Functional and behavioral consequences of DRD4 genetic variation in macaques

Drug and Alcohol Dependence 156 (2015) e183–e245 e229 Functional and behavioral consequences of DRD4 genetic variation in macaques Eric J. Vallender2,4,∗, Lisa M. Ogawa3,4, Laurie J. Lynch4, Jessica L. Schimmel4, Gregory M. Miller1,4 1 Pharmaceutical Sciences, Northeastern University, Boston, MA, United States 2 Psychiatry and Human Behavior, University of Mississippi Medical School, Jackson, MS, United States 3 Yale University, New Haven, CT, United States 4 New England Primate Research Center, Harvard Medical School, Southborough, MA, United States Aims: The dopamine D4 receptor (DRD4) associates with addiction-related phenotypes, including novelty seeking, risk taking, and stress resilience. This study aimed to investigate the consequences of genetic variation in DRD4 in macaques, model species often utilized in substance abuse research. We sought to identify genetic variation and characterize haplotypes across the genus, then test the functional effects of this variation on cAMP signaling and initiate studies on the behavioral impact of these polymorphisms in rhesus macaques. Methods: The DRD4 receptor was resequenced in 64 rhesus, 32 cynomolgus, and 32 Japanese macaques. HEK293 and AV12 cells with CRE-responsive firefly luciferase reporters were transfected with macaque and human DRD4 alleles. Cells were treated with forskolin and variable concentration of DRD4 agonist (A-412997). Luciferase fluorescence was measured and quantified. Genotyped rhesus macaques were presented with novel and familiar fruit stimuli. Latency to physical contact and ingestion served as a measure of novelty seeking. Results: 15 nonsynonymous SNPs were identified in multiple individual macaques; 11 localized to the third intracellular loop. C749T, (MAF>20%) showed an EC50 shift following agonist treatment (HEK293: 1.7nM vs. 2.6nM; AV12: 9.6nM vs. 3.1nM) comparable to that observed in humans comparing the 7-repeat and 4-repeat alleles (HEK293: 7.6nM vs. 3.2nM; AV12: 3.3nM vs. 5.0nM). In behavioral assays, animals harboring the T749 allele have a longer latency to both touch (1.5 s) and eat (6 s, n=9) the novel fruit. Conclusions: Inmacaques, a polymorphism (C749T) is observed that parallels the human repeat variation in both signaling and behavioral assays. This commonality may offer insights into both the natural history of the species and the genetics of noveltyseeking behavior associated with substance abuse. Financial support: AA019688 (EJV) and OD011103. http://dx.doi.org/10.1016/j.drugalcdep.2015.07.616 Predictors of agreement between hair analysis and self-report of drug use Paul VanVeldhuisen1,∗, Gaurav Sharma1, Michael Bogenschutz2 1 The EMMES Corporation, Rockville, MD, United States 2 University of NM, Albuquerque, NM, United States Aims: To examine predictors of agreement between hair analyses and self-report of drug use. Methods: This is a secondary analysis from the National Drug Abuse Treatment Clinical Trials Network (NIDA CTN) randomized trial “Screening, Motivational Assessment, Referral, and Treatment in EmergencyDepartments (CTN-0047; SMART-ED)”. Self-reported drug use during follow-up over a 90-day recall period on the Timeline Follow Back Instrument was compared to drug use from hair analysis for Cannabis, Cocaine, Prescribed Opioids [PO] and Street Opioids[SO]. Measures of agreement/disagreement, including under-reporting (self report negative when hair indicates drug use) and over-reporting (self-report positive when hair does not indicate drug use) were calculated. The following variables were examined as predictors of disagreement: source of hair (head vs. body), drug of choice, site, AUDIT-C score, DAST-10 score, visit, treatment arm, sex, race, ethnicity and age. Of the 1285 randomized participants, 1120 (87%), 875 (68%), 893 (69%) and 832 (65%) provided hair samples at baseline, 3-, 6-, 12-month visits, respectively. Results: The agreement between the hair sample results and TLFB was high for cannabis (cohen’s |=0.49–0.54) and SO (|=0.73–0.81), but lower for cocaine (|=0.31–0.35) and PO (|=0.18–0.30). Drug of choice, irrespective of being cannabis, cocaine, SO or PO, had statistically significantly lower underreporting of drug use compared with other self-reported drug use (all p-values <.01). Of note, females (p= .0085; F vs M; OR=1.33) and older age (p= .0076; 45–<55 vs 18–<25; OR=2.02) were associated with under-reporting of cannabis use. Few predictors of over-reporting were identified. Conclusions: Hair collection can be an important biological measure to assess drug use, and can be used to assist in corroborating self-report. From these analyses, there are a number of factors that impact agreement between drug use as measured by hair and drug use through self-report. Financial support: NIDA-HHSN271201400028C/N01DA-142237. http://dx.doi.org/10.1016/j.drugalcdep.2015.07.617 Exploring the feasibility to quantify prescription drug misuse and abuse: Focus group findings Deepthi S. Varma1,∗, Lee Hoffer2, Linda Cottler1 1 Epidemiology, University of Florida, Gainesville, FL, United States 2 Anthropology, Case Western Reserve University, Cleveland, OH, United States Aims: The epidemic of prescription drug misuse and addiction is increasing in the United States and researchers need to accuratelyquantifydrug intake to categorizeusers anddevelopeffective prevention interventions. This paper presents the findings on accurate quantification of prescription pill misuse from a series of focus groups (FGs) as part of a NIDA funded PrescriptionDrugMisuse and Abuse study in St. Louis. Methods: Fourteen FGs, seven each with older (aged 50–65 years) and younger adults (aged 18–25 years) were conducted using a semi-structured interview guide. All 38 participants were asked toestimate the totalnumberofpills theymisused in their lifetime. Sessions were audio taped, transcribed, and analyzed using the Atlas tiTM. Results: Though participants in all FGs reported misusing prescription drugs, only one older groupparticipant and three younger groupparticipantsmentioned that they could calculate the approximate number of lifetime prescription drugs they had misused or abused. Among older adults, barriers to calculating the number of pillsmisused includedavery longdurationof use, theuseof pills for both prescribed and non-prescribed motivations, and infrequent mixing of pills with alcohol and other illegal substances. Among