The 17th International Symposium of the Institute Network for
Biomedical Sciences International Symposium on Tumor Biology in Kanazawa 2022
2022 CRI & DUKE-NUS Joint Symposium

Fundamental
Biological
Principles and Cancer
Cancer Research Institute of Kanazawa University

October 13-14, 2022
Kanazawa University, Japan

The 17th International Symposium of the Institute Network for Biomedical Sciences
International Symposium on Tumor Biology in Kanazawa 2022
2022 CRI & DUKE-NUS Joint Symposium

Fundamental Biological Principles and Cancer
Cancer Research Institute of Kanazawa University
October 13-14, 2022
Kanazawa University, Japan

Thursday, October 13, 2022
12:30-13:00 Registration
13:00-13:05 Opening Remarks
Kunio Matsumoto

Session 1
13:05-13:30(Director of Cancer Research Institute, Vice president, Kanazawa University)(Chair Noriko Goto, Masaya Ueno )Ｓ-01: Tetsuro Yoshimaru(Institute of Advanced Medical Sciences, Tokushima University)

・・・1

The plasma membrane BIG3-PHB2 complex contributes to the acquisition of trastuzumab-resistance in
HER2-positive breast cancer
13:30-13:55

Ｓ-02: Kozo Tanaka(Institute of Development, Aging and Cancer, Tohoku University)

・・・3

Chromosomal instability induced by chromosome dynamics in cancer cells
13:55-14:07

Ｙ-01: Yutaka Kasai(The Institute of Medical Science, The University of Tokyo)

・・・5Trans-homophilic interaction of CADM1 promotes organ infiltration of T-cell lymphoma by adhesion tovascular endothelial cells
14:07-14:19

Ｙ-02: Shunya Tsuji(Research Institute for Microbial Diseases, Osaka University)・・・7

SARS-CoV-2 and cellular senescence
14:19-14:31

Ｙ-03: Taisho Yamada(Institute for Genetic Medicine, Hokkaido University)

・・・9

RIG-I restrains SARS-CoV-2 replication in human lung cells without activation of innate immune
signaling
14:31-14:45 BreakInternational Symposium (Chair Masanobu Oshima, Eishu Hirata)
14:45-14:50 Opening Remarks
Shinichi Nakamura(Trustee(Research, Social Co-creation, and Graduate School Support),Kanazawa University）

14:50-15:20

Ｉ-01: Koji Itahana(DUKE-NUS, Singapore)

The potential role of multi-drug resistance protein ABCB1 in tumor suppression in bats・・・57

15:20-15:32

Ｙ-04: Yasuto Takeuchi(Cancer Research Institute, Kanazawa University)

・・・59

The membrane-linked adaptor FRS2・ fashions a cytokine-rich microenvironment that promotes breast
cancer carcinogenesis
15:32-16:02

Ｉ-02: S. Tiong Ong

FS(DUKE-NUS, Singapore)

・・・61

Predicting primary resistance to cancer targeted therapies at diagnosis: lessons from chronic myeloid

9leukemia

9

16:02-16:10

Break

16:10-16:35

Ｓ-03: Yoshikazu Johmura(Cancer Research Institute, Kanazawa University)

・・・63

Identification and functional analysis of senescent cells in tumor microenvironment

16:35-17:05

Ｉ-03: Enrico Petretto(DUKE-NUS, Singapore)

・・・65

9

1

Systems Genetics identifies WWP2 as a new target for fibrotic disease in fibroblasts and macrophages

1
17:05-17:20

Session 2
17:20-17:45

Break

1

(Chair Atsushi Hirao)
Ｓ-04: Kosuke Hashimoto(Institute for Protein Research, Osaka University)

・・・11Single-cell transcriptome analysis of human immune cells and early embryos
17:45-17:57

Ｙ-05: Ryoji Kawakami(Institute for Life and Medical Sciences, Kyoto University)

1
・・・13

Coordinated activation of enhancer elements for thymic Treg development and immunological self-

1

tolerance
17:57-18:09

Ｙ-06: Akiko Satoh(Institute of Development, Aging and Cancer, Tohoku University)

The role of hypothalamic neurons in sleep, aging and longevity

S

・・・15

1

1

1

P

1

9

1

3

5

1

Friday, October 14, 2022
Sesson 3
9:00-9:25(Chair Dominic Voon, Kazuhiro Murakami)
Ｓ-05: Kazuhiro Suzuki

9:25-9:50

Ｓ-06: Tetsuro Izumi

・・・17(Institute for Molecular and Cellular Regulation, Gunma University)

・・・19

Roles of the GDF3-ALK7 axis in adiposity
9:50-10:02

Ｙ-07: Junko Sasaki(Medical Research Institute, Tokyo Medical and Dental University)

