INTERNATIONAL INSTITUTE FOR INTEGRATIVE SLEEP MEDICINEUNIVERSITY OF TSUKUBAIIISDIR EC TOR ʼS MESSA GE

Director’s message

We aim to solve one of the biggest black boxesof today’s neuroscience
Our discovery of the neuropeptide orexin and its
prominent role in sleep/wake regulation has generated ahighly active research field in neurobiology of sleep.
However, the fundamental governing principle for the
regulation of sleep pressure, e.g., the question of (1)what the neural substrate for “sleepiness” is, (2) why wehave to sleep, remains a mystery. Indeed, this is one of
the biggest mysteries in today’s neuroscience.
Based on my own 24-year experience as a PI in the US,
and by learning from the merits of US academia while
retaining the merits of Japanese traditions, IIIS provides
a scientific culture and environment that strongly
encourages all members, regardless of career stage, toinitiate and continue truly groundbraking studies.

Masashi Yanagisawa
Director

1

University of Tsukuba

A B OUT IIIS / FEA TUR ES OF THE INSTITUTE

World-class institute for sleep medicine,
aiming to solve the mechanism of sleep/wakefulness
by conducting basic to translational research
About IIIS
We spend nearly one-third of our lives asleep. However, the regulation and function of
sleep remain unclear. The importance of sleep is very clear because the loss of soundsleep impairs daytime performance and increases the risk of physical and mental
disorders. Deficiencies of sleep cause a huge economic loss, burdening the Japanesesociety about 1.5 billion yen per year. IIIS approaches this social problem throughinterdisciplinary sleep science, combining with basic neurobiology, pharmaceuticalchemistry, and experimental medicine.
IIIS is the world’s premier institute aiming to elucidate the fundamental principles of
sleep/wake biology, develop new strategies to diagnose, prevent sleep disorders, andcontribute to promoting well-being health through research activities.

Feature of the Institute
Based on Dr. Yanagisawa’s more than 20 years experience as a PI at the University ofTexas Southwestern Medical Center, IIIS has established the best and unique sleep
research center in Japan, by learning from the merits of the U.S.-style academic“department.” IIIS has created a free and vigorous atmosphere emphasizing: (i) flexible
and timely appointments of independent PIs regardless of their age and career stage,
with a necessary startup package (e.g. funding, personnel and space); (ii) a flexible anddynamic allocation of floor space for each laboratory to facilitate free and open
communications; and (iii) sharing of major facilities and capital equipment amonglaboratories. IIIS manages the organization so that all researchers and students canvigorously communicate and maximize their potentials.

M ISSION

To uncover the mystery of sleep, and to solve sleep-related social problems

Neuroscience

Our missions
mission1

To elucidate the function of sleep and thefundamental mechanisms of sleep/wake regulation

mission2

To elucidate molecular pathogenesisof sleep disorders and related diseases

mission3

To develop preventive measures, diagnostic methods,
and treatments for sleep

Experimental
Medicine

Establish
sleep science

Pharmaceutical
Science

To accomplish the missions above, IIIS has established a highly ambitious and novel research field, “sleep sciences”,
by integrating basic neurobiology, experimental medicine and pharmaceutical science.

International Institute for Integrative Sleep Medicine

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A B OUT LA B S

About Labs
There are seventeen core labs including twenty-three PIs, and fifteen collaborative groups with domestic andoverseas labs in IIIS, covering various research fields e.g. molecular genetics, neuroscience, physiology, medicinal
chemistry, human sleep physiology, etc. Those labs collaborate with each other to drive forward innovative sleep
research.
MasashiYanagisawa●●● ●Hiromasa
FunatoTakeshi
Sakurai●● ●●Qinghua
Liu

●●● ●

Arisa
Hirano●●●●Neural circuit
related tosleep/wakeregulation

Katsuyasu
Sakurai●●●

Yu
Hayashi●●

Genes related
to sleep/wakeregulation

Masanori
Sakaguchi●●

Sleephomeostasis& circadianrhythmNeuroscience

SakikoHonjoh● ●●MichaelLazarus

Hirofumi
Toda

●●●

Yo
Oishi
●●

●●
Experimental
Medicine

Sleep
disordersEstablish
sleep science

Pharmaceutical
Science

Development of
new medicine
regulating
sleep/wake●●RobertGreeneNorikiKutsumura●●●Kaspar
Vogt

Data Science

Sleep & Physiology
Psychology
Sports Science

TakashiKanbayashi●●●

Hiroyuki
Kitagawa

Tomohiro
Okura

●●

●●

Masao
IwagamiShun
Nakajima

●●

●●

Shoi
Shi

Takashi
Abe

●●

●●

Satellite

Overseas
Satellite
GroupsCollaborative Research Group

Domestic
Satellite
Labs

Genshiro A. Sunagawa

Masayuki Matsumoto

Hitoshi Shimano

Haruka Ozaki

Morimitsu Kurino

Yuichi Inoue

Akiyoshi Fukamizu

Toshiyuki Amagasa

Satoru Takahashi

Shinji Saiki

Hiroyasu Ando

3

Tsuyoshi
SaitohUniversity of Tsukuba

MEMB ER S

Members

Aiming to uncover the reciprocal linkage between
intracellular events and sleep/wakeful behavior
Our lab has been conducting forward genetic research on sleep/wakefulness using randomly
mutagenized mice. Luckily, we succeeded in identifying a protein kinase SIK3, histone
deacetylase HDAC4, and a non-selective cation channel NALCN as novel molecules regulating
sleep. Further studies using gene-modified mice and AAV-based gene manipulation have revealed
that the LKB1-SIK3-HDAC4/5 pathway constitutes intracellular signaling that regulates sleep
need. To unravel an enigmatic molecular mechanism for sleep need, we employ a variety of
research techniques including multi-omics approaches, such as quantitative phosphoproteomics
and single-cell RNA-seq. In addition, the long sleeper phenotype of Sik3 mutant mice provides
us a unique opportunity to examine the benefit of long total sleep time in aging, metabolism, andother phenotypes relevant to human health and diseases.

