinoids with opioids has been
associated with a substantial increase in opioid related death, however, the molecular mechanismof this is still unclear. To determine if gabapentin and pregabalin modulate acute μ receptor signaling,
K channel activation (membrane potential assay), adenylyl cyclase (AC) modulation (BRET based
CAMYEL assay), and ERK phosphorylation (ELISA assay) were assessed in HEK 293 cells stablytransfected with human μ-receptor. Pregabalin or gabapentin (1 µM, 100 µM each) did not activateµ receptors or affect K channel activation or ERK phosphorylation produced by morphine. Neither
drug affected the desensitization of K channel activation produced by prolonged (30 min) applicationof morphine. Gabapentin (1 µM, 100 µM) and pregabalin (1 µM) did not affect inhibition of forskolinstimulated AC activity by morphine, however, pregabalin (100 µM) potentiated forskolin mediatedcAMP production in a Gs-independent manner, although morphine still inhibited cAMP levels with asimilar potency to control. Gabapentin and pregabalin efficacy in neuropathic pain and epilepsy isthought to be mediated through action on α2δ subunit of presynaptic Ca channels. We did not findeffects of pregabalin or gabapentin on µ receptor function in 3 different signalling assays. Our data
does not support the hypothesis that gabapentin or pregabalin augment opioid effects through direct
or allosteric modulation of the µ receptor.

95

Presenter – Martucci, Katherine
Abstract ID – W48
Examining the effects of opioid use on neural responses to monetary incentives in chronic
pain
Katherine T. Martucci 1,3, Kelly H. MacNiven 2, Nicholas Borg 2, Brian Knutson 2, Sean Mackey 1
1
Department of Anesthesiology, Perioperative and Pain Medicine, Division of Pain Medicine,
Stanford University School of Medicine, USA, 2Department of Psychology, Symbiotic Project on
Affective Neuroscience Laboratory, Stanford University, USA,3Department of Anesthesiology, Duke
University School of Medicine, USA
Neural responses to incentives are altered in chronic pain and by opioid use. To understand how
opioid use modulates brain reward/value response in chronic pain, we compared brain functional
magnetic resonance imaging (fMRI) responses to a monetary incentive delay (MID) task in patientswith fibromyalgia taking opioids (n = 17), patients with fibromyalgia not taking opioids (n = 17), andhealthy controls (n = 15). Both groups of patients with fibromyalgia taking and not taking opioids had
similar levels of pain, psychological measures, and clinical symptoms. Neural responses in the
nucleus accumbens to anticipated reward did not differ from healthy controls in either fibromyalgia
group. However, neural responses in the medial prefrontal cortex differed, such that patients with
non-opioid fibromyalgia demonstrated significantly altered responses to anticipated rewards andnon-loss outcomes compared to healthy controls, but patients with opioid fibromyalgia did not.
Despite limitations including the use of additional non-opioid medications by opioid fibromyalgia
patients, these preliminary findings suggest “normalized” neural responses to monetary incentives
in chronic pain patients who take opioids versus those who do not.
Presenter – Matsui, Aya
Abstract ID – M8
MOR in direct-pathway medium spiny neurons contribute to the preference for alcohol
drinking
Aya Matsui1, Veronica Alvarez1
1
National Institute on Alcohol Abuse and Alcoholism, USA
Endogenous opioid peptides are released in the nucleus accumbens (NAc) following alcohol
exposure. The opioid release is thought to be a critical step associating with the rewarding propertyof alcohol. My goal is to elucidate how alcohol exposure modulate mu-opioid receptor (MOR) functionin the NAc and how it contributes to the development of alcohol use disorder. The two subclasses ofneurons, direct- (dMSNs) and indirect-pathway medium spiny neurons (iMSNs), in the NAc express
opioid peptides and opioid receptors; however, it is unclear whether MOR is expressed in subclassesof MSNs. Here we evaluated functionally the expression pattern of opioid receptors. Our results
showed that both dMSNs and iMSNs express functional mu-, and kappa-opioid receptors, but notdelta-opioid receptors. One main effect of MOR and kappa receptor activation in these MSNs is todepress GABAergic synaptic transmission from these neurons into the target area in the ventral
pallidum. We generated a mouse lines with targeted deletion of MORs to dMSNs or iMSNs in orderto evaluate the role of MOR alcohol drinking behavior. The results show that mice lacking MOR indMSN have lower preference for alcohol drinking in two-bottle choice paradigm, suggesting a rolefor MORs specifically on dMSNs in alcohol drinking. However, mice lacking MOR in iMSNs did notreveal the difference in alcohol drinking behaviors. The results of these studies reveal that MOR
activation in dMSN might underlie the behavioral transition to alcohol abuse.

96

Presenter – Matyas, Gary
Abstract ID – M26
A stable heroin hapten (6-AmHap) vaccine formulation slated for a phase 1 clinical trial
induces long duration antibody titers that block the antinociceptive effects of heroin andother opioids
Gary Matyas1, Oscar B. Torres1,2, Agnieszka Sulima3,4, Connor Whalen1, Arthur E. Jacobson3,4, Zoltan
Beck1,2, Kenner C. Rice3,41Walter Reed Army Institute of Research, USA, 2Henry M. Jackson Foundation for the Advancement ofMilitary Medicine, USA, 3National Institute on Drug Abuse, USA,4National Institute on Alcohol Abuse andAlcoholism, USA
We have recently described a novel heroin hapten, 6-AmHap, when conjugated to tetanus toxoid (TT),and mixed with Army Liposome Formulation (ALF) as an adjuvant, induced protective efficacy against
heroin subcutaneous (SC) and intravenous (IV) challenge of immunized mice and rats. (Sulima et al., J.
Med Chem 2018). Vaccine-induced antibodies bound to heroin, its degradation products and crossreacted with other opioids. The aim of this study was to determine the duration of the protective efficacyof the vaccine. 6-AmHap was conjugated to TT, mixed with ALF and aluminum hydroxide. Rats and mice
were immunized 4 times. The sera were tested for antibody titer by ELISA and affinity to heroin, its
metabolites and other opioids. Efficacy against heroin or other opioid repeat-dose challenges wasassessed using antinociception assays. The vaccine elicited IgG levels to 6-AmHap of ~1.2 mg/mL thatpersisted over a year in both rats and mice. The vaccine prevented heroin-induced antinociception
following repeated IV heroin challenges over a year in rats. The rats typically had a 0% maximum possible
effect (MPE) from multiple heroin doses and reached 100% MPE with one more dose. The mean heroin
dose needed to reach 100% MPE in the thermal place preference assay was 4 mg/kg and the tailimmersion (TI) assay was 2 mg/kg. The Kd values of the antibodies to 6-acetylmorphine ranged from 0.10.9 nM. The average EC50 for SC challenge of the immunized mice was 1.7, 1.0 and 7.7 mg/kg for heroin,hydromorphone and hydrocodone, respectively in the TI assay.

Presenter – McCall, Nora
Abstract ID – M16
Mapping the affective component of pain: amygdalar nociceptive neurons project to the
nucleus accumbens
Nora M. McCall1, Jessica A. Pelland2, Lauren J. Marconi1, Kevin T. Baier3, Gregory Corder1
1
Depts. of Psychiatry and Neuroscience, Perelman School of Medicine, University of Pennsylvania, USA,
2
Biology Graduate Group, University of Pennsylvania, 3Department of Physiology and Biophysics,
University of California, Irvine, USA
The negative affective dimension of pain underlies the suffering of chronic pain patients. Thus, thediscovery of specific neural circuits responsible for the unpleasant quality of pain perception must be thefirst step in developing novel, efficacious pain treatments. We recently identified a critical cell population
within the brain’s pain pathway that specifically encodes the affective, rather than sensory, component ofchronic pain. This nociceptive-specific cluster of amygdalar neurons does not directly encode rewardingor other aversive experiences. This suggests that novel treatments disrupting the processing of pain inthis pathway may meet the long-sought criteria of high analgesic efficacy without addictive liability. To
determine other circuit elements of this pathway, we are identifying the brain-wide, input-out neuralarchitecture using a mouse driver line to genetically target pain-active neurons (painTRAP2) incombination with activity-dependent viruses. We find that amygdalar nociceptive neurons send longrange projections that preferentially innervate the posterior nucleus accumbens (NAc) core and shell
subnuclei, and encircle neurons co-expressing dopamine receptor 2 and the endogenous opioid
enkephalin. Preliminary in vivo calcium imaging illustrates that nociceptive neural activity is localized tothe caudal NAc core/shell. This suggests pain valence information transmitted from the BLA to NAc could
influence motivational and hedonic behavior, e.g., depression co-morbid with chronic-pain. Constructing
a brain-wide global map of this pain affect pathway, and the circuits in which it is embedded, is our firststep toward identifying a series of candidate brain regions for therapeutic targeting of chronic pain.

97

Presenter – McDermott, Max
Abstract ID – T20
Exploring GPR171 as a novel pain therapeutic with reduced abuse liability
Max McDermott1, Leela Afrose1, Erin Bobeck1
1
Utah State University, USA
G protein-coupled receptors (GPCRs) are the most druggable receptors in the human body, andnewly deorphanized GPCRs are a new and unexplored avenue towards the development of new
pain therapeutics. GPR171 is one such GPCR. It was recently discovered that BigLEN, a peptide
derived from the precursor protein ProSAAS, is the endogenous ligand for GPR171. Despite thefinding that ProSAAS is upregulated in the cerebrospinal fluid of fibromyalgia patients no researchhas explored BigLEN-GPR171 system in pain. Promising work from our lab has shown that an
agonist for GPR171, MS15203, can increase the pain-relieving properties of morphine while anantagonist causes a decrease. In order to further investigate this agonist as a potential pain
therapeutic we examined its reward profile using conditioned place preference (CPP). Mice received
MS15203 alone or with morphine. Our results show that while MS15203 does not alter morphine’s
rewarding properties, importantly it does not have rewarding properties on its own. Additionally,
MS15203 has no effect on locomotor activity, but reduces the psychomotor enhancement caused bymorphine. In contrast, the antagonist, MS21570, decreased morphine preference and increased
psychomotor activity. This alteration in behavior corresponds to expression changes within the
reward circuit. Specifically, morphine decreases ProSAAS in the Medial Prefrontal Cortex, andincreases GPR171 in the Basolateral Amygdala and Nucleus Accumbens, suggesting that morphine
administration is modulating the BigLEN-GPR171 system. Our results support further investigationof GPR171, especially towards the development of a therapeutic that enhances morphine
antinociception but does not increase its abuse liability.
Presenter – McElroy, Bryan
Abstract ID – T9
Impact of short acting synthetic kappa-opioid antagonists LY2444296 and LY2795050 on selfgrooming and anhedonic-like behaviors in male mice
Bryan D. McElroy1, Eduardo Butelman1, Thomas E. Prisinzano2, Mary Jeanne Kreek1
1
The Rockefeller University, 2University of Kansas, USA
The kappa (k) opioid receptor / dynorphin system is involved in the modulation of depression-likestates and anhedonia, and adaptations to stress and exposure to abused drugs. Several relatively
short-acting k-receptor selective antagonists have been synthesized; their behavioral profile has
been partially studied. We hypothesize that pharmacological manipulations of the k-receptor system
will result in changes in anhedonic-like behavior in mice. Adult male C57BL/6j mice (n=6-8 per group)
were examined for self-grooming behavior in the “splash test” (an ethologically relevant marker ofanhedonia, in which grooming deficits are observed in response to diverse stressors). The plantderived k-agonist salvinorin A (0.56-1.8 mg/kg), caused dose-dependent decreases in self-grooming
compared to vehicle. Two relatively short-acting k-antagonists (LY2444296: (S)-3-fluoro-4-(4-((2-(3fluorophenyl) pyrrolidin-1-yl)methyl)phenoxy)benzamide, and LY2795050: 3-chloro-4-(4-(((2S)-2pyridin-3-ylpyrrolidin-1-yl)methyl) phenoxy)benzamide) were studied for their effectiveness inpreventing salvinorin A-induced grooming deficits. k-selective doses of both LY2444296 andLY2795050 dose- and time-dependently prevented grooming deficits caused by salvinorin A (1.8mg/kg). A k-selective dose of each of these antagonists also decreased immobility in the forced swim
test (FST), an antidepressant-like effect. This study shows that the k-receptor system is involved inan ethologically relevant measure of anhedonia, and that k-selective doses of these antagonists can
produce effects consistent with rapid anti-anhedonia.

98Presenter – McPherson, Kylie
Abstract ID – M15
Diverse cell types within the vlPAG exhibit opioid-induced GIRK currents.
Kylie McPherson1, Katie Suchland1, Susan Ingram1
1
Oregon Health & Science University, Portland, USA
The periaqueductal grey (PAG) is an important integration site within the descending pain modulatory
pathway that receives diverse inputs—the ventrolateral region in particular (vlPAG) is a key target for
opioid induced analgesia. The disinhibition theory for descending pain modulation proposes that opioids
selectively inhibit GABAergic interneurons within the vlPAG, disinhibiting glutamatergic output neurons
that project to the rostral ventromedial medulla (RVM). However, the vlPAG is a highly heterogeneous
region with diverse cell types. Using whole-cell patch-clamp recordings of membrane firing properties innaïve rats we defined 5 distinct cell-types within the vlPAG: onset-firing, tonic firing, phasic firing, randomfiring, and pacemaker neurons. These cell types are observed throughout the rostral-caudal axis without
any obvious topographical distribution. We examined opioid activation of G-protein coupled inwardlyrectifying potassium channel (GIRK) currents using the selective mu opioid receptor agonist DAMGO.
Unexpectedly, opioid-induced GIRK currents were observed in all of the cell types, and opioid-insensitive
cells were also observed in all groups suggesting further heterogeneity within the vlPAG. Additional
studies will use retrograde fluorescent CTb labeling from the RVM to examine the intrinsic properties ofvlPAG neurons that project to this key downstream target for pain modulation and whether these cells
are directly affected by persistent inflammation. Collectively, these findings indicate that aspects of thedisinhibition theory are over-simplified and that careful delineation of the circuits between the vlPAG and
RVM will enhance our understanding of the descending control of pain.
Supported by Grant numbers NIDA DA042565-01 and NSF GRFP

Presenter – Milner, Teresa
Abstract ID – T23
Sex and chronic stress influence the distribution of delta opioid receptors within the rat
hippocampal CA1 region following oxycodone conditioned place preference
Batsheva Reich1, Megan Johnson1, Jared M. Berman1, Ellen Goldstein1, Vera Pertsovskaya1, Yan Zhou2,
Natalina Contoreggi1, Andreina G. Dyer1, Jason D. Gray3, Elizabeth M. Waters3, Bruce S. McEwen3, Mary
Jeanne Kreek2, Teresa A. Milner1,31Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, USA, 2The Laboratory of the
Biology of Addictive Diseases, The Rockefeller University, USA, 3Harold and Margaret Milliken Hatch
Laboratory of Neuroendocrinology, The Rockefeller University, USA
Although naïve female and male Sprague Dawley rats acquire oxycodone conditioned place preference(CPP), only females acquire oxycodone CPP following chronic immobilization stress (CIS). We examinedthe distribution of DORs in CA1 pyramidal cells using electron microscopy in these rats. All females werein estrus. CIS-Naïve CPP rats: Saline-injected (Sal) females compared to Sal-males had elevatedcytoplasmic and total dendritic DORs. Oxycodone-injected (Oxy) females had more near-plasmalemmal,
cytoplasmic, and total dendritic DORs compared to Oxy-males. Oxy-males compared to Sal-males had
less plasmalemmal DORs in large dendrites and more DOR-labeled dendritic spines. CIS only:
Unstressed (US) females compared to US-males had more near-plasmalemmal dendritic DORs. CISfemales compared to US-females had increased near-plasmalemmal, cytoplasmic and total dendritic
DORs. CIS-females compared to CIS-males had more near-plasmalemmal and total dendritic DORs. CIS
rats plus CPP: CIS Sal-females compared to CIS Sal-males had more plasmalemmal DORs in large
dendrites and DOR-labeled spines. CIS Oxy-females had fewer cytoplasmic and total DORs in large
dendrites compared to CIS Sal-females. CIS Oxy-males compared to CIS Sal-males had fewer total
dendritic DORs. Following oxycodone CPP DORs within CA1 pyramidal cells remain positioned in CISnaïve female rats to enhance sensitivity to DOR agonists but internalize in CIS-naïve males where they
are less available for ligand binding. Following CIS, DORs are redistributed within CA1 pyramidal cellsof females to prime for enhanced sensitivity to DOR agonists. Conversely, CIS reduces DORs in CA1
pyramidal cells in males, which may contribute to the inability of males to acquire oxycodone CPP.
Supported by Grant number NIH-DA08259

99

Presenter – Morgan, Michael
Abstract ID – M36
Microglial P2X4 receptors contribute to morphine tolerance mediated by the periaqueductal
gray in the rat
Richik Neogi1, Christa E. Müller2, Michael M. Morgan1
1
Washington State University Vancouver, USA, 2University of Bonn, Germany
Tolerance to the analgesic effects of opioids develops rapidly in humans and laboratory animals.
P2X4 receptors on spinally located microglia have been implicated in opioid tolerance. The present
study extends this finding by examining the contribution of P2X4 receptors on microglia in theventrolateral periaqueductal gray (vlPAG) to morphine tolerance. To test this hypothesis, maleSprague-Dawley rats (n = 28) received microinjections of morphine or saline co-administered with
the P2X4 receptor antagonist PSB15147 or vehicle into the vlPAG twice a day for two days. On Day
3, the antinociceptive potency of morphine was assessed using the hot plate test following
administration of cumulative third-log doses of morphine (2.2, 4.6, & 10 µg) into the vlPAG.
Microinjection of PSB15147 into the vlPAG on Day 1 had no effect on nociception and did not altermorphine antinociception. Repeated microinjections of morphine caused a rightward shift in themorphine dose-response curve compared to vehicle treated rats when tested on Day 3. Coadministration of PSB15147 with morphine on Days 1 & 2 prevented the rightward shift in themorphine dose-response curve. Morphine ED50 values for vehicle and morphine/PSB15147 treatedrats were nearly identical (6.5 µg) and statistically different than the ED50 value for morphine tolerant
rats (13.1 µg; F (3,80) = 3.706, p = 0.014). These data demonstrate that P2X4 receptors on vlPAG
microglia contribute to morphine tolerance without altering morphine antinociception.
Presenter – Moriya, Yuki
Abstract ID – W18
Opioid system is involved in modulating the development of behavioral sensitization to METH
Yuki Moriya1, F. Scott Hall2, Yoshiyuki Kasahara3, Yoko Hagino1, Brigitte L. Kieffer4, George R. Uh5,
Ichiro Sora6, Kazutaka Ikeda1
1
Addictive Substance Project, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan,
2
Department of Pharmacology and Experimental Therapeutics, University of Toledo, USA,
3
Advanced Interdisciplinary Biomedical Engineering, Tohoku University School of Medicine, Sendai,
Miyagi, Japan, 4Department of Psychiatry, Douglas Mental Health Research Institute, McGill
University, Montreal, Canada, 5Research Service, New Mexico VA Healthcare System, Albuquerque,
NM, USA, 6Department of Psychiatry, Kobe University, Kobe, Japan
Behavioral sensitization is a progressive increase in a behavioral response such as locomotor activity
that develops upon repeated exposure to a drug of abuse, such as methamphetamine (METH). It is
used as a measure indicative of the neuroplasticity underlying the addictive process. The number oftreatments, interval between treatments, dose, sex, age, and genetic differences affect the strengthof behavioral sensitization. Opioids exert pharmacological actions through three opioid receptor
subtypes: the mu opioid receptor (MOP), the delta opioid receptor (DOP) and the kappa opioid
receptor (KOP). The purpose of the present study was to test whether METH-induced behavioral
sensitization was altered in MOP, DOP and KOP knockout mice. We assessed whether repeatedinjections of 1 mg/kg METH produce sensitization in MOP, DOP, and KOP +/+, +/-, and -/- mice. This
procedure was conducted seven times; sessions from 1 to 6 were performed daily, while session 7
occurred 8 days after the last injection. Large baseline locomotor differences (e.g. after saline
treatment) were observed, and these baseline differences interacted with strain (comparing WT micefrom each strain), genotype, sex and repeated testing. Differences were also observed in both initial
responses and sensitization to METH, due to strain, genotype and sex. METH sensitization wasobserved in all genotypes for all strains. Both initial responses and sensitization to METH wereaffected by strain, genotype & sex. These effects were observed on top of substantial differences inbaseline locomotion seen in these strains, despite them being on a congenic C57BL/6J background.