・・・21

Premature ovarian insufficiency in mice lacking phosphoinositide-metabolizing enzymes
10:02-10:14

Ｙ-08: Kou Motani(Institute of Advanced Medical Sciences, Tokushima University)

・・・23

ACBD3 forms specialized ER-Golgi contact sites to drive the ER exit of STING
10:14-10:26

Ｙ-09: Macpherson Tom(Institute for Protein Research, Osaka University)

・・・25

Cell type-specific control of reward learning in the brain
10:26-10:40 BreakSession 4
10:40-11:05(Chair Toshinari Minamoto, Ryu Imamura)
Ｓ-07: Nobuo Noda

(Institute for Genetic Medicine, Hokkaido University)

・・・27

Molecular mechanisms underlying autophagosome biogenesis

35(Research Institute for Microbial Diseases, Osaka University)

Novel control mechanisms of lymphocyte trafficking

11:05-11:17

Ｙ-10: Eiji Miyauchi(Institute for Molecular and Cellular Regulation, Gunma University)・・・29

Gut microbiota modulates inflammation in the central nervous system
11:17-11:29

Ｙ-11: Minako Ito(Medical Institute of Bioregulation, Kyushu University)

・・・31

Immune cell dynamics and role in central nervous system diseases
11:29-11:41

Ｙ-12: Jun Hatakeyama(Institute of Molecular Embryology and Genetics, Kumamoto University)

・・・33

Strategies for the expansion of cerebral cortex in primates
11:41-13:00 Lunch
Poster Session & Coffee Break
13:00-14:00

Ｐ-01: Yukako Nishimura(Institute for Genetic Medicine, Hokkaido University)

・・・45

A cross-talk between microtubules and focal adhesions regulates dynamics of actin cytoskeleton

Ｐ-02: Madoka Kawaguchi(Institute of Development, Aging and Cancer, Tohoku University)

Extracellular Domains I and II of CD44 mediate its trans-homophilic dimerization and tumor cluster
aggregation

・・・45

Ｐ-03: Ryosuke Kobayashi(Institute for Molecular and Cellular Regulation, Gunma University)

・・・46

Epigenetic dysregulations in the lysine methyltransferase (KMT2) deficient endometrial cancer cells
Ｐ-04: Nichika Sato

(The Institute of Medical Science, The University of Tokyo)

・・・46

A novel E3 ligase is involved in Unfolded Protein Response through Ubiquitination of eS7A and upregulation of Hac1 production in yeast
Ｐ-05: Yoshimi Okamoto(Medical Research Institute, Tokyo Medical and Dental University)

Ｐ-06: Yuanyuan Zhang(Cancer Research Institute, Kanazawa University)

Molecular mechanism of YAP-dependent hepatocyte elimination

Application of CDK4/6 inhibition therapies to PDACs

・・・47

・・・47

Ｐ-07: Yoko Fujita-Fujiharu (Institute for Life and Medical Sciences, Kyoto University)

・・・48

Ｐ-08: Yuki Akieda(Research Institute for Microbial Diseases, Osaka University)

・・・48

Ｐ-09: Koki Sakurai(Institute for Protein Research, Osaka University)

Ｐ-10: Yosuke Matsushita(Institute of Advanced Medical Sciences, Tokushima University)

Structural basis of Marburg virus helical nucleoprotein-RNA complex formation

Abnormal pH environment interrupts cell competition-mediated developmental robustness

Altered behavior and increased neuroinflammation in Importin α4/KPNA4 KO mice

S

・・・49

・・・49

RHBDL2 has essential roles for glutaminolysis and chemoresistance in triple negative breast cancer
Ｐ-11: Akinobu Matsumoto(Medical Institute of Bioregulation, Kyushu University)

Ｐ-12: Ryuki Shimada(Institute of Molecular Embryology and Genetics, Kumamoto University)

Physiological functions of novel proteins encoded by hidden ORFs

The female-specific regulation of meiotic cell cycle in murine germ cells

・・・50

・・・50

1

1

1

1

S

1
Ｐ-13: Shinya Ohta

(Institute for Genetic Medicine, Hokkaido University)

Formation of pericentromeric heterochromatin via ZNF518s that link satellite DNA to heterochromatin

・・・51

1
Ｐ-14: Fang Zhenzhou(Institute of Development, Aging and Cancer, Tohoku University)

Aurora A-dependent polyubiquitination of OLA1 regulates centrosome number through controlling the・・・51

recruitment of PCM proteins
Ｐ-15: Ikuko Maejima(Institute for Molecular and Cellular Regulation, Gunma University)

Ｐ-16: Hirofumi Kosuge(The Institute of Medical Science, The University of Tokyo)