Masashi Yanagisawa
Director

Nalcn-mutant mice (Dreamless mutant mice) show a short total REM sleep and short duration of
REM sleep episode. We aim to understand how REM sleep episode is maintained and how total
REM sleep time is homeostatically regulated through our research focusing on NALCN.
Importantly, we continue the forward genetic screening to discover novel genes regulating
sleep/wakefulness.

Hiromasa Funato

Forward genetic studies with other state-of-the-art techniques have identified the LKB1-SIK3-HDAC4/5 pathway
regulates the quantity and quality of NREM sleep.

International Institute for Integrative Sleep Medicine

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MEMB ER S

Just enjoy journey of research!
~ There is always something beyond our expect
A brain is the device that processes huge amount of information via nerve cells called neurons. How
animal’s brain interprets information from the external world and trigger appropriate behaviors alongwith autonomic/neuroendocrine responses? Our group combines optogenetics/pharmacogenetics andcutting-edge genetics in mice to address this question at a high level of precision.
We try to identify substances, neurons and neuronal pathways that are involved in eliciting particularbehaviors by means of various histological andphysiological techniques applied in geneticallymodified mice. We manipulate synaptic transmissions
or neuronal excitability in a cell or pathway specificmanner to characterize the consequences of thosemanipulations

Takeshi Sakurai
Vice Director

on

the

behavior

and

other

physiological responses.
These techniques allow us to study the physiological
function of neuronal pathway and substances in theregulatory mechanisms of our behavior. Sleep/
wakefulness behavior, emotion, circadian rhythm andsocial behaviors are our current interest. Along with
its academic values, the elucidation of mechanisms ofthese functions may lead to treatments of various
diseases.

We examine function of neuronal pathways and neurotransimitters in theregulation of sleep/wakefulness states and behavior, by means of
optogenetics, which enables us to manipulate particular neuronal
circuits.

From molecular to behavior: Aiming to understandmysterious biological phenomenon “Sleep” at various levels
We aim to understand why and how we sleep at night and wake up in the morning. In almost organisms
living on the earth, many biological processes including sleep exhibit rhythms with a period of ~24 hours,
which is called as circadian rhythms. Well-coordinated rhythms by internal time-keeping system, circadianclock, is essential for physical and mental health. We previously identified several gene variants in clock
genes altering sleep/wake pattern in humans by forward genetics approach. It is thus well established thatthe circadian clock has a great impact on sleep/wake rhythms, while the neural pathway from the masterclock in the brain (suprachiasmatic nucleus, SCN) to sleep/wake-regulating regions is still elusive. To
understand the neural network controlling the circadian sleep behavior, we utilize gene-modified animals,
macro injection and opto/chemo-genetics technics to evaluate the effect of manipulation of neurons in the
SCN or clock-target regions on sleep/wake rhythms. In addition to the output pathway from the clock, weare also interested in oscillatory mechanism of the clock within the SCN neurons (intracellular oscillator)
as well as mechanism of entrainment of the clock to environmental cycle such as light-dark cycle (input
pathway), which are considered as three main factors of the circadian clock. We believe that
understanding of the whole picture of the circadian clock is key to advance sleep study.
The circadian clocks receive virous stimuli, such as light,
temperature and feeding to entrain to the environmental cycles.
The basic structure of the circadian clock is an intracellularmolecular oscillator composed of several clock genes. The
master clock (center clock) resides in the suprachiasmatic
nucleus (SCN) in the hypothalamus in mammals. Through neuralcircuits, various brain regions input time information from SCN
to achieve biological rhythms including sleep/wake rhythms,
while it has been unknown which circuit is actually involved in
the output pathway.

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University of Tsukuba

Arisa Hirano

MEMB ER S

Why do we fall asleep when bored?
~ The gating of sleep by motivated behavior
The investigative focus of our laboratory is the cellular and synaptic basis by which the brain regulates
sleep and wakefulness. Our experiments seek to link the activity of defined sets of neurons withneurobehavioral and electroencephalographic outcomes in behaving animals by using innovative
genetically or chemically engineered systems (optogenetics, chemogenetics or optopharmacology) in
conjunction with recording of the electrical activity
produced by the brain or in-vivo imaging (fiber-optic

endomicroscopy).
For example, we investigate the control of sleep andwakefulness by the mesolimbic pathway comprising
the ventral tegmental area and nucleus accumbens. As
the mesolimbic pathway is implicated in motivational

Michael Lazarus

and cognitive behaviors, changes in vigilant states arelikely associated with the motivational and cognitive
responses in animals. Moreover, our laboratory is

Motivation, reward, pleasure

Nucleusaccumbens(GABA)

Ventral pallidum of
the basal forebrain(GABA)
Ventral tegmental
area (dopamine)

exploring novel agents for the treatment of insomnia
by targeting mesolimbic cell populations that have theability to produce sleep.
We also employ single-cell or spatial gene expression
profiling to understand how the sleep/immune systemcrosstalk is regulated at cellular and molecular levels.