100

Presenter – Nakata, Eriko
Abstract ID – M6
Novel delta-opioid receptor agonist NC-2800 has a low risk of addiction
Eriko Nakata1, Yoshiyuki Iwase1, Takao Komatsu1, Junichi Sakai1, Masaaki Hirose1, Tomio
Yamakawa1
1
Discovery Research Laboratories, Nippon Chemiphar Co. Ltd., Saitama, Japan
NC-2800, a novel δ-opioid receptor (DOR) agonist identified by Nippon Chemiphar Co., Ltd., is apotential therapeutic agent for depression and anxiety. In our presentation in INRC 2017, we showedthat NC-2800 has anxiolytic/antidepressant-like effects in rat models with high efficacy and rapid
onset without producing adverse effects like benzodiazepines or serotonin-selective reuptake
inhibitors. In olfactory bulbectomy model rats, repeated-dose of NC-2800 showed a potent
antidepressant-like effects even on day 1 of administration and the effect was sustained for 14 days.
This suggests that tolerance in pharmacological behavior does not form by repeated use of NC2800. In this study, we investigated the effect of NC-2800 on dependence liability in animals. At first,
we investigated reinforcing effect of NC-2800 with intravenous self-administration study in Rhesus
monkeys. Dose per infusion was set by behavior observation and plasma concentration after single
intravenous dosing of NC-2800. Two monkeys were tested but they did not show active selfadministration of NC-2800 compared to positive control pentazocine under these conditions.
Second, to study physical dependence, naloxone-precipitated withdrawal test was performed in mice
after repeated administration of NC-2800 for 10 days. The NC-2800 group did not show any
withdrawal symptoms. Taken together these results suggest that the DOR agonist NC-2800 is
unlikely to cause addiction, making it an even more promising antidepressant/anxiolytic candidate.
Supported by the Japan Agency for Medical Research and Development (AMED) grants.
Presenter – Nilges, Mark
Abstract ID – T39
Effects of cannabinoid and opioid combinations on the learning and performance of responsesequences in rhesus macaquesMark R. Nilges1, Joseph A. Grace1, Peter J. Winsauer1,2
1
Department of Pharmacology and Experimental Therapeutics, Louisiana State University Health
Sciences Center, New Orleans, USA, 2Alcohol and Drug Abuse Center of Excellence, Louisiana
State University Health Sciences Center, New Orleans, USA
Cannabinoids can enhance the antinociceptive effects of opioids, but this potentially valuable effectcould be undermined if they also enhance opioid-induced cognitive disruption. In this experiment,
cannabinoids (THC and CP55940) and opioids (heroin and U50488) were examined alone, and insome cases, in combination in rhesus macaques responding in an acquisition (learning) andperformance procedure with a concurrent warm-water tail-withdrawal antinociception procedure.
Cumulative dosing was facilitated by examining four discrete cycles of acquisition and performanceof response sequences. Each cycle included a 10-min blackout period, a 15-min acquisition, and a
5-min performance component. Subjects learned a new 5-response sequence during acquisition oremitted a well-rehearsed 5-response sequence during performance. Responding was reinforced byfood pellets under a second-order fixed-ratio (FR4) schedule. Antinociception was measured after
each cycle. Heroin and U50488 dose-dependently decreased response rates in acquisition andperformance, and heroin, but not U50488, increased errors emitted during acquisition. THC and
CP55940 disrupted acquisition and performance response rates, but only CP55940 increased errors
emitted during acquisition. THC in combination with heroin, shifted the heroin dose-effect curves foracquisition, performance, and antinociception leftward. Therefore, mu and kappa agonists disrupted
learning and performance, but only the mu agonist (heroin) increased learning errors. The CB1/CB2
agonist (THC) disrupted learning and performance, and in combination with a mu agonist (heroin)shifted the learning, performance, and antinociceptive dose-effect curves leftward. Overall,cannabinoids may not only enhance the antinociceptive effects of opioids, but also the learning andperformance deficits produced by opioids.
101

Presenter – Nozaki, Chihiro
Abstract ID – M32
CB2 receptor deletion on monocytes enhances neuropathic pain development in mice
Chihiro Nozaki1, Elisa Nent1, Anne Schmöle1, David Otte1, Andreas Zimmer1
1
Institute of Molecular Psychiatry, University of Bonn, Germany
We previously reported that cannabinoid receptor 2 (CB2) enhances neuropathic pain development.
However, why and how CB2 receptors regulate the development of peripheral neuropathy still
remains unclear. In this study, we determined which neuronal region and cells contribute to the CB2regulated neuropathic pain modulation. We first generated the mouse line expressing the GFP
protein under the CB2 promoter, and found that nerve injury-induced enhancement of CB2 receptor
expression has been only seen in sciatic nerve and not in DRG or spinal cord. This CB2 expression
was majorly colocalized with microglial marker Iba1, but not with neuronal marker synapsin1. We
then examined which CB2 receptors on where are critical for the development of neuropathic pain.
Using conditional knockout mice specifically lacking CB2 receptor on myeloid cells (CB2-LysM) oron neurons (CB2-Syn), we found that CB2 receptors expressed on microglia or synapse differently
regulates neuropathic pain development. Thus, although both CB2-LysM and CB2-Syn micedisplayed hypersensitivity on the ipsilateral paw, only CB2-LysM mice showed a significant
contralateral hypersensitivity after the nerve injury, which was similar to constitutive CB2-KO
animals. Accordingly, we observed an increased Iba1 signal in lumbar spinal cord of ligated CB2LysM on the both ipsilateral and contralateral side as well as constitutive CB2-KO mice. Additionally,
we found a similar upregulation of the leptin receptor in the ipsilateral sciatic nerve of ligated CB2LysM mice as seen previously in CB2-KO animals. Overall, our results suggest that CB2 receptors
on myeloid cells but not on neurons are essential for neuropathic pain development.
Presenter – Olson, Keith
Abstract ID – W13
Naltrexone C7-esters with improved nociceptin agonism as potential analgesics with reduced
rewarding effects
Keith Olson2, Gerta Cami-Kobeci1, Stephen Husbands1, John Traynor2
1
University of Bath, Bath, UK, 2University of Michigan, Ann Arbor, USA
There is an urgent need for analgesics with reduced abuse liability, withdrawal, respiratory
depression and constipation to treat chronic pain. Clinical opiates produce analgesia and associatedside effects through Mu Opioid Receptor (MOR) agonist activity, preclinical evidence indicates
activity at the Kappa- (KOR), Delta- (DOR), and Nociception Receptors (NOP) subtypes canminimize side-effects. Compounds displaying NOP agonist and weak MOR partial agonist activity
produce antinociception with minimal abuse liability in primates. Here, we evaluated C7 naltrexone
esters for opioid receptor affinity and agonist activity at MOR, NOP, DOR and KOR. Compound
affinity and potency showed a ~1000-fold range at NOP, ranging from low nanomolar to micromolar.
One compound, containing a phenyl ester at C7, and lacking a keto group at C6, showed the highest
potency NOP activity (EC50=0.97 +/- 0.22 nM, EMAX=47% +/- 11) with weak partial agonist potency
at MOR (EC50=3.9 +/- 1.3 nM, EMAX=23% +/- 7). This compound has lower MOR efficacy thanbuprenorphine suggesting reduced abuse liability. The series also showed a synergistic relationshipin NOP activity between the C7 functional group and the C6 oxidation state. For example, thepresence of a keto group at C6 decreased NOP potency significantly (EC50=164 +/- 51 nM,
EMAX=47% +/- 9) and was observed with all C7 functional groups. All compounds in the series were
putative antagonists at DOR and KOR. This series of compounds provides varied MOR and NOP
efficacies to determine the minimal MOR activity necessary for antinociception and reduced abuseliability.
Supported by Grant numbers R01 DA07315 and 035316.

102

Presenter – Ozawa, Akihiko
Abstract ID – M31
Neuroanatomical analysis of chronic pain-dependently activated neurons in the brain andspinal cord
Akihiko Ozawa1, Gilles Zribi1, Katarzyna Targowska-Duda1, Tatiana Gonzalez1, Lawrence Toll1
1
Department of Biomedical Science, Charles E. Schmidt College of Medicine, Florida Atlantic
University, Boca Raton, USA
Pain consists of both sensory and emotional experience where noxious stimuli trigger intense
aversion. Neuroanatomical studies have allowed us to define the regions and subsets of neurons
that are important for pain. Yet, the mechanisms by which the pain circuits regulating both sensory
and pain-related emotions are not well-explored; especially how these subsets of neurons contributeto transmit a pain signal and to elicit distinct pain behaviors. In this study, we shed light on paindependently activated neurons, in order to better understand the circuits in the central nerves system
activated only under chronic pain conditions, induced by spinal nerve ligation (SNL), using Fos2AiCreER mice. We primarily focused on two representative regions: anterior cingulate cortex (ACC)
as a key brain structure in the emotional/affective component of pain; and spinal cord for sensory
pain. Neural activation was quantified by monitoring mCherry expression triggered by a normally
non-painful stimulus in SNL mice, as a reference for c-Fos expression. We found that there were alarger number of neurons activated in the ipsilateral side of the spinal cord in SNL mice comparedto those of sham. The majority of spinal neurons activated during chronic pain was observed inlaminae IIinner, III and more ventral, which are important for mechanical allodynia. Similarly, neurons
in the contralateral side of the ACC were selectively activated by neuropathic pain due to SNL. The
methods established here represent a unique opportunity to further understand the roles of paindependently activated neurons in regulating specific pain behaviors under chronic pain conditions.
Presenter – Pan, Ying-Xian
Abstract ID – T31
Biased signaling through Oprm1 full-length 7TM exon 7-associated variants: Insights into in vivo
mu opioid actions
Jin Xu1, Valerie P. Le Rouzic1, Tiffany Zhang1, Zhigang Lu2, Gavril Pasternak1, Ying-Xian Pan1
1
Memorial Sloan-Kettering Cancer Center, New York, USA, 2Nanjing University of Chinese Medicine,
Nanjing, China
Extensive alternative pre-mRNA splicing of the mu opioid receptor gene (OPRM1) creates an array ofsplice variants that are conserved from rodents to humans. One type of the splice variants is full-length
7-transmembrane (TM) C-terminal variants that are identical except for the sequences at the tip of theintracellular C-terminal tail. Growing evidence supports the functional significance of these 7TM Cterminal variants. Particularly, exon 7 (E7)-associated C-terminal truncation in C57BL/6J mice (mE7MB6) diminished morphine tolerance and reward without altering physical dependence, like those seen ina β-arrestin2 KO mouse. Together with the results from cell-based studies, it suggests a physical andfunctional interaction of E7-associated C-terminal tails with β-arrestin2 that contributes to morphinedesensitization and tolerance. In the current study, we further examine the effect of E7-truncation on
fentanyl actions in mE7M-B6 mice. We observed that in mE7M-B6 mice fentanyl tolerance measured bydose-response curves before and after chronic fentanyl administered by an osmotic pump and fentanyl
reward determined by CPP were significantly reduced, while fentanyl analgesia was not affected, ascenario similar to those seen in morphine actions. On the other hand, fentanyl-induced physicaldependence determined by naloxone-precipitated withdrawal was slightly increased. Fentanyl-induce
locomotor activity was not significantly changed in mE7M-B6 mice. Furthermore, we deciphered aphosphorylation code in E7-encoded sequence that is important for β-arrestin binding using mutagenesis
studies with a Nanobit-based β-arrestin recruitment assay, providing new insights into the mechanisms
underlying fentanyl actions.
Supported in part by grants from NIH (DA042888, DA046714, DA007242, and CA08748)

103

Presenter – Peles, Einat
Abstract ID – W1
Polymorphisms in three stress-related genes are associated with reduced cocaine abuse andlonger retention in methadone maintenance treatment (MMT)
Peles E1,2, Levran O3, Randesi M3, Ott J4, Kreek MJ3, Adelson M.1,3,5
1
Dr. Miriam and Sheldon G. Adelson Clinic for Drug Abuse Treatment and Research, Tel Aviv Sourasky
Medical Center, Tel Aviv, Israel, 2Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel, 3The
Laboratory of the Biology of Addictive Diseases, The Rockefeller University, New York, USA, 4The
Laboratory of Statistical Genetics, The Rockefeller University, New York, USA, 5Dr. Miriam and Sheldon
G. Adelson Clinic for Drug Abuse Treatment and Research, Las Vegas, USA
We have recently reported an association between CRHBP SNP rs1500 and reduced cocaine abuse
after one year in MMT. To evaluate the relationships between 28 selected SNPs in 17 stress-related
genes and MMT patient's outcome (substance abuse and retention). From a non-selective sample of 405
MMT patients, close relatives and ancestry outliers (<50% Europeans/Middle Easterns, based on AIMs
markers) were excluded and 379 patients were analyzed for associations with addiction characteristics
and substance abuse. Comparisons under the dominant and recessive models were performed using chi
square for categorical and ANOVA for continues variables. Cumulative retention was studied using
Kaplan Meier analyses. Of the 379 patients, 330 (87.1%) stayed at least one year in treatment. Three
SNPs were associated with lower proportion of cocaine abusers after one year in treatment. The
percentage of cocaine abusers was lower in the AVP rs2282018 CC genotype (9.3%) compared to theother genotype groups combined (TT/TC) (22.6%, p=0.0455, pB=0.109, recessive model), in the CRHBP
SNP rs7728378 TT genotype (9.3%) compared to the other genotype groups combined (CC/CT) 25.2%,
p=0.002, pB=0.015 dominant model), and in the GAL rs3136541 TT/TC combined genotype (17.9%)
compared to CC (32.4%, p=0.0120, pB=0.051; recessive model). Cumulative retention (up to 25 years)
was longer in patients with the AVP rs2282018 CC genotype (13.3 years, 95% CI 10.8-15.8) than inpatients with the TT/TC genotypes (10.3, 95%CI 9.3-11.3, p=0.0384). These findings suggest an
association between specific SNPs in stress-related genes, reduced cocaine abuse and longer retention
in MMT.
Presenter – Pick, Chaim (Chagi)
Abstract ID – T30
Mirtazapine and Trazodone: A possible opioid involvement in their use (at low dose) for sleep?
S. Schreiber1, Chaim (Chagi) Pick2
1
Department of Psychiatry, Tel Aviv Sourasky Medical Center; Dr. Miriam and Sheldon G. Adelson Clinic
for Drug Abuse Treatment and Research, Tel Aviv and Israel; 2Department of Anatomy, Tel Aviv
University Sackler Faculty of Medicine; Dr. Miriam and Sheldon G. Adelson Center for the Biology of
Addictive Diseases, Tel Aviv University, Tel Aviv, Israel
The efficacy of each antidepressant available has been found equal to that of amitriptyline in double-blindstudies. However, a few of them are being prescribed (at very low, non-therapeutic doses) for sleep, in
non-depressed persons, when there are relative contraindications for hypnotics (i.e., history of addiction).
Following studies regarding the antinociceptive mechanisms of various antidepressants, we speculatethat the involvement of the opioid system in some of the antidepressants’ mechanism of action may
contribute to these medications use for the induction and maintenance of sleep. The mostly prescribed
antidepressants for sleep are trazodone (a weak, but specific inhibitor of the synaptosomal uptake ofserotonin, that also binds to alpha-1 and alpha-2 adrenoreceptor sites) and mirtazapine (a postsynaptic
drug which enhances noradrenergic and 5-HT1A-mediated serotonergic neurotransmission via
antagonism of central alpha-2-auto- and hetero-adrenoreceptors). When mice were tested with a hotplateanalgesia meter, both trazodone and mirtazapine induced, naloxone-reversible antinociceptive effectfollowing i.p administration. Summing up the various interactions of trazodone and mirtazapine with
opioid, noradrenergic and serotonergic agonists and antagonists, we found that the antinociceptive effectof trazodene is influenced by the opioid receptor subtypes μ and lass d (and a clear 5-HT mechanism ofantinociception), whereas the antinociceptive effect of mirtazapine is mainly influenced by κ and d opioid
receptor subtype (combined with both serotonergic and noradrenergic receptors). This opioid profile ofthe two drugs may be one of the explanations to their efficacy in the treatment of insomnia, when
sedatives (either benzodiazepines or the non-benzodiazepines “Z-compounds) cannot be prescribed.
104

Presenter – Polt, Robin
Abstract ID – M37
Glycosylation of opioid neuropeptides to produce brain-penetrant drugs
John Streicher1, Michael Heien1, Torsten Falk1, Lajos Szabo1, Chris Apostol1, Rachel K. Rowe2, Meredith
Hay1, Robin Polt1
1
The University of Arizona, 2Barrow Neurological Institute, USA
Peptide-based drugs offer significant advantages in that they bind with exquisite specificity to invivo targets (GPCRs), resulting in exceptionally high potencies and few off-target side effects. However,they also possess the disadvantages of very poor metabolic stability, low membrane permeability, andnon-linear pharmacokinetics and pharmacodynamics (PK/PD). For peptide drugs that target the central
nervous system (CNS), dynamic measurements of the drug’s blood-brain barrier (BBB) permeability, theirconcentrations at the site of action, and their metabolites are elusive due to their low concentration anddue to sampling difficulty. Applying a “biousian approach” (glycosylation strategy) which increases peptide
stability and half-life in vivo, and promotes penetration of the BBB, our research team has synthesized anarray of structurally diverse glycopeptide drug candidates that persist in serum long enough to penetrate
the blood-brain barrier (BBB). The peptide hormones targeted for conversion to glycopeptide drugs
include small water soluble peptides such as Angiotensin and Leu-Enkephalin (M.W.'s < 700), as well aslarger Dynorphin\Endorphin hormones (M.W. ~1800), as well as cyclic derivatives of Oxytocin (M.W.
~1000) and much larger peptide hormones such as PACAP\VIP (M.W. > 3,300). Despite their structuraldifferences all of these peptides were amenable to the biousian approach, with clinically-relevant amountsof glycopeptide being delivered across the BBB regardless of M.W. This emergent class of glycopeptides
may be able to combine the best features of small molecule drugs with the specificity and selectivity of
biologics by overcoming the PK/PD issues associated with many peptide drugs.

