Contents
03........ Message from Our Chairperson
04........ Message from Our Director
05........ History • Our Mission
06........ Our Strategies • Our Goal
07........ Organizational Chart • Our People at a Glance

Research Activities
Department of Genome Medicine
09........Genome Dynamics Project......................Hisao Masai
11........Mammalian Genetics Project...................Yoshiaki Kikkawa
13........Viral Infectious Diseases Project..............Fumihiko Yasui
15........Neurovirology Project...............................Satoshi Koike
17........Allergy and Immunology Project..............Takachika Hiroi
19........Molecular Medical Research Project.......Futoshi Shibasaki

Department of Dementia and Higher Brain Function
21........Dementia Research Project.....................Masato Hasegawa
23........Learning and Memory Project.................Minoru Saitoe
25........Neural Prosthesis Project.........................Yukio Nishimura
27........Parkinson’s disease Unit..........................Makoto Hashimoto

Department of Brain Development and Neural Regeneration
29........Developmental Neuroimmunology Project....... Hiroshi Sakuma
31........Synaptic Plasticity Project........................Kanato Yamagata
33........Neural Development Project....................Haruo Okado
35........Neural Network Project.............................Nobuaki Maeda

Department of Psychiatry and Behavioral Sciences
37........Mental Health Promotion Project..............Atsushi Nishida
39........Schizophrenia Research Project..............Makoto Arai
41........Affective Disorders Research Project......Yoshitaka Tatebayashi
43........Sleep Disorders Project............................Makoto Honda
45........Addictive Substance Project....................Kazutaka Ikeda
01

Department of Sensory and Motor Systems
47........Motor Disorders Project............................Shinji Kakei
49........Visual Research Project............................Takayuki Harada
51........ALS Nursing Care Project........................Yuki Nakayama
53........Diabetic Neuropathy Project....................Kazunori Sango

Department of Advanced Science for Biomolecules
55........Calpain Project..........................................Yasuko Ono
57........Ubiquitin Project........................................Noriyuki Matsuda
59........Stem Cell Project......................................Takahiko Hara
61........Regenerative Medicine Project................Yuichiro Miyaoka
63........Stroke Renaissance Project.....................Takashi Shichita
65........Protein Metabolism Laboratory................Yasushi Saeki
67........Biomembrane Unit....................................Kohji Kasahara

Research Support
69........ Center for Basic Technology Research
70........ Technology Licencing Office

71........ Center for Medical Research Cooperation
72........ Neuropathology Laboratory
73........ TMIMS Programs
74........ Access Map

02

Message from Our Chairperson
Chairperson Keiji Tanaka
The metropolis of Tokyo is the nerve center of Japan. Developing Tokyo into a
healthy welfare city will therefore go a long way toward building a prosperous
future for Japan. The mission of the Tokyo Metropolitan Institute of Medical
Science (TMIMS) is to promote research in the life and medical sciences toprotect the lives and health of Tokyo citizens, with the goal of improving thehealth, medical care, and welfare of this metropolitan city. It is well known thatJapan has the most rapidly ageing society in the world. Tokyo, which reflects
Japan itself, is undergoing a steady increase in cancer and infectious diseases,
which pose a threat to humanity, as well as lifestyle-related illnesses, neural and mental disorders,and various other health problems. Naturally, conquest of all of these diseases is a common goal of all
humankind, and considerable efforts have been made at the national level. Even so, because Tokyo is
one of the largest civilized cities in the world, it is also essential for the Tokyo Metropolitan Government
to take the initiative in this endeavor. Besides, Tokyo has numerous problems unique to megacities.
For instance, many people suffer from rare and intractable diseases that researchers often overlook.
TMIMS has been actively working on these important problems, promptly and practically addressing
health-related issues with the aim of protecting the health of numerous Tokyo citizens.
By the way, I am of the opinion that “research represents culture”. Accordingly, TMIMS aims to be
acclaimed both academically and culturally for the knowledge and wisdom of its excellent researchers,with the ultimate goal of becoming a symbol of the culture of Tokyo, the foremost megalopolis in theworld. Academic research is often roughly divided into top-down, exit-oriented applied research (ofimmediate use) and bottom-up, future-oriented fundamental research (seemingly not of immediateuse). Balancing these two research strategies, TMIMS endeavors to operate the organization in a
flexible manner in order to achieve additive and synergic effects. In fact, these two research strategies
are not incompatible, but can work in a cooperative and harmonious manner. Throughout the historyof science, we can find numerous examples of seemingly useless research suddenly becoming useful,resulting in great service to society.
Our medical researchers are energetically pursuing their research day and night to develop preventive
medicine and new therapeutic methods to protect citizens’ health in the future. While TMIMS naturally
takes on the role of educating young researchers who will help develop human knowledge andwisdom, it also seeks to generate concrete research findings that contribute to social prosperity.
Accordingly, all the staff members of TMIMS are working on life science research, ranging fromfundamental to practical, making the most of cutting-edge technologies to achieve their specific goals.
It is vitally important that TMIMS grow to be the world’s top-notch research institute, and advancing
and enriching its research power will eventually create an institute capable of rendering wide-ranging
services to society. To this end, the entire staff of TMIMS will strive as one to help pursue incomparable
fundamental research and pass the benefit of its research findings on to society, while recruiting andeducating talented people to build up momentum.
We look forward to your further guidance and encouragement, which are indispensable for the further
development of TMIMS. Thank you in advance for your continued support.

03

Message from Our Director
Director Hisao Masai
The Tokyo Metropolitan Institute of Medical Science (TMIMS) was establishedin April 2011 as a result of the merger of three institutes; the Tokyo Metropolitan
Institute for Neuroscience, the Tokyo Metropolitan Institute of Psychiatry, and theTokyo Metropolitan Institute of Medical Science, all of which had been foundedin early- to mid-1970s with the support of the Tokyo Metropolitan Governmentand had been present at different locations in Tokyo. The scientists from threedifferent disciplines came together in a new research building in a quiet residential
area at Kamikitazawa in Setagaya-ku, about 15 minutes by train from Shinjuku.
The institute is under the continuous support of the Tokyo Metropolitan Government, and our aim isto advance medical research and improve the health and welfare of people living in the metropolises
through collaborative research in basic life sciences, medical sciences, social medicine, and nursing.
In a metropolis such as Tokyo, people suffer from various diseases and other health-related problems
unique to urban life-styles. These include mental disturbances caused by stresses associated withdaily life, diseases associated with longevity, and diseases caused by complicated genetic traits aswell as infectious diseases caused by viruses such as hepatitis, influenza, and other outbreaks. We
are studying these problems using a unique combination of disciplines, technology and expertise.
We conduct forefront basic research in various biological fields including genome and protein functions,stem cells and development, genetic diseases, brain functions, neurobiology, neurodegeneration, stem
cells and development, virus infection, allergy, schizophrenia, and depression. Using the state-of-art
and newest technology and equipment, we are identifying molecules and mechanisms responsible forvarious biological phenomena as well as for disease progression. These knowledges and technologies
will be used to predict and prevent diseases, and develop new drugs and therapies that can be tailored
to individual patients.
We also emphasize importance of sociomedical approaches, including large scale cohort studies
aimed at identifying social and environmental factors associated with mental health of youths. We
develop effective care and nursing systems for elderly people suffering from dementia, and provide
those suffering from progressive and currently incurable diseases such as ALS (amyotrophic lateralsclerosis) with innovative care systems to improve QOL of these patients.
We will pursue research that will contribute to prediction, prevention, diagnosis, and treatment ofvarious diseases, will improve the care of patients, and will help realize longer healthy life. We will also
keep the people of the Tokyo Metropolises as well as those from other areas informed of our latest
progress by having various outreach activities including public lectures, science café , and lectures/classes to students. Through these efforts, we hope to serve as a leader and model institute for thelife/medical science in the coming decades.

04

History
The Tokyo Metropolitan Institute of Medical Science was established in April 2011 as a resultof the merger of three institutes; the Tokyo Metropolitan Institute for Neuroscience, the Tokyo
Metropolitan Institute of Psychiatry, and the Tokyo Metropolitan Institute of Medical Science, allof which had been founded in mid-1970s with the support of Tokyo Metropolitan Government andhad been located at different locations in Tokyo. The scientists from three different disciplines got
together in a new research building in a quiet residential area at Kamikitazawa in Setagaya-ku,
about 15 minutes by train from Shinjuku. The institute is under the continuous support from theTokyo Metropolitan Government, and our aim is to contribute to medical advances and improve thehealth and welfare of people living in the metropolises through collaborative research in basic life
sciences, medical sciences, social medicine, and nursing.

Our Mission
The mission of our institute is to pursue research that will lead tothe development of solutions for health-related problems commonly
observed in large urban areas and developed countries. Utilizing
results from the forefront basic research on molecular and cellularmechanisms of biological pathways and disease pathology, we will
collaborate with municipal hospitals and clinics to predict, prevent,and treat health problems. We will also identify causes of unsolved
diseases and develop drugs and therapies for them. Our missionis also to provide help and care with those suffering from serious
diseases such as ALS to better the patients’ quality of life. It is also
our mission to analyze the mental disturbances of the public and provide care and treatment.

05

Our Strategies
In a metropolis such as Tokyo, people suffer from various diseases and other health-related
problems unique to urban life-styles. These include mental disturbances caused by stresses
associated with daily life, diseases associated with longevity, and diseases caused by complicatedgenetic traits. We are studying these problems using a unique combination of disciplines,
technology and expertise.
We conduct forefront basic research invarious biological fields including genome
and protein functions, genetic diseases,
brain functions, neurobiology,
neurodegeneration, stem cells anddevelopment, virus infection, allergy,
schizophrenia, and depression. Using thestate-of-art and newest technology andequipment, we are identifying moleculesand mechanisms responsible for diseaseprogression and biological phenomena.
This knowledge and technology will be
used to predict and prevent diseases, anddevelop new drugs and therapies that canbe tailored to individual patients.
We are also combining sociomedical approaches with molecular and genomic approaches todiscover unique and effective treatments for mental disorders. Our institute takes advantage of amulti-disciplinary structure to provide novel solutions to various health-related issues. Located in
one of the biggest cities in the world, we hope to be a role model for medical research institutes in
the coming decades.

Our Goals
To pursue research that will help prediction, prevention,
diagnosis, and treatment of various diseases andimprove the care of patients, leading to longer healthy life.
To serve as a leader and model institute for the life/medical
science in the coming decades.

06

Organizational Chart

Our People at a Glance

07

08

Department of Genome Medicine
Project
LeaderHisao Masai Genome Dynamics Project

Genome Replication and Maintenance:

In search of unexplored messages in the genome

Masai H. et al. (2018) “Moleculararchitecture of G-quadruplex
structures generated on duplex Rif1
binding sequences.” J. Biol. Chem.
in pressMoriyama K, Yoshizawa-Sugata,N, and Masai H. (2018) “Oligomer
formation and G-quadruplex bindingby purified murine Rif1 protein, a key
organizer of higher-order chromatinarchitecture.” J. Biol. Chem.
293:3607-3624.
You Z, and Masai H. (2017) “Potent
DNA strand annealing activity
associated with mouse Mcm2~7
heterohexamer complex.” Nucleic
Acids Res. 45: 6495-6506.
Yang C-C, Suzuki M, Yamakawa S,
Uno S, Ishii A, Yamazaki S, Fukatsu R,
Fujisawa R, Sakimura K, Tsurimoto T,
and Masai H. (2016) “Claspin recruits
Cdc7 kinase for initia-tion of DNA
replication in human cells.” Nature
Commun. 7: 12135.
Kanoh Y, Matsumoto S, Fukatsu R,
Kakusho N, Kono N, Renard-Guillet
C, Masuda K, Iida K, Nagasawa K,
Shirahige K, and Masai H. (2015)“Rif1 binds to G-quadruplexes andsuppresses replication over long
distances.” Nature Struct. Mol. Biol.
22: 889-897.
Yamazaki S, Ishii A, Kanoh Y, Oda M,
Nishito Y, and Masai H. (2012)“Rif1 protein is a key regulator of
the genome-wide DNA replication
timing in human cells.” EMBO J. 31:
3167-3177.
Hayano M, Kanoh Y, Matsumoto S,
Shrahige K, and Masai H. (2012)“Rif1 is a global regulator of timingof replication origin firing in fission
yeast.” Genes and Development 26:
137-150.
Matsumoto S, Hayano M, Kanoh Y,
and Masai H. (2011) “Multiple pathways can bypass the essential roleof fission yeast Hsk1 kinase in DNA
replication initiation.” J. Cell Biol. 195:387-401.
Masai H, Matsumoto S, You Z,
Yoshizawa-Sugata N, and Oda M.
(2010) “Eukaryotic DNA replication;
where, when and how?” Annual Rev.
Biochem. 79: 89-130.

Precise duplication of genetic materials is central to the stable
maintenance of genomes through generations. Defects in the genome
copying processes would generate genomic instability which could
ultimately result in various diseases including cancer. The goal of ourstudies is to understand the molecular basis of how the huge genomes
are accurately replicated and the precise copies of the genetic materials
are inherited to the next generation. Three billion base pairs of thehuman genome (2 meter long) are replicated with almost no errors
during the 6-8 hr time span of the cell cycle. This requires an extreme
level of coordination of temporal and spatial arrangements of chromatinorganization and signaling events for initiation of DNA replication.

“We are trying to decipher ‘unexploredmessages’ of the genome that are crucial
for shaping the chromosomes, copying andreading out genetic information, and even for
causing detrimental diseases.”

We recently discovered novel and crucial roles of non-standard DNA
structures in regulation of DNA replication and transcription. Notably,
we found that G-quadruplex structures, which are widely present on
genomes (estimated to be present at more than 370,000 locations on
the human genome), regulate organization of chromatin architecture
and initiation of DNA replication. Our major goal is to establish a novel
principle of the genome by elucidating the fundamental and universal
functions of G-quadruplex andother non-B type DNA structuresin regulation of various genome
functions. Through these efforts,
we will also explore the possibility
that mutations found in various
diseases including cancer andneurodegenerative diseases are related
to alteration and mal-formation of these
non-B DNA structures, which are likely
to be essential components of genomes
but somehow have been disregarded in
the past.

Genome Dynamics
09

Department of Genome Medicine

Genome Dynamics
10Department of Genome Medicine
Project
LeaderYoshiaki Kikkawa Mammalian Genetics Project

Gene discovery:
Phenotype- and gene-driven
approaches to identify
disease-associated genes in mice
Yasuda SP, Miyasaka Y, andKikkawa Y. (2018) “Effects of genetic
background on susceptibility and theacceleration of hearing loss in mice.”
An Excursus into Hearing Loss
3–23.
Wada K, Saito J, Yamaguchi M, Seki
Y, Furugoria M, Takahashi G, Nishito
Y, Matsuda H, Shitara H, and Kikkawa
Y. (2018) “Pde6brd1 mutation modifies
cataractogenesis in Foxe3 rct mice.”
Biochem. Biophys. Res. Commun.
496: 231-237.

Mouse disease models have contributed to the study of pathogenic
mechanisms. The demand for mouse disease models will continue
to increase because the genetic factors and molecular mechanisms
behind many human genetic diseases are still unknown. Mouse diseasemodels are important tools for identifying genes that are responsible forgenetic diseases. They are also important for studying the processes
that regulate the onset of genetic diseases and for evaluating theeffectiveness of new drugs. We aim to develop novel mouse diseasemodels via forward and reverse genetics for the phenotypic analysis ofhuman genetic diseases and for the study of pathogenic mechanisms.