In the absence of motivational stimuli, a tonic sleep drive by the NAc
may allow the brain to fall asleep by depressing arousal circuits in thebasal forebrain.

To elucidate the mechanism and necessity of sleep
with mice like “short sleepers”
People spend approximately one-third of their life sleeping. Why sleep is compulsory for human life andcannot be avoided, however, remains unclear. It is important to understand why sleep is necessary and how
sleep is controlled – not only to elucidate physiologic behavior but also to enhance our quality of life.
Recently, in the course of analyzing sleep-wake regulation by the reward system, we happened to create
mice that require an extremely low amount of sleep. Surprisingly, these mice do not show an increase in asleepiness marker despite of the less sleep amount. In addition, they do not show sleep rebound after sleep
deprivation. Therefore, the behavior is very similar to that of so-called “short sleepers” – people that canfunction for long periods of time on little sleep without exhibiting excessive sleepiness. Using multiple“short sleep” models, including this novel mouse model, we study the neural mechanisms of short sleep toclarify the control mechanisms of sleep. We also study the effect of short sleep on other physiologic
functions in the body to understand the necessity of sleep.
In close collaboration with Prof. Lazarus, we use many neurobehavioral and biochemical techniques to

Yo Oishi

perform this research.

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%

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%


%%Amount of sleep and wakefulness ofnormal and “short sleeper” mice during
1 day. “Short sleeper” mice exhibit alower amount of sleep.

International Institute for Integrative Sleep Medicine

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MEMB ER S

How are memories organized during sleep?
~ The mystery of sleep and memory
During sleep, a phenomenon called "memory consolidation" occurs, in which we unconsciously organize
memories and reinforce necessary memories. There are various states of sleep, such as non-REM sleep andREM sleep, and each of these states plays a different role in memory consolidation. Our research team ischallenging to clarify the mechanisms of memory consolidation during sleep and to apply the results todevelop a novel therapy for PTSD.
For example, we have developed an AI that analyzes non-REM and REM sleep in real time. We also useda small, light microscope that can be placed on an animal's head to observe the activity of neurons in thebrain during sleep. Combining these studies, we are investigating the function of manipulating specificneuronal activity during sleep in memory consolidation.
From these studies, we have found that the activity of newly generated neurons in the adult hippocampus
is essential for memory consolidation during sleep. We have also shown that memory consolidation can be

Masanori Sakaguchi

manipulated by playing memory-linked sounds during specific phases of sleep. We are conducting clinicalresearch to apply this technology to realize a new home treatment for PTSD patients suffering fromtraumatic memories.
In the future, we hope that thesestudies will lead to better sleep and thedevelopment of new therapies that
apply this technology by clarifying themechanisms of memory processing
during sleep.

Mini-microscope on the hand andfull-scale microscope (behind)

Why we sleep?
~ Addressing the functional roles and evolutionary origin of sleep
Our goal is to elucidate what the function of sleep is. Why sleep is needed to maintain our health still
remains largely unknown. To address this large mystery, we focus on two animal species, the roundworm
Caenorhabditis elegans and mouse. The nervous system of C. elegans consists of merely 302 neurons. We
previously obtained evidence suggesting that sleep in this simple animal and mammalian sleep are
evolutionarily conserved. Furthermore, in mice, we successfully identified neurons that are crucial for the
regulation of REM (rapid eye movement) sleep, a major source of vivid dreams, and established mice
where REM sleep could be efficiently inhibited. Using these two model animals, we aim to elucidate theroles of sleep at multiple levels from molecular and cellular to individual levels. Moreover, we aim to apply
the obtained knowledge to understand and develop treatments for neurological disorders such as
dementia.

Yu Hayashi

7

University of Tsukuba

MEMB ER S

What does the cortex do during slow wave sleep?
~ To elucidate its mechanisms and functions
We are interested in cortical activity during deep, so-called non-REM or slow wave sleep, and during
waking. Slow wave sleep restores brain function from the reduced state after long waking to the refreshedstate after waking up. It also improves memory and has many other beneficial effects. How slow wave sleep
does this things is currently not known.
The cortex is the outermost part of the brain; higher functions such as perception, consciousness andlearning and memory depend on it. We record electrical signals from individual neurons or groups of themin freely behaving mice and also use in-vivo imaging to study the activity of these neurons. In slow wave
sleep cortical neurons show distinct

Robert Greene

rhythmic activity, different fromwaking. The more an animal needs tosleep, the stronger this rhythm
becomes.
We want to understand what controls
and creates this rhythm and how itchanges the ways in which neurons
interact. For this we develop new
algorithms to analyze the activity ofneurons and use molecular tools toactivate or silence certain types ofKaspar Vogt

neurons with high precision.

(1) Schematic of a brain with the postiton of the light stimulus and the electrodes that record the
response. With this experiment we can stimulate the brain in all states waking and sleeping andcan measure the response. We have found that the responses are larger in sleep than in waking.
(2) With this microscope we can look into the brain of a living mouse. We examine special mice,
which express a protein in their brain that changes its visibility, depending on how active thenerve cells in the brain are. We want to understand how the activity in a sleeping brain differs fromwaking.