Presenter – Poras, Hervé
Abstract ID – T5
DENKI (Dual ENKephalinase Inhibitors), a new class of painkillers devoid of abuse potential
and of opiate side-effects
Hervé Poras1, Gwendolyn E Burgess2, Bryan F Sears2, Tanja Ouimet1, Michel Wurm1, Marie-Claude
Fournie-Zaluski1, Bernard P Roques1, Emily M Jutkiewicz2
1
Pharmaleads SA, Paris, France, 2Department of Pharmacology, University of Michigan, Medical
School, Ann Arbor, USA
Pharmaleads develops two Dual Enkephalinase Inhibitors (DENKI®), PL37 and PL265, active on
different types of pain models and devoid of opiate side effects. Enkephalins are penta-peptides
produced, e.g. during extreme physical workout or during long lasting painful stimuli, in the central
nervous system and in peripheral tissues. Their release elicits a “physiological” analgesia, providing
the body with its own pain modulation system. Once released, enkephalins are rapidly inactivatedby two membrane-bound exo-metalloproteases, neprilysin (NEP) and aminopeptidase N (APN).
DENKI® specifically protect enkephalins from NEP and APN in those areas where pain is perceived,transmitted and integrated. Unlike exogenous opiates, naturally secreted enkephalins do not induce
rewarding effects, reinforcing effects, withdrawal symptoms or dependence. In animal models of
pain, DENKI® induce antinociceptive responses similar to those of morphine. They remain active
after chronic treatment with morphine, indicating that DENKI®-induced analgesia persists inmorphine-tolerant mice. DENKI® do not induce constipation in animals or humans. PL37 and PL265
do not produce morphine-like discriminative stimulus effects. They fail to maintain responding ormaintain significantly less responding in rats trained to self-administer remifentanil and morphine. In
addition, they induce delta-opioid receptor-mediated antidepressant-like effects in the forced swim
test and do not induce convulsions. PL37 and PL265 appear to be devoid of abuse liability even athigh doses and after repeated administration and could represent a new class of painkillers in all
types of severe pain as an affordable solution to the opioid crisis.

105

Presenter – Pradhan, Amynah
Abstract ID – M9
The critical role of central delta opioid receptors in models of migraine and opioid-induced
hyperalgesia
Isaac J. Dripps1, Zachariah Bertels1, Laura S. Moye1, Alycia F. Tipton1, Serapio M. Baca2, Amynah
Pradhan1
1
University of Illinois at Chicago, Chicago, USA, 2University of Colorado Denver, Denver, USA
The delta opioid receptor (DOR) shows promise as a target for novel migraine medications as DOR
agonists inhibit migraine-induced pain and negative affect in rodents without the abuse potential andhyperalgesia associated with µ opioid receptor agonists such as morphine. DORs are expressed inseveral peripheral and central regions important for pain processing and mood regulation, and it
remains unclear which receptors are critical for regulating these anti-migraine effects. To this end,
we examined the effects of the DOR agonist SNC80 in multiple headache models in conditional
knockout mice with DORs missing from specific central or peripheral regions. Periorbital mechanicalallodynia was measured following treatment with chronic nitroglycerin (NTG), a known migraine
trigger. Periorbital and hindpaw allodynia was also measured following repeated treatment withmorphine, a model of opioid-induced hyperalgesia. Migraine-associated negative affect wasassessed using conditioned place aversion to NTG. We also evaluated the effects of SNC80 on KClevoked cortical spreading depression. Conditional knockout of DORs deleted from peripheral
voltage-gated sodium channel Nav1.8-expressing neurons (Nav1.8-DOR) only inhibited the effectsof SNC80 on opioid-induced hyperalgesia in the hindpaw. Loss of DORs in GABAergic forebrainneurons (Dlx-DOR) significantly diminished the effects of SNC80 in all observed outputs. NTG alsoenhanced DOR expression in multiple brain regions and measured by DOR-eGFP fluorescence.
Taken together, these data further support the potential of the delta opioid receptor in the treatmentof headache disorders and suggest that central DORs are important mediators of the anti-migraine
effects of DOR agonists.
Presenter – Presker, Mark
Abstract ID – W37
Morphine and cocaine cues differentially control natural reward motivated behavior: role oflocus coeruleus.
Mark A. Presker Jr.1, David Sosidko1, Samuel Tombokan1, Gary Aston-Jones1
1
Brain Health Institute - Rutgers the State University of New Jersey, USA
A major obstacle in treating addiction is the high propensity for relapse. Stimulus-driven drug seeking
occurs despite efforts to remain abstinent and competes with ongoing alternative preferred behaviors
such as occupational or social activities. Noradrenergic locus coeruleus (LC) neurons projectthroughout the neuraxis and modulate sensory processing and attention to orient organisms tosensory stimuli. Previous work shows that LC activity is altered acutely and chronically by bothmorphine and cocaine. This supports a role for LC in mediating stimulus control of behavior by drugcues. However, few studies have examined whether morphine- or cocaine-associated stimuli
activate LC or exert control over non-drug seeking behaviors or how this may differ between drug
types. Here we implement a behavioral paradigm specifically designed to examine the effects ofmorphine-paired, cocaine-paired, and saline-paired auditory stimuli (tones) on operant responding
for a natural reinforcer (water) in rats. We found that morphine- and cocaine-paired tones exert
control over multiple behaviors and these effects differ between drugs. We also collected tissue toassess Fos expression in LC of rats exposed to each tone type. LC Fos expression will be comparedwith behavioral measures to determine whether they correlate. We previously found a relationshipbetween LC activity and behavioral response to cocaine tones. This study will establish whether
control of behavior by an opioid-associated cue is also encoded in LC activity. Given the role of cuesin relapse to opioids and cocaine, identifying a common neural substrate of attentional control may
provide a target for therapeutic intervention.

106

Presenter – Pryce, Kerri
Abstract ID – W42
Oxycodone-induced gene expression adaptations in the brain reward center in a murine
model of neuropathic pain
Kerri D. Pryce1, Hope Kronman1, Angelica Torres Berrio1, Sevasti Gaspari1, Aarthi Ramakrishnan1,
Li Shen1, Eric Nestler1, Venetia Zachariou1
1
Icahn School of Medicine at Mount Sinai, New York, USA
Long-term use of opioids for the treatment of chronic pain can lead to physical dependence andtransition to addiction. Recent studies from our group revealed that chronic neuropathic pain
promotes several adaptations in the brain reward regions (Descalzi et al., Sci. Signaling, 2017). Here,
we are using Next Generation RNA-sequencing to gain insight on gene expression adaptations
triggered by persistent exposure to oxycodone in the presence and in the absence of pain. We
employed the spared nerve injury model (SNI) of neuropathic pain in adult C57Bl/6 mice andassessed peripheral neuropathy symptoms for ~9 weeks. Mice were then treated daily withoxycodone, for two weeks and gene expression was monitored three weeks after oxycodone
withdrawal. Oxycodone treatment induced thermal hyperalgesia regardless of pain states, while
withdrawal produced significant mechanical allodynia and deficits in social interaction. Using RNA
Sequencing, we monitored changes in gene expression in the medial prefrontal cortex (mPFC), thenucleus accumbens (NAc) and the ventral tegmental area (VTA). Although oxycodone treatment
promotes mostly unique transcriptome profile across brain regions, we observed similar downstream
effectors and transcription factors to be affected in pain states when compared with non-pain states.
Our pathway analysis predicted that several targets of histone deacetylase 6 are differentiallyexpressed in oxycodone withdrawal as well as in SNI groups. Indeed, treatment with HDAC6
inhibitors prevents oxycodone-induced hyperalgesia and alleviates mechanical allodynia in SNI
mice. Overall this work will provide new insights on the influence of long term pain in oxycodoneinduced plasticity in the brain reward center.
Presenter – Rae, Mariana
Abstract ID – W41
Effects of environmental enrichment on alcohol and social reward and modulation of theoxytocinergic system
Mariana Rae1, Panos Zanos2, Polymnia Georgiou2, Alexis Bailey3, Achla Gupta4, Ivone Gomes4, Lakshmi
Devi4, Rosana Camarini1
1
Pharmacology Department, University of São Paulo, Brasil, 2Department of Psychiatry, University of
Maryland Baltimore, USA, 3Institute of Medical and Biomedical Education, St George's University of
London, UK, 4Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, USA
Environmental enrichment (EE) is hypothesized to protect against drug addiction-related behaviors.
However, the mechanisms remain unknown. The aim of this study was to evaluate the effect of EE on
alcohol reward. Adult Swiss male mice were distributed in control and EE groups. We characterized theeffects of EE on alcohol (2.0 g/Kg) conditioned-place preference (CPP). Since one of the main pillars of
EE is increased social interaction, we accessed the effects of EE on oxytocin (OT) and OT receptor(OTR), via ELISA, qPCR and autoradiography. EE group exhibited higher ethanol preference, suggestinga sensitization of the reward system induced by EE. EE animals had higher OT mRNA and peptide levelsin the hypothalamus and lower OTR binding in the olfactory nuclei and piriform cortices. These results
indicate that EE modulates the OT system by increasing OT synthesis, leading to OTR downregulationin specific brain regions. To assess the role of the OT in the observed EE-induced enhancement of
ethanol CPP, we treated animals with the OTR antagonist, L,369-899 (5 mg/kg/day, i.p) in that paradigm.
Treatment with the antagonist during the EE exposure blocked this enhancement in ethanol CPP. Thesefindings indicate a direct modulation of the OT system on ethanol rewarding effects. Finally, we tested if
confrontation between alcohol and social reward could reverse alcohol increased preference on EE but
found no effect. In conclusion, OT-based therapies regarding alcohol addiction should be considered
cautiously, particularly in the early stages of alcohol use.
Supported by CAPES, CNPq, FAPESP and Santander.

107

Presenter – Randesi, Matthew
Abstract ID – T25
Sex differences in opioid, plasticity and stress-related genes in rat caudate putamen following
oxycodone conditioned place preference
Matthew Randesi1, Yan Zhou1, B. Reich2, Bruce S. McEwen3, Teresa Milner2, Mary Jeanne Kreek1
1
The Rockefeller University, New York, USA, 2Feil Family Brain and Mind Research Institute, Weill
Cornell Medicine, New York, USA, 3The Laboratory of Neuroendocrinology, Rockefeller University,
New York, USA
This study was conducted to determine if there are sex differences in gene expression in adult rats
following oxycodone (oxy) conditioned place preference (CPP). RNA was isolated from both thecaudate putamen and nucleus accumbens of 24 rats (6 saline + 6 oxy treated females and an equal
number of males). cDNA was synthesized and gene expression levels measured using SYBR®
Green and a custom RT2-PCR array (Qiagen, Inc.), containing genes for opioid peptides/receptors,
genes related to synaptic plasticity, involved in stress-responses, and kinase and signaling genes.
Expression levels were compared to saline controls and statistically significant changes were foundin caudate putamen, but not nucleus accumbens. Two-way ANOVA showed an interaction effect(Sex x Condition) on expression of three genes: cadherin (Cdh2), mitogen-activated protein kinase
1 (Mapk1) and nociceptin receptor (Oprl1) (p=0.0056, p=0.0138, p=0.0142, respectively). Significant
main effect from sex was also found for Cdh2, Mapk1 and neuropeptide Y (Npy) gene expression
levels (p=0.0020, p=0.0017, p=0.0419, respectively). Post-hoc analysis showed that the saline males
had significantly greater expression of Cdh2 than both saline females and oxy males (p=0.001 and
p=0.005, respectively). While oxy females had significantly greater expression than the oxy males
for both Mapk1 and Npy (p=0.001 and p=0.034, respectively). Oxy females also had significantly
greater Oprl1 expression than saline females (p=0.026). In conclusion, we found modest sex- anddrug-differences in gene expression in the rat caudate putamen following oxycodone conditioned
place preference.
Supported by NIH grant DA08259 & Dr. Miriam and Sheldon Adelson Medical Research Foundation.
Presenter – Reiner, David
Abstract ID – M10
Role of the orbitofrontal cortex in relapse to fentanyl seeking after choice-induced voluntary
abstinenceDavid J Reiner1, Olivia M Lofaro1, Marco Venniro1, Carlo Cifani1, Jennifer Bossert1, Yavin Shaham1
1
Behavioral Neuroscience Research Branch, NIDA-IRP, NIH, USA
There are few preclinical studies of prescription opioid relapse, and these studies have used animalmodels in which the abstinence period is experimenter-imposed. However, in humans, abstinence
is often voluntary with drug available in the former drug user’s environment but forgone in favor ofother non-drug alternative rewards. We recently developed a rat model of food choice-induced
voluntary abstinence. Here we used the model to study the neurobiological mechanisms of relapse
to fentanyl seeking. We first trained male and female rats to self-administer palatable food pellets for
6 days (6-h/day) and fentanyl (2.5 microgram/kg/infusion, i.v.) for 12 days (6-h/day). We assessed
relapse to fentanyl seeking after 14 days of voluntary abstinence, which was achieved through a
discrete choice procedure between drug and palatable food (20 trials/day). In both sexes, relapse tofentanyl seeking was associated with increased expression of the neuronal activity marker Fos inboth the lateral and ventral orbitofrontal cortex (OFC). Reversible inactivation of either region with
GABA-A and GABA-B agonists muscimol and baclofen (50+50 ng/side) decreased relapse tofentanyl seeking. These data, along with previous data on the role of OFC in relapse to heroin
seeking after forced abstinence, indicate that OFC activity is critical for opioid relapse, independentof the opioid or the abstinence procedure.

108

Presenter – Retzlaff, Cassandra
Abstract ID – M45
Investigating the role of striatal neuropeptide Y expressing interneurons in opioid action
Cassandra L. Retzlaff1, David Leipold1, Quinlan Kile1, Patrick Rothwell1
1
University of Minnesota, Minneapolis, USA
Mu opioid receptor (MOR) agonists such as morphine are known to regulate the activity ofGABAergic interneurons in the ventral tegmental area and hippocampus, both areas known to beimportant for reward processing. Though the nucleus accumbens (NAc) is a critical structure in the
reward circuit, the effects of MOR agonism on interneurons in this brain region are not wellunderstood. While interneurons comprise roughly 5% of total neurons in the striatum, they can exert
significant control of the primary output cells in the striatum, the medium spiny projection neurons(MSNs). Using fluorescent in situ hybridization to identify neuropeptide Y (NPY) expressing
interneurons, a subpopulation of interneurons not well characterized in the NAc, we find that many
(68%) co-express MOR. Opioid modulation of NPY interneurons is further supported by slice
electrophysiological data, which show DAMGO (a MOR specific agonist) can reduce firing andhyperpolarize the membrane potential of NPY interneurons. To explore the contribution of NPY
interneurons to opioid-induced behaviors, we used Cre-dependent viral expression of potassiumchannel KIR2.1 in NPY-Cre mice. Preliminary data suggest this manipulations alters locomotor
sensitization following repeated opioid exposure, suggesting that NPY interneurons robustly
modulate the output of the NAc. Ongoing studies will examine changes in intrinsic excitability andsynaptic output of NPY interneurons in the NAc after repeated opioid exposure.
Presenter – Rivera Irizarry, Jean K.
Abstract ID – T41
Anterior paraventricular thalamus to nucleus accumbens projection is involved in rewardseeking behaviors
J.K. Rivera Irizarry1,2, J.D. Miller1, K.E. Pleil1,2
1
Department of Pharmacology Weill Cornell Medicine, Cornell University, New York, USA,
2
Neuroscience Graduate Program Weill Cornell Medicine, Cornell University, New York, USA
Glutamatergic signaling in the nucleus accumbens shell (NAcSh) promotes reward-seeking
behaviors including drug self-administration. The NacSh receives dense glutamatergic input fromthe paraventricular thalamus (PVT), a midline thalamic region also involved in these behaviors andimportant for the integration of reward and aversion. Recent experimental evidence indicates thatprojection neurons in the anterior PVT (aPVT) preferentially innervate the dorsomedial NAcSh(dmNAcSh), a subregion of the NAcSh associated with positive hedonic experiences. However, therole of the aPVT-dmNAcSh pathway on these motivated behaviors remains unknown. Here, we
evaluated the hypothesis that activity of the aPVT-dmNAcSh pathway is rewarding and sufficient todrive reward-seeking behaviors and mitigate behavioral responses to aversive stimuli using apathway-specific chemogenetic approach. To determine whether activation of the aPVT inputs to the
dmNAccSh are reinforcing or aversive, we measured home cage CNO self-administration andsucrose preference using a limited access two-bottle choice paradigm in male and female miceexpressing the excitatory Gq-DREADD or control vector in the aPVT-dmNAcSh pathway. GqDREADD mice displayed greater CNO consumption, higher CNO preference, along with increased
sucrose intake and preference than controls; demonstrating that this thalamo-limbic circuit promotes
reward-seeking behavior. Additionally, CNO activation of the Gq-DREADD prior to Pavlovian fear
conditioning significantly delayed the acquisition of fear conditioning in females but not males. These
results suggest that stimulating the aPVT-dmNAcSh pathway is reinforcing in male and female mice,with possible sex-specific effects in modulating behavioral outputs in aversive contexts. Future
experiments will examine whether this circuit competes with parallel aversion-promoting PVT
outputs.