Ｐ-17: Yoshitaka Murota(Medical Research Institute, Tokyo Medical and Dental University)

The role of small GTPase protein Rab35 in brain development

Identification and validation of multi-specific interactions of a tumor suppressor protein PRELP

Establishment of a high-content polymer array screening system to explore niche mimicries for

1
・・・52

・・・52

・・・53

cancerous cells
Ｐ-18: Masaya Ueno

(Cancer Research Institute, Kanazawa University)

Ｐ-19: Kazunari Aoki

(Institute for Life and Medical Sciences, Kyoto University)

Lysosomes regulate oncogenic signals in acute myelogenous leukemia

Identification of factors essential for CXCR4 signaling in leukemic cells using CRISPR dropout screens

・・・53

・・・54

6

Ｐ-20: Liu Yafei(Research Institute for Microbial Diseases, Osaka University)

6

Ｐ-21: Sho Tabata(Institute for Protein Research, Osaka University)

Ｐ-22: Motonori Matsusaki(Institute of Advanced Medical Sciences, Tokushima University)

An infectivity-enhancing site on the SARS-CoV-2 spike protein targeted by antibodies

Induction of cellular senescence by sustained NFκB activation

Ｐ-23: Kazuhiko Kawata(Medical Institute of Bioregulation, Kyushu University)

Ｐ-24: Yasushi Yabuki(Institute of Molecular Embryology and Genetics, Kumamoto University)

Essential role of ER membrane complex subunit 1 (EMC1) in B cell development

7

RNA phase transition disrupts α-Synuclein proteostasis

8

9

9

0

0

Session 5(Chair Chiaki Takahashi)

14:00-14:25

Ｓ-08: Masaki Takahashi

2

2

3

3

4

・・・55

(The Institute of Medical Science, The University of Tokyo)

・・・56

・・・56

・・・35

Generation of RNA aptamers and their applications in molecular biology
14:25-14:50

Ｓ-09: Itoshi Nikaido(Medical Research Institute, Tokyo Medical and Dental University)

・・・37

Identification of chromatin instability at the single-cell level from transcriptome data
14:50-15:15

Ｓ-10: Yasuyuki Ohkawa(Medical Institute of Bioregulation, Kyushu University)

・・・39

Chromatin regulation during skeletal muscle regeneration
15:15-15:25 BreakSession 6(Chair Takashi Suda)

15:25-15:50

Ｓ-11: Takahiro Ito(Institute for Life and Medical Sciences, Kyoto University)

15:50-16:15

Ｓ-12: Minetaro Ogawa(Institute of Molecular Embryology and Genetics, Kumamoto University)

1

1

・・・55

Endoplasmic Reticulum Stress Sensor IRE1 Directly Detects Misfolded Insulin by Oligomerization

7

8

・・・54

・・・41

Regulation on stem cells in myeloid leukemia: a metabolic perspective

Modeling hematopoietic stem cell development in a dish16:15-16:30 Award Ceremony & Closing Remarks
Kunio Matsumoto(Director of Cancer Research Institute, Vice president, Kanazawa University)

・・・43

Ｓｅｓｓｉｏｎ1

8

Ｓ－０１

The plasma membrane BIG3-PHB2 complex contributes to the acquisition of
trastuzumab-resistance in HER2-positive breast cancer
Yoshimaru1, Tetsuro; Matsushita1, Yosuke; Kosako2, Hidetaka; Sasa3, Mitsunori; Miyoshi4,
Yasuo; Katagiri1, Toyomasa
Division of Genome Medicine, Institute of Advanced Medical Sciences, Tokushima University, Japan
Division of Cell Signaling, Fujii Memorial Institute of Medical Sciences, Tokushima University, Japan3
Department of Surgery, Tokushima Breast Care Clinic, Japan
4
Division of Breast and Endocrine Surgery, Department of Surgery, Hyogo College of Medicine, Japan
1
2