How to generate mice who never sleep?
~ To fully understand and regulate sleep
We are interested in homeostatic sleep/wake regulation in behavioral vigilance states (e.g., mice sleep
longer after when they stayed awake longer) and in cortical neural activity (e.g., a cortical region sleeps
deeper when the region has been active during prior waking).
It is becoming clearer that brain regions contribute to sleep/wake regulation to a different extent. Several
wake, and nonREM sleep and REM (REM; rapid eye movement) sleep-promoting or -inhibiting areas
have been identified. To understand the nature of the homeostatic regulation, using mice, we investigate
what kind of changes the critical sleep/wake regulating centers undergo during persistent waking andsubsequent sleep.
As for the cortical activity, we focus on slow wave, a hallmark of NREM sleep EEG. Though it is the best
marker for sleep need so far and it plays important roles in sleep-dependent memory consolidation, theunderlying neural circuits remain largely unknown. Currently we investigate the role of thalamic matrix

Sakiko Honjoh

cells in slow wave generation.

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International Institute for Integrative Sleep Medicine

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MEMB ER S

Elucidating the Neural Mechanisms
involved in Sleep drive and Appetite
Living organisms, including humans, survive by adapting to ever-changing internal conditions (physical,
physiological, mental, etc.), environment, and stimuli. For example, instinctive behaviors such as appetiteand sleep drive are greatly influenced by the internal state of mind, body, and environment. Theirdisruption can have a profoundly negative impact on healthy daily and social life. Therefore, acomprehensive understanding of the neural mechanisms involved in appetite and desire for sleep is crucial
for both individuals and society.
Detailed studies have been conducted on the neural mechanisms of appetite, from the molecular to theneural circuit level. On the other hand, the neural mechanisms of anorexia, a state of decreased appetite,

Qinghua Liu

remain largely unknown. Therefore, our research aims to comprehensively understand the neuralmechanisms of anorexia from the molecular level to the neuralcircuit level. To understand the neural circuits, we are focusing
on brain regions that monitor brain-organ connections and thestate of the body and mind.
Concerning sleep research, we are focusing on the effects offood on sleep. One of these studies seeks to elucidate the neuralmechanisms of postprandial sleepiness. Although we frequently
experience postprandial sleepiness, its mechanism has not beenclarified. We are trying to elucidate the brain-organ coupled

Katsuyasu Sakurai

signaling mechanisms involved in postprandial sleepiness fromthe molecular to the neural circuit.

How is the sleepiness represented inside a brain?
~ The power of using fruit flies in sleep research
The main goal of our lab is to understand the molecular and cellular mechanisms of sleep. Sleep isimportant and conserved in all animals from humans to insects. Yet, the function of sleep: why we need it,
and the mechanisms of sleep: how it occurs remains largely mystery.
In order to understand the mechanisms of sleep, we use fruit flies as a model system to tackle suchquestions because flies are fantastic genetic model organisms especially amenable for genetic screening.
For example, to identify molecular factors that induces sleep, we carried out a genomewide behavioral
screen, evaluating over 12,000 fly lines, and found a novel sleep inducing factor named “nemuri”. Nemuri
functions as an anti-microbial peptide that is essential for bacterial infection-induced sleep, suggesting
Nemuri served as a key link between sleep and immunity.
Our group combines genetic, biochemical, behavioral and imaging techniques to elucidate themechanisms of how Nemuri functions inside neurons to induce sleep. Moreover, we aim to identify the
mammalian homologue of nemuri through behavioral screening by expressing human candidate genes infly brain. These studies will improve our understanding of how the sleepiness is represented inside thebrain.

9

University of Tsukuba

Hirofumi Toda

MEMB ER S

Solving mysteries of life through organic chemistry
We are engaged in drug discovery research and drug discovery-related research activities. Medicinal
chemistry is very challenging research that is based on many disciplines, including medical science,
pharmacology, biochemistry, molecular biology, pharmacology, pharmacokinetics, and drug formulation,with a focus on synthetic organic chemistry. Due to the nature of the discipline, it may be said that one of
the characteristics of medicinal chemistry research is that it often develops into collaborative research withcompanies and other universities.
Our main research activities can be broadly categorized into "chemical synthesis of small moleculecompounds that act in vivo" and "development of chemical reactions useful for synthesizing suchbiologically active molecules". In the synthesis of small molecules acting in vivo, we aim to create "superiormolecules" through structure-activity relationship study and structure optimization study. Research
themes, for example, include the synthesis of compounds that specifically act on protein phosphatases andNoriki Kutsumura

structure-activity relationship studies based on natural products acting on orexin receptors, both of whichlead to the elucidation of sleep/wake mechanisms. On the other hand, in the development of chemical
reactions, we aim to develop novel chemical reagents and/or synthetic methods that anyone can easily
utilize. Specifically, we are interested in thedevelopment of sulfurizing reagents without badodor or chemoselective halogenation methods.
Our goal is to develop innovative chemical
reactions that can be used by pharmaceutical andchemical companies in near future.