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Presenter – Roques, Bernard P.
Abstract ID – W45
Exaltation of natural opioids enkephalins functions by inhibiting their synaptic peptidases
inactivation in painful and depressive conditions. Extension to migraine
Bernard P. Roques1, Marie-Claude Fournié-Zaluski2
1
University Paris Descartes, France, 2Pharmaleads SAS, France
Leu- and Met- enkephalins are the main effectors of the opioid Mop and Dop receptors. Their very
short lifetime, due to their rapid physiological catabolism by two ectopeptidases Neprilysin (NEP)and Aminopeptidase N (APN), prevents their potency as pain killer to be evaluated directly.
Therefore, potent and selective inhibitors of these peptidases have been developed leading to an
innovative approach to pain relief through the concept of “Dual ENKephalinase Inhibitors” DENKIs.
Among them, two molecules have been selected PL37 and PL265. They are prodrugs of two different
chemical series. These compounds protect in vivo the enkephalins released during physical or
psychic painful events after i.v. and/or oral routes. Moreover, PL265, orally administered does notenter the brain, leading to pure peripheral effects. Tested on animal models of pain, these molecules
are highly efficient in protecting the enkephalins released during all types of pain, inflammatory andocular injury, migraine, post-operative nociception, or neuropathy, without all drawbacks induced byexogenous opioids i.e. tolerance, dependence, respiratory depression, constipation. Moreover, they
are efficient to regulate affective disorders and depressive illness. Preclinical studies of PL37 andPL265 in human have shown the absence of toxicity. Clinical Phase 2 for PL37 in neuropathic pain
(capsaicin test and diabetic neuropathy) has demonstrated the efficacy of this strategy. DENKis
effects are enhanced after association with drugs acting on other mechanisms than opioids. This
could prevent or attenuate the « crisis » occurring by use of morphine derivatives.
Roques et al, Nature Rev. Drug Discovery, 2012, 11, 292-311.
Presenter – Rossi, Grace
Abstract ID – M47
The pharmacological and molecular characterization of mu opioid receptor splice variants in
the rat using gene targeting models generated by Easi-CRISPR
Grace C. Rossi1, Jin Xu2, Valerie P. Le Rouzic2, Gavril W. Pasternak2, Miyasaka Yoshiki3, Tomoji
Mashimo3, Thom Saunders4, Channabasavaiah B. Gurumurthy5, Ying-Xian Pan2
1
Department of Psychology, Long Island University, Post Campus, Brookville, USA, 2Department of
Neurology and Molecular Pharmacology, Memorial Sloan Kettering Cancer Center, USA, 3Instituteof Experimental Animal Sciences, Graduate School of Medicine, Osaka University, Osaka, Japan,
4
Transgenic Animal Core, University of Michigan, Ann Arbor, USA, 5Mouse Genome Engineering
Core Facility, University of Nebraska Medical Center, Omaha, USA
A single-copy mu opioid receptor (OPRM1) gene generates a vast array of mu opioid receptor splicevariants through extensive alternative splicing that is conserved from rodent to human. Thesevariants can be divided into two sets: exon 11 (E11) and exon 1 (E1) associated variants. The
functional differences between the E1 and E11 sets of variants have been demonstrated using genetargeted mouse models. While mouse models are valuable, rats have many advantages both inbehavioral modeling and in vivo manipulation. We have generated two conditional Oprm1 knockout(KO) models, E11-KO and E1-KO, in Sprague Dawley rats using a novel Easi-CRISPR approach.
Cre-mediated E11 and E1 deletions was confirmed through breeding a CAG-Cre rat. Initial
behavioral studies showed that in both E11 and E1 KO rats, buprenorphine analgesia was lost,
Buprenorphine CPP was significantly reduced in E11 KO rats. Morphine analgesia was completely
lost in the E1-KO rat, but intact in the E11-KO rat. Morphine CPP was also eliminated in E1-KO
rat. Our results suggest that buprenorphine analgesia is dependent on both E11 and E1-associatedvariants, a similar scenario seen in the E11-KO and E1-KO mouse models. E11-associated variants
also involved in buprenorphine reward in rat. These conditional Oprm1 KO rat models will provide
valuable resources for the research community and would expand our ability explore mu opioid
pharmacology with approaches and techniques not feasible in mice.
Supported in part by grants from NIH (DA040858, DA042888, DA46714, DA007242 and CA08748).
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Presenter – Rubaharan, Myurajan
Abstract ID – T45
The impact of age, sex, and chronic pain on microglia-related neuroinflammation in the rat
periaqueductal gray
Myurajan Rubaharan1, Evan F. Fullerton1, Meira B. Robbins1, Anne Z. Murphy1
1
Georgia State University, USA
Approximately 50 million adults in the United States experience chronic pain on a daily basis, withan increased prevalence observed in people aged ≥65. Opioid-based medications are the mostcommon treatment used for the management of chronic pain, however there is a dearth of
information regarding the impact of advanced age on central opioid signaling and pain. Using amodel of persistent inflammatory pain, our studies show that morphine efficacy is significantly
attenuated in aged male and female rats. Microglia, the resident immune cells of the brain, have theability to modulate the analgesic effects of morphine. Specifically, our lab has previously shown that
activated microglia within the midbrain periaqueductal gray (PAG), a key site for morphine action,
stimulate cytokine release that directly oppose the antinociceptive and antihyperalgesic effects ofmorphine. As aging is associated with increased levels of inflammation, we hypothesized that
increased levels of neuroinflammation/microglia activation contribute to the attenuated response tomorphine observed in aged rats. Adult and aged male and female rats received an intraplantar
injection of Complete Freund’s adjuvant and were perfused 1 or 3 days later. Microglia activation
within the PAG was assessed via morphometric and inflammatory protein marker analyses.
Collectively, our data indicate that sex, age, and chronic pain may lead to differential changes inbasal microglial activity and provide a mechanism for the attenuated effect of morphine in chronic
pain management in the aged population.
Presenter – Ryan, James
Abstract ID – M12
Interrogating the role of brain-derived neurotrophic factor and the ventral hippocampus inmediating fear generalization
James D. Ryan1,2, Nathaniel Tse1,2, Deqiang Jing1, Francis S. Lee1
1
Department of Psychiatry, Weill Cornell Medicine, New York, USA, 2Weill Cornell Graduate Schoolof Medical Sciences, New York, USA
Fear generalization is a maladaptive behavior central to several psychiatric disorders that manifests
from exposure to particularly traumatic situations via underlying neural and cellular mechanisms thatremain largely unclear. Here, we address this question by first establishing a behavioral model offear generalization induced by high-intensity foot shocks during auditory fear conditioning in C57BL/6
mice. We then assessed the role of brain-derived neurotrophic factor (BDNF) in fear generalization
by exposing knock-in mice with deficient activity-dependent BDNF secretion (BDNF Val66Met) to
the fear generalization protocol as well as by prophylactic treatment with ketamine, a drug that exerts
effects on synaptic plasticity and remodeling in part by a BDNF-dependent mechanism. Our results
indicate that BDNF Val66Met mice exhibit fear generalization even when conditioned with the lowintensity foot shock, and further, that prophylactic ketamine treatment accelerates the extinction ofgeneralized fear in wildtype mice but is less effective in BDNF Val66Met mice. To then parse out theneural circuits that mediate fear generalization, we used in vivo calcium imaging via fiber
photometry to measure population-level signaling activity in the vCA1 during fear and safety
memory recall. We found robust, tone-evoked and genotype-dependent changes in activity in mice
conditioned with a high-intensity foot shock that is distinct from the signaling patterns seen in lowintensity foot shock-conditioned mice. Overall, our work demonstrates the significant role of the vCA1and BDNF signaling in mediating generalized fear while also suggesting the potential therapeuticutility of prophylactic ketamine for the treatment of PTSD.

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Presenter – Saberi, Shahin
Abstract ID – W16
Small molecule inhibitors of PSD95–nNOS protein–protein interactions suppress opioid-induced
reward in rats
Shahin A. Saberi1, Tannia Gutierrez1,2, Yvonne Y. Lai1, Pushkar M. Kulkarni3, Sumanta Garai3, Ganesh
A. Thakur3, Jonathon D. Crystal1, Andrea G. Hohmann1,2,4
1
Department of Psychological and Brain Sciences, Indiana University, Bloomington, USA, 2Program in
Neuroscience, Indiana University, Bloomington, USA, 3Center for Drug Discovery and Department ofPharmaceutical Sciences, Northeastern University, Boston, USA, 4Gill Center of Biomolecular Science,
Indiana University, Bloomington, USA,
Excessive glutamate signaling through N-methyl-D-aspartate receptors (NMDARs) is implicated in alteredforms of neuronal plasticity including opioid reward and dependence. NMDAR antagonists can blockthese effects, but their clinical use is limited by adverse side-effects. Excessive NMDAR activity activatesthe enzyme neuronal nitric oxide synthase (nNOS) to produce the signaling molecule nitric oxide (NO).
nNOS is tethered to NMDARs via the scaffolding protein postsynaptic density 95 kDa (PSD95). Thus,
disruption of the protein-protein interactions between PSD95 and nNOS may provide a viable therapeuticroute to inhibit NMDAR-dependent opioid reward without unwanted side-effects of NMDAR antagonists.
We previously reported that small-molecule inhibitors of PSD95-nNOS protein-protein interactions(IC87201, ZL006) inhibited NMDAR-dependent NO formation and suppressed pathological pain without
impairing memory or motor function (Neuropharm 97 (2015) 464-75; Behav Brain Res (2016) 305:23-29).
However, whether PSD95-nNOS inhibitors themselves lack abuse liability or interfere with NMDARdependent opioid reward remains unknown. We used a three-chamber conditioned place preference(CPP) assay to evaluate the impact of IC87201 and ZL006 on opioid reward. Morphine produced robust
CPP whereas IC87201 and ZL006 failed to induce either place preference or aversion. Both IC87201 and
ZL006 blocked CPP to morphine at doses that lacked intrinsic rewarding or aversive properties. Similar
results were observed using CPP paradigms that involved either repeated or single pairings. Our results
support the hypothesis that PSD95-nNOS inhibitors exhibit limited abuse liability. Our findings also
highlight a previously unrecognized role for PSD95-nNOS disruptors as a novel non-opioid therapeuticstrategy that shows promise for suppressing opioid reward.
Presenter – Saitoh, Akiyoshi
Abstract ID – W23
KNT-127, but not SNC80, facilitates extinction learning of contextual fear memory via delta opioid
receptor in mice
Akiyoshi Saitoh1, Daisuke Yamada1, Shoko Yanagisawa11, Shinya Yanagita2, Jun-Ichiro Oka1, Hiroshi
Nagase3
1
Lab Pharmacol, Fac Pharm Sci, Tokyo Univ of Science, Chiba, Japan, 2Dept Liberal Arts, Fac Sci andTech, Tokyo University of Science, Chiba, Japan, 3IIIS, Univ of Tsukuba, Ibaraki, Japan
In the present study, we evaluate the effects of selective delta opioid receptor (DOP) agonists KNT-127
and SNC80, which are compounds of different structures, on the contextual fear memory in the contextual
fear conditioning test. Male C57BL/6J mice were used. On the conditioning day, the mice were trainedwith eight conditioning trials (0.8 mA x 1s, inter-trial interval 30 s). Twenty-four h following conditioning,the mice were injected subcutaneous administration with drugs. After 30 min, the mice were reexposedto the same apparatus for 6 min (extinction training session). Twenty-four hours after extinction, eachmouse was placed back in the conditioning chamber and a 6-min test was performed (memory testing
session). During each session, no footshock was given, and the mouse was observed every 1 min. In
extinction training session, KNT-127 and SNC80 significantly reduced contextual conditioned freezingtime in mice when compared to saline, respectively. In memory testing session, KNT–127 significantly
reduced contextual conditioned freezing time, while SNC80 treated-freezing time produced no significant
effects. Further, the effects of KNT-127 were reversed by pretreatment with a DOP antagonist naltrindole.
These results suggested that KNT-127 produced the anxiolytic-like effects in extinction training session
and facilitated the fear extinction learning in memory testing session though the DOP. On the other hand,
it was suggested that SNC80 produced the anxiolytic-like effects but produced no facilitation effects on
fear extinction leaning. We proposed that KNT-127, but not SNC80, facilitates extinction learning ofcontextual fear memory via DOP in mice.
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Presenter – Sakloth, Farhana
Abstract ID – M41
Inhibition of HDAC6 activity alleviates symptoms of chronic pain
Farhana Sakloth1, Olivier Berton1, Lefteris Manouras1, Kleopatra Avrampou1, Vasiliki Mitsi1, Valeria
Cogliani1, Kerri Pryce1, Randal Serafini1, Mathew Jarpe2, Venetia Zachariou1
1
Nash Family Department of Neuroscience and Department of Pharmacological Sciences, Icahn
School of Medicine at Mount Sinai, New York, USA, 2Regenacy Pharmaceuticals, 303 Wyman St,
Suite 300, Waltham, USA
HDAC6 is a class II histone deacetylase, expressed at many points of the pain matrix. Previous
studies have indicated that pharmacological modulation of histone deacetylases (HDACs) may
provide a possible avenue for the development of new treatments for pain. In this study, we assessed
the ability of two selective HDAC6 inhibitors, ACY-738 and ACY-257, to alleviate symptoms of
peripheral inflammation and peripheral nerve injury. Using the spared nerve injury (SNI) mousemodel of peripheral neuropathy we showed that ACY-738 abolishes mechanical allodynia andthermal hyperalgesia in male and female mice. The peripherally acting ACY-257 compound also
attenuated mechanical allodynia in the SNI model. The two compounds showed antiallodynic
properties in the CFA (Complete Freud’s Adjuvant) models of hindpaw inflammation. To gain insighton the physiological role of HDAC6 on chronic pain-related behaviors, we used conditional knockout
models and downregulated HDAC6 in the periaqueductal gray (PAG) of adult male mice. Specifically,
we infected the PAG of HDAC6fl\fl mice with viral vectors expressing Cre recombinase (AAV-Cre) orcontrol AAV-GFP to locally downregulate HDAC6 in the adult life. Using the CFA model, we foundthat downregulation of HDAC6 in the PAG does not affect the onset or the intensity of mechanicalallodynia, but it has a prominent effect on the maintenance of allodynia. Our findings suggest that
inhibition of HDAC6 may provide a new therapeutic avenue for the treatment of chronic pain
conditions.
Presenter – Sears, Bryan
Abstract ID – W10
In vivo evaluation of AMB67 and AAH8, mixed efficacy MOR agonist/DOR antagonist ligands:
An alternative approach for pain management
Bryan F Sears1, Aaron M Bender1, Jessica P Anand1, Anthony F Nastase1, Aubrie H Harland1,
Henry I Mosberg1, Emily M Jutkiewicz1
1
University of Michigan, USA
Opioids, such as morphine, have long been used to treat pain and exert their effects through the muopioid receptor (MOR). While effective, these MOR agonists produce on-target adverse effects suchas tolerance, physical dependence and euphoria. Previous studies show co-administration of a MOR
agonist and delta-opioid receptor (DOR) antagonist provide antinociception with diminished adverse
effects. AMB67 is a MOR agonist/DOR antagonist with low nanomolar affinity at both receptors with
24-fold greater affinity for MOR over DOR. To evaluate acute and chronic antinociceptive effects, I
utilized models of thermal (warm water tail withdrawal (WWTW)), chemical (acetic acid-induced
writhing assay), and mechanical (von Frey assay) nociception. In C57BL/6 mice, AMB67 produced
dose-dependent antinociceptive effects in all three nociception assays. Following 5 days ofadministration with increasing doses, the ED50 for the AMB67 dose effect curve in the WWTW was
unchanged; however, following the same chronic dosing regimen, the dose effect curve for AMB67
in the von Frey assay was shifted 2.5-fold to the right. Naltrexone precipitated more jumping behavior
in mice treated with chronic morphine as compared with chronic AMB67. After 5 days of conditioning,
AMB67 appeared to be less potent than morphine in a conditioned place preference paradigm andexhibited less morphine-like discriminative stimulus effects. Overall, these studies show AMB67
produces similar antinociceptive effects as morphine but produces less tolerance, physicaldependence, and rewarding effects. These data suggest DOR antagonism may play a key role indiminishing some of the on-target adverse effects while preserving the analgesic properties of MOR
agonists.
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Presenter – Séguéla, Philippa
Abstract ID – M42
Mu opioid receptor expression and function in mouse ACC versus S1 cortex
Maria Zamfir1,2, Behrang Sharif1,2, Samantha Locke1,3, Aliza Ehrlich4, Brigitte Kieffer4, Alfredo Ribeiro-daSilva1,3, Philippe Séguéla1,2
1
Alan Edwards Centre for Research on Pain, Canada, 2Montréal Neurological Institute, Dept. Neurology& Neurosurgery, McGill University, Canada, 3Dept. Pharmacology & Therapeutics, McGill University,
Canada, 4Douglas Mental Health University Institute, Dept. Psychiatry, McGill University, Canada,
The analgesic effect of morphine and its derivatives relies on the presence of the mu opioid receptors(MOR) in pain pathways. MORs are expressed in several cortical areas including the anterior cingulate
cortex (ACC) and primary somatosensory cortex (S1). The ACC is involved in generating the affective
component of the pain experience whereas S1 encodes the discriminative component. Our aim is todetermine both the function and localization of MOR in ACC and S1. We performed whole-cell patch
clamp recordings of cells in acute slices from both wild type and Oprm1-mCherry transgenic mice. We
bath applied DAMGO, a selective mu opioid receptor agonist, and observed the effect of MOR activation
on cellular excitability and potassium current downstream of MOR activation (GIRK). Based on ourelectrophysiological data, we found that in ACC mostly ACC interneurons (regular spiking non-pyramidalcell) express MOR whereas in S1 mostly excitatory neurons express MOR. We took advantage of theOprm1-mCherry line for tracking MOR distribution using IHC. We performed immunostaining for SATB2,
a transcription factor that is a marker of pyramidal neurons, and for typical interneuron markers such as
somatostatin (SOM). Our IHC results suggest that different proportions of inhibitory and excitatoryneurons express MOR in ACC versus S1. Recent studies have suggested that cortical microcircuitry
differs between 6- layered primary sensory areas. Our results show that the expression of MOR follows
distinct patterns in two somatosensation-related cortical areas, ACC and S1, processing distinct
components of the pain perception.
Presenter – Severino, Amie
Abstract ID – M18
Kappa opioid receptors on dopaminergic projections from the ventral tegmental area mediate
distinct aversion behaviors in chronic pain.
Amie L Severino1,3, Shiwei Liu1,2,3, Isabel Bishop1,3, Sarah Pickens1,2,3, Ivy Carroll4, Caroline Bass5,
Frances Leslie2, Chris Evans1,3, Cathy Cahill1,2,3
1
University of California Los Angeles, Los Angeles, USA, 2University of California Irvine, Irvine, USA,
3
Shirley and Stefan Hatos Foundation Center for Neuropharmacology, Los Angeles, USA, 4Research
Triangle Institute, Research Triangle Park, USA, 5University at Buffalo, Buffalo, USA
Patients with chronic pain often present with comorbid mood disorders such as anxiety or depression.
Kappa opioid receptors (KOR) mediate dysphoric states and contribute to affective-like anxiogenic anddepressive behaviors. We hypothesized that KORs contribute to an aversive state associated with chronic
pain. We induced chronic neuropathic (NP) pain in adult male and female C57BL6/J mice using the sciatic
cuff model, which has established affective dysregulation. We observed that negative affective behaviors
in pain depend on KOR-signaling and were attenuated with pretreatment of selective KOR antagonist
JDTic. NP pain mice exhibited enhanced conditioned place aversion to U50,488. To investigate thecircuitry involved in KOR-mediated aversion in chronic pain, we used a viral approach in transgenic floxed
KOR mice. We bilaterally injected AAV2/10-TH-Cre virus or control virus into the ventral tegmental area
(VTA) to eliminate KOR from dopamine neurons. After 3 weeks, we assessed separate cohorts of animals
for U50,488-induced place aversion. Interestingly, elimination of KORs from VTA TH neurons enhanced
U50,488 place aversion in pain-naïve animals. To investigate the involvement of KOR in on-going pain,
we conducted single chamber conditioning, a model of assessing the ongoing pain aversion. Eliminationof KORs from VTA TH neurons prevented on-going pain aversion in NP pain mice. These data suggestthat KORs on VTA dopamine neurons mediate a tonic aversive state in chronic pain and blunt agonistinduced aversion in naïve animals.