Seki Y, Miyasaka Y, Suzuki S, WadaK, Yasuda SP, Matsuoka K, Ohshiba
Y, Endo K, Ishii R, Shitara H, Kitajiri
SI, Nakagata N, Takebayashi H, andKikkawa Y. (2017) “A novel splicesite mutation of myosin VI in miceleads to stereociliary fusion causedby disruption of actin networks in theapical region of inner ear hair cells.”
PLoS One 12, e0183477.
Miyasaka Y, Shitara H, Suzuki S,
Yoshimoto S, Seki Y, Ohshiba Y,
Okumura K, Taya C, Tokano H,
Kitamura K, Takada T, Hibino H,
Shiroishi T, Kominami K, Yonekawa H,
and Kikkawa Y. (2016) “Heterozygous
mutation of Ush1g/Sans in micecauses early-onset progressive
hearing loss, which is recoveredby reconstituting the strain-specificmutation in Cdh23.” Hum. Mol.
Genet. 25: 2045-2059.

“We are trying to identify genes associatedwith human diseases
using mutant mice and are aiming to develop
new mouse models for human disease.”

Kikkawa Y, and Miyasaka Y. (2016)“Genetic modifiers of hearing loss
in mice: The case of phenotypicmodification in homozygous Cdh23ahl
age-related hearing loss.” Monogr.
Hum. Genet. 20: 97–109.
Suzuki S, Ishikawa M, Ueda T,
Ohshiba Y, Miyasaka Y, Okumura K,
Yokohama M, Taya C, Matsuoka K,
and Kikkawa Y. (2015) “Quantitative
trait loci on chromosome 5 forsusceptibility to frequency-specificeffects on hearing in DBA/2J mice.”Exp. Anim. 64: 241-251.

Mammalian Genetics
11Department of Genome Medicine

Mammalian Genetics
12Department of Genome Medicine
Project
LeaderFumihiko Yasui Viral Infectious Diseases ProjectControl of viral infectious diseases:
Virology, immunology, vaccinology
and therapy

Tokunaga Y, Osawa Y, Ohtsuki T,
Hayashi Y, Yamaji K, Yamane D, Hara
M, Munekata K, Tsukiyama-Kohara K,
Hishima T, Kojima S, Kimura K, andKohara M. (2017) “Selective inhibitorof Wnt/β-catenin/CBP signalingameliorates hepatitis C virus-induced
liver fibrosis in mouse model.” Sci.
Rep. 7: 325.
Sanada T, Hirata Y, Naito Y,
Yamamoto N, Kikkawa Y, Ishida Y,
Yamasaki C, Tateno C, Ochiya T, andKohara M. (2016) “Transmission of
HBV DNA mediated by ceramidetriggered extracellular vesicles.”Cell Mol. Gastroenterol Hepatol. 3:
272-283.
Yasui F, Itoh Y, Ikejiri A, Kitabatake M,
Sakaguchi N, Munekata K, Shichinohe
S, Hayashi Y, Ishigaki H, Nakayama
M, Sakoda Y, Kida H, Ogasawara K,
and Kohara M. (2016) “Sensitization
with vaccinia virus encoding H5N1
hemagglutinin restores immune
potential against H5N1 influenza
against H5N1 influenza virus.” Sci.
Rep. 6: 37915.

Our project studies the virology, immunology, vaccinology and therapy
of incurable viral diseases. We currently focus on liver diseases,
influenza and dengue fever. However, the lack of suitable infection
models in in vitro and in vivo has hampered the clarification ofpathogenesis by these virus infections. To overcome the problems, wehave been developing various animal models including transgenic mice,
humanized mice with human liver cells, monkeys and tree shews. We
also investigate the precise mechanisms by which host factors regulate
viral growth.

“We are studying to clarify the mechanisms
underlying development of severe acuteinflammation and establishment of chronic
infection by viruses through the developmentof suitable animal models that are capable ofinfecting viruses.”

Sanada T, Tsukiyama-Kohara K,
Yamamoto N, Ezzikouri S, Benjelloun
S, Murakami S, Tanaka Y, Tateno C,
and Kohara M. (2016) “Property ofhepatitis B virus replication in Tupaia
belangeri hepatocytes.” Biochem.
Biophys Res. Commun. 469: 229235.
Yamamoto N, Sato Y, Munakata T,
Kakuni M, Tateno C, Sanada T, Hirata
Y, Murakami S, Tanaka Y, Chayama K,
Hatakeyama H, Hyodo M, HarashimaH, and Kohara M. (2016) “Novel
pH-sensitive multifunctional envelopetype nanodevice for siRNA-based
treatments for chronic HBV infection.”
J. Hepatol. 64: 547-555.
Ohtsuki T, Kimura K, Tokunaga Y,
Tsukiyama-Kohara K, Tateno C,
Hayashi Y, Hishima T, and Kohara M.
(2015) “M2 Macrophages Play CriticalRoles in Progression of Inflammatory
Liver Disease in Hepatitis C Virus
Transgenic Mice.” J. Virol. 90: 300307.

Viral Infectious Diseases
13Department of Genome Medicine

Members

Michinori Kohara
Tsubasa Munakata
Yasuyuki Miyazaki
Daisuke Yamane
Kenzaburo Yamaji
Naoki Yamamoto
Takahiro Ohtsuki
Yuko Tokunaga
Takahiro Sanada
Tomoko Honda

Viral Infectious Diseases
14Department of Genome Medicine
Project
LeaderSatoshi Koike Neurovirology ProjectProtect the Central Nervous
System from infectious
Diseases
Kobayashi K, et al. (2018) “Amino
Acid Variation at VP1-145 ofEnterovirus 71 Determines Attachment
Receptor Usage and Neurovirulencein Human Scavenger Receptor B2
Transgenic Mice.” J. Virol., 92:(15)e00681-18
Fujii K, et al. (2018) “VP1 Amino Acid
Residue 145 of Enterovirus 71 Is a Key
Residue for Its Receptor Attachment
and Resistance to Neutralizing
Antibody during Cynomolgus Monkey
Infection.” J. Virol., 92:(15)e00682-18
Fujii K, et al. (2013) “Transgenic
mouse model for the study ofenterovirus 71 neuropathogenesis.”
Proc. Natl. Acad. Sci. USA., 110:14753-14758
Yamayoshi S, et al. (2013) “Functional
Comparison of SCARB2 and PSGL1
as Receptors for Enterovirus 71.” J.
Virol., 87:3335-3347
Yamayoshi S, et al. (2012) “Human
SCARB2-dependent Infection by
Coxsackievirus A7, A14, A16 andEnterovirus 71.” J. Virol., 86:56865696
Abe Y, et al. (2012) “The Toll-likereceptor 3-mediated antiviral response
is important for protection against
poliovirus infection in poliovirus
receptor transgenic mice.” J. Virol.,
86:185-194“The development of vaccine and anti-viral drugs
and that of experimental models for the evaluation ofthese agents are important for controlling emerging
and re-emerging viral infections. We will study thebasic principles of neurotropic enterovirus infection
and provide knowledge and technologies to control
infectious diseases.”
Enterovirus 71 (EV71) is a human enterovirus species A of the genus
Enterovirus within the Picornaviridae family, and it is known to be one of
the causative agents of hand-foot-and-mouth disease (HFMD). HFMD
is considered to be a mild and self-limiting disease in general. However,in some infants and young children, HFMD caused predominantly by
EV71 can be complicated by neurological manifestations. Thus, EV71
infection is a serious public health concern. Unfortunately, there is still
very little information concerning EV71 pathogenesis, and vaccines oranti-EV71 drugs have yet to be developed.

Members

Yamayoshi S, Koike S. (2011)“Identification of the Human SCARB2
Region That Is Important forEnterovirus 71 Binding and Infection.”
J. Virol., 85: (10) 4937-4936
Yamayoshi S, et al. (2009) “Scavenger
receptor B2 is a cellular receptor forenterovirus 71.” Nature Medicine
15:789-801

Neurovirology
15

Kyosuke Kobayashi

Department of Genome Medicine

Research Topics

Neurovirology
16Department of Genome Medicine
Project
LeaderTakachika Hiroi Allergy and Immunology Project

Allergy and Mucosal Immunology:
Investigating molecular mechanisms ofsublingual immunotherapy (SLIT) anddeveloping therapeutic biomarkers
for allergic diseases.

Gotoh M, Kaminuma O, Nakaya A,
Katayama K, Motoi Y, Watanabe N,
Saeki M, Nishimura T, Kitamura N,
Yamaoka K,Okubo K, and Hiroi T.
(2017) “Identification of biomarker
sets for predicting the efficacy of
sublingual immunotherapy againstpollen-induced allergic rhinitis.”
International Immunology 29: 291300.
Nishimura T, Kaminuma O, Saeki M,
Kitamura N, Matsuoka K, Yonekawa
H, Mori A, and Hiroi T. (2016)“Essential contribution of CD4+
T cells to antigen-induced nasal
hyperresponsiveness in experimental
allergic rhinitis.” PLOS ONE 11:
e0146686.
Yokoyama S, Takada K, Hirasawa
M, Perera LP, and Hiroi T. (2011)“Transgenic mice that overexpress
human IL-15 in enterocytes
recapitulate both B and T cellmediated pathologic manifestations ofceliac disease.” J. Clin. Immunol. 31:
1038-1044.
Kaminuma O, Kitamura F, Miyatake
S, Yamaoka K, Miyoshi H, Inokuma
S, Tatsumi H, Nemoto S, Kitamura N,
Mori A, and Hiroi T. (2009) “T-box 21transcription factor is responsible for
distorted TH2 differentiation in humanperipheral CD4+ T cells.” J. Allergy
Clin. Immunol. 123: 813-823.

Japanese cedar pollen allergy is the major allergic
disease in Japan, and the approximately 35% ofJapanese people are affected. In recent years,
sublingual immunotherapy has been recognized as aneffective curative treatment for the allergic diseases.
However, the molecular mechanisms of mucosal
tolerance still remain unclear. In our laboratory, we
focus on the following subjects.

“We are developing new diagnostics
and treatments for allergies.”
1. Search for effective biomarkers of SLIT
・CNVs
・SNPs
・Epigenome
・Proteome, etc.

2. Elucidation of molecular mechanisms to induce
immunological tolerance by SLIT

Yokoyama S, Watanabe N, Sato N,
Filkoski L, Tanaka T, Miyasaka M,
Waldmann TA, Hiroi T, and Perera
PL. (2009) “Antibody-mediatedblockade of IL-15 signaling reverses
autoimmune intestinal damage ina mouse model of celiac disease.”Proc. Natl. Acad. Sci. USA 106:
15849-15854.

Allergy and Immunology
17・iTregs
・Apoptosis
・CTLA-4・TGF-β・IL-10 etc.

Department of Genome Medicine

Allergy and Immunology
18

Department of Genome Medicine
Project
LeaderMedical Research
Futoshi Shibasaki Molecular
Project

Translational Research for Cancer andInfectious Diseases: Basic to Applied Science

Li Q, et al. (2018) “Int6/eIF3e Silencing
Promotes Placenta Angiogenesis ina Rat Model of Pre-eclampsia.” Sci.
Reports 12, 8(1):8944

Recent discoveries of biomarkers and novel technologies have opened
the new aspects of the mechanisms and drug developments especiallyin cancer and infectious diseases. In basic research, we have been
focusing on the mechanisms of cancer angiogenesis and the drug
development using siRNA, and on malignant transformation andmetastasis caused by cell fusion. In addition, the novel mechanisms forH5 influenza virus entrance into cell surface would be a drug target.

Endo F, et al. (2017) “Development ofa simple and quick immun-chromatography method for detection of antiHPV-16/-18 antibodies.” PLoS One.
12(2):e0171314.
Sakurai A, et al. (2015) “Fluorescentimmunochromatography for rapid andsensitive typing of seasonal influenza
viruses.” PLoS One. 10(2):e0116715.
Nakano S, et al. (2015)“Immunochromato-graphic Detectionof Serum Anti-a-Galactosidase A
Antibodies in Fabry Patients after
Enzyme Replacement Therapy.”
PLoS One. 10(6):e0128351.
Hashimoto T and Shibasaki F. (2015)“Hypoxia-inducible factor as anangiogenic master switch.” Front.
Pediatr. 3:33.
Sakurai A, et al. (2014) “Multi-coloredimmunochromatography using
nanobeads for rapid and sensitivetyping of seasonal influenza viruses.”J. Virol. Methods. 209:62-68.
Sakurai A, et al. (2013) “Broadspectrum detection of H5 subtype
influenza A viruses with a new
fluorescent immunochromatographysystem.” PLoS One. 8(11):e76753.

In clinical and translational research, we focus on the establishment ofplatform to perform “Precision Medicine” by Whole genome analysis
with next generation sequence in collaboration with Metropolitan
Hospitals. For Private Public Partnership (3P), we have alreadyestablished the Bio-Consortium “Tokyo Biomarker Innovation Research
Association” (TOBIRA).
Our specific aims are to perform the basic science and be to develop
the new findings to the translational research.

Nakano S, et al. (2013) “Developmentof a highly sensitive immuno-PCR
assay for the measurement of
a-galactosidase A protein levels inserum and plasma.” PLoS One.
8(11):e78588.
Li Chen, et al. (2007) “Mammalian
Tumor Suppressor Int6 SpecificallyTargets HIF-2alfa To Degradation by
Hypoxia- And pVHL- Independent
Regulation.” J. Biol. Chem. 282.
12707.
Chen L, et al. (2010) “Int6/eIF3eSilencing Promotes Functional Blood
Vessel Outgrowth and Enhances
Wound Healing by Upregulating
HIF2-a Expression.” Circulation 122:
910-919.

Molecular Medical Research19Department of Genome Medicine

Molecular Medical Research
20Department of Dementia and Higher Brain Function
Project
LeaderMasato Hasegawa Dementia Research Project

Prion-like propagation of tau, α-synuclein
and TDP-43 in neurodegenerative diseases

Shimozawa A, Ono M, Takahara
D, Tarutani A, Imura S, MasudaSuzukake M, Higuchi M, Yanai K,
Hisanaga SI, and Hasegawa M .
(2017) “Propagation of pathological
α-synuclein in marmoset brain.” Acta.
Neuropathol. Commun. 5:12.
Hasegawa M, Nonaka T, and MasudaSuzukake M. (2016) “α-Synuclein:
Experimental Pathology.” Cold
Spring Harb Perspect Med. 6. pii:
a024273.
Tarutani A, Suzuki G, Shimozawa
A, Nonaka T, Akiyama H, Hisanaga
S, and Hasegawa M. (2016) “The
Effect of Fragmented Pathogenic
α-Synuclein Seeds on Prion-like
Propagation.” J. Biol. Chem.
291:18675-18688.
Tanaka Y, Nonaka T, Suzuki G,
Kametani F, and Hasegawa M. (2016)“Gain-of-function profilin 1 mutations
linked to familial amyotrophic lateralsclerosis cause seed-dependent
intracellular TDP-43 aggregation.”
Hum. Mol. Genet. 25:1420-1433.
Shimonaka S, Nonaka T, Suzuki G,
Hisanaga S, and Hasegawa M. (2016)“Templated Aggregation of TAR DNAbinding Protein of 43 kDa (TDP-43) bySeeding with TDP-43 Peptide Fibrils.”
J. Biol. Chem. 291:8896-8907.
Taniguchi-Watanabe S, Arai T,
Kametani F, Nonaka T, MasudaSuzukake M, Tarutani A, Murayama
S, Saito Y, Arima K, Yoshida M,
Akiyama H, Robinson A, Mann D,
Iwatsubo T, and Hasegawa M.
(2016) “Biochemical classification oftauopathies by immunoblot, protein
sequence and mass spectrometricanalyses of sarkosyl-insoluble
and trypsin-resistant tau.” Acta.
Neuropathol. 131: 267-280.

