The 3rd KULOS Symposium

Brain Research for Well-Being
Date: Jan 26, 2024, 1:30 pmʛ
Venue: Cascade Room in Burke Museum (University of Washington)
12:00 pm

Networking Lunch- Introduction of Kobe University and its Project

1:30 pm

Opening Remarks
Tamotsu NAKAMURA (Kobe University)
Cecilia GIACHELLI (University of Washington)

Session 1

Role of Kinases in Brain Function
Session Chair : Toru TAKUMI (Kobe University)
Smita YADAV (University of Washington)
Yasuhito SHIRAI (Kobe University)
Kimberly ALDINGER(Seattle Children’s Research Institute/University of Washington)

1:50 pm
2:20 pm
2:50 pm
3:10 pm

BreakSession 2

Technology in Brain Function
Session Chair : Ayaka BOTA (Kobe University)
Nobuhiro NAKAI (Kobe University)
Garret STUBER (University of Washington)
Cindy van VELTHOVEN (Allen Brain Institute)

3:30 pm
4:00 pm
4:30 pm
5:00 pm

Closing Remarks
John SCOTT (University of Washington)
Junko TANAKA (Kobe University)

For information andregistration, visit
https://www.office.kobe-u.ac.jp/ipiep/events/20240126_en.html

Institute for Promoting International Partnerships,
Kobe University
E-mail:plan-global@office.kobe-u.ac.jp

Image:freepik.com

Mechanisms Underlying TAOK1 Kinase Dysfunctionin Neurodevelopmental Disorders
Dr. Smita YADAVUniversity of Washington

TAOK1 encodes for a serine-threonine protein kinase and is strongly associated with both neurodevelopmental
delay and autism spectrum disorder. The molecular function of this evolutionarily conserved kinase, and how
TAOK1 mutations lead to neuropathology is unknown. We have found that TAOK1 is highly expressed in both
cortical and hippocampal neurons in the mammalian brain. TAOK1 kinase binds and deforms the plasma
membrane in an activity dependent fashion. Using domain dissection and biochemical assays we mapped theminimal domain required for plasma membrane association, and show that the C-terminal helical coiled coil bundle
can directly bind phospholipids enriched in the plasma membrane. Expression of the membrane binding domain
is necessary and sufficient to bind PM and induce filopodial protrusions in neurons. We characterized four autismassociated TAOK1 mutations and found that these point mutations render TAOK1 catalytically dead, lead toaberrant membrane association and tubulation. Exogenous expression of the kinase domain alone is sufficient torescue the localization and phenotype of these gain of function TAOK1 autism mutations.

Finally, using

proteomics we have identified critical residues in the helical bundle that are phosphorylated by TAOK1 toautoregulate its plasma membrane association. This work defines the molecular function of a plasma membranetubulating kinase and reveals the molecular mechanisms underlying TAOK1 dysfunction in neurodevelopmental
disorders.

Dr. Smita YADAV
Biography:
Dr. Smita Yadav is an Associate Professor in the Department of Pharmacology at theUniversity of Washington, and her research is focused on elucidating how mutations inprotein kinases lead to abnormal brain development. Her laboratory utilizes proteomics,biochemistry, stem cell technology and brain organoids to understand neuropathology
caused by kinase dysfunction.
Research keyword:
Protein Kinases, Phosphorylation, Autism, Neurodevelopmental Disorders, Stem Cells

The role of protein kinase C in impairments of memory, emotion, andcoordinated movement associated with aging.
Yasuhito Shirai, Ph.D
Graduate School of Agricultural Science, Kobe University

Aging has become a serious issue recently. Specifically, it is predicted that by 2065, approximately one infour Japanese individuals will be 75 or older. Medical costs are consequently expected to rise, and issues
such as locomotive disorders (locomotive syndrome) and dementia are projected to become increasingly
severe in near future. Aging often coincides with a combination of disorders affecting memory, movement,and emotion. Improvement of these motor, memory, and emotional disorders is necessary if we hope tobuild a healthy and long-lived society. Our research found that knockout (KO) mice that lack theDGKgamma lipid-metabolizing enzyme exhibit memory, motor and emotional deficits as a result of aging.
By using these aged model mice, we discovered that the memory, motor, and emotional deficits were causedby an increase in the activity of PKCgamma in specific regions of the brain. We also found an increased
activity of PKCgamma in the brains of SAM mice, a well-known mouse model for human aging. Moresurprisingly, the memory, emotional, and motor deficits were ameliorated after two weeks of oral
administration of a certain flavonoid, along with normalization of PKCgamma activity. These findings
suggest that a relatively short-term intake of functional food targeting on PKCgamma can improve thememory, emotional, and motor impairments associated with aging.