114

Presenter – Sharma, Indrajeet
Abstract ID – T38
Structure–activity relationships on collybolide: Discovery of a potent kappa-opioid agonist
with enhanced metabolic stability
Nicholas Massaro2, Adrien Cavé3, Heidi E. Hamm3, Joseph Parello3, Vsevolod Katritch4, Lakshmi
Devi1, Indrajeet Sharma2
1
Icahn School of Medicine at Mount Sinai, USA, 2University of Oklahoma, Norman, USA,3
Department of Chemistry, Vanderbilt University, Nashville, USA, 4Department of Pharmacology,
University of Southern California, USA
Collybolide, a non-nitrogenous sesquiterpene natural product extracted from the mushroom Collybia
maculata was found to be a highly selective and potent KOR agonist (EC50 ~ 1 nM) having in vivo
analgesic and antipruritic activities. Collybolide has a furyl-δ-lactone core similar to that of salvinorinA, however, differs from the latter in exhibiting biased agonistic activity. Collybolide shows noconditioned place aversion unlike classical kappa agonists at doses at which it blocks non-histaminemediated itch, suggesting the possibility of separating kappa mediated liabilities from its ability toblock itch with this novel natural product based template. However, there are liabilities associatedwith use of collybolide as a drug candidate such as aqueous solubility, efficacy, and metabolicstability. These liabilities can be overcome by designing new collybolide analogues guided by thecomputational docking studies. We performed semi-synthesis of collybolides and discovered ananalogue MN-074, which was found to have similar potency and selectivity to collybolide. Microsomal
stability studies demonstrated that MN-074 was more metabolically resistant than collybolide. In
addition, MN-074 had better drug-like properties than collybolide. The goal is to comprehensively
examine the SAR of collybolide, which will yield a set of selective potent KOR ligands with thepotential of becoming lead drug candidates for itch and pain with reduced side effects. Further, highthroughput screening at the NIMH-PDSP against 50 CNS receptors could find new hits, which could
be optimized further to explore new therapeutic opportunities.
Presenter – Shibasaki, Masahiro
Abstract ID – T44
Identification of cells in the ventral tegmental area activated by systemic treatment with
opioids and methamphetamine
Masahiro Shibasaki1, Daisuke Sato1, Ryousuke Wakai1, Moe Watanabe3, Michiko Narita1, Hiroyuki
Tezuka4, Yusuke Hamada1, Naoko Kuzumaki1,2, Minoru Narita1,2
1Department of Pharmacology, Hoshi University School of Pharmacy and Pharmaceutical Sciences,
Japan, 2Life Science Tokyo Advanced Research Center (L-StaR), Hoshi University School of
Pharmacy and Pharmaceutical Sciences, Japan, 3Department of Pharmacology, University of
Arizona, USA, 4Department of Cellular Function Analysis, Fujita Health University, Japan
Opioids and methamphetamine cause psychological dependence via activation of the dopaminergic
system from the ventral tegmental area (VTA). In the present study, we characterized the cells thatare activated by opioids or methamphetamine by analyzing Thy1+/GFP+ (positive) and Thy1+/GFP(negative) cells in the ventral tegmental area using cFos-GFP mice. The mRNA levels of tyrosinehydroxylase, dopamine transporter and mammalian target of rapamycin (mTOR) in cFos-GFPpositive cells were much higher than those in cFos-GFP-negative cells. Generally, mTOR signaling
has emerged as a critical integrator of neuronal activity and synaptic inputs that in turn can affectmany cell biological processes. A growing body of evidence suggests that mTORC1 and mTORC2
play crucial roles in drug addiction. Taken together, these findings suggest that addictive drugs,
such as opioids and methamphetamine, activate the dopaminergic neurons containing enriched
mTOR in the VTA and may modulate the function of dopamine neurons. We propose here thatmTOR could be the biomarker that predict the liability of drug dependence and may be a therapeutictarget for drug of abuse.

115

Presenter – Simmons, Sarah
Abstract ID – W5
Endogenous kappa opioid system regulation of discrete populations of lateral habenula
neurons.
Sarah C. Simmons1, Ryan D. Shepherd1, Shawn Gouty1, Robert Hammack1, Ludovic Langlo1, Brian
M. Cox1, Fereshteh S. Nugent1
1
Uniformed Services University of the Health Sciences, Department of Pharmacology, Bethesda,
USA
The lateral habenula (LHb) is an epithalamic brain region associated with value-based decision
making and stress evasion. Increased activity of the LHb is associated with drug addiction andstress-related mood disorders. Previously, we have shown that maternal deprivation (MD), a severe
early life stress, increases LHb intrinsic excitability while blunting its response to central corticotropin
releasing factor (CRF). While kappa opioid receptor (KOR) signaling may mediate part of central
CRF response in reward circuitry, little is known on KOR regulation of the LHb despite its distinctrole in stress, reward, and aversion processing. Here, we show that maternal deprivation significantly
increases immunolabeled dynorphin (DYN) fibers (endogenous KOR agonist) and significantly
decreases mRNA levels of KORs in LHb tissue compared to non-stress controls. To elucidate thefunctional role of a DYN-KOR signaling in the LHb, we utilized ex-vivo electrophysiology combinedwith pharmacological tools in rat LHb slices. We found that under non-stress control conditions, tonic
KOR stimulation may contribute to basal LHb neuronal excitability which can be uncovered using
KOR antagonist, norBNI. Additionally, we show that KOR agonists (U50,488, and dynorphin) havedistinct effects on LHb neuronal excitability through changes in passive and active membrane
properties in distinct neuronal populations based on the presence of Ih current. Additionally, KOR
activation alters both glutamatergic mEPSC and GABAergic mIPSC synaptic transmission onto LHb
neurons. To our knowledge this is the first demonstration of the existence of DYN-KOR signaling in
the LHb which can be modulated by severe early life stressors such as MD.
Presenter – Stam, Frida
Abstract ID – W31
Commonly used prescription opioids induce membrane damage and mitochondrial
dysfunction in the neuronal-like NG108-15 cells
Frida Stam1, Jennifer Birgersson1, Erik Nylander1, Sofia Zelleroth1, Fred Nyberg1, Alfhild Grönbladh1,
Mathias Hallberg1
1
Uppsala University, Uppsala, Sweden
Non-medical opioid use is increasing world-wide, along with an increased number of opioid-related
overdose deaths. The consequence of this over consumption is devastating, affecting both health,social, and economic welfare. Chronic use of opioids is reported to induce several dysfunctions, suchas cognitive impairments. These effects are suggested to partly be caused by induced neuronal cell
death and inhibited neurogenesis. However, further evaluation of the neurotoxic effects ofprescription opioids is needed in order to gain a deeper understanding of both long- and short- term
effects of opioid use and how to counteract these problems. In the present study, the toxic effects ofthe commonly used opioids methadone, morphine, oxycodone, hydromorphone, ketobemidone, andfentanyl were studied in the neuronal-like NG108-15 cells. The toxic effects were assessed bymeasuring mitochondrial activity and membrane integrity after an acute 24h treatment inconcentrations ranging between 1 and 1000 µM. The results demonstrate that commonly used
opioids induce toxic effects in neuronal-like cell cultures to different extent, and that the toxicity
seems to be mainly, but not solely, dependent on treatment dose. These results support previous
findings and highlights important differences between opioid analgesics.

116

Presenter – Standifer, Kelly
Abstract ID – W51
Sex differences in nociceptin/orphanin FQ peptide (NOP) receptor knockout rat response totraumatic stress: Tumor necrosis factorα (TNFα) and prepronociceptin (PNOC) mRNA
Kelly M Standifer1,2,3,4, Ian Schalo1,3, Yong Zhang1,3
1
University of Oklahoma Health Sciences, USA, 2Oklahoma Center for Neuroscience, USA, 3Departmentof Pharmaceutical Sciences, University of Oklahoma College of Pharmacy, USA, 4Harold Hamm Diabetes
Center, USA
Nociceptin/Orphanin FQ (N/OFQ) modulates pain transmission, learning/memory, stress and anxiety via
activation of the NOP (OPRL1) receptor. We previously reported that single-prolonged stress (SPS)model of PTSD produces allodynia and anxiety-like behaviors and elevates N/OFQ in male and female
wild type (WT) Wistar and in female NOP receptor knockout (KO; OPRL1-/-) rats. However, male NOP
receptor KO rats never developed allodynia or anxiety-like behaviors. The objective of this study was to
examine acute differences in TNFα response to traumatic stress between male and female NOP receptor
knockout rats. TNFα was found to increase N/OFQ expression (Buzas et al., 2002), and elevated serum
TNFα correlates with allodynia. We examined changes in expression of TNFα and PNOC mRNA 2 hr
post-SPS in circulating peripheral blood mononuclear cells (PBMCs) and several brain regions. Sexdependent differences were noted in TNFα mRNA in PBMCs, hypothalamus (HYP), periaqueductal gray
(PAG), and hippocampus (HC); a sex-dependent effect also was noted for PNOC mRNA in PBMCs. Twoway ANOVA also revealed specific interactions between sex and SPS for PNOC mRNA in PAG andamygdala (AMY). TNFα treatment produced time- and dose-dependent changes in PNOC mRNA in
cultured neuroblastoma cell lines. Studies in WT rats and of differential PNOC peptide expression are
underway as we continue to explore the role that endogenous N/OFQ-NOP receptor signaling plays inmales and females following traumatic stress, and its regulation.
Support provided by the Oklahoma Center for the Advancement of Science and Technology (OCAST
HR17-041) and the Richard T. Anderson Neuroscience Chair Endowment (KMS).
Presenter – Streicher, John
Abstract ID – M38
The mu-delta opioid receptor heterodimer acts as a negative feedback brake to reduce opioid antinociception by repression of CaMKII and Src signaling
Attila Keresztes1, Keith Olson1,2, Paul Nguyen1, Natalie Barker3, Zekun Liu2, Paul Langlais3, Victor
Hruby2, John Streicher1
1
Department of Pharmacology, College of Medicine, University of Arizona, USA, 2Department ofChemistry and Biochemistry, College of Science, University of Arizona, USA, 3Department of Medicine,College of Medicine, University of Arizona, USA,The mu-delta opioid receptor heterodimer (MDOR) has been hypothesized to act as a negative feedback
brake on the opioid system to repress analgesia while enhancing side effects. However, evidence for the
MDOR in vivo is limited and controversial, due to the lack of selective tools, with no MDOR-selective
antagonists. We thus designed and synthesized a series of heterobivalent linked compounds with mu
and delta opioid antagonist pharmacophores. We screened these compounds in vitro and in vivo (CD-1
mice) and identified a lead compound (D24M) with high potency (<1 nM) and selectivity (~90 fold) andselective for the MDOR in vivo for doses up to 10 nmol. We next found that D24M strongly enhanced
oxymorphone anti-nociception by ~50-600% in models of acute and chronic pain (tail flick, post-surgicalpaw incision, chemotherapy-induced peripheral neuropathy). D24M also strongly reduced naloxoneprecipitated withdrawal in acute and chronic morphine dependence. Seeking a molecular mechanism, we
performed phosphoproteomic analysis on brain stems of treated mice, finding that CaMKII and Src
kinases were repressed by oxymorphone and restored by D24M treatment. Experiments with the CaMKII
inhibitor KN-93 and the Src inhibitor SrcI-1 in mice suggested that these kinases are repressed by the
MDOR and restored by D24M treatment, leading to enhanced anti-nociception. Our findings establish a
novel selective antagonist for the MDOR with therapeutic potential to improve opioid therapy; our findings
further support the MDOR as a negative feedback brake on the opioid system acting in part via repressionof CaMKII and Src.

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Presenter – Targowska-Duda, Katarzyna
Abstract ID – W36
NOP receptors system in migraine
Katarzyna M Targowska-Duda1, Zachariah Bertels2, Akihiko Ozawa1, Hanna Mielke-Maday1, Gilles
Zribi1, Amynah A Pradhan2, Lawrence Toll1
1
Department of Biomedical Science, Florida Atlantic University, Boca Raton, USA, 2Department of
Psychiatry, University of Illinois at Chicago, Chicago, USA
Migraine is an extraordinarily prevalent and disabling headache disorder that affects one billion
people worldwide. Throbbing pain is one of several migraine symptoms including nausea, andvomiting as well as sensitivity to light (photophobia), sometimes to sounds, smell and touch. The
basic mechanisms underlying migraine remain inadequately understood, and current treatments(with triptans being the primary standard of care) are not well tolerated by some patients. NOP(Nociceptin Opioid) receptors (originally called ORL1) are the fourth member of the opioid receptor
family that are highly expressed in dorsal root ganglia (DRG), spinal cord, and in brain regions
involved in pain and drug reward. In addition, some of the highest expression levels of NOP receptors
were found in the trigeminal ganglia and trigeminal nucleus caudalis (TNC). The aim of our study is
to assess NOP receptor agonists influence on parameters of sensory (sensitivity of paw and head)
and affective (conditioned place avoidance) migraine pain as well as photophobia in mice. In
addition, we also examine NOP receptors in trigeminal ganglia and TNC after an acute migraine andchronic migraine models. Immunohistochemical studies with NOP-eGFP knock-in mice demonstratethat NOP receptors are on the vast majority of neurons in the trigeminal ganglia and very highly
expressed in the TNC. We observed very limited co-localization of NOP receptors and calcitonin
gene-related peptide (CGRP). Our findings, using a nitroglycerin-induced migraine model, haveclearly demonstrated that the selective and potent NOP receptor agonist, Ro 64-6198, cancompletely eliminate nitroglycerin-induced paw and head allodynia.
Presenter – Tejada, Frederick
Opioid awareness among UMES pharmacy students
Frederick Tejada1, Khaled Hasan1, Nkem Nonyel1
1
University of Maryland Eastern Shore School Pharmacy, USA

Abstract ID – T40

The aim of this study was to assess the knowledge and opinion of University of Maryland Eastern
Shore School Pharmacy (UMES-SOP) students towards opioid use and misuse. This cross-sectional
study was approved by the UMES Institutional Review Board. The 1st, 2nd, and 3rd year pharmacy
students were surveyed through Google form, and the responses were analyzed using Excel 2016
and StatView 5.0. Ninety-seven students completed the survey. Ninety-seven percent of students
reported their knowledge and awareness about opioids use, misuse and risk of addiction. The
students’ response to the time needed to be addicted was not statistically significant (p-value 0.93)
across three periods [3 days (n=24); two weeks (n=28); one month (n=26)]. Pain management was
identified as the main reason for addiction (50%), followed by escape from stress and depression(29%). Reasons for addiction includes lack of knowledge about the complications of drugs (34%),
easy access to opioids (34%), and teenage curiosity (29.9%). Students suggested that the most
likely cause of addiction include lack of knowledge about complications of drugs (34.0%), easily
access to the drugs (34.0%), and teenagers’ curiosity (29.9%). Students believed that physicians
are mainly responsible for opioid addiction. Students agreed that education on safe opioid use is thebest way to fight opioid addiction. To conclude, pharmacists can play important roles in combating
the opioid crisis, and students require adequate opioid education on opioid use and misuse.