The human epidermal growth factor receptor 2 (HER2) amplification is linked to aggressive tumor behavior
and a poorer prognosis in breast cancer. Although the anti-HER2 antibody, trastuzumab, have greatly
improved outcomes of HER2-positive breast cancer, resistance acquisition remains a severe clinical problem.
Several resistance mechanisms against trastuzumab, such as proteolytic cleavage of the extracellular domain
of HER2, alternative activation of downstream signaling, and EGFR overexpression, have been proposed, but
a conclusive trastuzumab-resistant acquisition mechanism still has not been revealed. Therefore, the mosturgent issue is to develop a novel therapeutic approach targeting resistance acquisition mechanisms.
We previously reported that the cancer-specific A-kinase anchoring protein BIG3 (brefeldin A-inhibited
guanine nucleotide exchange protein 3) acts as a critical modulator of estrogen signaling in breast cancer cells
by inhibiting the tumor-suppressive activity of prohibitin 2 (PHB2). Interestingly, RNA-seq data in public
databases revealed high expression of the BIG3 in refractory HER2-positive breast cancer patients.
Interestingly, BIG3-PHB2 complexes showed different intracellular localization between trastuzumabsensitive and -resistant cells. Here, we report the pathophysiological significance of the BIG3-PHB2
complexes in HER2-positive breast cancer, especially in trastuzumab-resistant cells. BIG3 forms a complex of
PHB2 in the trans-Golgi network (TGN) of trastuzumab-sensitive cells, suppressing the tumor-suppressive
activity of PHB2 as well as estrogen receptor-positive breast cancer cells. Notably, BIG3-PHB2 complexes
regulate secreting the extracellular proteins associated with HER2-positive breast cancer progression.
Furthermore, we found that BIG3-PHB2 complexes are transported to the plasma membrane from the TGN by
EGFR overexpression via its cargo-X protein, contributing to the HER2-EGFR heterodimerization involved in
trastuzumab resistance.
More importantly, treatment with peptide inhibitor ERAP targeting the BIG3-PHB2 interaction led to theremarkable inhibition of transcriptional activity, extracellular protein secretion, and HER2-EGFR
heterodimerization, resulting in suppression of NF-κB signaling and trastuzumab-sensitive/resistant cellproliferation. Furthermore, intravenous injection of this peptide inhibitor rapidly exhibited the long-term
antitumor effect in vivo mice with HER2-positive breast cancer cells, especially trastuzumab-resistant cells.
These findings suggest that the BIG3-PHB2 complexes have pivotal roles in the progression of HER2-positive
breast cancer, especially in acquiring trastuzumab resistance.

11References(Related articles)

1. Cancer Science, 2021, 112, 4208‒4219
2. Journal of Human Genetics, 2021, 66, 927‒935
3. International Journal of Oncology, 2020, 56, 581‒595
4. Biochemical and Biophysical Research Communications, 2019, 518, 183‒189
5. Cancer Research, 2018, 78, 2233‒2247, 2018
6. Nature Communications, 2017, 8, 15427
7. Scientific Reports, 2017, 7, 1821
8. Cancer Science, 2017, 108, 785‒794
9. PLoS One, 2015, 10, e0127707
10. Cancer Science, 2015, 106, 550‒558
11. Scientific Reports, 2014, 4, 7355
12. BMC Res Notes, 2014, 7, 435
13. Nature Communications, 2013, 4, 2443

CV
Education:
1997 - 2000 : Department of Food Sciences and Technology, Graduate School of Agriculture, Kyushu
University, Awarded the degree of Ph.D (Agricultural science)
1995 - 1997 : Department of Food Sciences and Technology, Graduate School of Agriculture, Kyushu
University, Awarded the degree of Master of Science
1990 - 1994 : Faculty of Agriculture, Kyushu University, Awarded the degree of Bachelor of Science
Professional carriers:
2019
Associate Professor, Tokushima University
2016
Lecturer, Tokushima University
2012
Assistant Professor, Tokushima University
2010
Designated Assistant Professor, Tokushima University
2009
Academical Researcher, Tokushima University
2000 - 2009 Post-doctoral Fellow, Advanced Medical Research Center,
Graduate School of Medical Sciences Nihon University
Specialties:
Molecular oncology
Personality:
I can promote good working relationships with any persons due to my cheerful character and cooperative
personality. I believe that “communication with people” is the best shortcut and solution to “research
expansion”, that’s why I will grasp the objective of research progress through regular communication. In
addition, I am always honest, enthusiastic, patient, and a very creative hard worker for scientific research.

2

2

Ｓ－０２

Chromosomal instability induced by chromosome dynamics in cancer cells

Iemura1, Kenji; Tanaka1, Kozo

1

Department of Molecular Oncology, Institute of Development, Aging and Cancer, Tohoku University, Japan

Most cancer cells show chromosomal instability, a condition in which chromosome missegregation occurs at
high rates. Among the causes of chromosomal instability, insufficient correction of erroneous kinetochoremicrotubule attachments plays pivotal roles in various situations. We recently found the previously
unappreciated role of chromosome oscillation, an iterative chromosome motion during metaphase, in thecorrection of erroneous kinetochore-microtubule attachments. Chromosome oscillation facilitates themetaphase phosphorylation of Hec1, a kinetochore protein that binds to microtubules, by Aurora A kinase on
the spindle. The phosphorylation of Hec1 reduces its affinity to microtubules, which promotes the correctionof erroneous kinetochore-microtubule attachments. Our data showed that the Aurora A–dependent Hec1
phosphorylation and the chromosome oscillation amplitude facilitate each other. Importantly, chromosome
oscillation is attenuated in cancer cell lines compared with non-transformed cell lines. The Hec1
phosphorylation in metaphase was reduced in cancer cells, which was restored by enhancement ofchromosome oscillation. Enhancement of chromosome oscillation in cancer cells also reduced the number of
erroneous kinetochore–microtubule attachments and chromosome missegregation, whereas inhibition of
Aurora A during metaphase increased such errors. We propose that Aurora A–mediated metaphase Hec1
phosphorylation through chromosome oscillation ensures mitotic fidelity by eliminating erroneous
kinetochore-microtubule attachments. Attenuated chromosome oscillation and the resulting reduced Hec1
phosphorylation may be a cause of chromosomal instability in cancer cells.