Neurodegenerative diseases are characterized by progressive
degeneration of subsets of neurons and gliosis. Many of the diseases
are accompanied with intracellular amyloid-like protein pathologies,
such as tau in Alzheimer’s disease (AD), α-synuclein in dementia with
Lewy bodies (DLB) and TDP-43 in amyotrophic lateral sclerosis (ALS)
and frontotemporal dementias (FTLD). Importantly, the distributions andspread of these proteins are closely correlated with clinical presentationand disease progression. However, little attention had been given to
the questions of why these diseases are progressive, and why thepathologies spread to different brain regions during the course of the

Tau in AD

α-synuclein in DLB“Emerging evidence indicates that intracellularamyloid-like proteins have prion-like properties andpropagate from cell to cell by converting normal
proteins into abnormal forms.
We are trying to elucidate the molecular mechanisms of‘prion-like propagation of these proteins.”
diseases.We have been investigating these intracellular abnormal
proteins in brains of patients, proteinchemically using LC/MS/MS,
immuno-histochemically with specific antibodies and ultrastructuraly.
And we found that all of these proteins are accumulated in brains ofpatients as fibrous or filamentous forms in hyperphosphorylated andpartially ubiquitinated states.

Schematic diagrams of human tau, α-synuclein and TDP-43.

Dementia Research
21

TDP-43 in ALS

Department of Dementia and Higher Brain Function

To investigate the molecular mechanisms of aggregation of these proteins, we established seed-induced
aggregation model which recapitulate the pathological protein aggregation in vitro, cultured cells and inbrains of animals (mouse and marmoset) and proposed a hypothesis “prion-like propagation of these
intracellular pathological proteins in brain”. These models are highly useful not only for clarifying themolecular mechanisms involved in the pathogenesis and progression of neurodegenerative diseases
but also for the development of disease modifying drugs and therapy.

Senior Research Scientist

Takashi Nonaka

Molecular mechanisms of cell-to-cell
propagation of aggregated proteins

Nonaka T, et al. Phosphorylationof TAR DNA-binding Protein of
43 kDa (TDP-43) by TruncatedCasein Kinase 1 Triggers
Mislocalization and Accumulationof TDP-43. J. Biol. Chem. 291:
5473-5483, 2016

I’m studying molecular mechanisms of cell-to-cell propagationof aggregated proteins (tau, α-synuclein and TDP-43) inneurodegenerative diseases. Also, I’m trying to make in vitro and invivo models recapitulating abnormal features found in cells of brainsof patients using cultured cells and mice. These models will contributenot only to a better understanding of the mechanisms involved in these
diseases, but also to the development of novel therapeutic strategies.

Nonaka T, et al. Prion-like
properties of pathological TDP-43
aggregates from diseased brains.
Cell Rep. 4: 124-134 , 2013
Nonaka T, et al. Seeded
aggregation and toxicity of alphasynuclein and tau: cellular modelsof neurodegenerative diseases. J.
Biol. Chem. 285: 34885-34898,
2010

Dementia Research22Department of Dementia and Higher Brain Function
Project
LeaderMinoru Saitoe Learning and Memory Project

Investigating the Molecular
Mechanisms that Generate Memoryencoding Neural Networks

Ueno K, Suzuki E, Naganos S, Ofusa
K, Horiuchi J, and Saitoe M. (2017).
“Coincident postsynaptic activity
gates presynaptic dopamine release
to induce plasticity in Drosophila
mushroom bodies.” eLife, 6: e21076.
Hirano Y, Ihara K, Masuda T,
Yamamoto T, Iwata I, Takahashi A,
Awata H, Nakamura N, Takakura
M, Suzuki Y, Horiuchi J, Okuno
H, and Saitoe M. (2016). “Shifting
transcriptional machinery is required
for long-term memory maintenanceand modification in Drosophila
mushroom bodies.” Nat. Commun.
7: 13471.
Matsuno M, Horiuchi J, Yuasa Y,
Ofusa K, Miyashita T, Masuda T, andSaitoe M. (2015). “Long-term memory
formation in Drosophila requires
training-dependent glial transcription.”
J. Neurosci. 35: 5557-5565.
Yamazaki D, Horiuchi J, Ueno K, Ueno
T, Saeki S, Matsuno M, Naganos
S, Miyashita T, Hirano Y, Nishikawa
H, Taoka M, Yamauchi, Y, Isobe T,
Honda Y, Kodama T, Masuda T, andSaitoe M. (2014). “Glial dysfunction
causes age-related memory
impairment in Drosophila.” Neuron
84: 753-763.
Hirano Y, Masuda T, Naganos S,
Matsuno M, Ueno K, Miyashita T,
Horiuchi J, and Saitoe M. (2013).
“Fasting Launches CRTC to Facilitate
Long-term Memory Formation in
Drosophila.” Science 339: 443-446.
Miyashita T, Oda Y, Horiuchi J, Yin
JC, Morimoto T, and Saitoe M. (2012).
“Mg2+ block of Drosophila NMDA
receptors is required for long-term
memory formation and CREBdependent gene expression.” Neuron
74: 887-898.
Yamazaki D, Horiuchi J, Nakagami Y,
Nagano S, Tamura T, and Saitoe M.
(2007). “The Drosophila DC0 mutation
suppresses age-related memory
impairment without affecting lifespan.”Nat. Neurosci. 10: 478-484.
Tamura T, Chiang AS, Ito N, Liu HP,
Horiuchi J, Tully T, and Saitoe M.
(2003) “Aging specifically impairs
amnesiac-dependent memory in
Drosophila.” Neuron 40: 1003-1011.

Memories mold our personalities to make us who we are. Using
powerful genetic tools, numbers of genes and neural substrates
underlying memory-associated behaviors have been identified in
Drosophila. Given these scientific backgrounds, we have investigatedwhen, where and how identified memory-associated gene products
function to produce memory-based behavior, and how the underlying
mechanism is changed in response to changes in physical condition
such as aging.
In addition to behavioral genetic approach, we employ in vivo and exvivo imaging techniques to characterize physiological properties ofmemory-associated neural networks, and understand how memoryassociated genes and neuromodulatory systems regulate functionof these networks; how sensory information is associated and how
memory information is stored in neural substrates and recalled uponreceiving test stimuli.

“Combining behavioral genetics and state-of-arts
imaging techniques, we aim to understanding how
our brains form, store and retrieve memory.”

Flies perform olfactory conditioning
behavior, avoiding conditioned odor that
had been paired with electrical shock (left)
in the teaching machine (right)

Left, in vivo imaging of fixed fly under microscope is used to investigate activity of identified
neurons and network during sensory association (odor and shock), reinforcement, storage andretrieval. Right, using ex vivo imaging we attempt to make artificial memory in cultured brain,
thereby elucidate the whole picture of the memory-associated networks.

Learning and Memory23

Department of Dementia and Higher Brain Function

Current Research Topics
Encoding and decoding of memory: In Drosophila,
the mushroom body (MB) is a neural center for olfactory
memory. As described by Hermann Ebbinghaus (1885),
repetitive olfactory conditioning with rest intervals, namely
spaced training, stabilizes labile short-term memory (STM)
into robust long-term memory (LTM), which requires
Structure of MBs and their lobetranscriptional activity of CREB. Interestingly, STM andLeft: MBs in the fly brain expressing GFP.
LTM are encoded in different subset of MB neurons. While
Right: Subdivision of MB lobes derived from eachtype of MB neurons.
aversive STM is encoded in gamma neurons, aversive LTM
is encoded in alpha/beta neurons. We are interested in
how such anatomical shifting is occurred during stabilization of STM to LTM, functional relationshipbetween STM and LTM.
Neuron-glia interactions: Recent researchdemonstrates that neuron-glia communication is also
important for memory formation. We have identified
a cell adhesion molecule Klingon (Klg) that mediates
neuron-glia communication required for LTM-based
behavior. Currently, we are studying how Klg-mediatedneuron-glia interaction regulates memory acquisition,
stabilization and retrieval. Also, we are interested in
how this mechanism is altered upon aging.
Klingon mediates neuron-glia interaction for LTM formation. A, B) Expression of Klingon (Klg) in the fly
brain. C) Expression of Klg in neurons. D) Expression of Klg in glial cells.
Spaced training increases Klg-mediated neuron-glia interaction, thereby induces Repo-dependent glialtranscription required for LTM.
Senior Research Scientist

Kohei Ueno

Exploring Physiological Systems
Underlying Learning and Memory
Neural plasticity in the MBs is believed to be a cellular basis of olfactory
memory. To understand how odor and shock information are associatedto produce plastic changes in the MB neurons, we developed ex vivo
brain imaging system. Using this system, we found that simultaneous
stimulation of odor and shock input pathways to the MBs produces
long-term enhancement (LTE) in MB neurons in a manner dependent
on activity of D1 receptor in the MBs. We further discovered a novel
mode of dopamine release locally evoked by postsynaptic MB neurons
which have been coincidentally activated by odor and shock input
pathways. We have investigated how coincidentally activated MB
neurons direct dopamine release and whether such on-demand releasemode also takes place for other neuromodulators and other animals.
If mushroom body neurons are activated by two inputs, namely odor
and somatosensory inputs, the activated mushroom body requires
dopamine release from dopaminergic neurons.

Learning and Memory24Department of Dementia and Higher Brain Function
Project
LeaderYukio Nishimura Neural Prosthesis Project

Restoring Lost Function
After Neural Damage
Our research goal is to conceive innovative neuro-rehabilitation to
restore lost functions after impairment of central nervous system, and totranslate our findings into clinical applications capable of improving thequality of life for individual with neural damages.
Kato K, Sasada S, and Nishimura
Y. (2016) “Flexible adaptation to an
artificial recurrent connection frommuscle to peripheral nerve in man.” J.
Neurophysiol. 115(2):978-991.
Sawada M, Kato K, Kunieda T, Mikuni
N, Miyamoto S, Onoe H, Isa T, andNishimura Y (2015) “Function of thenucleus accumbens in motor control
during recovery after spinal cord
injury.” Science. 350(6256):98-101.
Sasada S, Kato K, Kadowaki S, Groiss
SJ, Ugawa Y, Komiyama T, andNishimura Y. (2014) “Volitional walking
via upper limb muscle-controlled
stimulation of the lumbar locomotor
center in man.” J. Neurosci.
34(33):11131-11142.
Nishimura Y, Perlmutter SI, Eaton
RW, and Fetz EE. (2013) “Spiketiming-dependent plasticity in primate
corticospinal connections induced
during free behavior.” Neuron.
80(5):1301-1309.

“Bridging Damaged Neural Pathways
using a Neural Interface.”
Regaining the function of an impaired
limb is highly desirable in individuals
experiencing paralysis. Functional lossof limb control in individuals with spinal
cord injury or stroke can be caused bytransection of descending and ascending
pathways connecting cortical to spinal
network, although neural circuits thatlocate above and below the impaired site remains their function.

Translational Study
Artificial NeuralConnection

Neural Mechanism

Neuron 2013

Frontiers 2013

Nishimura Y, Perlmutter SI, and Fetz
EE. (2013) “Restoration of upper limb
movement via artificial corticospinal
and musculospinal connections in amonkey with spinal cord injury.” Front.
Neural Circuits. 7:57.
Nishimura Y, Morichika Y, and Isa
T. (2009) “A subcortical oscillatory
network contributes to recovery ofhand dexterity after spinal cord injury.”
Brain. 132(Pt 3):709-721
Nishimura Y, Onoe H, Morichika
Y, Perfiliev S, Tsukada H, and Isa
T. (2007) “Time-dependent centralcompensatory mechanisms of finger
dexterity after spinal cord injury.”
Science. 318(5853):1150-1155.

We are developing a neural interface which so-called “artificial neuronalconnection (ANC)”. The ANC bridges supra-spinal system andspinal network beyond the lesion site to restore lost function. We areconducting clinical trials to assess effectiveness of ANC in restoring
motor function in paralyzed patients. We investigate neural changes that
occur during recovery.

Neural Prosthesis
25

J Neurosci 2014

Department of Dementia and Higher Brain Function

Neural Mechanisms of Functional Recovery
Using a large scale multichannel recording, pharmacologicalintervention, neuroanatomy, viral tools, computation
and whole brain imaging, we seek to uncover the neuralmechanisms underlying the voluntary limb movement inintact animals, as well as the processes in which motor
functions are reestablished after neural damages such as
spinal cord injury and stroke. We are also performing clinicalstudies to test the efficacy of the ANC in human patients.

Psychological Effect on Motor Control
Mental Effort

Mesolimbic

Motor

Science. 2007, Brain 2009

Emotional states influence how we perform
motor activities and how we perceive errors.
Depression impedes and motivation enhances
functional recovery after neuronal damage.
However, the neuronal substrate underlying such
psychological effects on functional recovery
remains unclear. We investigate the neuralsubstrate underlying such psychological effects
on motor performance in human and animalmodel of neural damages.

Motor Control
PLoS ONE 2011, Science. 2015

Members
Yukio Nishimura
Toshiki Tazoe

Yoshihisa Nakayama
Hiroaki Ishida

Osamu Yokoyama Michiaki Suzuki
Nobuya Sano

Miki Kaneshige

Kei Obara

Naoya Kabe

Noboru Usuda
Yu Shimada

Ryoutaro Numata

Neural Prosthesis
26

Department of Dementia and Higher Brain Function

Unit Leader

Makoto Hashimoto Parkinson’s disease Unit

Protection ofneurodegenerative diseases
Research description

Hashimoto M, Ho G, Takamatsu Y,
Shimizu Y, Sugama S, Takenouchi
T, Waragai M, and Masliah E. (2018)“Evolvability and Neurodegenerative
Disease: Antagonistic Pleiotropy
Phenomena Derived from Amyloid
Aggregates.” J. Parkinsons Dis.
8(3):405-408.
Hashimoto M, Ho G, Sugama S,
Takamatsu Y, Shimizu Y, Takenouchi
T, Waragai M, and Masliah E. (2018)“Evolvability of Amyloidogenic Proteinsin Human Brain.” J. Alzheimers Dis.
62:73-83.
Waragai M, Ho G, Takamatsu Y,
Sekiyama K, Sugama S, Takenouchi
T, Masliah E, and Hashimoto M.
(2017) “Importance of adiponectin
activity in the Pathogenesis of
Alzheimer's Disease.” Ann. Clin.
Transl. Neurol. 4:591-600.
Takamatsu Y, Ho G, Koike W, Sugama
S, Takenouchi T, Waragai M, Wei J,
Sekiyama K, and Hashimoto M. (2017)“Combined immunotherapy with“anti-insulin resistance” therapy as
a novel therapeutic strategy against
neurodegenerative diseases.” NPJ
Parkinson’s Disease 3: 4.
Takamatsu Y, Koike W, Takenouchi
T, Sugama S, Wei J, Waragai
M, Sekiyama K, and Hashimoto
M. (2016) “Protection against
neurodegenerative disease on Earthand in space.” NPJ Microgravity 2:
16013.
Waragai M, Adame A, Trinh I,
Sekiyama K, Takamatsu Y, Une K,
Masliah E, and Hashimoto M. (2016)“Possible Involvement of Adiponectin,
the Anti-Diabetes Molecule, in the
Pathogenesis of Alzheimer's Disease.”
J. Alzheimers Dis. 52:1453-1459.
Sekiyama K, Takamatsu Y, Koike W,
Waragai M, Takenouchi T, Sugama
S, and Hashimoto M. (2016) “Insight
into the Dissociation of Behavior
from Histology in Synucleinopathies
and in Related Neurodegenerative
Diseases.” J. Alzheimers Dis. 52:831841.