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Presenter – Thorsell, Annika
Abstract ID – T51
The OPRM1-A118G modulates reward sensitivity to opiates as well as the biased MOR agonist
TRV130Annika Thorsell1, Lovisa Holm1, Markus Heilig1
1
CSAN, IKE, Linkoping University, Sweden
Both the therapeutic response to mu-opioid receptor (MOR) antagonists, as well as sensitivity to therewarding effects of MOR-agonists are moderated by a variation at the mu-opioid receptor gene
locus (OPRM1: Oprm-A118G). We have previously generated a “humanized” mouse carrying therespective human OPRM1 A118G alleles (i.e.118AA and 118GG). Here, we examine the role of theOPRM1 A118G variation in opiate reward and intake. Reward-related behavior for opiates(morphine, oxycodone, as well as the biased MOR agonist, TRV130) was measured using theConditioned Place Preference (CPP) model. Drug intake was measured using an oral limited accessmodel as well as during continuous access. Pain sensitivity was evaluated using tail-flick and hot
plate, and anxiety-related behavior was measured on the elevated plus-maze (EPM). The OPRM1A118G’s as well as C57Bl6/J(Rj) were used in the experiments. The OPRM1 118AA carriers
displayed a blunted anxiogenic behavioral response following acute withdrawal from opiatescompared to the 118GG’s. Additionally, the 118GG’s overall displayed more pronounced CPP, andat doses not inducing CPP in the C57Bl6/J for morphine and TRV130. Oxycodone conditioning
generated strong CPP in all animals used. A higher consumption of oral morphine was seen in the
118GG’s compared to the 118AAs. Pain responses were also differentially modulated by morphine,
but not by oxycodone. In conclusion, for morphine and oxycodone, the OPRM1 A118G modulates
reward sensitivity in a manner not easily related to the antinociceptive effects. Additionally, theA118G SNP seemingly contributes to the reward-value of the MOR biased agonist TRV130.
Presenter – Toddes, Carlee
Abstract ID – W49
The social brain and mu: Modulation of social behavior by mu opioid receptors in the brain
reward network
Carlee Toddes1, Patrick E. Rothwell1
1
Department of Neuroscience, University of Minnesota, USA
The rewarding aspects of social interaction require activation of mu opioid receptors within the brain
reward network. However, it is unclear where in this network mu opioid receptor expression isessential for the regulation same-sex affiliative social interaction. The goal of this study was todetermine how global and regional genetic knockout of the mu opioid receptor alters social behavior
in male and female mice, using a variety of behavioral assays. Our behavioral battery includes thesocial conditioned place preference test, the dyadic social interaction test, and the reverse threechamber social test. The reverse three-chamber test is a novel social assay that examines the social
preference of a wildtype “judge” between a “typical” wildtype mouse and an “atypical” mu opioid
receptor knockout mouse. These assays focus on different aspects of social behavior including social
reward, motivation, and preference. Our data demonstrates that constitutive homozygous andheterozygous deletion of the mu opioid receptor impairs social behavior across tests. Additionally,
preliminary data has shown a social impairment in mice following conditional knockout of the mu
opioid receptor from cortico-limbic regions that converge upon the nucleus accumbens. These
results highlight critical locations within the brain reward network where mu opioid receptor
expression is necessary for normal social behavior

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Presenter – Trieu, Brian
Abstract ID – T50
Modulation of accumbal synaptic transmission by the enkephalin heptapeptide MERF (TyrGly-Gly-Phe-Met-Arg-Phe)
Brian H Trieu1,2, Wei Xie3, Swati S More3, Patrick E Rothwell4
1
Graduate Program in Neuroscience, University of Minnesota, USA, 2Medical Scientist Training
Program, University of Minnesota, USA, 3Center for Drug Design, University of Minnesota, USA,
4
Department of Neuroscience, University of Minnesota, USA,
Enkephalins belong to a family of endogenous opioid peptides that are highly concentrated within
the striatum. In this region, they are exclusively released by a subtype of medium spiny neurons(MSNs) expressing the dopamine type-2 receptor (D2-MSNs). Little is known about the regulation or
physiological properties of Tyr-Gly-Gly-Phe-Met-Arg-Phe (MERF), a heptapeptide enkephalin, which
has the highest affinity toward all three opioid receptor subtypes. Its relatively low concentration andrapid degradation have made it an enigma since its discovery in the 1970’s. Through whole-cell patch
clamp recordings we show that exogenous MERF causes a large but transient presynaptic
depression of excitatory synaptic inputs into the nucleus accumbens core. Furthermore, blockade of
MERF degradation using an angiotensin-converting enzyme inhibitor potentiates the effect of this
peptide on excitatory synaptic transmission. We are currently adapting HPLC-MS/MS techniques tomeasure endogenous opioid peptides after in vitro optogenetic stimulation of D2-MSNs. Moreover,with this approach, we are screening an array of peptidase inhibitors that may elevate concentrationsof endogenous opioid peptides following evoked release. Insight into the physiological properties of
MERF and its regulation will aid in our understanding of how endogenous enkephalins modulate
local synaptic plasticity within the nucleus accumbens.
Presenter – Ueda, Hiroshi
Abstract ID – T42* Hot Topics Selection, Symposium 6, Thursday, July 11 AM
Chronic pain status and loss of morphine analgesia have independent brain mechanisms inmice with fibromyalgia-like generalized pain
Hiroshi Ueda1,2, Hiroyuki Neyama2,3
1
Kyoto University, Japan, 2Nagasaki University, Japan, 3Riken Kobe, Japan
Fibromyalgia (FM) is an unmet medical need in terms of poor understanding of pathogenesis. We
attempted to study the molecular mechanisms underlying the lack of morphine analgesia, which isone of FM-specific pharmacotherapeutic features, by using the intermittent cold stress (ICS)-induced
generalized pain disease model in mice, which show fibromyalgia-like pathophysiological andpharmacotherapeutic features. First we confirmed the lack of morphine analgesia in ICS-treated miceby measuring mechanical somatic and muscular hyperalgesia and electrical stimulation-induced
nociceptive responses. Next, we obtained the recovery of morphine analgesia in NMDA receptor
subunit NR2A-deficient (KO) mice, which blocks the morphine analgesic tolerance, possibly througha blockade of anti-opioid counterbalance mechanisms. These results suggest that the lack ofmorphine analgesia in ICS mice is presumably mediated by μ-opioid receptor (MOPr)
desensitization/tolerance through a repeated stimulation by stress-induced endogenous opioid
release. The involvement of NR2A was supported by the successful recovery of morphine analgesia
by microinjection of NR2A siRNA into the PAG, but not VTA of ICS mice. In accordance, thepretreatment with dextromethorphan with NMDAr antagonist activity recovered morphine analgesia,
and single administration of methadone with NMDAr antagonist and MOPr agonist activities showed
potent analgesia. The ICS-induced hyperalgesia and lack of morphine analgesia remain unchangedin MOPr- and LPA1-KO mice, respectively. These results suggest that both mechanisms underlying
chronic pain and lack of morphine analgesia are independent to each other.
Supported by JSPS KAKENHI Grants JP17H01586 (H.U.) and JP26253077 (H.U.).

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Presenter – van Rijn, Richard
Abstract ID – W29
Structural analysis of the G protein-biased delta opioid peptide rubiscolin to probe the receptor’s‘biased’ binding pocket
Robert J. Cassell1, Breanna L. Zerfas1, Benjamin R. Cummins1, Haoyue Cui1, Kendall L. Mores1, Darci
J. Trader1, Richard M. van Rijn1
1
Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, West Lafayette,
USA
A major question pursued in the field of chemical and molecular pharmacology of G protein-coupled
receptors pertains to what conformation allows receptors to selectively activate G-proteins but not recruit
beta-arrestins. We recently identified rubiscolin-6 (YPLDLF) as a delta opioid receptor (DOR) selective
peptide with very low β-arrestin recruitment efficacy. Here we utilized rubiscolin-6 as a starting point toaddress two questions. 1) What aspect of rubiscolin-6 provides the G-protein bias? 2) Can rubiscolin-6
be modified to increase its potency without increasing beta-arrestin 2 recruitment efficacy? To interrogate
our questions, we primarily exchanged the proline with residues of varying flexibility and lipophilicity.
Peptides were synthesized using standard Fmoc-based solid-phase synthesis with HBTU. Sequenceswere derived from four different series: YPLDLF, YPLDLV, YPMDLV as well as YGGFL (leu-enkephalin)
representing. In vitro characterization of G-protein (cAMP Glosensor), beta-arrestin 2 recruitment(PathHunter) and radioligand binding (3H-DPDPE competition) was performed for each peptide at the
DOR. Our results reveal that the proline residue plays an important role in the apparent bias of therubiscolin peptides. Specifically, at this second position it appears that increasing residue flexibility
correlates with increased beta-arrestin 2 recruitment, but that lipophilicity is not. Increased potency wasoften accompanied by increased beta-arrestin 2 efficacy but not always. These findings should provide
opportunities to use computational analysis to model the DOR in conformations that are either G-proteinbiased or that promote beta-arrestin recruitment.
Supported by a grant from the National Institutes on Drug Abuse (R03DA045897 to RMvR).
Presenter – Varshneya, Neil
Abstract ID – W44
The effects of N-phenyl-fluorinated butyrylfentanyl regioisomers in mice
Neil B. Varshneya1, Sherif H. Hassanien2, Donna M. Iula2, D. Matthew Walentiny1, Lea T. Moisa1, Teneille
D. Walker3, Luli R. Akinfiresoye3, Patrick M. Beardsley1
1
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, USA,
2
Division of Forensic Chemistry, Cayman Chemical Company, Inc., Ann Arbor, USA, 3Diversion Control
Division, DEA, United States Department of Justice, Springfield, USA
In 2017, drug overdose resulted in 70,237 fatalities in the United States, 47,600 of which were opioid-related.
Contributing to these opioid-related fatalities was an 816% increase in deaths between 2013 and 2017
involving synthetic narcotics, principally fentanyl and its analogs, which prominently include butyrylfentanyl
isomers. In the present study, regioisomers of N-phenyl-fluorinated butyrylfentanyl were evaluated in adultmale Swiss Webster mice for classical opioid effects including antinociception, hyperlocomotion, andrespiratory depression and compared to those of the archetypical opioids, fentanyl and morphine. In warmwater tail-withdrawal tests, fentanyl (0.1-1 mg/kg), morphine (10-32 mg/kg), ortho-fluorobutyrylfentanyl (0.0321 mg/kg), meta-fluorobutyrylfentanyl (0.1-1 mg/kg), and para-fluorobutyrylfentanyl (1-10 mg/kg) produced
significant (p<0.05) dose-dependent increases in antinociception with increasing ED50s (CI) for doing so of
fentanyl [0.08 (0.04-0.16)] ≈ ortho-fluorobutyrylfentanyl [0.09 mg/kg (0.06-0.14)] > meta-fluorobutyrylfentanyl[0.25 mg/kg (0.18-0.33)] > para-fluorobutyrylfentanyl [0.91 mg/kg (0.46-1.58)] > morphine [7.82 mg/kg (5.4211.02)]. Naltrexone pretreatment (1 mg/kg) increased antinociceptive ED50s with decreasing potency ratios ofmeta-fluorobutyrylfentanyl (65.77x) > para-fluorobutyrylfentanyl (10.87x) > ortho-fluorobutyrylfentanyl (9.55x)> fentanyl (3.95x) > morphine (1.48x). In locomotor activity tests, fentanyl (1-10 mg/kg), morphine (100 mg/kg),
ortho-fluorobutyrylfentanyl (1-10 mg/kg), meta-fluorobutyrylfentanyl (10 mg/kg), and para-fluorobutyrylfentanyl(10-100 mg/kg) produced significant (p<0.05) dose-dependent increases in locomotion. In respiration tests,
fentanyl (0.1-3.2 mg/kg), morphine (1-32 mg/kg), ortho-fluorobutyrylfentanyl (0.01-0.32 mg/kg), metafluorobutyrylfentanyl (0.32-1 mg/kg), and para-fluorobutyrylfentanyl (0.32-10 mg/kg) produced significant
decreases in respiration. The results indicate that the potencies of N-phenyl-fluorinated butyrylfentanyl

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regioisomers to elicit antinociception, hyperlocomotion, and respiratory depression are separable, not only as
expected between drugs, but also within individual drugs.

Presenter – Wang, Yun-Jun
Abstract ID – T7
Discovery of a highly selective and potent kappa opioid receptor agonist with reduced CNS
side effects
Yu-Jun Wang1, Yan Ma1, Xue-Jun Xu1, Wei Li2, Jing-Gen Liu1
1
Key Laboratory of Receptor Research, Shanghai Institute of Materia Medical, Chinese Academy of
Sciences, Shanghai, China, 2Department of Medicinal Chemistry, School of Pharmacy, Fudan
University, Shanghai, China
Opioid analgesics such as morphine and fentanyl cause severe side effects including respiratory
depression and dependence liability, which limits the clinical use. k Opioid receptor agonists havelong been recognized as potent analgesic with low abuse potential, but their use can be associatedwith motor incoordination, sedation and dysphoria-like states. We recently discovered a new highly
selective k agonist SLL-039. SLL-039 showed a high affinity to k receptor with Ki value of 0.47nM
and low affinity to m and d receptor (κ/μ=682, κ/δ=284). It stimulated [35S]GTPgS binding to G proteinwith an EC50 value of 2.0nM. In vivo, SLL-039 was more potent than U50,488H and morphine with
the ED50 values of 0.4 mg/kg and 0.1 mg/kg in the hot plate and the abdominal constriction tests,
respectively. SLL-039 produced a relatively long-term action with peak effect at 2h after the injection.
SLL-039-induced antinociception was significantly inhibited by k antagonist nor-BNI, but not m
antagonist b-FNA pretreatment. SLL-039 also produced significant antipruritic effects with ED50 valueof 0.2 mg/kg. In addition, at the doses of antinociceptive ED50 values, SLL-039 (0.4mg/kg) did not
induce sedation, motor incoordination and dysphoria (conditioned place aversion behavior, CPA). At
the dose up to 1.2 mg/kg (3×ED50 values in the antinociceptive test), SLL-039 still did not. SLL-039
at 2.7 mg/kg (6.75×ED50 values) caused significant CPA. Taken together, our results show that SLL039 is a novel selective k-opioid receptor agonist with potent antinociception effects and low CNS
side effects.
Presenter – Webb, Chase
Abstract ID – T17
Computational discovery of novel µ receptor agonists
Chase Webb1,2, Tao Che3, Brian Roth3, Aashish Manglik1,4, Brian Shoichet1
1
Department of Pharmaceutical Chemistry, UCSF, USA, 2Pharmaceutical Sciences andPharmacogenomics Graduate Program, UCSF, USA, 3Department of Pharmacology, University ofNorth Carolina, USA, 4Department of Anesthesiology, UCSF, USA
Opioids confer pain relief by stimulating neuronal µ opioid receptors (µOR). The µOR, part of the
GPCR family of proteins, are allosteric transmembrane machines that transduce extracellular stimuli
into intracellular signals by coupling to both ligands and myriad downstream effectors, including the
eponymous G-proteins and β-arrestins. Despite an abundance of µOR modulators, the chemical
space explored by these ligands is severely limited. Indeed, most µOR agonists are morphinans,
diphenylheptanes, benzomorphans, or phenylpiperidine derivatives. Accordingly, knowledge of thepharmacological properties of the receptor and their relation to analgesia and side effects is lacking.
In this project, DOCK3.7 was employed to prosecute a structure-based virtual screen against theµOR to discover novel chemotypes that can activate the receptor. To this end, 282 million
compounds were docked against the orthosteric site of the agonist bound crystal structure of theµOR (PDB:5C1M). Through retrospective screening the receptor was prepared to optimize the recallof known opioid agonists over inactive ‘decoy’ molecules. In a prospective screen, the top 5,000
chemically novel scaffolds were manually examined for their interactions with key polar residues andconformational strain. Ultimately, 75 high-scoring molecules were selected for biochemical testing.
This testing yielded 26 binders, all of which are chemically novel, and 15 of which strongly activate
G proteins with minimal recruitment of β-arrestin.

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Presenter – Whistler, Jennifer
Abstract ID – W50
Balanced opioids provide excellent analgesia and reduced side effects
Li He1,2, Sarah Gooding1, Anirudh Gaur1, Lindsey Felth1, Jennifer Whistler1,2
1Center for Neuroscience, University of California, Davis, USA, 2Department of Physiology and
Membrane Biology, University of California, Davis, USA
There remains an ongoing debate as to whether “biased” or “balanced” agonists at the mu opioid
receptor (MOR) will produce analgesia with reduced side effects. Endorphins and enkephalins are“balanced” agonists at the MOR, signaling to G-protein while effectively engaging beta-arrestins andpromoting substantial MOR endocytosis and recycling/resensitization. Conversely, morphine and allits derivatives are significantly biased for G protein, because they promote incomplete MOR
phosphorylation, and only low efficacy arrestin recruitment unless GRKs are overexpressed. Recent
efforts to identify new opioid ligands have focused on MOR agonists even more G-biased than our
existing drugs (e.g. TRV130, PZM21). We have previously described the creation of knock-in miceexpressing a modified MOR (RMOR), which signals in a “balanced” way when activated by morphine,
due to enhanced phosphorylation and fully efficacious arrestin recruitment. These mice show
excellent analgesia with reduced tolerance, reduced dependence and reduced compulsive drug
seeking behavior. Here we report that RMOR mice also display fewer deficits in cognitive flexibility
after morphine use and long-term abstinence. We further demonstrate using WT, RMOR mice,Barr2-KO and RMOR/Barr2-KO mice that MOR engagement with Barr2 does not increased opioid
induced respiratory depression. Rather, we show that the best therapeutic window for analgesia isachieved with balanced opioid receptor signaling, and that opioid dependence and drug seeking are
likewise reduced with balanced signaling. Taken together, these data suggest that “balanced” opioid
agonists, that engage arrestins similarly to the endogenous ligands could provide excellent
analgesia, no increase in respiratory effects and reduced abuse liability.
Presenter – Wiese, Beth M.
Abstract ID – M2
The role of cannabinoid receptor mediation of opioid induced respiratory depression in the
Prebötzinger complex
Beth M. Wiese2, Erika Liktor-Busa2, Alexandros Makriyannis1, Todd W. Vanderah2, Tally M. LargentMilnes2
1
Northeastern University, USA, 2University of Arizona, USA
The impact of the opioid epidemic remains an escalating problem in the United States and North
America with fatalities from accidental overdoses at an all-time high despite investigation on
alternative means to address opioid use disorder (OUD). A fatal overdose occurs following activationof mu opioid receptors (MOR) within the preBötzinger complex of the medulla causing respiratory
depression and resulting in death. Considering that opioids are prescribed at alarmingly high rates
for pain, and 80% of global opioids are consumed by Americans, it is essential that research is
conducted on new medications that do not induce respiratory depression for pain relief. Recent
evidence has shown that cannabis and cannabinoids may substitute as a viable analgesic, on theirown or in combination with opioids. As more states liberalize cannabis laws, it is imperative weunderstand the mechanisms of action on respiratory depression of cannabinoids. In this study wesought to investigate the role of cannabinoid 1 receptor (CB1R) and cannabinoid 2 receptor (CB2R)
activation on respiratory depression from both peripherally restricted or brain-penetrant CB1/CB2
agonists and in coadministration with morphine. Results suggest CB1 and CB2 activation does not
induce respiratory depression. In addition, CB2 activation can weaken opioid induced respiratory
depression. Lastly, these findings suggest that alleviation of morphine induced respiratory
depression with a cannabinoid requires a brain penetrant compound. These data support additional
research of cannabinoids as potential adjuncts and therapeutic analgesics in the wake of the opioid
epidemic.
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Presenter – Wilson, Lisa L.
Abstract ID – T34
Kratom tea as a therapeutic agent for opioid dependence
Lisa L. Wilson1, Hannah M. Harris3, Ariana C. Brice-Tutt1, Thomas J. Cirino1, Shainnel O. Eans1,
Heather M. Stacy1, Chloe Simons1, Juan F. León2, Christopher R. McCurdy2, Jay P. McLaughlin1
1
Department of Pharmacodynamics, University of Florida, USA, 2Department of Medicinal Chemistry,
University of Florida, USA, 3Department of Psychology, University of Mississippi, USA
Mitragyna speciosa Korth (Rubiaceae) is a Southeast Asian tree whose leaves are the source of theThai traditional drug “Kratom”. Kratom extract reportedly possess pharmacological actions that
include both a coca-like stimulant and opium-like depressant effects. The extract of this plant has
been used as a substitute for opioids in dependent subjects, and recent studies have reported that
constituents in Kratom possesses affinity for the mu opioid receptor (MOR). Its primary
pharmacological activity is thought to be attributed to its main alkaloids, Mitragynine and 7-OHMitragynine. The objective of this study was to evaluate the antinociceptive and physical dependence
effects of Kratom tea using neuropharmacology assays of antinociception and dependence. We
hypothesize that Kratom Tea will significantly decrease opioid withdrawal and exhibit antinociceptive
properties with fewer liabilities. Oral administration of Kratom extract to C57BL/6J mice resulted indose-dependent antinociception in the 55 °C warm-water tail-withdrawal assay at higher doses (≥1
g/kg, p.o.). However, these doses of Kratom extract did not alter coordinated locomotion on a rotorod
test, suppress respiration, or induce conditioned place preference. Chronic orally administered
Kratom Tea did not produce signs of physical dependence, but significantly decreased naloxoneprecipitated withdrawal in subjects chronically treated with morphine. Taken together the results
produce a better understanding of the mechanisms of action of Kratom tea and may lead to a newclass of therapeutics; therefore, suggest a new treatment for opioid dependence.
Presenter – Windisch, Kyle
Abstract ID – M29
Investigating the effect of Nalmefene on extended access cocaine self-administration in malemice
Kyle Windisch1, Michelle Morochnik1, Brian Reed1, Mary Jeanne Kreek1
1
Rockefeller University, New York, USA
The mu opioid receptor antagonist (MOR)/kappa opioid receptor (KOR) partial agonist naltrexone(NTX) has been shown preclinically and clinically to reduce seeking and intake of cocaine. No studies
have examined the effects of nalmefene (NMF), a close structural analog of NTX, on cocaine
reinforcement. Given its greater KOR potency, improved bioavailability, and lack of dose-dependent
liver toxicity, NMF may be a promising pharmacotherapeutic for cocaine use disorder (CUD). Here
we examine the effects of NMF pretreatment on chronic daily extended access (4-hr) cocaine
intravenous self-administration (IVSA) in male C57Bl/6J mice. Following recovery from jugularcatheterization, separate groups of mice had daily 4-hr cocaine IVSA sessions (0.25 or 0.5 mg/kg/inf,
FR1) for 14 days. During the second week of IVSA (days 8-14), mice were pretreated with NMF (0,1, or 10 mg/kg) 30 minutes prior to each session. Very nominal reductions in cocaine selfadministration, if any, were observed following pretreatment with NMF for either dose of NMF (1 or
10 mg/kg) with either unit dose of cocaine (0.25 or 0.5 mg/kg/inf). NMF did not significantly attenuatecocaine self-administration in mice with extended access following either acute or repeatedadministration suggesting that NMF may not be suitable for non-abstinent CUD patients. Researchis needed with primarily KOR selective partial or full agonists, with or without MOR antagonism, todetermine their effect on cocaine reinforcement.
Support by the Gary R. Helman Postdoctoral Fellowship (KAW) and Dr. Miriam and Sheldon Adelson
Research Foundation (MJK).