Yasuhito Shirai
Biography:
Dean, Graduate School of Agricultural Science, Kobe University (2023-present)
Vice Dean, Graduate School of Agricultural Science, Kobe University (2022-present)
Director of Americas Division, Institute of Promoting Partnership (2021-present)
Professor of Graduate School of Agricultural Science, Kobe University (2010-present)
Visiting scholar, Department of Chemistry, University of California, San Diego(2000-2001)
Research keyword:
PKC, DGK, aging, diabetic disfunction, functional foodExpanding the role of MAST kinases in brain development and epilepsy:
identification of de novo pathogenic variants in MAST4
Kimberly ALDINGER
Seattle Children’s Research Institute/University of Washington

The MAST family of microtubule-associated serine-threonine kinases has been implicated inneurodevelopmental disorders, including developmental brain abnormalities (MAST1) and epilepsy(MAST3). Through an international collaboration, we identified 7 individuals with de novo
heterozygous variants in MAST4, including a recurrent missense variant associated with a consistent
brain malformation in two unrelated patients. All MAST4 patients presented with developmental
delay and vision abnormalities. Neuroimaging findings included periventricular leukomalacia,
cerebellar atrophy, polymicrogyria, and mega corpus callosum. A developmental and epileptic
encephalopathy was diagnosed in 4 individuals. Temporal and spatial gene expression analyses ofthe developing human brain show that MAST4 is prominently expressed in excitatory cells of thedeveloping thalamus. Our investigations into the molecular functions through which MAST4variants could impact cellular functions demonstrate one variant abolishes MAST4 microtubulebinding and mislocalizes to the nucleus, potentially due to formation of an alpha helix that could
stabilize the protein. Further investigation is ongoing to decipher how MAST4 mutations impact
brain development and function.

Kimberly ALDINGER
Biography:
Dr. Aldinger is an Assistant Professor in the departments of pediatrics and neurology at
the University of Washington and a Principal Investigator at Seattle Children's ResearchInstitute. She has over 20 years of research experience using neuroscience and genomics
tools to study identify genetic changes that cause neurodevelopmental disorders and theimpact of these changes to brain development and function. She is also a rare diseaseadvocate, raising awareness and aiding treatment development for rare genetic epilepsy.
Research keyword:rare diseaseVR-based real-time imaging of cortical networks in voluntary behavior
NAKAI Nobuhiro
Department of Physiology and Cell Biology, Graduate School of Medicine, Kobe University

The cerebral cortex plays a critical role in integrating sensory information from both the external
environment and the body, and in generating appropriate behavioral responses. Complex information
processing within the cerebral cortex is achieved through the cooperative activity of various
functional areas. However, the overall implications of this process for brain network activity remain
elusive. To analyze the expansive network activity of the cerebral cortex during voluntary behavior
in mice, we designed an experimental platform that integrates wide-field calcium imaging with
virtual reality (VR). This system enabled us to visualize the cooperative activity between corticalareas as a functional network. In this discussion, I would like to explore how network patterns are
rapidly reorganized in response to shifts in behavioral states. Locomotion induces a swift decouplingof sensory regions and an increased coupling between motor regions, leading to a more distinct
modular network organization compared to the rest state. Using a mouse model of autism spectrum
disorder (ASD), I unveiled specific abnormalities in the cortical network during the start and stop
phases of locomotion indicative of ASD. Looking towards future research directions, the potentialof using the VR system to study brain functional network dynamics associated with social behavior,and the possibility of predicting behavior using brain information via machine learning, are topics
that warrant further exploration.

NAKAI Nobuhiro, Ph.D. (email: nnakai@med.kobe-u.ac.jp)