The number of patients with ageassociated neurodegenerative diseases,
such as Alzheimer’s disease (AD) andParkinson’s disease (PD), is rapidly
increasing worldwide. Consequently, hugecosts for medical treatment and nursing
care for these patients have becomea serious socioeconomic dilemma.
Nonetheless, extensive studies of amyloid
immunotherapy in AD have been so far
unsatisfactory. Thus, the development ofan effective disease-modifying therapy is
the highest priority in neurodegenerative
disease research.

In our laboratory, we seek to exploit a mechanism-based diseasemodifying strategy for a-synucleinopathies, such as PD and dementia
with Lewy bodies. In this context, we have a particular interest in the
suppressive effect of adiponectin on neurodegeneration (Fig.1). We
will also attempt to identify new molecules that could be useful for theprevention of neurodegenerative diseases. For such purposes, wecurrently perform the Drosophila molecular genetics (Fig.2) in addition
to cell biological and transgenic mice studies. Apparently, the results will
be applicable to other diseases, including AD and Huntington’s disease.

Members
Yoshiki Takamatsu
Masaaki Waragai
Hiromu Sugino
Ryoko Wada

Parkinson’s disease
27

Fig. 1
Effect of adiponectin on
neurodegeneration in tg mice

Fig. 2 Drosophila molecular genetics

28

Department of Brain Development and Neural Regeneration
Project
LeaderHiroshi Sakuma Developmental
Neuroimmunology Project

Towards a Better Understanding of
Neuro-immune Interactionsin the Developing Brain

Saika R, Sakuma H, Noto D,
Yamaguchi S, Yamamura T, andMiyake S. (2017) “MicroRNA-101aregulates microglial morphology andinflammation.” J. Neuroinflammation
14:109Nakahara E, Sakuma H, KimuraKuroda J, Shimizu T, Okumura A,
and Hayashi M. (2015) “A diagnostic
approach for identifying anti-neuronalantibodies in children with suspectedautoimmune encephalitis.” J.
Neuroimmunol. 285:150-155.
Sakuma H, Tanuma N, Kuki I,
Takahashi Y, Shiomi M, and Hayashi
M. (2015) “Intrathecal overproductionof proinflammatory cytokines andchemokines in febrile infectionrelated refractory status epilepticus.”
J. Neurol. Neurosurg. Psychiatr.
86:820-822.
Noto D & Sakuma H (double firstauthors), Takahashi K, Saika R, SagaR, Yamada M, Yamamura T, andMiyake S. (2014) “Development of a
Culture System to Induce Microglialike Cells from Haematopoietic Cells.”
Neuropathol. Appl. Neurobiol.
40:697-713.

Our research focuses on the role ofimmune system in the developing brain.
Immune and inflammatory responses
not only combat pathogens but also
play a variety of physiological roles in
the central nervous system.
Microglia are brain-resident immune
cells and play multiple roles in theprotection from pathogens and the clearance of debris. In addition,
recent studies have shed light on unexpected functions of microglia in
the physiological condition. For example, microglia actively participate
in the brain development by modulating synapses.

“We are investigating the mechanisms bywhich microglia maintain homeostasis in
the developing brain.”
Our main research areas include:
1)
2)
3)
4)
5)

Development and differentiation of microglia
Neuron-microglia interaction
In-vitro differentiated myeloid cells for cell therapy
Autoantibodies associated with neurological diseases
New biomarkers for pediatric immune-mediated neurological diseases

Sakuma H, Awaya Y, Shiomi M,
Yamanouchi H, Takahashi Y, Saito Y,
Sugai K, and Sasaki M. (2010) “Acuteencephalitis with refractory, repetitive
partial seizures (AERRPS): a peculiar
form of childhood encephalitis.” Acta
Neurol. Scand. 121:251-256.

Developmental Neuroimmunology
29

Department of Brain Development and Neural Regeneration

Developmental Neuroimmunology
30Department of Brain Development and Neural Regeneration
Project
LeaderKanato Yamagata Synaptic Plasticity Project

Synaptic Plasticity and Brain Diseases:
Elucidating the mechanism fordevelopmental epilepsy,
intellectual disability and autism

Shimada T, and Yamagata K. (2018)“Pentylenetetrazole-Induced Kindling
Mouse Model.” JoVE (136).
Shimada T, Yoshida T, and Yamagata
K. (2016) “Neuritin Mediates ActivityDependent Axonal Branch Formationin Part via FGF Signaling.” J.
Neurosci. 36(16):4534-4548.
Sugiura H, Yasuda S, Katsurabayashi
S, Kawano H, Endo K, Takasaki K,
Iwasaki K, Ichikawa M, Kobayashi
T, Hino O, and Yamagata K.
(2015) “Rheb activation disrupts
spine synapse formation throughaccumulation of syntenin in tuberous
sclerosis complex.” Nat. Commun.
6:6842.
Masui K, Tanaka K, Ikegami S, Villa
GR, Yang H, Yong WH, Cloughesy TF,
Yamagata K, Arai N, Cavenee WK,
and Mischel PS. (2015) “Glucosedependent acetylation of Rictor
promotes targeted cancer therapy
resistance.” Proc. Natl. Acad. Sci.
USA 112(30):9406-9411.
Shimada T, Takemiya T, Sugiura
H, and Yamagata K. (2014) “Roleof Inflammatory mediators in thepathogenesis of epilepsy.” Mediators
Inflamm. 2014:901902.
Yasuda S, Sugiura H, Katsurabayashi
S, Shimada T, Tanaka H, Takasaki
K, Iwasaki K, Kobayashi T, Hino O,
and Yamagata K. (2014) “Activationof Rheb, but not of mTORC1, impairs
spine synapse morphogenesis in
tuberous sclerosis complex.” Sci.
Rep. 4:5155.
Kim SY, Yasuda S, Tanaka H,
Yamagata K, and Kim H. (2011) “Nonclustered protocadherin.” Cell Adh.
Migr. 5(2):97-105.

Our project examines the molecular basis of activity-dependent synaptic
plasticity. In particular, we have cloned a set of immediate early genes(IEGs) that are rapidly transcribed in neurons involved in information
processing, and that are essential for long term memory. IEG proteinscan directly modify synapses and provide insight into
cellular mechanisms that support synaptic plasticity.
Furthermore, these IEG products have been shown
to be involved in developmental brain disorders,
including refractory epilepsy, intellectual disability and/or autism.

“We have clarified mechanisms ofrefractory epilepsy, intellectual
disability and/or autism caused by
impaired synaptic plasticity.
Based on the novel mechanisms
we found, we are trying to findnew treatments for developmental
brain disorders”

For example, COX-2 and mPGES-1 are prostaglandin synthases andexacerbate neuronal cell death after seizures, leading to intractableepilepsy. Arcadlin is a protocadherin and induces spine shrinkages after
seizures, resulting in developmental delay or amnesia. Rheb regulates
excitatory synapse formation via syntenin.
Its constitutive activation causes TSC (tuberous sclerosis complex),
which is accompanied by epilepsy, mental retardation and autism.
Finally, neuritin is a secreted or membrane-anchored protein andinduces neurite branching. It may be involved in temporal lobe epilepsy.
Thus, rapid de novo transcription provides novel insights into the cellularand neural network basis of behavioral plasticity.
We will also explore the possibility thatthese IEG products could be therapeutictargets for developmental disorders. We
are making genetic mouse models ofdevelopmental disorders and are testing
several drug inhibitors against IEGs forsuch diseases.

Synaptic Plasticity
31

Department of Brain Development and Neural Regeneration

Synaptic Plasticity
32

Department of Brain Development and Neural Regeneration
Project
LeaderHaruo Okado Neural Development Project

Brain Development and Maintenance:

Hirai S, Hotta K, and Okado H.
(2018) “Developmental Rolesand Evolutionary Significanceof AMPA-Type Glutamate
Receptors.” Bioessays. 2018 2018
Sep;40(9):e1800028.

Various factors control differentiation of neural stem cells and survival of
the resulting neurons and aberrancy of these processes are involved in
the incidences of intellectual disability and age-related brain disorders
and brain tumors.
We aim to elucidate the mechanisms of the development andmaintenance of brain functions and ultimately to develop methods forprevention and treatment of intractable cranial nerve diseases.

Hirai S, Hotta K, Kubo Y, Nishino A,
Okabe S, Okamura Y, and Okado H.
(2017) “AMPA glutamate receptors
are required for sensory-organ
formation and morphogenesis in thebasal chordate.” Proc. Natl. Acad.
Sci. USA. 114: 3939-3944.
Nakajima K, Hirai S, Morio T, andOkado H. (2015) “Benzodiazepines
induce sequelae in immature micewith inflammation-induced statusepilepticus.” Epilepsy & Behavior 52:
180-186.
Ohtaka-Maruyama C, Hirai S, Miwa
A, Heng JI, Shitara H, Ishii R, Taya
C, Kawano H, Kasai M, Nakajima K,
and Okado H. (2013) “RP58 regulates
the multipolar-bipolar transition of
newborn neurons in the developing
cerebral cortex.” Cell Rep. 3: 458-471.
Hirai S, Miwa A, Ohtaka-Maruyama
C, Kasai M, Okabe S, Hata Y, andOkado H. (2012) “RP58 controls
neuron and astrocyte differentiation
by downregulating the expression of
Id1-4 genes in the developing cortex.”
EMBO J. 31: 1190-1202.

Various gene-targeted mice

“We are studying the effects of various
genetic and environmental factors on themolecular mechanisms of brain
development and maintenance,with the ultimate goal of developing newtreatments mental diseases.”

Ohtaka-Maruyama C, Hirai S, Miwa
A, Takahashi A, and Okado H. (2012)“The 5’-flanking region of the RP58
coding sequence shows prominentpromoter activity in multipolar cells
in the sub- ventricular zone duringcorticogenesis.” Neuroscience 201:
67-84.
Okado H, Ohtaka-Maruyama C,
Sugitani Y, Fukuda Y, Ishida R, Hirai S,
Miwa A, Takahashi A, Aoki K, MochidaK, Suzuki O, Honda T, Nakajima K,
Ogawa M, Terashima T, Matsuda
J, Kawano H, and Kasai M. (2009)“Transcriptional repressor RP58 is
crucial for cell-division patterning andneuronal survival in the developing
cortex.” Dev. Biol. 331: 140-151.

Neural Development
33

in utero electroporation

Laboratory Members

Department of Brain Development and Neural Regeneration

Shinobu Hirai

Tomoko Tanaka

Yoshie Matsumoto

Seigi Kanzaki

Tomoko Fukuoka

Neural Development
34

Department of Brain Development and Neural Regeneration
Project
LeaderNobuaki Maeda Neural Network ProjectMechanisms of Neural Network Formation:
Neuronal migration and synapse formation

Ohtaka-Maruyama C, Okamoto M,
Endo K, Oshima M, Kaneko N, YuraK, Okado H, Miyata S, and Maeda
N. (2018) ”Synaptic transmission
from subplate neurons controls
radial migration of cortical neurons.”
Science 360: 313-317.
Maeda N. (2015) “Proteoglycans andneuronal migration in the cerebral
cortex during development anddisease.” Front Neurosci. 9: 98.
Ohtaka-Maruyama C, and Okado
H. (2015) “Molecular pathways
underlying projection neuron
production and migration during
cerebral cortical development.” Front
Neurosci. 9: 447.
Kamimura K, Ueno K, Nakagawa J,
Hamada R, Saitoe M, and Maeda
N. (2013) “Perlecan regulates
bidirectional Wnt signaling at the
Drosophila neuromuscular junction.”
J. Cell Biol. 200: 219-233.
Ohtaka-Maruyama C, et al. (2013)“RP58 regulates the multipolar-bipolartransition of newborn neurons in thedeveloping cerebral cortex.” Cell Rep.
3: 458-471.

In the developing nervous system, diverse types of neurons are
generated from neural progenitors, and migrate to specific destinations.
Then, neurons extend axons toward specific target cells and form
synapses on them. These developmental processes require complex
cell-cell and cell-extracellular matrix interactions. The extracellularmatrix is an intricate network of molecules composed of proteoglycans,hyaluronan, and fibrous glycoproteins, which fill up the extracellularspace. In the meshwork of extracellular matrix, various signal molecules
such as growth factors and chemokines are stored.

“We are interested in the roles of theextracellular matrix in neuronal network
formation. In the developing nervous system,
the extracellular matrix plays a dynamic role
in regulating the behaviors of diverse types ofneurons.”
To explore the functions of extracellular matrix in the developing neuralnetworks, we adopted two animal models: mouse cerebrum andDrosophila neuromuscular junction (NMJ). Using in utero eletroporationand live cell imaging techniques, we are investigating the migrationof excitatory neurons in the mouse neocortex. Drosophila NMJ is areadily accessible model of excitatory synapses, which resemble theglutamatergic synapses of vertebrate central nervous system. By using
the sophisticated genetic tools of Drosophila, it is possible to unravel
the complicated roles of extracellular matrix in the synapse formation.

Kamimura K, Maeda N, and Nakato
H. (2011) “In vivo manipulation of
heparan sulfate structure and its
effect on Drosophila development.”
Glycobiology 21: 607-618.
Nishimura K, Ishii M, Kuraoka M,
Kamimura K, and Maeda N. (2010)“Opposing functions of chondroitin
sulfate and heparan sulfate during
early neuronal polarization.”
Neuroscience 169: 1535-1547.
Ishii M, and Maeda N. (2008)“Oversulfated chondroitin sulfate plays
critical roles in the neuronal migrationin the cerebral cortex.” J. Biol. Chem.
283: 32610-32620.

Neural Network
35

Department of Brain Development and Neural Regeneration

Senior Research Scientist

Chiaki Ohtaka-Maruyama

Subplate layer in development andevolution of neocortex

How does mammalian neocortex acquire a unique six-layered structurethat is a structural basis for the complex neural circuits, the remarkableproduct of evolution? To approach this question, we are focusing on the
subplate (SP) neurons in the developing neocortex: one of the first bornand matured cortical neurons that disappear postnatally. Recently, we
found that SP neurons play critical roles in radial neuronal migration
by forming transient glutamatergic synapses on migrating young neurons. Moreover, SP layer isrich in extracellular matrix, which may play important regulatory roles in the neuronal polarization.
Functional elucidation of SP layer should lead to the better understanding of brain development andevolution.

Neural Network36

Department of Psychiatry and Behavioral Sciences
Project
LeaderAtsushi Nishida Mental Health Promotion Project

No health without Mental Health:Mental Health promotion as the first priority in our society

Nakanishi M, Niimura J, Yamasaki
S, and Nishida A. (2017) “Death ofdementia inpatients in Japanese
psychiatric hospitals accounts for
one-fifth of discharge destination innational data from 1996 to 2014” J.
Alzheimers Dis. 56, 817-824.
Ando S, Koike S, Shimodera S, Fujito
R, Sawada K, Terao T, Furukawa TA,
Sasaki T, Inoue S, Asukai N, Okazaki
Y, and Nishida A. (2017) “Lithium
levels in tap water and the mental
health problems of adolescents:
an individual level cross-sectional
survey.” J. Clin. Psychiatry.
78(3):e252-e256.
Nakanishi M, Nakashima T, Shindo
Y, Niimura J, and Nishida A. (2016)“Japanese care location and medical
procedures for people with dementia
in the last month of life.” J. Alzheimers
Dis. 51, 747-755

While life in big cities, microcosms of today’s stressful societies, is afull of risk factors for mental health, interpersonal bonds that support
individuals are increasingly weakening. Hence, multifaceted researchof mental health promotion via clinical medicine and sociomedical
methodologies needs to be promoted.
Mental Health Promotion Project engages in promoting mental wellbeing in big cities through: empirical findings from large-scale birthcohort studies conducted in partnership with municipalities of Tokyo
which is experiencing increasingly aging population and low birthrate;and developing programs in collaboration with clinical forefront of care.