124

Presenter – Yamuda, Daisuke
Abstract ID – W3
KNT-127, a selective agonist for delta-receptors, modulates excitatory synaptic transmission
in the medial prefrontal cortex in mice
Daisuke Yamada1, Jun-Ichiro Oka1, Hiroshi Nagase2, Akiyoshi Saitoh1
1
Laboratory of Pharmacology, Faculty of Pharmaceutical Sciences, Tokyo University of Science,
Chiba, Japan, 2International Institute for Integrative Sleep Medicine, University of Tsukuba, Ibaraki,Japan
It has been reported that delta-opioid receptor (DOP), one of the types of opioid receptors, regulates
emotional behaviors in rodents. We previously reported that local perfusion of KNT-127, a selective
agonist for DOP, in the prelimbic subregion of the medial prefrontal cortex (PL-PFC) decreased
anxiety-like behavior induced by veratrine, a sodium channel activator, in mice, and simultaneously
caused attenuation of veratrine-induced increases in extracellular glutamate levels in PL-PFC using
a in vivo reverse microdialysis study. These results suggested the possibility that KNT-127 exerted
anxiolytic-like effect through modulation of glutamatergic excitatory synaptic transmission via DOP
in the PL-PFC. To clarify these points, we investigated the effect of KNT-127 on excitatory synaptictransmission in the PL-PFC. Brain slices including PL-PFC were prepared from male C57BL/6J miceusing a vibratome. We performed whole-cell patch-clamp recordings from pyramidal shaped neurons
in layer 2/3 of the PL-PFC. Spontaneous excitatory postsynaptic current (sEPSC) and evoked EPSC(eEPSC) induced by electrical stimulation in layer 5 of the PL-PFC were recorded. After baseline
recordings of sEPSC and eEPSC in artificial cerebrospinal fluid, KNT-127 was perfused for at least
15 min, and then sEPSC and eEPSC were recorded again. As a result, the frequency of sEPSC wassignificantly decreased by application of KNT-127. In addition, paired-pulse ratio of eEPSC wassignificantly higher in the recordings after KNT-127 application than that in baseline recordings.
These results indicated that the reduction of glutamate release from presynaptic site by activation of
the DOP in the PL-PFC in mice.
Presenter – Suda, Yukari
Abstract ID – W30
Parkinson’s disease-related overexpression of COMT in nigrostriatal dopaminergic neurons
inhibits μ-opioidergic systems
Yukari Suda2, Naoko Kuzumaki1,2,3, Michiko Narita1, Tomohisa Mori1, Nobutaka Hattori4, Hideyuki
Okano2,3, Minoru Narita1,2
1
Hoshi University, Tokyo, Japan, 2Life Science Tokyo Advanced Research Center, Japan, 3Keio
University School of Medicine, Japan, 4Juntento University, Tokyo, Japan
Parkinson’s disease (PD) is a neurodegenerative disorder associated with the progressive loss of
dopaminergic neurons in substantia nigra area. The mechanism by which dopaminergic neurons are
selectively lost in PD is still unclear. In the present study, to identify susceptibility genes related to
dopaminergic (DAnergic) cell vulnerability underlying PD, we first performed the gene expression
analysis of DAnergic neurons derived from PD-specific induced pluripotent stem cells (iPSCs). We
found a drastic increase in the expression of catechol-O-methyltransferase (COMT) in PD-specific
iPSC-derived DAnergic neurons generated from patients carrying parkin gene (PARK2) mutations
compared to those from healthy controls. Furthermore, overexpression of COMT in the DAnergic
neurons derived from iPSCs leads to cell vulnerability due to a significant increase in ROS
production. Considering these backgrounds, we next performed the in vivo experiments using
Cre/loxP system and found that overexpression of COMT in DAnergic neurons of the substantia
nigra significantly suppressed morphine-induced hyperlocomotion as well as DA elevation in the
striatum. These findings suggest that overexpression of COMT in nigrostriatal DAnergic neurons
may reduce the extracellular release of DA and hyperlocomotion induced by μ-opioids.

125

Presenter – Zaidi, Saheem
Abstract ID – T8
Designing bitopic opioids: Activity modulation in targeting sodium binding pocket of the
opioid receptors
Saheem A. Zaidi1, Tao Che2, Chad Kormos3, Abdelfattah Faouzi4, Susruta Majumdar4, Ivy F. Carroll3,
Bryan Roth2, Vsevolod Katritch1
1
University of Southern California, USA, 2University of North Carolina at Chapel Hill School of
Medicine, USA, 3Research Triangle Institute, USA, 4Washington University at St. Louis, USA
Design and development of functionally biased opioid receptor ligands, able to selectively recruit Gprotein over β-arrestin, offers a very promising approach towards development of limited liability
analgesics. Engagement and stabilization of specific sub-pockets within the receptor by ligands can
induce desirable functional response. The “sodium binding” pocket could be a very effectual receptor
sub-pocket to induce such functional profile. The sodium binding pocket consists of 16 highly
conserved amino acid residues in the center of the transmembrane bundle of class A GPCRs andundergoes major conformational changes upon activation. The coupling of the conformationalchanges between the sodium pocket and the functionally relevant helices 6 and 7 highlights thepocket as a compelling target for modulation of agonist action and functionality. We hypothesizedthat displacement of sodium and stabilization of the pocket in a specific conformational state bybitopic ligands targeting the pocket could lead to distinct functional profiles, including biased Gprotein recruitment profile of the ligands. Two bitopic analogs for nalfurafine scaffold were designed
via ligand docking and ligand-receptor optimization protocols using the conformationally optimized
active/inactive state kappa opioid receptor crystal structures. Pharmacological evaluation of thesynthesized ligands revealed them to be indeed G-protein biased in kappa opioid receptor, thus
validating our hypothesis. Moreover, the highly conserved constitution of the sodium binding pocket
grants a generic receptor sub-pocket to be targeted to control functional profile in other class A
GPCRs.
Supported by P01 DA035764 and R21/R33 DA038858 from NIDA.
Presenter – Zelleroth, Sofia
Abstract ID – T32
Anabolic androgenic steroids affect weight gain, behavior and brain gene expression in malerats
Sofia Zelleroth1, Erik Nylander1, Fred Nyberg1, Alfhild Grönbladh1, Mathias Hallberg1
1
Uppsala University, Sweden
Anabolic androgenic steroids (AAS) are used by certain body builders, elite athletes, andadolescents in order to gain muscle mass and strength. Due to the high doses several AAS-users
experience psychiatric and behavioral changes after long-term abuse, where the severity of theadverse effects seems to depend on the specific AAS used, concentration, and exposure time.
Concomitant use of other drugs is common and several heroin-abusers for example report a historyof previous AAS misuse. In fact, AAS have been suggested as a gateway to abuse of other drugs,
such as opioids. The present study investigated the effects of the two commonly used AAS,
nandrolone decanoate and testosterone undecanoate, on behavioral profile, body weight, and gene
expression in the brain of the male rat. The animals received 15 mg/kg of AAS every third day
repeatedly for 18 days. The effects on behavior were studied using the multivariate concentric square
fieldÔ test (MCSF-test) and the gene expression of relevant neurochemical systems were analyzed
by qPCR. The results demonstrate that nandrolone decanoate cause a reduction in body weight gain
in comparison with testosterone undecanoate and control. In addition, the nandrolone decanoate
treated rats expressed an anxiety-like behavior in the MCSF, and both AAS treatments induced adiverse risk-taking behavior. The qPCR analysis furthermore revealed changes in mRNA expressionof genes that may contribute to the changes in behavior detected in the MCSF.
126

Presenter – Zhang, Li
Abstract ID – T29
Morphine tolerance is attenuated in germ-free mice and reversed by probiotics implicating
the role of gut microbiome
Li Zhang1,2, Jingjing Meng2, Yuguang Ban2, Richa Jalodia2, Irina Chupikova2, Irina Fernandez2, Nivis
Brito2, Umakant Sharma2, Maria T. Abreu2, Sundaram Ramakrishnan2, Sabita Roy2
1
University of Minnesota, USA, 2University of Miami, USA
Prolonged exposure to opioids results in analgesic tolerance, drug overdose, and death. The
mechanism underlying morphine analgesic tolerance still remains unresolved. We show thatmorphine analgesic tolerance was significantly attenuated in germ-free (GF) and in pan-antibiotictreated mice. Reconstitution of GF mice with naïve fecal microbiota reinstated morphine analgesic
tolerance. We further demonstrated that tolerance was associated with microbial dysbiosis withselective depletion in Bifidobacteria and Lactobacillaeae. Probiotics, enriched with these bacterial
communities, attenuated analgesic tolerance in morphine-treated mice. These results suggest thatprobiotic therapy during morphine administration may be a promising, safe, and inexpensive
treatment to prolong morphine’s efficacy and attenuate analgesic tolerance. We hypothesize avicious cycle of chronic morphine tolerance: morphine-induced gut dysbiosis leads to gut barrier
disruption and bacterial translocation, initiating local gut inflammation through TLR2/4 activation,
resulting in the activation of pro-inflammatory cytokines, which drives morphine tolerance.
Presenter – Zhang, Yong
Abstract ID – W33
Effects of nalfurafine on oxycodone-induced conditioned place preference and locomotor
activity in adolescent male and female C57BL/6J mice
Yong Zhang1, Mary Jeanne Kreek2
1
The Rockefeller University, 2Rockefeller University, New York, USA
Prescription opioid oxycodone is abused by millions of adolescents and young adults in the UnitedStates. Kappa opioid agonists can antagonize the euphoria effects–induced by mu opioid agonists,such as oxycodone. Nalfurafine is a clinically used kappa agonist in Japan for the treatment ofpruritus. However, its impact on the rewarding effects of oxycodone in adolescents has not beenstudied. The current study examined the effects of nalfurafine on oxycodone-induced conditioned
placed preference (CPP) in adolescent male and female mice. Male and female adolescent mice (5week old) were trained with saline, oxycodone (3 mg/kg) or with nalfurafine (30 ug/kg) pretreatment
and oxycodone (3 mg/kg) in the CPP apparatus (n=8). There was a total of 4 oxycodone sessions
alternating with 4 saline sessions. Locomotor activities were recorded during each conditioning
session. Time spent on oxycodone-paired compartment was compared between pre- and postconditioning sessions to determine whether a mouse developed CPP to oxycodone. Mice trainedwith oxycodone (3 mg/kg) significantly increased time spent in the oxycodone-paired
compartment. Pretreatment with nalfurafine attenuated oxycodone-induced CPP in both male andfemale mice. Oxycodone also increased locomotor activities; such effect was blocked by
pretreatment of nalfurafine. This is the first study to examine the effects of nalfurafine on oxycodone–
induced CPP in adolescent mice. Nalfurafine was found to decrease the rewarding effect of
oxycodone in adolescent mice as measured by CPP.

127

Presenter – Zhou, Yan
Abstract ID – T4
Combination of clinically utilized kappa opioid receptor agonist nalfurafine with low-dose
naltrexone reduces excessive alcohol drinking in male and female mice
Yan Zhou1, Mary Jeanne Kreek1
1
Rockefeller University, New York, USA
Nalfurafine is the first clinically approved (Japan) kappa-opioid receptor (KOP-r) agonist, as an antipruritus drug, with few side effects in humans. No study has been done using nalfurafine on alcoholin rodents or humans. We investigated whether nalfurafine alone or in combination with mu-opioid
receptor [MOP-r] antagonist naltrexone changed excessive alcohol drinking in male and female
C57BL/6J (B6) mice subjected to a chronic intermittent access drinking paradigm for 3 weeks.
Neuronal proopiomelanocortin enhancer (nPE) knockout mice with brain-specific deficiency of betaendorphin (endogenous ligand of MOP-r) were used as a genetic control for the naltrexone
effects. Single administration of nalfurafine dose-dependently decreased alcohol intake andpreference in both sexes. Pretreatment with nor-BNI, selective KOP-r antagonist, blocked thenalfurafine effect on alcohol drinking, indicating a KOP-r mediated mechanism. Pharmacologicaleffects of nalfurafine (10ug/kg, five-doses) were further evaluated: the repeated nalfurafine
administrations decreased alcohol consumption without showing any blunted effects, suggesting
nalfurafine did not develop tolerance. Nalfurafine did not produce any sedation (locomotor activity),anhedonia-like (sucrose preference), anxiety-like (elevated plus maze), or dysphoria-like(conditioned place aversion) behaviors, suggesting that nalfurafine had few side
effects. Investigating synergistic effects between low-dose naltrexone and nalfurafine, we found thatsingle combinations of nalfurafine and naltrexone, at doses lower than individual effective dose,
profoundly decreased excessive alcohol intake in both sexes. The effect of nalfurafine was confirmed
in nPE -/- mice, suggesting independent mechanisms by which nalfurafine and naltrexone reduced
alcohol drinking. The clinically utilized KOP-r agonist nalfurafine in combination with low-dose
naltrexone has potential in alcoholism treatment.

128

Index by Last Names
Last nameFirst name

Adhikary
AfroseAiravaara
Alder
Amgott-Kwan
Anand
Arguelles
AtwoodBaileyBaloghBarnett
Bart
BediniBeier
BeierleBelliaBen-Ezra
Bertels
Birdsong
BobeckBorrelli
BouchetBrice-Tutt
Brown
BryantBrynildsen
Burgess
Cami-Kobeci
Campos-Jurado
Chan
Chen
Chiou
Cippitelli
Collins
ConibearCox
De Marco
Degrandmaison
Dorval
Dunn
Effah

Sweta
Leela
Mikko
Amy
Ariel
Jessica
NicoleBrady
AlexisMihályMiriamGavin
Andrea
Kevin
Jacob
Fabio
Ariel
Zachariah
WilliamErin
Kristyn
Courtney
ArianaKate
CamronJulia
Gwendolyn
Gerta
YolandaWallace
Carina
Lih-Chu
AndreaDevon
Alexandra
Brian
Rossella
Jade
LoubenAmeliaFelix

Abstract ID
T11
W9
T16
M49
T21
T52
T13
W7
M20
T43
T1
W47
M44
W32
M3
T15
W38
T46
T12
M24
M19
W15
W12
M33
T49
T35
W39
M17
T3
M30
T14
W2
T48
T22
M51
W17
M22
T33
M40
M46
M11
W28
129

EmeryFakiraFang
Faouzi
FujitaFullerton
Ghosh
Gillis
Gomes
Groom
Gupta
HamadaHe
He
Hearing
Hearing
Hill
Hipólito
Holm
Hsu
Inan
Inan
Iyer
JullieKandasamy
Kapoor
Karkhanis
KirályKliewer
Knapp
KochanKuo
KuzumakiLee
Leff
Lemos Duarte
LevineLevranLiliusLiu
Lueptow
Lutfy
Lutfy
Lutfy

MichaelAmandaSophiaAbdelfattah
WakakoEvan
Nillu
Alexander
Ivone
Sam
Achla
YusukeYing
Jenny
Matthew
Matthew
Rob
Lucia
LovisaWen-YuSaadetSaadetVishakhDamienRam
AbhijeetAnushreeKornélAndreaBrian
KelseyLawrence
Naoko
Cynthia Wei-Sheng
Emily
Mariana
OliviaOrna
TuomasJing-Gen
Lindsay
Kabirullah
Kabirullah
Kabirullah

W14
T27
M7
M5
W24
M52
W27
T47
T24
M25
W22
W52
T28
W34
T10
W6
M23
M28
M50
M1
W20
W21
W8
W19
M21
M35
W26
M39
T2
M48
W11
T19
T37
W40
W46
M43
M14
T6
W48
W4
M4
M34
T36
W35
130

Mack
MaguireMajumdar
Manandhar

SeshatDavid
Susruta
Preeti

T26
T18
M13
W25

Martucci

Katherine

W48

Matyas
McCallMcDermottMcElroy
McPherson
MilnerMorganMoriyaNakataNilges
Nozaki
Olson
Ozawa
Pan
Peles
Pick
Polt
Poras
PradhanPresker
Pryce

Gary
Nora
Max
Bryan
Kylie
TeresaMichael
Yuki
Eriko
Mark
ChihiroKeith
AkihikoYing-Xian
Einat
Chaim (Chagi)
Robin
Hervé
Amynah
Mark
Kerri

M26
M16
T20
T9
M15
T23
M36
W18
M6
T39
M32
W13
M31
T31
W1
T30
M37
T5
M9
W37
W42

Rae
Randesi
ReinerRetzlaff
Rivera Irizarry
RoquesRossi
Rubaharan
Ryan
Saberi
Saitoh
SaklothSears
SéguélaSeverino
SharmaShibasaki

Mariana
MatthewDavid
Cassandra
Jean K.
Bernard P.
Grace
MyurajanJames
ShahinAkiyoshi
Farhana
Bryan
Philippe
Amie
Indrajeet
Masahiro

W41
T25
M10
M45
T41
W45
M47
T45
M12
W16
W23
M41
W10
M42
M18
T38
T44
131

Simmons
StamStandifer
Streicher
Targowska-Duda
TejadaThorsell
ToddesTrieu
Ueda
van Rijn
Varshneya
Wang
Webb
Whistler
Wiese
WilsonWindisch
YamadaYukari
Zaidi
Zelleroth
Zhang
Zhang
Zhou

Sarah
Frida
Kelly
John
Katarzyna
Frederick
AnnikaCarleeBrian
HiroshiRichardNeil
Yu-JunChase
Jennifer
Beth
Lisa
Kyle
Daisuke
Suda
Saheem
Sofia
Li
Yong
Yan

W5
W31
W51
M38
W36
T40
T51
W49
T50
T42
W29
W44
T7
T17
W50
M2
T34
M29
W3
W30
T8
T32
T29
W33
T4

132

Thank you to our generous SPONSORS!