Innovative new drugs from IIIS
Our group is the only chemistry-based group in IIIS engaged in drug discovery research with the goal of'New Drugs from IIIS!'.
Our group focuses on a class of membrane proteins called G-protein coupled receptors (GPCRs). GPCRs
are the largest protein family in the human genome and are involved in a wide variety of diseases.
Therefore, approximately 30% of all market drugs target GPCRs, making them the biggest group of drug
targets. Our group is mainly targeting orexin receptors involved in arousal and emotion, opioid receptors
involved in pain and itch, and adenosine receptors involved in sleep and inflammation to serve new drugs
for sleep disorders such as narcolepsy and insomnia, pain, drug dependence, depression, anxiety, andother psychiatric disorders. For example, in 2015, we discovered the first orexin 2 receptor (OX2R)
selective agonist, YNT-185, from Hit compounds obtained through a high-throughput screening (HTS)
campaign and demonstrated that OX2R agonists are effective for the treatment of narcolepsy.
Recently, we have been challenging the Structure-Based Drug Design (SBDD) of GPCR-targeting drugs

Tsuyoshi Saitoh

by incorporating the latest technologies such as cryo-EM SPA and wide-range signal analysis toward thelogical design of drugs without side effects.
We have also been developing innovative
chemical tools that contribute to biology andmedical science by combining our drugs andchemical reactions with the latest biological
techniques.

International Institute for Integrative Sleep Medicine

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MEMB ER S

Understanding the human mind and behavior
related to sleep
Our laboratory aims to understand human mind and behavior related to sleep. Currently, we are focusing
on two main areas of research. (1) Our laboratory is conducting research to understand the role of human
REM sleep in waking emotional processing. We have discovered a brain potential that occurs before rapid
eye movements during REM sleep, which we have named the pre-rapid-eye-movement negativity (PRN).
Our research also found that the PRN originates from the ventromedial prefrontal cortex. This brain
region is involved in emotional regulation, so we believe that the PRN reflects emotional processes during
REM sleep. We are using the PRN as a marker to investigate the psychological functions of REM sleep.
(2) Our laboratory is conducting research to understand and mitigate
performance deficits caused by sleep loss in order to prevent accidents
and human errors related to sleep loss. One aspect of this research
involves developing a new system for evaluating and predicting declines

Takashi Abein vigilant attention due to sleep loss using an accurate and unobtrusive
measure. We have identified several new markers for vigilant attention
and developed a new algorithm for detecting the level of vigilant
attention by integrating these markers. We are conducting research
towards its practical implementation. In addition, we are also exploring
ways to prevent performance deficits caused by sleep loss. Ourlaboratory is developing non-invasive methods for promoting sleep.
Through these research and technological developments, we aim toprevent accidents and human errors related to sleep loss.

Using a laboratory that can simultaneously
measure the sleep and wakefulness of four
people, we are conducting research how sleep
and sleepiness relate to the human mind andbehavior.

Elucidating the pathogenesis of central hypersomnia andschizophrenia through research on neural autoantibodies
It has already been 20 years since orexin was discovered, but it is still the only biochemical marker for
diagnosis of sleep wake disorders. Other than orexin, diagnosis is made only by sleep polygraphexaminations and clinical symptoms. Since orexin is decreased in narcolepsy, it has been measured from2000 for a definitive diagnosis for narcolepsy and about 3000 samples have been measured. Orexin is also
an important biomarker of symptomatic hypersomnia due to hypothalamic lesions. We have reported
symptomatic narcolepsy due to AQP4 antibody as the first sleep wake disorder involving an autoimmune
mechanism. In addition to cases with hypothalamic lesion, it has been reported that orexin decreases andsymptomatic narcolepsy occurs in hereditary diseases and neurodegenerative diseases. One of the goals of
our group is to clarify the pathophysiology
of hypersomnia through the study ofsymptomatic narcolepsy.
Similar to AQP4 antibody, NMDA receptor
antibody that acts on the surface of cell
membranes

causesNMDA

receptor

encephal it is with psychosis. Schizophrenia
is a disease of unknown cause, but webelieve that it may also develop due to an
Left: An MRI image of NMO with hypersomnia. Hypothalamus and orexin system are
impaired by AQP4 antibody. Right: The classification symptomatic narcolepsy withdecreased orexin levels. NMO, MS, ADEM, Ma2 encephalitis are caused due to the
autoimmune mechanism.

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University of Tsukuba

autoimmune mechanism, such as NMDA
receptor antibody. We are continuing toconsider this direction of research.

Takashi Kanbayashi

MEMB ER S

Supporting people's lives through psychotherapy
and digital technology
Globally, one in seven individuals experiences sleep problems. While pharmacological therapy might be
the common perception for those with insomnia, cognitive-behavioral therapy (CBT-I), a form of
psychotherapy, is advocated as the first-choice treatment for insomnia. However, there remains a lack ofwidespread access to cognitive-behavioral therapy for insomnia (CBT-I). In response to this issue, ourresearch laboratory is conducting studies to provide effective psychotherapy to individuals of all ages whoare suffering from sleep and mental health problems.
Furthermore, we have recently initiated research on digitizing therapists' empathic abilities andimplementing empathy into AI therapists. Additionally, we are developing psychotherapy based on
objective markers derived
fromShun Nakajima

variousdigitaldevices. We believe that
advancing research thatcombines subjective andobjective data could leadto breakthroughs in sleep
healthcare.