“We are trying to elucidate preventive factors tomental health problems and enhancing factors tomental well-being, and to improve care for people
living in the community and their families.”
Our goals are as follows: 1) Elucidate preventive factors to mental healthproblems and enhancing factors to mental well-being in adolescence.
2) Improve care for people with dementia living in the community andtheir families. 3) Develop transition support program connecting acutephase hospital treatment and post-discharge outpatient treatment.

Yamasaki S, Ando S, Koike S, Usami
S, Endo K, French P, Sasaki T,
Furukawa TA, Hasegawa-Hiraiwa
M, Kasai K, and Nishida A. (2016)“Dissociation mediates the relationshipbetween peer victimization andhallucinatory experiences among
early adolescents.” Schizophr. Res.
Cogn. 4, 18–23● Increase of dementia people: Est. number in 2025 is 7 million(MHLW, 2014)● The biggest cause of health damage among young people ismental illnesses and suicide (Patton, Lancet, 2009)

Mental Health Promotion37

Department of Psychiatry and Behavioral Sciences

Mental Health Promotion38

Department of Psychiatry and Behavioral Sciences
Project
LeaderMakoto Arai Schizophrenia Research Project

Identifying Biomarkers of Schizophrenia

Itokawa M, Miyashita M, Arai M, Dan
T, Takahashi K, Tokunaga T, Ishimoto
K, Toriumi K, Ichikawa T, Horiuchi
Y, Kobori A, Usami S, Yoshikawa T,
Amano N, Washizuka S, Okazaki Y,
and Miyata T. (2018) ‘’Pyridoxamine:
A novel treatment for schizophrenia
with enhanced carbonyl stress.’’
Psychiatry Clin. Neurosci. 72: 35-44.
Miyashita M, Watanabe T, Ichikawa
T, Toriumi K, Horiuchi Y, Kobori A,
Kushima I, Hashimoto R, Fukumoto
M, Koike S, Ujike H, Arinami T,
Tatebayashi Y, Kasai K, Takeda M,
Ozaki N, Okazaki Y, Yoshikawa T,
Amano N, Washizuka S, Yamamoto
H, Miyata T, Itokawa M, Yamamoto Y,
and Arai M. (2016) “The regulation ofsoluble receptor for AGEs contributes
to carbonyl stress in schizophrenia.”
Biochem. Biophys. Res. Commun.
479: 447-452.
Arai M, Miyashita M, Kobori A, Toriumi
K, Horiuchi Y, Hatakeyama S, andItokawa M. (2014) “Carbonyl stressand schizophrenia.” Psychiatry Clin.
Neurosci. 68: 655-665.

The profiling of the peripheral metabolic system is a viableschizophrenia research strategy that can lead to earlier diagnostic
methods, elucidation of its molecular mechanisms, and novel strategies
for the prevention and treatment.
We focus on 1) developing individualized medicine for treating
schizophrenia, 2) investigating factors involved in disease onset, and
3) understanding the molecular pathology by using biomarkers to
overcome the barrier of heterogeneity. Our research outcomes will
be applied to drug development by establishing a new biomarkerbased field of research in molecular psychiatry. Data obtained frommetabolomics, genomics, induced pluripotent stem (iPS) cell models,animal models, post-mortem brain analyses, neuropsychology, andgenetic counseling research will be consolidated to elucidate thegenetic and environmental factors relevant to psychiatric disorders suchas schizophrenia.

“Identifying biomarkers will allow us to classify
schizophrenia into different types, and aid in
earlier diagnoses and better treatments, leading to
improvements in patients’ quality of life.”

Miyashita M, Arai M, Kobori A,
Ichikawa T, Toriumi K, Niizato K,
Oshima K, Okazaki Y, Yoshikawa
T, Amano N, Miyata T, and Itokawa
M. (2014) “Clinical Features of
Schizophrenia With Enhanced
Carbonyl Stress.” Schizophr. Bull.
40: 1040-1046.
Arai M, Nihonmatsu-Kikuchi N,
Itokawa M, Rabbani N, and Thornalley
PJ. (2014) “Measurement ofglyoxalase activities.” Biochem Soc.
Trans. 42: 491-494.
Miyashita M, Arai M, Yuzawa H,
Niizato K, Oshima K, Kushima I,
Hashimoto R, Fukumoto M, Koike S,
Toyota T, Ujike H, Arinami T, Kasai
K, Takeda M, Ozaki N, Okazaki Y,
Yoshikawa T, Amano N, Miyata T,
and Itokawa M. (2014) “Replicationof enhanced carbonyl stress in a
subpopulation of schizophrenia.”
Psychiatry Clin. Neurosci. 68: 83-84.
Arai M, Koike S, Oshima N, Takizawa
R, Araki T, Miyashita M, Nishida A,
Miyata T, Kasai K, and Itokawa M.
(2011) “Idiopathic carbonyl stress ina drug-naive case of at-risk mental
state.” Psychiatry Clin. Neurosci. 65:
606-607.
Arai M, Yuzawa H, Nohara I, Ohnishi
T, Obata N, Iwayama Y, Haga S,
Toyota T, Ujike H, Arai M, Ichikawa
T, Nishida A, Tanaka Y, Furukawa
A, Aikawa Y, Kuroda O, Niizato
K, Izawa R, Nakamura K, Mori N,
Matsuzawa D, Hashimoto K, Iyo M,
Sora I, Matsushita M, Okazaki Y,
Yoshikawa T, Miyata T, and Itokawa
M. (2010) “Enhanced Carbonyl Stressin a Subpopulation of Schizophrenia.”
Arch. Gen. Psychiatry. 67: 589-597.

This biomarker-based approach is anticipated to become an
innovative and creative strategy for elucidating the metabolic system
of schizophrenia disease expression independently of conventional
pathological hypotheses. Verification in cellular and animal models
can shed light on the molecular mechanisms underlying the utility ofnaturally-derived substances, and is expected to lead to the future
development of much safer treatments and prophylactic methods.

Schizophrenia Research
39Department of Psychiatry and Behavioral Sciences

Schizophrenia Research40Department of Psychiatry and Behavioral Sciences
Project
Leader

Disorders
Yoshitaka Tatebayashi Affective
Research Project

Our Goal is to Decipher the
Neurobiological Bases ofAffective Disorders.

Bauer M, (64 co-authors), Tatebayashi
Y et al. (2014) “Relationship between
sunlight and the age of onset ofbipolar disorder: an international
multisite study.” J. Affect. Disord.
167:104-111.
Nihonmatsu-Kikuchi N, Hayashi Y,
Yu XJ, and Tatebayashi Y. (2013)“Depression and Alzheimer’s disease:novel postmortem brain studies reveal
a possible common mechanism.” J.
Alzheimers Dis. 37: 11-21.
Tatebayashi Y, Nihonmatsu-Kikuchi
N, Hayashi Y, Yu XJ, Soma M, andIkeda K. (2012) “Abnormal fatty acidcomposition in the frontopolar cortex
of patients with affective disorders.”
Transl. Psychiatry 2:e204.

Major depressive disorder (MDD) and bipolar disorder (BD), collectively
known as affective disorders, are essentially relapsing and remitting
disorders of affect with nearly full recovery between episodes. We
use human postmortem brains of these disorders and animal and cell
culture models to identify the processes in which stress or aging causeschanges in brain to induce these disorders. A major focus of our work is
stress-induced or age-related changes in cellular structure, especiallythat of oligodendrocyte lineage cells and lipid, within the brain’s mood
circuitry. We are also interested in the biological relationship between
affective disorders and dementias such as Alzheimer’s disease.

“Our human postmortem brain studies reveal
oligodendroglial reductions and myelindependent
fatty acid abnormalities in thefrontopolar cortex in affective disorders.”

Hayashi Y, Nihonmatsu-Kikuchi N,
Hisanaga S, Yu XJ, and Tatebayashi
Y. (2012) “Neuropathologicalsimilarities and differences between
schizophrenia and bipolar disorder:
a flow cytometric postmortem brain
study.” PLoS One. 7: e33019.
Hayashi Y, Nihonmatsu-Kikuchi N,
Yu XJ, Ishimoto K, Hisanaga SI,
and Tatebayashi Y. (2011) “A novel,
rapid, quantitative cell-counting
method reveals oligodendroglial
reduction in the frontopolar cortex
in major depressive disorder.” Mol.
Psychiatry. 16: 1155-1158.

The exact functions of the human frontopolar cortex (BA10)
remain enigmatic. Given that the BA10 is thought to be the mostevolutionarily recent expansion of the primate prefrontal cortex, its
function may uniquely reflect human adaptations in the context of
selecting and updating models of reward contingency in dynamic
environments. As adulthood cortical myelination is an essential
process for the establishment of efficient neuronal signaling networks,
any abnormalities in this process may have important roles in thepathophysiology of affective disorders.

Affective Disorders Research41

Department of Psychiatry and Behavioral Sciences

Affective Disorders Research42Department of Psychiatry and Behavioral Sciences
Project
LeaderMakoto Honda Sleep Disorders Project

Narcolepsy and Hypersomnia:

Find the causes to develop better treatments

Shimada M, Miyagawa T, Toyoda H,
Tokunaga K, and Honda M. (2018)“Epigenome-wide association studyof DNA methylation in narcolepsy: an
integrated genetic and epigenetic
approach.” Sleep 41:zsy019
Toyoda H, et al. (2017) “Narcolepsy
susceptibility gene CCR3 modulates
sleep-wake patterns in mice.” PLoS
ONE 12:e0187888
Miyata R, Hayashi M, KohyamaJ, and Honda M.(2017) “Steroid
therapy ameliorated cataplexy inthree children with recent-onset of
narcolepsy.” Sleep Med. 29:86-87.
Tanaka S, Honda Y, Honda M,
Yamada H, Honda K, and Kodama
T. (2017) “Anti-tribbles pseudokinase
2 (TRIB2)-immunization modulate
Hypocretin/Orexin neuronal functions.”
Sleep 40:zsw036.
Miyagawa T, et al. (2015) “Newsusceptibility variants to narcolepsy
identified in HLA class II region.”
Hum. Mol. Genet. 24:891-898.
Miyagawa T, et al. (2013) “Effectsof oral L- carnitine administration in
narcolepsy patients: a randomized,
double-blind, cross-over and placebocontrolled trial.” PLoS ONE 8:e53707.

Narcolepsy is a sleep disorder with abnormal intrinsic sleep-wakeregulation, resulting in unique symptoms including frequent lapses into
sleep, nocturnal sleep instability and REM sleep related manifestations
such as cataplexy (abrupt loss of muscle tone triggered by emotion),sleep paralysis and hypnagogic hallucination.
Narcolepsy is associated with a deficiency of wake-promoting orexin/
hypocretin producing neurons localized in hypothalamus, and virtually
all the patients carry human leukocyte antigen (HLA)-DQB1*06:02.

“We are trying to solve the mystery of narcolepsy:
Listen to the patients, get the whole picture, andimprove their lives”

Narcolepsy is associated with a variety of physical and psychiatriccomorbid conditions. Since appropriate wakefulness is essential forhigher brain functions, abnormal sleep-wake regulation can lead tovarious associated features. Despite the progress in sleep research
fields, we currently have inadequate symptom-based-treatments forsleep disorders, including narcolepsy. We are trying to elucidate thepathophysiology of narcolepsy with multifaceted problems to improve
the QOL of hypersomnia patients.

Research Strategy

Miyagawa T, et al. (2011) “Abnormally
low serum acylcarnitine levels innarcolepsy patients.” Sleep 34:349353.
Tanaka S, Honda M (2010) “IgG
abnormality in narcolepsy and idiopathic hypersomnia.” PLoS ONE 5:
e955.
Toyoda H, et al. (2010) “Anti-tribbles
homolog 2 autoantibodies inJapanese patients with narcolepsy.”
Sleep 33:875-878.
Honda M, et al. (2009) “IGFBP3
colocalizes with and regulates
hypocretin(orexin).” PLoS ONE
4:e4254.
Honda M, Arai T, et al. (2009)“Absence of ubiquitinated inclusions
in hypocretin neurons of narcolepsy
patients.” Neurology 73:511-517.

Sleep Disorders
Tanaka S, Honda Y, Inoue Y, andHonda M. (2006) “Detection of
autoantibodies against hypocretin,
hcrtrl, and hcrtr2 in narcolepsy: antiHcrt system antibody in narcolepsy.”
Sleep 29:633-638.

43

Department of Psychiatry and Behavioral Sciences
Research Interests

Hypothalamus works as a center for sleep-wake switch in coordination with the integrated informationfrom the body. Among them, we have particular interests in immune and metabolic status, which can be
the key to understand altered sleep-wake regulation in narcolepsy.

1. HLA association and immune abnormality

In addition to the tight association with HLA, narcolepsy is also associated with T cell receptor(TCR) alpha locus, indicating the HLA-TCR mediated immunological alterations. Both genetic andenvironmental factors are implicated in narcolepsy predisposition. We have reported the alteredimmune status in narcolepsy, but so far could not confirm the leading hypothesis that orexin neurons aredestructed by direct autoimmune attacks. Immune mechanisms other than autoimmunity might lead tostop producing orexin neuropeptide.

2. Metabolic aspect of narcolepsy and related hypersomnia

Through genome-wide association studies (GWAS), we have identified novel narcolepsy (and otherhypersomnia) related genes. They are key enzymes located in the pathway of fatty acid metabolism. We
confirmed their functional relevance, performed the clinical trials, and analyzing the potential efficacy ofthe novel therapy (promoting metabolism) in hypersomnia patients.

Sleep Disorders44

Department of Psychiatry and Behavioral Sciences
Project
LeaderKazutaka Ikeda Addictive Substance Project

Addictive Drugs are Double-edged Swords:
They can become both harmful and beneficial
depending on how they used

Ide S, and Ikeda K. “Mechanisms of
the antidepressant effects of ketamine
enantiomers and their metabolites.”
Biol. Psychiatry. in press
Sugaya N, Ogai Y, Aikawa Y, Yumoto
Y, Takahama M, Tanaka M, Haraguchi
A, Umeno M, and Ikeda K. (2018) “A
randomized controlled study of theeffect of ifenprodil on alcohol use inpatients with alcohol dependence.”
Neuropsychopharmacology Rep.
38(1):9-17.
Ide S, Takahashi T, Takamatsu Y, Uhl
G, Niki H, Sora I, and Ikeda K (2017)“Distinct roles of opioid and dopamine
systems in lateral hypothalamic
intracranial self-stimulation.“ Int.
J. Neuropsychopharmacol. 20:
403-409.
Fujita M, Hagino Y, Takeda T, Kasai S,
Tanaka M, Takamatsu Y, Kobayashi
K, and Ikeda K. (2017) “Light/darkphase-dependent spontaneous
activity is maintained in dopaminedeficient mice.“ Mol. Brain. 10: 49.
Nishizawa D, Fukuda K, Kasai S,
Hasegawa J, Aoki Y, Nishi A, Saita
N, Koukita Y, Nagashima M, Katoh R,
Satoh Y, Tagami M, Higuchi S, Ujike
H, Ozaki N, Inada T, Iwata N, Sora
I, Iyo M, Kondo N, Won MJ, Naruse
N, Uehara K, Itokawa M, Koga M,
Arinami T, Kaneko Y, Hayashida M,
and Ikeda K. (2014) “Genome-wide
association study identifies a potent
locus associated with human opioid
sensitivity.” Mol. Psychiatry. 19:
55-62.
Sato A, Kasai S, Kobayashi T,
Takamatsu Y, Hino O, Ikeda K, andMizuguchi M. (2012) “Rapamycin
reverses impaired social interaction inmouse models of tuberous sclerosis
complex” Nat. Communun. 3: 1292.