33References(Related articles)

1. Iemura K, Natsume T, Maehara K, Kanemaki MT, Tanaka K. Chromosome oscillation promotes Aurora Adependent Hec1 phosphorylation and mitotic fidelity. J Cell Biol (2021) 220, e202006116.
2. Campos Medina M, Iemura K, Kimura A, Tanaka K. A mathematical model of kinetochore-microtubule
attachment regulated by Aurora A activity gradient describes chromosome oscillation and correction oferroneous attachments. Biomed Res (2021) 42, 203-219.
3. Iemura K, Yoshizaki Y, Kuniyasu K, Tanaka K. Attenuated chromosome oscillation as a cause ofchromosomal instability in cancer cells. Cancers (2021) 13, 4531.

CV
Education

1985.4~1991.3
1993.4~1997.3

Faculty of Medicine, University of Tokyo
Graduate School of Medicine, University of Tokyo

Appointments
1991.6~1992.5 Resident, Internal Medicine, The University of Tokyo Hospital
1992.6~1993.5 Junior Resident, Internal Medicine, Jichi Medical School Hospital
1997.4~1998.3 Senior Resident, Third Department of Internal Medicine, Faculty of Medicine, University ofTokyo
1998.4~2001.12 Research Associate, Department of Molecular Pathology, Research Institute for Radiation
Biology and Medicine, Hiroshima University
2002.1~2007.2 Postdoctoral Research Assistant, School of Life Sciences, University of Dundee
2007.3~2011.2 Assistant Professor, Institute of Development, Aging and Cancer, Tohoku University
2011.3~
Professor, Institute of Development, Aging and Cancer, Tohoku University

4

4

Ｙ－０１

Trans-homophilic interaction of CADM1 promotes organ infiltration of T-cell
lymphoma by adhesion to vascular endothelial cells
Kasai1, Yutaka; Funaki1, Toko; Kumagai1, Yuki; Tominaga1, Mizuki; Matsubara1,2, Daisuke;
Ito1, Takeshi; Murakami1, Yoshinori
1Division of Molecular Pathology, The Institute of Medical Science, The University of Tokyo, Tokyo,

Japan

2Department of Diagnostic Pathology, University of Tsukuba, Tsukuba, Japan

The initial step of organ infiltration of malignant cells is the interaction with host vascular endothelial cells,
which is often mediated by specific combinations of cell adhesion molecules. Cell adhesion molecule 1(CADM1) is a single-pass membranous protein of immunoglobulin superfamily (IgSF) and participates in theformation of various types of cell adhesion, including epithelial cell- and synaptic cell-adhesion throughhomophilic and heterophilic interactions1. CADM1 is overexpressed in adult T-cell leukemia/lymphoma(ATL) and provides a cell-surface diagnostic marker, and also promotes the adhesion of ATL cells to vascularendothelial cells and multiple organ infiltration in mice. However, its binding partner on host cells has not yet
been identified. In this study, we show that CADM1 promotes transendothelial migration of ATL cells in
addition to the adhesion to vascular endothelial cells. Moreover, CADM1 enhances liver infiltration of mouseT-cell lymphoma cells, EL4, after tail vein injection, whereas a CADM1 mutant lacking adhesive activity did
not. Among the known CADM1-binding proteins expressed in primary endothelial cells, only CADM1 andCADM4 could induce morphological extension of ATL cells when plated onto glass coated with these
proteins. Furthermore, CADM1-mediated liver infiltration of EL4 cells was canceled in conventional Cadm1
knockout mice, whereas it was not canceled in Cadm4 knockout mice. Finally, CADM1-mediated liver
infiltration was cancelled in Tie2-Cre; Cadm1fl/fl mice, vascular endothelium-specific Cadm1 knockout
mice2. These results suggest that CADM1 on host vascular endothelial cells is required for organ infiltrationof ATL and other T-cell lymphomas expressing CADM1. Possible involvement of CADM1 in metastasis ofsmall cell lung cancercancer, another tumor expressing high amount of CADM1, will be also discussed.