Exploring safe and enjoyable exercise conditions
to improve sleep
Methods of improving sleep are divided into drug therapy and non-pharmacological therapy. Because
various side effects have been reported in the elderly, American Geriatrics Society recommends thatsleeping pills not be prescribed. We have focused on the study regarding exercise therapy for good sleep.
Few previous studies examine the effects of exercise on sleep in the elderly, and it is unknown what kindof exercise may be effective in improving sleep. Also, most of the previous studies examined on simple
exercise by using a treadmill or cycling ergometer. In order to improve sleep by exercise, it is important tocontinue doing exercise, but the aforementioned exercise is easy to get tired of and has a high risk of injury.
So, we put safe and enjoyable exercise first and comprehensively reviewed the intensity, type,
implementation time, and form of exercise. As a result, we confirmed that performing low-intensity
aerobic exercise for 30 minutes at night is effective for improving sleep. In addition, compared to simple
exercise multi-tasking exercise with stimulating frontal lobe showed the effect of increasing deep sleep
along with activation of the frontal lobe immediately after exercise. In other words, appropriately

Tomohiro Okura

stimulating frontal lobe
and going to bed may beeffective in improving
sleep for the elderly.

International Institute for Integrative Sleep Medicine

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MEMB ER S

Sleep data science and engineering~ Big data analysis, databases, data mining
Data exploration is recognized as the fourth paradigm of science after experimentation, theory, andsimulation. Data-intensive research has been more and more important in science, and sleep medicine is
no exception. In the past, special measuring equipment and hospitalization were required to collect sleep
data for analysis and diagnosis. Advances in IT have made it possible to easily measure sleep data at home.
We are approaching the science of sleep from a data science and engineering perspective.
One of our recent research topics is automatic sleep stage scoring. Sleep is classified into stages such as
REM sleep, non-REM sleep, and wake, and the analysis of stage transitions is essential in understanding
sleep. Conventionally, sleep stages were determined by experts, but this requires a high level of expertiseand a lot of time. By utilizing machine learning technology such as deep learning, we have developed asystem that can automatically determine sleep stages with the same accuracy as experts and can present
the basis for the determination. Other research themes include improving the robustness of analysis

Hiroyuki Kitagawa

against noise, technology to flexibly respond to thecharacteristics of each subject, and real-time analysis
of electroencephalogram data acquired during sleep.
To strengthen our backbone technologies, we are also
actively engaged in research on innovative technologies
in data engineering and data analysis such as big data
analysis, databases, data mining, and machine learningin collaboration with the Center for Computational
Sciences.

Establishing “Sleep epidemiology” as a branchof epidemiology
Epidemiology is the study of diseases in populations and its scope ranges from disease causation andprevalence to disease prevention. Environmental epidemiology, nutritional epidemiology, genetic
epidemiology, among others, have been established as the main subfields of epidemiology. Althoughepidemiological research on sleep has been conducted as part of various cohort studies, “sleep
epidemiology” is not yet a fully established branch of epidemiology. This is mainly due to the difficulty ofmeasuring sleep objectively.
However, recent advances in technology enable at-home monitoring of brain waves during sleep andrecording of sleep parameters using smartphones. The opportunity for the development of sleep
epidemiology arises when large-scale data acquisition becomes possible. Our goal is to collectobjective sleep data on a large scale, analyze it in conjunction with various health information, clarify
the true relationship between sleep and disease, and extend people's healthy life expectancy throughimproved sleep.
Furthermore, routinely collected clinical data haverecently been accumulated and made available forclinical research in Japan as well, including records ofconsultation and prescriptions for narcolepsy, sleep
apnea syndrome, and insomnia. We will also contributeto the establishment of sleep epidemiology by
InSomnograf, a device to measure brain waves during sleep at home
(S'UIMIN Inc., a start-up company from the University of Tsukuba)

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University of Tsukuba

promoting epidemiological research on sleep-relateddiseases utilizing these records.

Masao Iwagami

Comparative neuroscience has a long tradition of studying the structure and function of the central
nervous system. Advances in imaging technologies have enabled the examination of structural andfunctional properties at the single-spine level in some model organisms. However, it is undeniable thatneural structures in a given species are constrained by the unique evolutionary history and characteristicsof that species. As such, we propose a new approach, referred to as Comparative Neuroscience 2.0, in
which model and non-model organisms are compared at the same resolution to distinguish species-specificfeatures from those that are evolutionarily conserved. This approach allows for the identification of thesmallest structural and functional units that are shared across organisms.
Sleep is a physiologic process that is conserved across a diverse array of organisms, including mammals,
birds, and reptiles. Our research group aims to understand the mechanisms and functions of sleep, as wellas its evolutionary significance. To this end, we study

Shoi Shi

mice as a representative vertebrate and ants as arepresentative arthropod. We also collaborate withresearchers studying other non-model animals anddevelopingnewtechnologies

forcomparative

neuroscience. Our team comprises members withdiverse expertise, including neurosciences, genetics,
computational science, and other fields, and we
employ a range of techniques to realize quantitative
comparisons between species.

Satellite and University of Tsukuba
Collaborative Research GroupWe are creating novel imaging and phenotyping techniques specificallydesigned for non-model organisms. By integrating these technologies
with data science and mathematical techniques including clustering,machine learning, and mathematical modeling, we are able to investigate
and expand the field of comparative neuroscience.

Our sleep research network is also spreading outside of IIIS. Throughcollaboration with fifteen satellite laboratories and collaborative groups in
University of Tsukuba, we are powerfully pushing ahead with unique sleep
research including basic and translational research.