Addiction to substances (e.g. drugs, alcohol, tobacco) and behavior (e.g.
internet, gambling) is a serious public health problem. Moreover, use oflegal drugs has been increasing inJapan in recent years. It is importantto prevent and solve problems of
addictions.
Addiction
On the other hand, some addictive
drugs are also widely used as
analgesics and treatment ofdevelopmental disorders. Thus, it
Developmental
Pain
disorderstreatment
is considered that some molecules
involved in action of addictive drugs
are commonly related to analgesics
and developmental disorders.
The goals of our project are as
follows: 1) Development of novel treatment and prevention of addiction.
2) Improvement of personalized pain treatment. 3) Development ofnovel treatment against developmental disorders.

“We are trying to improve treatment, prevention,and understanding of addiction, pain, anddevelopmental disorders by revealing the mechanisms
underlying addiction.”
All goals can make significant contribution to the society. We aim tothose goals through studying the action mechanisms of addictive drugs
using molecular biological approach, behavioral pharmacologicalapproach, human genome analysis, and clinical approach.

Addictive Substance45

Department of Psychiatry and Behavioral Sciences

Addictive Substance46

Department of Sensory and Motor Systems
Project
LeaderShinji Kakei Motor Disorders Project

From Neuron to Action and its Disorders

Tomatsu S, Ishikawa T, Tsunoda Y,
Lee J, Hoffman DS, and Kakei S.
(2016) “Information processing in thehemisphere of the cerebellar cortrex
for motor control of wrist movement.”
J. Neurophysiol. 115:255-270.
Ishikawa T, Tomatsu S, Izawa J,
and Kakei S. (2016) “The cerebrocerebellum: Could it be loci of forward
models?” Neurosci. Res. 104:72-79.
Lee J, Kagamihara Y, and Kakei S.
(2015) “A new method for functional
evaluation of motor commands inpatients with cerebellar ataxia.”PLoS
One 10:e0132983.
Ishikawa T, Tomatsu S, Tsunoda
Y, Lee J, Hoffman DS, and Kakei
S. (2014) “Releasing dentate
nucleus cells from Purkinje cell
inhibition generates outputs fromthe cerebrocerebellum.” PLoS One
9:e108774 (pp.1-16).

We try to understand how the brain controls our movements in thereal world. We study the process of action generation at a single
neuron level using animal models to understand how the movement
is processed in the brain. We also study actions of healthy people, as
well as those with neurological disorders, such as cerebellar disorders,Parkinson’s disease or strokes. We look for building-blocks of motor
control with multidisciplinary approaches. We employ both invasive
and non-invasive approaches to achieve the deepest understandingof our brain. Our tools include various neurophysiological recording
techniques (single unit recording, electromyography(EMG) and electroencephalography (EEG)), brain stimulation, neuroimaging, analysis of
movement kinematics and a large-scale modeling. We have two longterm goals: 1) to understand the basic function of the motor structuresof the brain including the cerebellum, the basal ganglia, and the motor
cortex; and 2) to understand how our brain controls our movements on
the basis of the findings in 1).

“Through our research, we are trying tounderstand the brain.
The brain was first created to control movement
and extended to control higher brain functions.”
CG by Katsunori Goto

“The brain mechanism
for motor control must
provide a basic framework
to understand higher
brain functions.”
The brain is an assembly of neural networks.
CG by Katsunori Goto

Motor Disorders47

Department of Sensory and Motor Systems

Motor Disorders48

Department of Sensory and Motor Systems
Project
LeaderTakayuki Harada Visual Research Project

Elucidation of Pathology and Developmentof Therapeutic Strategies for Retinal
Neurodegenerative Diseases

Kimura A, Namekata K, Guo X, Noro
T, Harada C, and Harada T. (2015)“Valproic acid prevents NMDAinduced retinal ganglion cell death
via stimulation of neuronal TrkB
receptor signaling.” Am. J. Pathol.
185: 756-764.
Noro T, Namekata K, Kimura A, Guo
X, Azuchi Y, Harada C, Nakano T,
Tsuneoka H, and Harada T. (2015)“Spermidine promotes retinal
ganglion cell survival and optic nerve
regeneration in adult mice following
optic nerve injury.” Cell Death Dis.
6, e1720.
Namekata K, Kimura A, Kawamura
K, Harada C, and Harada T. (2014)“Dock GEFs and their therapeuticpotential: Neuroprotection and axon
regeneration.” Prog. Retin. Eye Res.
43: 1-16.
Namekata K, Harada C, Guo X,
Kimura A, Kittaka D, Watanabe
H, and Harada T. (2012) “Dock3
stimulates axonal outgrowth via GSK3β-mediated microtubule assembly.”
J. Neurosci. 32: 264-274.

More than 1.6 million people in Japan are visually impaired,representing economic social losses estimated at more than 8 trillion
yen. In the particular context of the increased penetration of Western
lifestyles and an aging society, the increase in the number of patientswith conditions such as glaucoma and diabetic retinopathy, which could
be called “adult eye diseases,” has become a major social issue.
To achieve improved quality of life (QOL) for the visually impaired in an
increasingly aging population, we seek to elucidate detailed pathogenic
mechanisms and develop new therapies through the development of amodel of intractable eye disease.

“We are focusing on elucidating the molecularmechanisms of neuroprotection andneuroregeneration, and our final goal is theprevention or treatment of blindness in retinal
neurodegenerative disorders such as glaucoma andtraumatic injury.”

Harada C, Guo X, Namekata K,
Kimura A, Nakamura K, Tanaka K,
Parada LF, and Harada T. (2011)“Glia- and neuron-specific functionsof TrkB signalling during retinaldegeneration and regeneration.”
Nature Commun. 2: 189.
Guo X, Harada C, Namekata K,
Matsuzawa A, Camps M, Ji H,
Swinnen D, Jorand-Lebrun C,
Muzerelle M, Vitte P, Ruckle T,
Kimura A, Kohyama K, MatsumotoY, Ichijo H, and Harada T. (2010)“Regulation of the severity ofneuroinflammation and demyelination
by TLR-ASK1-p38 pathway.” EMBO
Mol. Med. 2: 504-515.
Namekata K, Harada C, Taya C, Guo
X, Kimura H, Parada LF, and Harada
T. (2010) “Dock3 induces axonal
outgrowth by stimulating membrane
recruitment of the WAVE complex.”
Proc. Natl. Acad. Sci. USA 107:
7586-7591.
Harada T, Harada C, Nakamura K,
Quah HA, Okumura A, Namekata
K, Saeki T, Aihara M, Yoshida H,
Mitani A, and Tanaka K. (2007)“The potential role of glutamate
transporters in the pathogenesis ofnormal tension glaucoma.” J. Clin.
Invest. 117: 1763-1770.

Visual Research49

Our objectives

Department of Sensory and Motor Systems

Our major aim• To develop a neuroprotective retinal therapy using animal disease models• To elucidate the mechanisms involved in the onset of optic neuritis• To establish a method to promote regeneration of the optic nerve

Senior Research Scientist

Kazuhiko Namekata

Dock family proteins

Namekata K, Kimura A,
Kawamura K, Harada C,
Harada T. (2014) “Dock GEFs
and their therapeutic potential:
Neuroprotection and axon
regeneration.” Prog. Retin. Eye
Res. 43: 1-16,

The dedicator of cytokinesis (Dock) family is composed of atypicalguanine exchange factors (GEFs) that induce actin polymerization.
To date, 11 Dock family members have been identified. Dock3 is
predominantly expressed in the central nervous system. In the growthcone, Dock3 induces actin polymerization by activating WASP family
verprolin-homologous protein (WAVE) and modulates microtubule
dynamics through inactivation of GSK-3β, leading to axon elongation.
In addition, Dock3 plays a role in protecting retinal ganglion cells fromneurotoxicity and oxidative stress. Dock3 may be a therapeutic target
for optic neuropathy including glaucoma.

Namekata K, Harada C, Guo X,
Kimura A, Kittaka D, Watanabe
H, Harada T. (2012) “Dock3
stimulates axonal outgrowth via
GSK-3β-mediated microtubule
assembly.”
J. Neurosci. 32: 264-274,
Namekata K, Harada C, Taya
C, Guo X, Kimura H, Parada LF,
Harada T. (2010) “Dock3 induces
axonal outgrowth by stimulating
membrane recruitment of theWAVE complex.” Proc. Natl.
Acad.Sci. USA 107: 7586-7591,

Axonal regeneration was enhanced
in Dock3 overexpressing mouse (Tg)
(Arrow heads indicate regenerating axons)

Visual Research
50Department of Sensory and Motor Systems
Project
LeaderYuki Nakayama ALS Nursing Care Project

Improving the Quality Of Life of Patientswith Amyotrophic Lateral Sclerosis

Matsuda C, Shimizu T, Nakayama
Y, and Haraguchi M. (2018) “Coughpeak flow decline rate predicts
survival in patients with amyotorophic
lateral sclerosis.” Muscle Nerve. 2018
Aug 14.
Shimizu T, Bokuda K, Kimura H,
Kamiyama T, Nakayama Y, Kawata
A, Isozaki E, and Ugawa Y.(2018)“Sensory cortex hyperexcitability
predicts short survival in amyotrophic
lateral sclerosis.” Neurology 1;90(18):e1578-e1587.
Nakayama Y, Shimizu T, Matsuda C,
Haraguchi M, Hayashi K, Mochizuki
Y, Nagao M, Kawata A, and Isozaki
E.(2018) “Non-motor manifestations in
ALS patients with tracheostomy andinvasive ventilation.” Muscle Nerve.
57(5):735-741.
Nakayama Y, Shimizu T, Mochizuki
Y, Hayashi K, Matsuda C, Nagao
M, Watabe K, Kawata A, Oyanagi
K, Isozaki E, and Nakano I.
(2016) “Predictors of impaired
communication in amyotrophic lateralsclerosis patients with tracheostomy
invasive ventilation.” Amyotroph.
Lateral Scler. Frontotemporal
Degener. 17(1-2):38-46.

“Our mission is to establish the best practices
for respiratory and communication management
for ALS patients in a community-based setting .
We have established a multidisciplinary researchteam to develop a Brain Machine Interface
for ALS patients.”

Matsuda C, Shimizu T, Nakayama
Y, Haraguchi M, Hakuta C, Itagaki
Y, Ogura A, Murata K, Taira M,
Numayama T, and Kinoshita
M.(2016) “Macroglossia in advanced
amyotrophic lateral sclerosis.” Muscle
Nerve. 54:386-390.

ALS Nursing Care51

Department of Sensory and Motor Systems

ALS Nursing Care52

Department of Sensory and Motor Systems
Project
LeaderKazunori Sango Diabetic Neuropathy Project

Pathogenesis-based Therapeutic
Approaches to Diabetic Neuropathy

Takaku S, Yako H, Niimi N, Akamine
T, Kawanami D, Utsunomiya K, andSango K. (2018) “Establishment of amyelinating co-culture system with amotor neuron-like cell line NSC-34 andan adult rat Schwann cell line IFRS1.”
Histochem. Cell Biol. 149: 537-543.
Yoshida S, Hasegawa T, Suzuki M,
Sugeno N, Kobayashi J, Ueyama
M, Fukuda M, Ido-Fujibayashi A,
Sekiguchi K, Ezura M, Kikuchi A, Baba
T, Takeda A, Mochizuki H, Nagai
Y, and Aoki M. (2018) “Parkinson's
disease-linked DNAJC13 mutation
aggravates alpha-synuclein-induced
neurotoxicity through perturbation ofendosomal trafficking.” Hum. Mol.
Genet. 27: 823-836.
Niimi N, Yako H, Takaku S, Kato H,
Matsumoto T, Nishito Y, Watabe K,
Ogasawara S, Mizukami H, Yagihashi
S, Chung SK, and Sango K. (2018) “A
spontaneously immortalized Schwann
cell line from aldose reductasedeficient mice as a useful tool forstudying polyol pathway and aldehyde
metabolism.” J. Neurochem. 144:
710-722.

Peripheral neuropathy is one of the most common complicationsof Diabetes Mellitus, and its irritating symptoms such as pain andnumbness can be the cause of insomnia and depression, and whenallowed to progress to more advanced disease stages can result
in serious consequences such as lower limb amputation and lethal
arrhythmia. In addition, recent studies have indicated that diabetes is amajor risk factor for cognitive disorders such as Alzheimer’s disease.

“We are trying to improve QOL
for diabetics and help them tolive longer lives by elucidating
the pathogenesis of neurologicaldisorders and establishingeffective treatments.”
The goals of our project are as follows: 1) Establishing effective
pathogenesis-based treatments for diabetic peripheral neuropathy.
2) Elucidating mechanistic link between metabolic dysfunction andneurodegenerative diseases.

Tomoyo Akamine
Masaki Oba

Naoko Niimi

Shizuka Takaku

Sango K, Mizukami H, Horie H, andYagihashi S. (2017) “Impaired axonal
regeneration in diabetes. Perspectiveon the underlying mechanism from invivo and in vitro experimental studies.”
Front. Endocrinol. 8: 12.
Niimi N, Yako H, Tsukamoto M,
Takaku S, Yamauchi J, Kawakami E,
Yanagisawa H, Watabe K, Utsunomiya
K, and Sango K. (2016) “Involvementof oxidative stress and impaired
lysosomal degradation in amiodaroneinduced schwannopathy.” Eur. J.
Neurosci. 44: 1723-1733.

Mari Suzuki

Diabetic Neuropathy
53

Hideji Yako

Department of Sensory and Motor Systems

Wild-type

MutantDiabetic Neuropathy
54

Department of Advanced Science for Biomolecules
Project
LeaderYasuko Ono Calpain Project

Calpain: Structure-Function Relationships
Exploring calpain-mediated biological modulation

Hata S, Kitamura F, Yamaguchi M,
Shitara H, Murakami M, and Sorimachi
H. (2016) “A Gastrointestinal Calpain
Complex, G-calpain, Is a Heterodimerof CAPN8 and CAPN9 Calpain
Iso-forms, Which Play Catalytic andReg-ulatory Roles, Respectively.” J.
Biol. Chem., 291: 27313-27322.

Proteins are chains of amino acids, and their functions change bypartial cuts. Calpains are enzymes that perform such “cuts” or “limited
proteolytic processing” in cooperation with calcium.
Humans have 15 calpain species. Defects of either calpain cause various
deficiencies, such as muscular dystrophy, stomach ulcer, and embryoniclethality.

Ono Y, Saido TC, and Sorimachi H.
(2016) “Calpain research for drug
discovery: challenges and potential.”
Nature Reviews: Drug Discovery,
15: 854-876.
Shinkai-Ouchi F, Koyama S, Ono Y,
Hata S, Ojima K, Shindo M, duVerle D,
Ueno M, Kitamura F, Doi N, Takigawa
I, Mamitsuka H, and Sorimachi H.
(2016) “Predictions of cleavability of
calpain proteolysis by quantitative
structure-activity relationship analysis
using newly determined cleavage
sites and catalytic efficiencies ofan oligopeptide array.” Mol. Cell.
Proteomics, 15: 1262-1280.
Ojima K, Ono Y, Hata S, Noguchi
S, Nishino I, and Sorimachi H.
(2014) “Muscle-specific calpain-3 isphosphorylated in its unique insertion
region for enrichment in a myofibril
fraction.” Genes Cells, 19: 830-841.
Ono Y, Shindo M, Doi N, Kitamura
F, Gregorio CC, and Sorimachi
H. (2014) “The N- and C-terminal
autolytic fragments of CAPN3/p94/
calpain-3 restore proteolytic activity
by intermolecular complementation.”
Proc. Natl. Acad. Sci. USA, 111:
E5527-5536.
Tonami K, Hata S, Ojima K, Ono
Y, Kurihara Y, Amano T, Sato T,
Kawamura Y, Kurihara H, andSorimachi H. (2013) “Calpain-6
deficiency promotes skeletal muscle
development and regeneration.”
PLoS Genet., 9: e1003668.