55References(Related articles)
1.

Ito T†, Kasai Y†, Kumagai Y, Suzuki D, Ochiai-Noguchi M, Irikura D, Miyake S and Murakami Y.
Quantitative analysis of interaction between CADM1 and its binding cell-surface proteins using surface
plasmon resonance imaging. Front Cell Dev Biol, 2018;6:86.

2. Kasai Y, Gan SP, Funaki T, Ohashi-Kumagai Y, Tominaga M, Shiu SJ, Suzuki D, Matsubara D, Sakamoto
T, Sakurai-Yageta M, Ito T and Murakami Y. Trans-homophilic interaction of CADM1 promotes organ
infiltration of T-cell lymphoma by adhesion to vascular endothelium. Cancer Sci, 2022;113(5):16691678.

CV

Education:2014 BSc, Faculty of Science and Technology, Tokyo University of Science
2016 MSc, Graduate School of Frontier Sciences, the University of Tokyo
2019 PhD, Graduate School of Frontier Sciences, the University of Tokyo
Position:
2019-2021 Postdoctoral Fellow, Division of Molecular Pathology, The Institute of Medical Science,
The University of Tokyo
2022- Assistant Professor, Division of Molecular Pathology, The Institute of Medical Science, The
University of Tokyo

6

6

Ｙ－０２

SARS-CoV-2 and cellular senescence
Tsuji1, Shunya; Minami2, Shohei; Hashimoto3, Rina; Suzuki4, Tatsuya; Sasai5, Miwa; Torii6,
Shiho; Ono6, Chikako; Shichinohe7, Shintaro; Sato2, 8, Shintaro; Yamamoto5, 9, 10, Masahiro;
Watanabe7, 10, Tokiko; Matsuura6, 10, Yoshiharu; Takayama3, Kazuo; Kobayashi2, 10, Takeshi;
Okamoto4, Toru; Hara1, 9, 10, Eiji;
Department of 1Molecular Microbiology, 2Virology, 5Immunoparasitology, 7Molecular Virology, 6Laboratory ofVirus Control, Research Institute for Microbial Diseases, Osaka University
3
Center for iPS Cell Research and Application, Kyoto University
4
Division of Infectious Diseases, Institute for Advanced Co-Creation Studies, Osaka University
8
Osaka City University Graduate School of Medicine
9
Immunology Frontier Research Center, Osaka University
10
Center for Infectious Disease Education and Research, Osaka University

Reports of post-acute COVID-19 syndrome, in which the inflammatory response persists even after SARSCoV-2 has no longer detected, are increasing1, but the underlying mechanisms of post-acute COVID-19
syndrome remain unknown. Here, we show that SARS-CoV-2-infected cells trigger senescence-like cell-cycle
arrest2,3 in neighboring uninfected cells in a paracrine manner via virus-induced cytokine production. In
cultured human cells or bronchial organoids, these SASR-CoV-2 infection-induced senescent cells express
high levels of a series of inflammatory factors known as senescence-associated secretory phenotypes(SASPs)4 in a sustained manner, even after SARS-CoV-2 is no longer detectable. We also show that theexpression of the senescence marker CDKN2A (refs. 5,6) and various SASP factor4 genes is increased in thepulmonary cells of patients with severe post-acute COVID-19 syndrome. Furthermore, we find that mice
exposed to a mouse-adapted strain of SARS-CoV-2 exhibit prolonged signs of cellular senescence and SASPin the lung at 14 days after infection when the virus was undetectable, which could be substantially reducedby the administration of senolytic drugs7. The sustained infection-induced paracrine senescence describedhere may be involved in the long-term inflammation caused by SARS-CoV-2 infection.

7

7

References(Related articles)

1. Nalbandian, A. et al. Post-acute COVID-19 syndrome. Nat. Med. 27, 601–615 (2021).
2. He, S. & Sharpless, N. E. Senescence in health and disease. Cell 169, 1000–1011 (2017).
3. Gorgoulis, V. et al. Cellular senescence: defining a path forward. Cell 179, 813–827 (2019).
4. Coppé, J.-P., Desprez, P.-Y., Krtolica, A. & Campisi, J. The senescence- associated secretory phenotype:the dark side of tumor suppression. Annu. Rev. Pathol. Mech. Dis. 5, 99–118 (2010).
5. Serrano, M., Hannon, G. J. & Beach, D. A new regulatory motif in cell-cycle control causing specificinhibition of cyclin D/CDK4. Nature 366, 704–707 (1993).
6. Hara, E. et al. Regulation of p16 expression and its implications for cell immortalization and senescence.
Mol. Cell. Biol. 16, 859–867 (1996).
7. Di Micco, R., Krizhanovsky, V., Baker, D. & d’Adda di Fagagna, F. Cellular senescence in ageing: frommechanisms to therapeutic opportunities. Nat. Rev. Mol. Cell Biol. 22, 75–95 (2021).