133

G R A N D H Y A T T N E W Y O R K
P a r k A v e n u e a t G r a n d C e n t r a l
N e w Y o r k , N Y 10 0 17 , U S A+ 1 2 12 8 8 3 12 3 4+ 1 2 12 6 9 7 3 7 7 2

T E L E P H O N E
F A X

g r a n d n e w y o r k .h y a t t .c o m

F L O O R P L A N
L o b b y L e v e lN

G A L L E R YIR E S T R O O M S

II

O N

L E X
III

M A R K E T

M U S E U M
M A N H A T T A N
B A L L R O O M
F O Y E RF R O N T D E S K

M A N H A T T A N
B A L L R O O M

L O B B Y

T O

S T R E E T L O B B Y

M E Z Z A N IN E F O Y E R

1 1 .1 2

134

PARTICIPANTS
Last NameFirst Name

Email

Adhikary
AfroseAiravaara
Akil
Akinfiresoye
Alder
AldrichAl-Hasani
Amgott-Kwan
Anand
Arguelles
Arttamangkul
AtwoodBabey
BagleyBaileyBaileyBallasBaloghBarnett
Bart
BayoumyBediniBeier
BeierleBelliaBen-Ezra
Benmerzouga
Bertels
Bidlack
Birdsong
Blais
BlendyBobeckBohn
Borrelli
BouchetBowes Lawlor

Sweta
Leela
Mikko
Huda
Luli
Amy
Jane
Ream
Ariel
Jessica
NicoleSeksiri
Brady
Anna-Marie
Elena
Chris
AlexisSamir
MihályMiriamGavin
Shariff
Andrea
Kevin
Jacob
Fabio
Ariel
Imaan
Zachariah
Jean
WilliamVeronique
Julie
Erin
Laura
Kristyn
CourtneyCarolsweta.adh@gmail.com
leela.afrose@gmail.com
mikko.airavaara@helsinki.fi
akil@umich.edu
luli.r.akinfiresoye@usdoj.gov
amy.alder@vuw.ac.nz
janealdrich@ufl.edu
al-hasanir@wustl.edu
aamgottk@tulane.edu
janand@umich.edu
nicole.arguelles@mail.utoronto.ca
arttaman@ohsu.edu
bkatwood@iu.edu
ababey@une.edu.au
elena.bagley@sydney.edu.au
cb304@bath.ac.uk
abailey@sgul.ac.uk
samir.ballas@jefferson.edu
balogh.mihaly@med.semmelwes-univ.hu
miriam_barnett@urmc.rochester.edu
bartx005@umn.edu
shariff.bayoumy@mebiasdiscovery.com
andrea.bedini@unibo.it
kbeier@uci.edu
jbeierle@bu.edu
fabio.bellia91@gmail.com
aben@rockefeller.edu
ibenmerzouga@osteo.wvsom.edu
zbertel2@uic.edu
jean_bidlack@urmc.rochester.edu
wtbird@umich.edu
veronique.d.blais@usherbrooke.ca
blendy@pennmedicine.upenn.edu
erin.bobeck@usu.edu
lbohn@scripps.edu
kristynb@bu.edu
bouchet@ohsu.edu
carol.lawlor@alkermes.com

135

Last NameFirst Name

EmailBrice-Tutt
Brown
BrowneBruchasBruinsBryantBrynildsen
Burgess
Butelman
CahillCami-Kobeci
Campos-Jurado
CanalsCarroll
Chan
Chavkin
Chen
Chiu
Christie
Cippitelli
Comer
Conibear
CorderCowan
Cox
Degrandmaison
Devi
DorvalDubroff
Dunn
Effah
Emery
Evans
FakiraFan
Fang
FaouziFilizola
Fricker
FujitaFullertonGellerArianaKate
CalebMichaelJohn
CamronJulia
Gwendolyn
EduardoCathy
Gerta
YolandaMeritxell
F. IvyWallaceCharlesCarinaIsaac
Mac
AndreaSandraAlexandra
Gregory
Alan
Brian
Jade
LakshmiLoubenJacob
AmeliaFelix
Michaelchris
AmandaJun
SophiaAbdelfattah
Marta
Lloyd
WakakoEvan
Ellen Bariana.brice@ufl.edu
kbrown@rockefeller.edu
caleb.browne@mssm.edu
mbruchas@uw.edu
john.bruins@sandoz.com
camron@bu.edu
jbryn@pennmedicine.upenn.edu
burgessg@umich.edu
butelme@rockefeller.edu
cmcahill@ucal.edu
chpck@bath.ac.uk
yolanda.campos@uv.es
m.canals@nottingham.ac.uk
fic@rti.org
wallakin@umich.edu
cchavkin@u.washington.edu
cchen@rockefeller.edu
isaac_chiu@hms.harvard.edu
mac.christie@sydney.edu.auacippitelli@health.fau.edu
sdc10@cumc.columbia.edu
alex.conibear@bristol.ac.uk
gregory.corder@pennmedicine.upenn.edu
acowan@temple.edu
brian7580@verizon.net
jade.degrandmaison@usherbrooke.ca
lakshmi.devi@mssm.edu
louben_dorval@urmc.rochester.edu
jacob.dubroff@pennmedicine.upenn.edu
adunn@rockefeller.edu
p1805034@sgul.ac.uk
emeryma@med.umich.edu
cevans@ucla.edu
amanda.fakira@mssm.edu
jfan@neurocrine.com
sophiamfang@gmail.com
afaouzi@wustl.edu
marta.filizola@mssm.edu
Lloyd.Fricker@einstein.yu.edu
wakakofu@nagasaki-u.ac.jp
evanffullerton@gmail.com
ebgeller@temple.edu
136

Last NameFirst Name

Email

GendronGhosh
Gillis
GOMES
Groom
GUPTA
Hallberg
HamadaHassanien
He
He
Hearing
Henderson
Hill
Hipólito
Hohmann
Hornyák
Hsu
Hsu
Husbands
Inan
IngramInturrisi
Iyer
Javitch
Ji
Jin
Jones
JULLIE
Jutkiewicz
Kandasamy
Kapoor
Karkhanis
Katritch
Kelly
Khan
KimmeyKirályKliewer
Knapp
KochanLouis
Nillu
Alexander
IVONE
Sam
ACHLA
MathiasYusukeSherifYing
Jenny
Matthew
GraemeRob
Lucia
AndreaHajnalka
Wen-YuDavid
StephenSaadetSusanCharles
VishakhJonathanRu-Rong
ChunyangSara
DAMIEN
Emily
Ram
AbhijeetAnushreeVsevolod
Eamonn
TabibulBlake
KornélAndreaBrian
Kelseylouis.gendron@Usherbrooke.ca
nxg460@med.miami.edu
agil7104@uni.sydney.edu.au
ivone.gomes@mssm.edu
S.Groom@bath.ac.uk
achla.gupta@mssm.edu
Mathias.hallberg@farmbio.uu.se
y-hamada@hoshi.ac.jp
shassanien@caymanchem.com
jspain@rx.umaryland.edu
xih174@ucsd.edu
matthew.hearing@marquette.edu
g.henderson@bristol.ac.uk
rob.hill@nottingham.ac.uk
lucia.hipolito@uv.es
hohmanna@indiana.edu
hornyaakhajnalka@gmail.com
cchhsuwenyu@gmail.com
david.hsu@stonybrookmedicine.edu
s.m.husbands@bath.ac.uk
sinan@temple.edu
ingrams@ohsu.edu
ceintur@med.cornell.edu
vishiyer@indiana.edu
jaj2@cumc.columbia.edu
ru-rong.ji@duke.edu
cjin@rti.org
srjones@wakehealth.edu
damien.jullie@ucsf.edu
ejutkiew@umich.edu
ramka@umich.edu
abhijeet.kapoor@mssm.edu
akarkhan@binghamton.edu
katritch@usc.edu
e.kelly@bristol.ac.uk
tabibulk@hotmail.com
bkimmey@pennmedicine.upenn.edu
kiraly.kornel@med.semmelweis-univ.hu
andrea.kliewer@med.uni-jena.de
brian_knapp@urmc.rochester.edu
kkochan@umich.edu

137

Last NameFirst Name

Email

Koob
KozellKreek
Kuo
Kuzumaki
LändinLee
Leff
Lemos Duarte
LevineLevran
Li
Li
lin
Liu
Liu-Chen
Lueptow
Lutfy
Lutz
Mack
Maguiremajumdar
Manandhar
Manglik
Mao
MarconiMargolis
Martucci
MatsuiMatyas
Mazei-Robison
McCallMcDermottMcElroy
McLaughlin
McPherson
MilnerMorganMoriyaMorochnik
MurphyGeorgeLaura
Mary Jeanne
Lawrence
Naoko
Inger
Cynthia Wei-Sheng
Emily
Mariana
OliviaOrna
Jun-XuYangmei
XiaoyanJing-Gen
Lee-Yuan
Lindsay
Kabirullah
Pierre-Eric
SeshatDavidsusruta
Preeti
AashishLimin
LaurenElyssaKatherine
Aya
Gary
Michelle
Nora
Max
Bryan
Jay
Kylie
TeresaMichael
Yuki
MichelleAnnekoobgf@mail.nih.gov
kozellla@ohsu.edu
kreek@rockefeller.edu
lawrence.kuo@mebiasdiscovery.com
n-kuzumaki@hoshi.ac.jp
inger.landin@nordicdrugs.se
cindywslee@gmail.com
leff@ohsu.edu
mariana.lemosduarte@mssm.edu
OLL2004@med.cornell.edu
levrano@rockefeller.edu
junxuli@buffalo.edu
liyangme@cop.sc.edu
XL28@indiana.edu
jgliu@simm.ac.cn
lliuche@temple.edu
llueptow@mednet.ucla.edu
klutfy@westernu.edu
pierreeric.lutz@gmail.com
seshatmack@gmail.com
maguired@uthscsa.edu
susrutam@email.wustl.edu
preeti.manandhar@hdr.mq.edu.au
Aashish.Manglik@ucsf.edu
maol@umkc.edu
lauren.marconi@pennmedicine.upenn.edu
elyssa.margolis@ucsf.edu
katherine.martucci@duke.edu
aya.matsui@nih.gov
gmatyas@hivresearch.org
mazeirob@msu.edu
nmmccall@gmail.com
maxvanmcd@gmail.com
bmcelroy@rockefeller.edu
jmclaughlin@cop.ufl.edu
kyliemcpherson3@gmail.com
tmilner@med.cornell.edu
mmmorgan@wsu.edu
moriya-yk@igakuken.or.jp
mmorochnik@rockefeller.edu
amurphy@gsu.edu

138

Last NameFirst Name

EmailNakataNaritaNaritaNatesan
Negus
Neogi
Nilges
NozakiNyberg
Oliva
Ozawa
Pan
Pang
Peles
PellandPick
PintarPolt
PORAS
PradhanPresker
Provasi
Pryce
Puthenveedu
Rae
Randesi
Reed
ReinerRetzlaff
Riley
Rivera Irizarry
ROQUES
Rossi
RothwellRubaharan
Ryan
Saberi
Saitoh
SaklothSaleemSamad

Eriko
MinoruMichikoSenthilSteve
RichikMark
ChihiroFred
IdairaAkihikoYing-Xian
Zhiping
Einat
Jessica
Chaim (Chagi)
John
Robin
Hervé
AmynahMarkDavideKerri
Manoj
Mariana
Matthew
BrianDavid
Cassandra
Andrew
Jean K.
Bernard P.
Grace
Patrick
MyurajanJames
ShahinAkiyoshi
Farhana
Ameen
Marielle

e-nakata@chemiphar.co.jp
narita@hoshi.ac.jp
michiko.t.narita@gmail.com
senthil.natesan@wsu.edu
sidney.negus@vcuhealth.org
richik.neogi@wsu.edu
mnilge@lsuhsc.edu
chihiron@uni-bonn.de
fred.nyberg@farmbio.uu.se
idaira.oliva@gmail.com
aozawa@health.fau.edu
pany@mskcc.org
pangzh@rwjms.rutgers.edu
einatp@tlvmc.gov.il
jpelland@sas.upenn.edu
PICKC@TAUEX.TAU.AC.IL
pintar@rwjms.rutgers.edu
polt@u.arizona.edu
herve.poras@pharmaleads.com
pradhan4@uic.edu
mark.presker@rutgers.edu
davide.provasi@gmail.com
kerri.pryce@mssm.edu
puthenve@umich.edu
marianarae@hotmail.com
randesm@rockefeller.edu
reedb@rockefeller.edu
david.reiner@nih.gov
retzl016@umn.edu
apriley@uic.edu
jkr2004@med.cornell.edu
bernard.roques@parisdescartes.fr
grace.rossi@liu.edu
rothwell@umn.edu
mrubaharan@gsu.edu
jdr2003@med.cornell.edu
ssaberi@indiana.edu
akiyoshi_saitoh@rs.tus.ac.jp
farhana.sakloth@mssm.edu
ameen.saleem@alkermes.com
marielle.samad@canada.ca

139

Last NameFirst Name

Email

Sarne
SchulzSearlesSears
SéguélaSeverino
ShahamSharmaShibasaki
Simmons
Snyder
SnyderSombers
StamStandifer
Stauffer
Stevens
Streicher
Targowska-Duda
TawfikTaylor
TEIXEIRA
TejadaToddesToll
ToungeTrang
Traynor
Trieu
Tsai
Ueda
Unterwald
van Rijn
Varshneya
Volkowvon Zastrow
WackerWalker
Wang
Wang
Webb

Yosef
StefanChris
Bryan
Philippe
Amie
Yavin
Indrajeet
MasahiroSarah
GretchenSolomonLeslieFrida
Kelly
JosephCraig
John
Katarzyna
VivianneAnna
ALVARO
FrederickCarleeLawrence
Brett
Tuan
John
Brian
Anne
HiroshiEllen
Richard
Neil
Nora
Mark
DanielEllen
Yu-JunQiang
Chasesarney@post.tau.ac.il
stefan.schulz@med.uni-jena.decsearles3@student.gsu.edu
searsbry@umich.edu
philippe.seguela@mcgill.ca
amieseverino@g.ucla.edu
yshaham@intra.nida.nih.gov
isharma@ou.edu
m-shibasaki@hoshi.ac.jp
sarah.cooper.ctr@usuhs.edu
gsnyder@intracellulartherapies.com
smcteer1@jhmi.edu
leslie_sombers@ncsu.edu
frida.stam@farmbio.uu.se
Kelly-standifer@ouhsc.edu
acrusco@nektar.com
cw.stevens@okstate.edu
jstreicher@email.arizona.edu
ktargowskaduda@health.fau.edu
vivianne@stanford.edu
ataylor1@ualberta.ca
vcarregais@osiris.pt
frtejada@umes.edu
todde001@umn.edu
ltoll@health.fau.edu
brett.tounge@mebiasdiscovery.com
trangt@ucalgary.ca
jtraynor@umich.edu
trieu016@umn.edu
stsai@nih.gov
ueda.hiroshi.8e@kyoto-u.ac.jp
ellen.unterwald@temple.edu
rvanrijn@purdue.edu
varshneyan@vcu.edu
nvolkow@nida.nih.gov
mark@vzlab.org
daniel.wacker@mssm.edu
ellen.walker@temple.edu
yjwang@mail.simm.ac.cn
wangjq@umkc.edu
chase.webb@ucsf.edu

140

Last NameFirst Name

Email

Wiese
WilcoxWilsonWimmerWindischWingerWoods
Wu
XU
YamadaYuferov
Yukari
Zachariou
ZadinaZaidi
Zaveri
Zelleroth
Zhang
Zhang
Zhang
Zhou
Zhu

Beth
GeorgeLisa
Mathieu
Kyle
Gail D.
James H
Chung-Yu
JIN
Daisuke
Vadim
Suda
VenetiaJames
SaheemNurulain
Sofia
Li
Yong
Yanan
Yan
Hu

bmwiese@email.arizona.edu
george@umn.edu
lisawilson@ufl.edu
mathieu.wimmer@temple.edu
kwindisch01@rockefeller.edu
winger@uthscsa.edu
woodsjh@uthscsa.edu
peterwu@mail.nctu.edu.tw
xuj1@mskcc.org
yamadada@rs.tus.ac.jp
yuferov@rockefeller.edu
y-suda@hoshi.ac.jp
venetia.zachariou@mssm.edu
jzadina@tulane.edu
saheemza@usc.edu
nurulain@astraeatherapeutics.com
sofia.zelleroth@farmbio.uu.se
zhan1999@umn.edu
zhangyo@rockefeller.com
yzhang@rti.org
zhouya@rockefeller.edu
hu.zhu@nih.gov

141

NOTES142

NOTES143

ADVANCES IN OPIOID“NEWRESEARCH: FROM BENCH
TO THE BEDSIDE
”

INRC
2020

See you in Valencia!
VALENCIA (SPAIN)
6-10 July 2020 | Hotel Meliá Valencia

144