Calpain55“Translational research involving calpains is stillat the development stage. We need to learn more
about the calpains themselves, as well as their impact
on various physiological systems and molecularpathways.” (Nat. Rev. Drug Discov. 2016).
In this project, we aim to understand biology of calpains with wide
scope of interest, and translate the knowledge to the development of
our health as well as science.

Department of Advanced Science for Biomolecules

Calpains in health and disease
Some calpains predominantly expressed in specific tissue(s) are responsible for genetic
diseases; e.g., defects in CAPN3 cause muscular dystrophy. Other calpains with rather
ubiquitous expression pattern lead to lethality if deficient. It is also important to realize thatsome calpain species express their activity through unique and unexpected mechanisms,
such as intermolecular complementation (CAPN3), heterodimerization (CAPN8/9), etc.
To explore how calpains protect our health, analyses of cells/mice lacking the function of
specific calpain species or its expected targets are being performed. We are also improving
research platform for studying calpains by biochemistry including proteomics, genetics, andbioinformatics.

Protection of epithelial cells byheterodimeric calpain, G-calpain

Characterization of calpainsubstrate interface

Strategy for activity regulation of
CAPN3

Multiplicity of calpain actions

Shoji Hata, Ph.D.

Fumiko Shinkai-Ouchi, Ph.D.

Aya Noguchi, Ph.D.

Calpains in epithelial function andtissue development

Proteomic analysis of muscular
dystrophy and calpain substratespecificities

Cross talk of calpain and otherproteolytic systems

Calpain56

Department of Advanced Science for Biomolecules
Project
LeaderNoriyuki Matsuda Ubiquitin Project

Ubiquitin-Mediated Mitochondrial
Quality Control:
A shield against Parkinson's disease

Yamano K, Wang C, Sarraf S, Münch
C, Kikuchi R, Noda N, Hizukuri Y,
Kanemaki M, Harper W, Tanaka K,
Matsuda N, and Youle R. (2018)“Endosomal Rab cycles regulate
Parkin-mediated mitophagy.” eLife 7:
e31326
Okatsu K, Koyano F, Kimura M,
Kosako H, Saeki Y, Tanaka K, andMatsuda N. (2015) “Phosphorylated
ubiquitin chain is the genuine Parkin
receptor.” J. Cell Biology 209:111128
Koyano F, Okatsu K, Kosako H,
Tamura Y, Go E, Kimura M, Kimura Y,
Tsuchiya H, Yoshihara H, Hirokawa
T, Endo T, Fon E-A, Trempe J-F,
Saeki Y, Tanaka K, and Matsuda N.
(2014) “Ubiquitin is phosphorylated by
PINK1 to activate Parkin.” Nature 510,
162-166.
Okatsu K, Oka T, Iguchi M, Imamura
K, Kosako H, Tani N, Kimura M,
Go E, Koyano F, Funayama M,
Shiba-Fukushima K, Sato S, Shimizu
H, Fukunaga Y, Taniguchi H, Komatsu
M, Hattori N, Mihara K, Tanaka K,
and Matsuda N. (2012) “PINK1
autophosphorylation upon membranepotential dissipation is essential
for Parkin recruitment to damaged
mitochondria.” Nature Commun. 3 :e1016 (10 pages).
Matsuda N, Sato S, Shiba K, Okatsu
K, Saisho K, Gautier C, Sou Y-S,
Saiki S, Kawajiri S, Sato F, Kimura M,
Komatsu M, Hattori N, and Tanaka
K. (2010) “PINK1 stabilized by
mitochondrial depolarization recruits
Parkin to damaged mitochondria andactivates latent Parkin for mitophagy.”
J. Cell Biology 189: 211-221.

Ubiquitin
57

Ubiquitin is well-known as a signal for proteasome-dependentdegradation; however, it also functions in autophagic degradation.
Increasing evidence indicates that selective autophagy functionsin intracellular quality control by using ubiquitin tags to delineate
aggregated proteins and damaged organelles for degradation.
In 2000, Dr. Mizuno and Dr. Hattori (Juntendo Univ.) collaboration with
Dr. Suzuki and Dr. Tanaka (TMIMS) reported for the first time that
Parkin, which is a causative gene product of familial Parkinson’s disease(PD), is an ubiquitin-protein ligase (Nat. Genet. 2000). In addition, theidentification of another gene PINK1 that linked to the familial forms ofhereditary recessive early-onset PD (Science 2004) has revealed that
phosphorylation, ubiquitylation, and mitochondrial integrity are key
factors in disease pathogenesis. Nevertheless, the exact mechanism
underlying the functional interplay between Parkin and PINK1 remained
an enigma. We thus have investigated how PINK1 and Parkin cooperate
to keep mitochondrial integrity against mitochondrial stresses.

“We found that low-quality mitochondria aremarked with ubiquitin for selective degradation,and the key factors in this process are PINK1(a mitochondrial kinase) and Parkin (a ubiquitin
ligase), two proteins implicated in Parkinson’s
disease.”PINK1 is a mitochondrial Ser/Thr kinase whereas Parkin is a ubiquitinprotein ligase that catalyzes ubiquitylation of diverse mitochondrial
outer membranous proteins (J. Cell Biol. 2010). We revealed that PINK1
is rapidly and constitutively degraded under steady-state conditions ina mitochondrial membrane potential-dependent manner but that a loss
in mitochondrial membrane potential stabilizes PINK1 mitochondrial
accumulation (J. Cell Biol. 2010). Previously our and other groups
found that PINK1 acts as an upstream factor for Parkin, but how PINK1
activates latent Parkin and recruits cytoplasmic Parkin to damaged
mitochondria were still obscure.

Department of Advanced Science for Biomolecules

Ubiquitin58

Department of Advanced Science for Biomolecules
Project
LeaderTakahiko Hara Stem Cell Project

Blood regeneration from ESC/iPSC anddevelopment of novel anti-cancer drugs

Tanegashima K, Takahashi R, Nuriya
H, Iwase R, Naruse N, Tsuji K,
Shigenaga A, Otaka A, and Hara T.
(2017) “CXCL14 acts as a specificcarrier of CpG DNA into dendritic
cells and activates Toll-like receptor
9-mediated adaptive immunity.”
EBioMed. 24: 247-256.
Tanegashima K, Sato-Miyata Y,
Funakoshi M, Nishito Y, Aigaki T, andHara T. (2017) “Epigenetic regulationof the glucose transporter geneSlc2a1 by β-hydroxybutyrate underlies
preferential glucose supply to thebrain of fasted mice.” Genes Cells
22: 71-83.
Suzuki T, Kazuki Y, Oshimura M,
and Hara T. (2016) “Highly efficienttransfer of chromosomes to a broad
range of target cells using Chinese
hamster ovary cells expressing murine
leukemia virus-derived envelope
proteins.” PLoS ONE 11: e0157187.
Kodaka Y, Tanaka K, Kitajima,Tanegashima K, Matsuda R, and Hara
T.(2015) “LIM homeobox transcription
factor Lhx2 inhibits skeletal muscle
differentiation in part via transcriptional
activation of Msx1 and Msx2.” Exp.
Cell Res. 331: 309-319.
Tanaka K, Kondo K, Kitajima K,
Muraoka M, Nozawa A, and Hara T.
(2013) “Tumor-suppressive function ofprotein-tyrosine phosphatase nonreceptor type 23 in testicular germ celltumors is lost upon overexpressionof miR142-3p microRNA.” J. Biol.
Chem. 288: 23990-23999.
Kitajima K, Minehata K, Sakimura
K, Nakano T, and Hara T. (2011)“In vitro generation of HSC-like cells
from murine ESCs/iPSCs by enforced
expression of LIM-homeobox
transcription factor Lhx2.” Blood 117:3748-3758.

Stem Cell
59

Dr. Yamanaka’s inducible pluripotent stem cell (iPSC) technology
has opened a new avenue to overcome incurable diseases bytransplantation of missing cells. In 2011, we discovered thatoverexpression of Lhx2 in hemogenic mesodermal cells resulted in exvivo expansion of transplantable hematopoietic stem cells (HSCs) frommouse embryonic stem cells (ESCs) and iPSCs. Since then, we aremaking great efforts to improve the system and apply this method tohuman iPSCs. We believe that comparison of the in vitro differentiation
capacity of hematopoietic cells between mouse and human iPSCs
would uncover novel and fundamental aspects of human HSC
development.

“We are making efforts to derive HSCs from human
iPSCs in vitro. We are also challenging to develop
novel anti-leukemia drugs and chemokine-based
anti-cancer drugs.”
The presence of cancer stem cells has been proposed in various typesof human cancer. As with tissue stem cells, cancer stem cells residein a niche and stay dormant, thereby surviving chemotherapy andradiotherapy. Presumably, both tissue and cancer stem cells commonly
express critical transcriptional regulators and signal transducers. We
have already identified DDX1 and PTPN23 as essential molecules forthe onset of testicular tumors.

In 2007, we discovered that CXCL14, a CXC-type chemokine, is oneof the causative factors for obesity-associated diabetes. In contrast,
CXCL14 is known to possess tumor-suppressive activity against lungand oral carcinomas. Recently, we discovered that CXCL14 binds
to CXCR4 with high affinity, thereby inhibiting the CXCL12-mediatedcell migration. This could be one of the underlying mechanisms of
the CXCL14’s anti-tumor function. We are vigorously investigating
physiological roles of CXCL14 and its action mechanisms. CXCL14 is apromising tool for developing novel anti-cancer and anti-diabetes drugs.

Department of Advanced Science for Biomolecules

Stem Cell60

Department of Advanced Science for Biomolecules
Project
LeaderYuichiro Miyaoka Regenerative Medicine Project

Genome Editing in Human iPS Cells:
To study and cure genetic disorders

Kato-Inui T, Takahashi G, Hsu S, andMiyaoka Y. (2018) “Clustered regularlyinterspaced short palindromic repeats(CRISPR)/CRISPR-associated protein
9 with improved proof-reading
enhances homology-directed repair.”
Nucleic Acids Res. 46: 4677-4688.
Miyaoka Y, Mayerl SJ, Chan AH, andConklin BR. (2018) “Detection and
Quantification of HDR and NHEJ
Induced by Genome Editing at
Endogenous Gene Loci Using Droplet
Digital PCR.” Methods Mol. Biol.
1768: 349-362.
Workman MJ, Mahe MM, Trisno S,
Poling HM, Watson CL, Sundaram
N, Chang CF, Schiesser J, Aubert P,
Stanley EG, Elefanty AG, Miyaoka Y,
Mandegar MA, Conklin BR, Neunlist
M, Brugmann SA, Helmrath MA, andWells JM. (2017) “Engineered humanpluripotent-stem-cell-derived intestinal
tissues with a functional entericnervous system.” Nat. Med. 23: 49-59.
Miyaoka Y, Chan AH, and Conklin
BR. (2016) “Using Digital Polymerase
Chain Reaction to Detect SingleNucleotide Substitutions Induced by
Genome Editing.” Cold Spring Harb.
Protoc. 2016:688-692.
Miyaoka Y, Berman JR, Cooper SB,
Mayerl SJ, Chan AH, Zhang B, KarlinNeumann GA, and Conklin BR. (2016)“Systematic quantification of HDR
and NHEJ reveals effects of locus,
nuclease, and cell type on genomeediting.” Sci. Rep. 6: 23549.
Miyaoka Y, Chan AH, Judge LM, Yoo
J, Huang M, Nguyen TD, Lizarraga
PP, So PL, and Conklin BR. (2014)“Isolation of single-base genomeedited human iPS cells without
antibiotic selection.” Nat. Methods 11:
291-293.

Genome editing technology allows us to rewrite the genetic informationin virtually any species and any cell type including human cells. To
study the pathogenesis of human diseases at the molecular level, andto develop new therapies using genome editing, we need appropriate
human cellular models. Our focus is on human iPS (induced pluripotentstem) cells, a type of pluripotent stem cell that can be generated frompatients’ cells by introduction of specific transcription factors, anddifferentiated into other cell types. Our goal is to use genome editing of
iPS cells to both model human diseases, and develop new therapies.

“Our goal is to develop methods to precisely andefficiently edit the genome in human iPS cells toallow us to develop disease models using humancells, and develop new therapies for these diseases.”

We developed an efficient method to isolate iPS cell lines containing asingle nucleotide substitution. The nucleotide substitution is created bygenome editing based on digital PCR, and isolation is accomplishedby repeated limited dilutions in the absence of selection markers(Miyaoka, Nat. Methods 2014). Using this method, we are analyzing thepathogenesis of cardiomyopathy caused by point mutations of RBM20(RNA-binding motif protein 20) in isogenic cardiomyocytes derived
from genome-edited iPS cells. We are also improving the precisionand efficiency of genome
editing technology, anddeveloping new therapies
based on correcting
mutations in iPS cells frompatients. In addition, weare developing a strategy
to directly edit the genome
in cells in the human body.

Regenerative Medicine
61

Department of Advanced Science for Biomolecules

Members

Tomoko Kato-Inui

Gou Takahashi

Szuyin Hsu

Regenerative Medicine
62Department of Advanced Science for Biomolecules
Project
LeaderTakashi Shichita Stroke Renaissance Project

Sterile Inflammation
After Ischemic Stroke

Tsuyama J, Nakamura A, Ooboshi
H, Yoshimura A, and Shichita T.
(2018) “Pivotal role of innate myeloid
cells in cerebral post-ischemic
sterile inflammation.” Semin.
Immunopathol.
Shichita T, Ito M, Morita R, Komai
K, Noguchi Y, Ooboshi H, Koshida
R, Takahashi S, Kodama T, andYoshimura A. (2017) “Mafb prevents
excess inflammation after ischemic
stroke by accelerating clearance of
danger signals through MSR1.” Nat.
Med. 23(6): 723-732.
Shichita T, Hasegawa E, Kimura
A, Morita R, Sakaguchi R, Takada
I, Sekiya T, Ooboshi H, Kitazono
T, Yanagawa T, Ishii T, Takahashi
H, Mori S, Nishibori M, Kuroda K,
Akira S, Miyake K, and Yoshimura A.
(2012) “Peroxiredoxin family proteins
are key initiators of post-ischemic
inflammation in the brain.” Nat. Med.
18(6): 911-917.
Shichita T, Sugiyama Y, Ooboshi H,
Sugimori H, Nakagawa R, Takada
I, Iwaki T, Okada Y, Iida M, Cua DJ,
Iwakura Y, and Yoshimura A. (2009)“Pivotal role of cerebral interleukin17-producing gammadelta T cells inthe delayed phase of ischemic brain
injury.” Nat. Med. 15(8):946-950.

“What triggers neural
repair after stroke?”We have identified peroxiredoxin family proteins as DAMPs (damage
associated molecular patterns) which trigger the post-ischemic
inflammation (Nat. Med. 2012). DAMPs induce IL-23 production
from infiltrating macrophages and neutrophils, and this sustains the
inflammation after ischemic stroke by promoting IL-17 production of
gdT lymphocytes (Nat. Med. 2009). Cerebral
post-ischemic inflammation resolves several days
after the stroke onset. The clearance of DAMPs
from ischemic brain through MSR1, a scavenger
receptor, plays a pivotal role in the resolution ofsterile inflammation after ischemic stroke (Nat.
Med. 2017). Now our question is how the cerebral
post-ischemic inflammation switches into theprocess of neural repair.