photoBiography:
I am a project assistant professor at Kobe University. I obtained a Ph.D. in biostudies
from Kyoto University in 2017. From 2013 to 2019, I worked in Dr. TAKUMI Toru's
laboratory at the RIKEN Brain Science Institute and joined Kobe University together
with the laboratory when it moved here in 2020.
Research keywords:
Cortical functional network, calcium imaging, VR, mouse behavior, machine learningNeural Circuits for Motivation and Reward
Dr. Garret STUBER
Professor – University of Washington
Center for the Neurobiology of Addiction, Pain, and Emotion (NAPE)
Department of Anesthesiology and Pain Medicine
Department of Pharmacology
In the quest to thrive within a dynamic and often unpredictable environment, animals are tasked with thecontinual adaptation of their behaviors to effectively procure essential resources while evading potentiallyperilous situations. This delicate balance of attraction and aversion is orchestrated by intricate neural circuitry,
which is dynamically influenced by both environmental stimuli and internal physiological states to elicit
appropriate behavioral responses. The research conducted in my laboratory is focused on unraveling thecomplex neural circuits that govern both reward-seeking and aversion-driven behaviors. Employing cuttingedge methodologies—optogenetics, calcium imaging, and single-cell sequencing—our goal is to elucidate thenuanced functional interactions among molecularly distinct neuronal populations. These interactions arepivotal for the genesis of these critical behavioral states. Through this approach, we aim to construct acomprehensive map of the neural circuitry underpinning diverse motivational behaviors, shedding light on themultifaceted networks that drive decision-making processes in response to both positive and negative stimuli.
A deep understanding of these neural networks and their interplay is not just an academic pursuit; it holds
profound implications for the field of medicine, particularly in understanding and potentially mitigatingcomplex neurological and neuropsychiatric conditions. Insights gleaned from our research could significantly
advance our knowledge of disorders such as addiction, chronic pain, and emotional disturbances. By decoding
the neural substrates of motivation and behavioral regulation, we pave the way for novel therapeutic strategies
and interventions, potentially transforming the landscape of treatment for these pervasive and challenging
conditions.

Dr. Garret STUBER
Biography:
Professor of Anesthesiology and Pain Medicine, University of Washington (2018-)
Professor of Pharmacology, University of Washington (2018-)
Associate Professor, Department Psychiatry, Department of Cell Biology andPhysiology, Neuroscience Center of North Carolina, Chapel Hill (2015-2018)
Assistant Professor, Department Psychiatry, Department of Cell Biology andPhysiology, Neuroscience Center of North Carolina, Chapel Hill (2010-2015)
Research keyword:
neurocircuits, motivation, reward, behavior, mouseUnderstanding Cell Type Diversity in the Brain
Cindy van VELTHOVEN
Allen Institute for Brain ScienceThe mammalian brain is composed of millions to billions of cells that are organized intonumerous cell types with specific spatial distribution patterns and structural and functionalproperties. An essential step towards understanding brain function is to obtain a parts list, i.e., acatalog of cell types, of the brain. We recently created a comprehensive and high-resolution
transcriptomic and spatial cell type atlas for the whole adult mouse brain. This cell type atlascombines two single-cell-level, whole-brain-scale datasets: a single cell RNA-sequencing datasetof ~4.0 million cells passing QC, and a spatially resolved transcriptomic dataset of ~4.3 million
cells using MERFISH. The atlas is hierarchically organized into four nested levels ofclassification: 34 classes, 338 subclasses, 1,201 supertypes and 5,322 clusters. We systematicallyanalyzed the neuronal, non-neuronal, and immature neuronal cell types across the brain andidentified a high degree of correspondence between transcriptomic identity and spatial specificity
for each cell type. The results reveal unique features of cell type organization in different brain
regions, the extraordinary diversity and heterogeneity in neurotransmitter and neuropeptide(co-)expression patterns in different cell types across the brain, and the combinatorialtranscription factor code that defines cell types across all parts of the brain. With the completionof this benchmark reference atlas, we are now starting to explore what cell type characteristics
we can use to gain genetic access and develop new cell type targeting tools, how these cell types
arise from common progenitors during development, and which cell types are susceptible toprocesses like aging. Finally, we can integrate all these data from mouse to define the cell typelandscape over the life span of a mouse as well integrate with data from other species to answer
questions about evolution and cell type specification.

Cindy van VELTHOVEN
Biography:
Cindy van Velthoven is an Associate Investigator at the Allen Institute for Brain Science
where she is part of a large effort to define cell types in the developing and adult mousebrain. Here she develops and applies computational tools to identify cell types in various
mouse brain regions and characterize changes in gene regulation associated withdevelopment or experimental perturbations.
Prior to joining the Allen Institute, she completed her postdoctoral training in the lab ofTom Rando at Stanford University where she studied post-transcriptional mechanisms
involved in regulating stem cell quiescence. She used various transcriptional andtranslational profiling techniques to probe the earliest activation changes in quiescentstem cells in vivo. Cindy received her Ph.D. in neuroscience from Utrecht University,
The Netherlands, where she examined the use of stem cells to repair the neonatal brain
after ischemic injury.
Research keyword:
Single cell genomics