CV

8

8

Ｙ－０３

RIG-I restrains SARS-CoV-2 replication in human lung cells without
activation of innate immune signaling
Yamada1,2, Taisho; Sato1,2, Seiichi; Orba3,4, Yasuko; Sawa,3,4,5, Hirofumi; Sasaki3,4,Michihito; Takaoka1,2, Akinori
Division of Signaling in Cancer and Immunology, Institute for Genetic Medicine, Hokkaido University,
Japan. 2Molecular Medical Biochemistry Unit, Biological Chemistry and Engineering Course, Graduate
School of Chemical Sciences and Engineering, Hokkaido University, Japan.

1

Division of Molecular Pathobiology, International Institute for Zoonosis Control, Hokkaido University,

3

Japan.
International Collaboration Unit, Research Center for Zoonosis Control, Hokkaido University, Japan.

4
5

COVID-19, caused by SARS-CoV-2, which is a positive-sense, single-stranded RNA betacoronavirus,
remains an ongoing global pandemic. Although most people infected with SARS-CoV-2 show a mild andself-limited course, there are also a few severe and critical patients with exacerbated inflammatory response.
The wide spectrum of clinical manifestations of COVID-19 suggests that individual immune responses to theunderlying pathogen may play some crucial roles in determining the clinical course. However, the innate
immune responses against SARS-CoV-2 remains poorly understood. Here we show that retinoic acidinducible gene-I (RIG-I) but not MDA5 or TLR3 sufficiently restrains SARS-CoV-2 replication in humanlung cells in a type I/III interferon (IFN)-independent manner.
RIG-I inhibits viral RNA-dependent RNA polymerase (RdRp)-mediated first step of replication through thepreferential binding of RIG-I helicase domain to the 3- untranslated region of the viral genomic positive-senseRNA. This new mode of RIG-I recognition fails to activate the conventional downstream MAVS/IPS-1dependent signaling, which is in accordance with lack of cytokine induction after SARS-CoV-2 infection.
Consistently, genetic ablation of RIG-I expression allows lung cells to produce viral particles. On the other
hand, SARS-CoV-2 can replicate with innate cytokine induction in cells with low levels of RIG-I expression.
In such situation where negative-sense RNA initiates to be transcribed, MDA5 in turn senses the negativesense RNA to induce type I/III IFNs and other cytokines.
Our data have defined RIG-I expression levels as one of the intrinsic determinants for the defense in humanlung cells at least during the initial process of SARS-CoV-2 infection. It has been reported that nearly 40-45%
people infected with SARS-CoV-2 show asymptomatic with no robust upregulation of innate cytokines. In
this respect, the balance between RIG-I expression levels and the amount of invading virus would regulate thefate of viral replication, which might determine cytokine responses and the related patient outcomes.

99References(Related articles)

Yamada T., Sato S., Sotoyama Y., Orba Y., Sawa H., Yamauchi H., Sasaki M., and Takaoka A. RIG-I
triggers a signaling-abortive anti-SARS-CoV-2 defense in human lung cells. Nat. Immunol., 22, 820-828,
2021.
Takaoka A. and Yamada T. Regulation of signaling mediated by nucleic acid sensors for innate interferonmediated responses during viral infection. Int. Immunol., 31, 477-488, 2019.
Yamada T., Horimoto H., Kameyama T., Hayakawa S., Yamato H., Dazai M., Takada A., Kida H., Bott D.,
Zhou AC., Hutin D., Watts TH., Asaka M., Matthews J. and Takaoka A. Constitutive aryl hydrocarbon
receptor signaling constrains type I interferon-mediated antiviral innate defense. Nat. Immunol., 17, 687-694,
2016.

CV

2006-2010

Bachelor of Medical Sciences, College of Nursing and Medical Technology, School of
Medicine and Health Sciences, University of Tsukuba

2010-2012

Master of Medical Sciences, Graduate School of Medicine, Hokkaido University

2012-2016

Doctor of Medical Sciences, Graduate School of Medicine, Hokkaido University

2016-present

Assistant Professor, Institute for Genetic Medicine, Hokkaido University

Awards2016

ICI-FIMSA Travel Award (Australia)