Stroke Renaissance63

Department of Advanced Science for Biomolecules

Stroke Renaissance64Department of Advanced Science for Biomolecules
Laboratory
Head

Yasushi Saeki Protein Metabolism Laboratory

The Ubiquitin-Proteasome System“Elucidation of Fundamental andPathophysiological Mechanisms”

Tsuchiya H, Burana D, Ohtake F, AraiN, Kaiho A, Komada M, Tanaka K,
and Saeki Y. (2018) “Ub-ProT reveals
global length and composition ofprotein ubiquitylation in cells.” Nature
Commun. 9, 524.
Ohtake F, Tsuchiya H, Saeki Y, andTanaka K. (2018) “K63 ubiquitylation
triggers proteasomal degradation
by seeding branched chains.”
Proc. Natl. Acad. Sci. USA. 115,
E1401-E1408.
Xie X, Matsumoto S, Endo A,
Fukushima T, Kawahara H,
Saeki Y, and Komada M. (2018)“Deubiquitylases USP5 and USP13
are recruited to and regulate heatinduced stress granules throughdeubiquitylating activities.” J. Cell.
Sci. 131, jcs201856
Tsuchiya H, Ohtake F, Arai N,
Kaiho A, Yasuda S, Tanaka K, andSaeki Y. (2017) “In vivo ubiquitin
linkage-type analysis reveals that the
Cdc48-Rad23/Dsk2 axis contributes
to K48-linked chain specificity of theproteasome.” Mol. Cell 66, 485-502.
Ohtake F, Saeki Y, Ishido S, Kanno J,
and Tanaka K. (2016) “The K48-K63
branched ubiquitin chain regulates
NF-κB signaling.” Mol. Cell 64, 251266.
Yoshida Y, Saeki Y, Murakami A,
Kawawaki J, Tsuchiya H, Yoshihara
H, Shindo M, and Tanaka K. (2015) “A
comprehensive method for detecting
ubiquitinated substrates using TRTUBE.” Proc. Natl. Acad. Sci. USA.
112, 4630-4635.
Pack Chan-Gi, Yukii H, Toh-e A,
Kudo T, Tsuchiya H, Kaiho A, Sakata
E, Murata S, Yokosawa H, Sako Y,
Baumeister W, Tanaka K, and Saeki Y.
(2014) “Quantitative live-cell imaging
reveals molecular dynamics andcytoplasmic assembly of the 26S
proteasome.” Nature Commun. 5,4396
Saeki Y, Toh-e A, Kudo T, Kawamura
H, and Tanaka K. (2009) “Multiple
proteasome-interacting proteinsassist the assembly of the yeast 19S
regulatory particle.” Cell 137, 900-913.

The ubiquitin-proteasome system (UPS) plays a pivotal role inproteostasis and controls almost all of cellular functions by selective
protein degradation. As the maintenance of protein homeostasis isessential to human health, dysfunction of the UPS due to stresses,
age-associated changes, or gene mutations causes various diseases
such as cancers, inflammation, and neurodegeneration. However, we
do not yet know the overall principle of the ubiquitin signaling, decoding
mechanism, and the proteasome. We aim to elucidate the fundamental
mechanisms of the ubiquitin code as well as proteasome function andto integrate it into pathophysiology, and then to develop therapeuticstrategies for UPS-related diseases.

Research Projects
1. Proteasome Dynamics and Pathophysiology
The proteasome is a highly organized proteolytic machinery that
degrades ubiquitylated proteins in an ATP-dependent manner. We havecharacterized the structure, assembly pathway, and substrate targetingmechanism of the proteasome. We also found that the proteasome
dynamically changes its intracellular localization and its accessory
proteins under various stresses to restore proteostasis. Currently,
we are generating knock-in mice to visualize proteasome localization
and activity to analyze physiological changes of the proteasomeaccompanying stress and aging. Furthermore, we have generated
model mice of proteasomal gene mutation derived from patients withneurodevelopmental disorders. Using the mutant mice, we will elucidate
the pathophysiology of the proteasome mutation at the whole-bodylevel.

Protein Metabolism
65Department of Advanced Science for Biomolecules
2. Roles of Specialized Proteasomes in Cell-Mediated Immunity
The proteasome has acquired diversity of the catalytic β subunits, whichhave evolved during the acquisition of adaptive immunity. To date, wehave discovered the vertebrate-specific alternative proteasomes, which
we named the "immunoproteasome" and the “thymoproteasome”.
Whereas the immunoproteasome plays a specialized role as aprofessional antigen-processing enzyme in cell-mediated immunity,
the thymoproteasome is involved in the development of CD8+T cells inthymus; i.e., it has a key role in the generation of MHC class I-restricted
CD8+T cell repertoire during thymic selection called “positive selection”.
Currently, we are conducting a deep proteomic screen to validate thepositive selection model.

Keiji Tanaka(The chairperson of TMIMS)
Murata S, Takahama Y, Kasahara
M, and Tanaka K. (2018)“The immunoproteasome andthymoproteasome: functions, evolution
and human disease.” Nature
Immunol. 19, 923-931.
Murata S, Sasaki K, Kishimoto T,
Niwa S, Hayashi H, Takahama Y, andTanaka K. (2007) “Regulation of CD8+
T cell development by thymus-specificproteasomes.” Science 316, 13491353.

3. Deciphering the Ubiquitin CodeDifferent polyubiquitin chain linkages direct substrates to distinct pathways, as referred to as ‘ubiquitin
code’. We have developed a highly sensitive MS/MS-based quantification method for ubiquitin chains.
The method allows us to analyze linkage-type selectivity of ubiquitin decoder proteins at endogenous
experimental setting. We recently identified the main pathway targeting the K48-linked ubiquitylated
substrates for proteasomal degradation. We also identified more complexed ubiquitin chains branched
at K48 and K63, which act as a unique coding signal to enhance NF-κB signaling. We are further
analyzing the decoder proteins throughout the ubiquitin-mediated pathways to reveal the ubiquitin
network.

Fumiaki Ohtake

Hikaru Tsuchiya

Members
Keiji Tanaka
Ai Kaiho
Yasushi Saeki Naoko Arai
Fumiaki Ohtake Yuko Okamoto
Hikaru Tsuchiya Arisa Kawano
Sayaka Yasuda Marcel Diallo

Protein Metabolism
66Department of Advanced Science for Biomolecules

Unit Leader

Kohji Kasahara Biomembrane Unit

Physiological Functions of Lipid Rafts /Glycosphingolipid Microdomains inTransmembrane Signaling

Kasahara K, Kaneda M, Miki T, Iida
K, Sekino-Suzuki N, Kawashima I,
Suzuki H, Shimonaka M, Arai M,
Ohno-Iwashita Y, Kojima S, Abe M,
Kobayashi T, Okazaki T, Souri M,
Ichinose A, and Yamamoto N. (2013)“Clot retraction is mediated by factor
XIII-dependent fibrin-αIIbβ3-myosin
axis in platelet sphingomyelin-richmembrane rafts.” Blood 122, 33403348.
Sekino-Suzuki N, Yuyama K, Miki T,
Kaneda M, Suzuki H, Yamamoto N,
Yamamoto T, Oneyama C, Okada M,
and Kasahara K. (2013) “Involvementof gangliosides in the process of
Cbp/PAG phosphoryla-tion by Lyn indeveloping cerebellar growth cones.”
J.Neurochem. 124, 514-522.
Kasahara K, Souri M, Kaneda M,
Miki T, Yamamoto N, and Ichinose
A. (2010) “Impaired clot retraction
in factor XIII A subunit-deficientmice.”Blood 115, 1277-1279.
Yuyama K, Sekino-Suzuki N, Sanai Y,
and Kasahara K. (2007) “Translocationof activated heterotrimeric G protein
Gαo to ganglioside-enriched
detergent-resistant membrane rafts
in developing cerebellum.” J.Biol.
Chem. 282, 26392-26400.
Kasahara K, Watanabe K, Takeuchi
K, Kaneko H, Oohira A, Yamamoto
T, and Sanai Y. (2000) “Involvementof gangliosides in GPI-anchoredneuronal cell adhesion molecule
TAG-1 signaling in lipid rafts.” J.Biol.
Chem. 275, 34701-34709.
Kasahara K, Watanabe Y, Yamamoto
T, and Sanai Y. (1997) “Associationof src family tyrosine kinase Lyn withganglioside GD3 in rat brain. Possible
regulation of Lyn by glycosphingolipid
in caveolae-like domains.” J.Biol.
Chem. 272, 29947-29953.

Lipid rafts are dynamic assemblies of glycosphingolipids, sphingomyelin,cholesterol, and proteins that can be stabilized into microdomains
involved in the regulation of a number of cellular processes.
We have been investigating the association of glycosphingolipids withspecific proteins in the nervous system and blood platelets.
We demonstrated that anti-ganglioside GD3 antibody co-precipitates
GPI-anchored neural cell adhesion molecule TAG-1, src-family kinase
Lyn, its substrate Cbp, trimeric G protein Goα of cerebellar granule cells.
TAG-1 plays roles in axonal
guidance, and cellularmigration. GPI anchors haveno direct contact with thecytoplasm. We demonstrated
that TAG-1 transduces signal
Ganglioside GD3-rich rafts
via Lyn/Cbp in gangliosideof cerebellar granule cells
GD3-rich rafts of cerebellar
granule cells. Chemokine SDF-1α triggers the chemoattraction of
cerebellar granule cells during cerebellar development.
We demonstrated that SDF-1α stimulates GTPγS binding to Goα, andcauses Goα translocation to lipid rafts, leading to growth cone collapseof cerebellar granule cells.

“We found that glycosphingolipids function
as platforms in transmembrane signaling
for the attachment of various signaling moleculesof neurons and platelets.”
Fibrin associates with lipid rafts on theplatelets and raft integrity is requiredfor clot retraction. We propose thatclot retraction is mediated by factor
XIII-dependent fibrin-integrin αIIbβ3myosin axis in sphingomyelin-richmembrane rafts.

Sphingomyelin-rich rafts of platelets

Members: Ikuo Kawashima, Kiyoshi Ogura,
Tomohiro Iguchi, Keisuke Komatsuya

Biomembrane67

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Center for Basic Technology Research
The Basic Technology Research Center provides multiple resources
to assist the researchers of the institute conduct researches with the
maximum efficiency. The services provided include state-of-the art
technologies required for biomedical and life science researches at theirhighest level, and maintenance of various facilities which are routinely
used by the researchers. Details of the services provided and specificactivities of related facilities are given below.

General Manager

Minoru Saitoe

1. The Animal Research Division maintains the animal facility that isused by all the researchers who use animals for their research. It assists
researchers generate knock-out or transgenic animals as well as maintain
sperms and eggs of various lines of mutant animals. It also provides
maximal care for the welfare of the animals used in the research.
2. The Advanced Technical Support Department consists of Protein
Analyses, FACS, Microarray, Confocal Microscopy, Electron Microscopy,
and Histology Laboratories etc. and offers state-of-the-art technology to
the researchers.
3. The Information Support Department consists of Library, Information
Technology Section, Media Technology Laboratory and Public Relation
Office. It assists researchers search for references, provides supports fordaily use of computer systems, and deals with the media.
4. The Authorized and General Core Facility Department consistsof Radioisotope Laboratory, Hazardous Chemical Control Room andGeneral Common Facility, and provides the researchers with various
special and common facility as well as safety regulations for accident-freedaily operation of the institute.

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Technology Licencing Office (TLO)
Who we are
・ The Technology Licensing Office (TLO) facilitates the conversion ofscientific discoveries to innovative technologies with the ultimate goalof improving public health and welfare.
・ We evaluate basic research findings (seeds) as intellectual
property assets, and license promising candidates to industries fordevelopment as medicines, diagnostics, medical devices, foods,
cosmetics and research tools.
General Manager

Futoshi Shibasaki, MD, PhD

Senior Manager

Kazumasa Aoki, PhD

What we do・ We manage intellectual properties from our institute including
patents, copyrights and materials in order to develop them forcommercialization.
・ To promote technology transfer, we introduce seeds andintellectual properties with potential commercial value to pharmaceutical, medical device, and startup
companies.
・ We attend business meetings such as the BIO international convention in the US, BIO-EUROPE, andBioJapan, to develop Public Private Partnership opportunities between our institute and industries.
We support collaborative research projects with industries by arranging Joint Research Agreements,
・ Material Transfer Agreements (MTA), and other contracts to protect and develop a wide range of
research discoveries.

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Center for Medical Research Cooperation

Head of Office

Masanari Itokawa MD, PhD

We provide---Consultation on statistical analyses-Ethical issues related to human specimens-A bridge between basic scientists and medical doctors

We provide tools for developing medical technology from
simple findings at the bench; the process similar to making abrilliantly sparkling jewelry by cutting a piece of crude stone.
We provide consultation on statistical analyses of raw data.
We manage ethical issues related to human specimens. We
facilitate the communication between the basic scientists andmedical doctors.

Conference with researchers and medical doctors

Most of discovery of scientific research is unexpectedly madeby scientists who still have very “flexible” scientific mind, theprivilege of young people.
A young scientist discussing with medical
doctors in a conference71

Neuropathology Laboratory
Laboratory Head

Nobutaka Arai

Translational Research using human materials
andManagement of Database for
Essential Brain Anatomy & Neuropathology

Laboratory of Neuropathology has more than 5,000 sets of human autopsied brain slides with a wide
variety of human neurological diseases. In recent years, we have been scanning these slides with virtual
slide instruments. Using those digital data and their derivatives, we construct digital neuropathology.

Neuropathology
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TMIMS Programs
Seminars/ lectures and related activities
Scientific seminars:

We have scientific seminars by renowned scientists from Japanand overseas almost every week in various subjects.

International Symposium:

We have three to four annual international symposium by inviting
top-flight scientists from all over the world in various subjects.

Science forum with industry:

We have annual forum hosted by our TLO office where scientists
at our institute introduce our discovery and discuss potential
applications with scientists from industry.

In-house and delivery lectures to high school students:
We give lectures to visiting high-school students or visit highschools to present lectures as a part of our effort to educate
youngsters and introduce them into this field.

Joint graduate school programme:

Many scientists at our institute have joint appointment as alecture or as a visiting professor at various universities. Many
undergraduate and graduate students are conducting their master/
Ph.D. researches at our institute under the guidance of out
scientists.

Open laboratory to graduate students:

Once a year we invite students to our institute, conduct
presentation and give them opportunities to experience thelaboratory.

Summer training courses:

We have annual summer training courses to provide in-housetraining on various experimental techniques.

Lectures to public:

We conduct public lectures(8 times a year) to let public know thescientific progress and our various activities made at our institute.

Science Café:

We have Café-style meetings (3-4 times a year) where we give
public the opportunities to learn, discuss and experience scienceand let them know the joy of science in a very casual setting.

Supports for young scientists
Research Associate Fellowship:

We provide graduate students who conduct their master/ Ph.D.
thesis research at our institute with research associate fellowship togive them financial support.

Travel support for young scientists attending aninternational meeting:

We provide students and young scientists of our institute with travel
fellowship with which they can attend international meetings where
they will present their latest findings.

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Access Map

AIRPORT to INSTITUTE

From Kamikitazawa Station to Institute
Walk (approx. 10 min From South entrance of Station).
From Hachimanyama Station to Institute74