MEMB ER S / SA TELLITE A ND UNIVER SITY OF TSUKUBA COL L ABORATI VE RE S E ARCH GROUP

Exploring the Conservation and Diversity of Sleep
by Comparative Neuroscience

Satellite
Overseas Satellite Groups

Domestic Satellite Labs

Genshiro A. Sunagawa Morimitsu Kurino

Hiroyasu Ando

Masayuki Matsumoto Yuichi Inoue

Center for Biosystems
Dynamics Research,
RIKEN

Advanced Institute forMaterials Research,
Tohoku University

Center for the
Evolutionary Origins
of Human Behavior,
Kyoto University

Faculty of Economics,
Keio University

Collaborative Research Group

Institute of
Neuropsychiatry

Hitoshi Shimano

Akiyoshi Fukamizu

Satoru Takahashi

Haruka Ozaki

Toshiyuki Amagasa

Shinji Saiki

Faculty of Medicine

Faulty of Life andEnvironmental Sciences

Faculty of Medicine

Faculty of Medicine

Center for Computational
Sciences

Faculty of Medicine

International Institute for Integrative Sleep Medicine

14

GLOSSA R Y

Glossary
Non-REM Sleep/REM Sleep(REM; Rapid Eye Movement)

Hours

  
 
Shallow

Deepabout 90 minutes

Our sleep can be classified into two stages by brain wave, non-REM
sleep and REM sleep. Just after falling asleep, a shallow non-REM
sleep begins and it gradually becomes deeper as time passes. As brain
activity decreases compared with that during arousal, non-REM sleep
is often considered as "sleep of the brain". Additionally, during deep
non-REM sleep, a characteristic brain wave called “slow wave” isobserved. This is caused by the synchronization of neuronal activity inthe brain. After about 90 minutes of non-REM sleep, REM sleep
occurs. During REM sleep, skeletal muscles relax while the brain
activity increases, and vivid dreams are typically experienced.
Although it is often mentioned that non-REM sleep is a deep sleep andthat REM sleep is a shallow sleep, this is not really correct. These twosleep stages are completely different. In overnight sleep, we regularly
repeat the periodic of non-REM sleep and REM sleep cycles, theneventually leading to wakefulness.

Orexin is a substance in the brain (neuropeptide) that regulates sleep and arousal, discovered in1998 by Dr. Masashi Yanagisawa and Dr. Takeshi Sakurai. It is secreted from neurons in the
hypothalamus and functions to keep the arousal state. It is known that orexin deficiency causes
narcolepsy, patients of which experience excessive daytime sleepiness, uncontrollable sleep attacks,
and sudden loss of muscle tone triggered by intense emotions such as surprise and amusement.
Orexin interacts with proteins called “receptors” on the surface of neurons. There are two types oforexin receptors, OX1R and OX2R, and these receptors are also research and development targets
for pharmaceuticals. "Antagonists", which compete with orexin to bind to receptors and inhibit its
effect, have been used in medicines such as Suvorexant, Lemborexant, and Daridorexant for thetreatment of insomnia. Conversely, "Agonists", which bind to the orexin receptors and cause thesame effect as orexin are expected to be applied to treatment for narcolepsy. Several compounds
have been found already, and some clinical studies have been conducted to confirm theireffectiveness and safety.

Q-neuron-induced
hypothermia andhypometabolism (QIH)

Normal mouse (left, body temperature 37°C) and QIH mouse (right, bodytemperature 23°C). QIH mouse

15

University of Tsukuba

OrexinComplex structure of Orexin B,
OX2R, and G proteins

QIH is an artificially induced hypothermic and hypometabolic state in mice orrat, which has similar characteristics to hibernation observed in some kinds ofspecies, such as Syrian hamsters and ground squirrels. In 2020, Dr. Takeshi
Sakurai, the vice director of IIIS, discovered that quiescence-inducing neurons(Q neurons) in a small population in the hypothalamus potentially induce thishibernation-like state, QIH. An excitation of Q neurons by chemogenetics or
optogenetics rapidly decreases mouse body temperature, while the homeostatic
regulation of the body temperature is still conserved. Oxygen consumption,
heart rate, locomotor activity and brain activity are also largely suppressed
during the excitation of Q neurons. However, once the neuronal manipulation
stops, these parameters including the body temperature return to normal levels
without any damages in the tissues. This artificial hibernation technique has
many potentials in clinical application. For example, in the emergency medicalcare or tissue transplantation, we are possibly able to prevent server tissue
damages by reducing the oxygen demand, leading to an increase of the survival
rate. Future applications could include manned space exploration.

Optogenetics・Chemogenetics
Optogenetics

Chemogenetics

Neurons expressing effector molecules that turn

Neurons expressing effector molecules that turnneural activity on and off with specific wavelengths of lightneural activity on or off depending on specific compounds

Proteomics is the method to comprehensively identify the proteins in cells or
tissues and elucidate their functions using a mass spectrometer. Allbiological phenomena are driven by various proteins encoded in the genome.
"Post-translational modifications" is one of the important processes toregulate the functions of various proteins. “Phosphorylation”, one of the
post-translational modifications, switches the activity of proteins by adding
a phosphate group to the proteins. The phosphoproteomic analysis enables
us to comprehensively reveal the phosphorylation status of all proteins.
While the genomic information in all cells is basically the same, theinformation on proteins varies among the cell types or tissue types, andchanges over time or depending on the environment, even in the same
animal. Proteomic and phosphoproteomic analyses allow researchers to
obtain more detailed and complex information about the molecularmechanisms of biological phenomena.

GLOSSA R Y

Forward Genetics is a research approach for exploring the genetic basis
responsible for a certain inherited characteristic (phenotype). For example,
IIIS researchers are searching for novel genes involved in sleep/arousal
regulation as follows.
1. Using a certain chemical, generate enormous numbers of animals thathave random mutations in their genome. (Usually, animals with short
generation time such as nematodes and Drosophila are used, but at IIIS,
forward genetics research is being conducted using mice, which isunprecedented in the world.)
2. Observe the brain waves of the offspring of mice obtained above to findmice with sleep abnormalities. Then, we examine whether the abnormality
is inherited by the next generation.
3. Analyze the genome of those mutants and find out gene mutations
responsible for inherited sleep abnormalities.
Although examining the brain waves of a very large number of mice istime-consuming and labor-intensive, this hypothesis-free approach maylead us to serendipitous discoveries of sleep-regulating genes andmolecular mechanisms of sleep regulation.

Forward Genetics

The sleep measurement facility for mice in IIIS. For forward genetics research, alarge-scale facility is necessary.

Neurons, the cells that make up the brain, use electricalsignals to process and transmit information. Changes
in the electrical potential between the outside andinside neurons generate these electrical signals.
Therefore, neural activity can be turned on or off if we
can artificially cause a change in the potential between
the inside and outside of a neuron.
Techniques that can artificially alter neural activity
include optogenetics and chemogenetics. Bothtechniques require that the neurons whose neuralactivity is manipulated express effector molecules,
using genetic engineering, to cause a change in the cell's
potential. Optogenetics and chemogenetics canartificially alter neural activity by activating effector
molecules through irradiating specific wavelengths oflight and introducing specific compounds, respectively.

Proteomics

Mass spectrometer for proteomics

International Institute for Integrative Sleep Medicine

16

P HOTO GA LLER YIIIS Parking

N
100m

University Hospital W.
cannot pass through by car

Gate

Oikoshi Gakusei Shukusha-mae

Nishi Odori

International Institute for Integrative
Sleep Medicine
University of Tsukuba

University of Tsukuba
Hospital

University Information Center

1-1-1 Tennodai, Tsukuba, Ibaraki, 〒305-8575
TEL/FAX +81-29-853-5857 / 3782

Kasuga 3 Chome
To TX Tsukuba Station

17

University of Tsukuba

The World Premier International Research Center Initiative (WPI) was launched in 2007 by the Japanese
Government’s Ministry of Education, Culture, Sports, Science and Technology (MEXT) with the aim ofbuilding globally visible research centers with high research standards and excellent research environments toaccommodate globally prominent researchers. The International Institute for Integrative Sleep Medicine(WPI-IIIS) was established on December 1, 2012, following the FY 2012 selection round of its application for
the WPI. The initiative seeks to create a vibrant environment where frontline researchers, regardless of nationality, are invitedfrom around the world to engage in research where they are accompanied by an administrative support system that allows thesescientists to fully devote themselves to their research.

A B OUT WP I

About WPI

Four Missions of WPI

1Advancing leading edge
research

3

Reforming researchorganizations

2

Establishing international
research environments

4

Creating interdisciplinary
domains

Hokkaido University
Institute for Chemical Reaction Design and Discovery (ICReDD)(Selected in 2018)

Tohoku University
Advanced Institute for Materials Research (AIMR)(Selected in 2007. WPI Academy)

Tohoku University
Advanced Institute for Marine Ecosystem Change(Selected in 2023)

High Energy Accelerator Research Organization (KEK)International Center for Quantum-field Measurement
Systems for Studies of the Universe and Particles（QUP）(Selected in 2021)

University of Tsukuba
International Institute for Integrative
Sleep Medicine (IIIS)(Selected in 2012)

Kanazawa University
Nano Life Science Institute (NanoLSI)(Selected in 2017)

Kyoto University
Institute for the Advanced Studyof Human Biology (ASHBi)(Selected in 2018)

Kyoto University
Institute for Integrated
Cell-Material Sciences (iCeMS)(Selected in 2007. WPI Academy)

Hiroshima University
International Institute for Sustainability
with Knotted Chiral Meta Matter（SKCM²）(Selected in 2022)

Kyushu University
International Institute for Carbon-Neutral
Energy Research (I²CNER)

The University of Tokyo
Kavli Institute for the Physics andMathematics of the Universe (Kavli IPMU)(Selected in 2007. WPI Academy)

National Institute for Materials Science
International Center for Materials
Nanoarchitectonics (MANA)(Selected in 2007. WPI Academy)

The University of Tokyo
International Research Center for
Neurointelligence (IRCN)(Selected in 2017)

Keio University
Human Biology-Microbiome-Quantum
Research Center（Bio2Q）(Selected in 2022)(Selected in 2010)

Osaka University
Immunology Frontier Research Center (IFReC)

Tokyo Institute of Technology
Earth-Life Science Institute (ELSI)(Selected in 2012)(Selected in 2007. WPI Academy)

Osaka University
Premium Research Institute for
Human Metaverse Medicine（PRIMe）(Selected in 2022)

Nagoya University
Institute of Transformative Bio-Molecules (ITbM)(Selected in 2012)*WPI Academy: WPI centers that have achieved “world-premier status”.

WebsiteInternational Institute for Integrative Sleep Medicine

18

IIIS

NIPPON