2017

Frate Reseach Award, Hokkaido University Medical School Alumni Association2017

Reseach Award, The Hokkaido Medical Society

2021

Reseach Award, Japanese Society of Interferon & Cytokine Research

1010Ｓｅｓｓｉｏｎ2

11

12

Ｓ－０４

Single-cell transcriptome analysis of human immune cells and early embryos

Hashimoto1, Kosuke

1Institute for Protein Research, Osaka University, Japan

Aging has a profound impact on the immune system. Supercentenarians, people who have reached 110 y ofage, are a great model of healthy aging. Their characteristics of delayed onset of age-related diseases andcompression of morbidity imply that their immune system remains functional. In the first part of my talk, I
present our single-cell transcriptome analysis of peripheral blood mononuclear cells (PBMCs), derived from 7
supercentenarians and 5 younger controls. In this study, we identified a marked increase of cytotoxic CD4 T
cells (CD4 cytotoxic T lymphocytes [CTLs]) as a signature of supercentenarians. Furthermore, single-cell T
cell receptor sequencing of 2 supercentenarians revealed that CD4 CTLs had accumulated through massive
clonal expansion, with the most frequent clonotypes accounting for 15 to 35% of the entire CD4 T cell
population.
In the second part, I present the single-cell analysis of human embryos. Long terminal repeat (LTR)
retrotransposons are widely distributed across the human genome. They have accumulated through retroviral
integration into germline DNA and are latent genetic modules. Active LTR promoters are observed in
germline cells; however, little is known about the mechanisms underlying their active transcription in somatic
tissues. We revealed that the LTR families MLT2A1 and MLT2A2 are primarily expressed in human fourcell and eight-cell embryos and are also activated in some adult somatic tissues, particularly pineal gland.

11

11References(Related articles)

1. Hashimoto K, Jouhilahti EM, Töhönen V, Carninci P, Kere J, Katayama S. Embryonic LTR
retrotransposons supply promoter modules to somatic tissues. Genome Res. 31, 1983-1993 (2021)

2. Hashimoto K, Kouno T, Ikawa T, Hayatsu N, Miyajima Y, Yabukami H, Terooatea T, Sasaki T, Suzuki T,
Valentine M, Pascarella G, Okazaki Y, Suzuki H, Shin JW, Minoda A, Taniuchi I, Okano H, Arai Y, Hirose
N, Carninci P. Single-cell transcriptomics reveals expansion of cytotoxic CD4 T cells in supercentenarians.
Proc Natl Acad Sci USA. 116, 24242-24251 (2019)

CV
Aug. 2020 – Present
Apr. 2016 – July 2020
Oct. 2011 – Mar. 2016
Apr. 2009 – Sep. 2011
Mar. 2009

Associate Professor, Osaka University
Senior Research Scientist, RIKEN, Japan
Research Scientist, RIKEN, Japan
Postdoctoral Visiting Fellow, NCBI, NIH, USA
Ph.D. Bioinformatics Center, Kyoto University, Japan

12

12

Ｙ－０５

Coordinated activation of enhancer elements for thymic Treg development andimmunological self-tolerance
Kawakami, Ryoji1; Sakaguchi, Shimon1

1

Institute for Life and Medical Sciences, Kyoto University, Japan

CD4+Foxp3+ regulatory T cells (Tregs) are one of the most significant cells in the maintenance of immune
tolerance. Treg cells work to maintain homeostasis of the body in various situations such as prevention of
autoimmune diseases, termination of immune responses, and damaging tissue repairing, by braking excessive
immune responses.
Here, we would like to discuss the differentiation mechanism of Treg cells. Many Tregs are generated from a
portion of self-antigen-stimulated CD4+ T cells during the thymic negative selection process. However, thedetailed mechanism, especially “How Treg-type epigenetic landscape is installed in immature T cells underselection”, is largely unknown. The research questions are;(i) The detailed mechanisms of Treg generation in the epigenetic and physical three-dimensional chromatincontexts(ii) Contribution/Significance of non-coding enhancer elements for Treg differentiation e.g. at thetranscription factor Foxp3 gene locus(iii) Kinetics of Treg differentiation in the thymic selection and identification of epigenetically-poised Tregprecursor cells
To elucidate these, we first described genome-wide DNA epigenetic landscape and their physical 3Dinteractions during thymic T cell differentiation (from DN-CD4SP-Treg) using ATAC-seq, anti-histone ChIPseq, and H3K27ac-HiChIP. This analysis revealed the T cell-specific activation of Treg-type enhancer
elements at the initial stages of T cell differentiation, prior to induction of Foxp3 expression. Using CRISPRCas9 technology in mice, we found that distinct two non-coding enhancer elements at the Foxp3 locus areindispensable for thymic Treg generation and immunological self-tolerance. These results demonstrate thesignificance of coordinated activation of enhancer elements in an epigenetic context for the establishment andmaintenance of immunological self-tolerance.

13

13

References(Related articles)

R. Kawakami et al. Immunity. 2021 May 11;54(5):947-961.e8.
doi: 10.1016/j.immuni.2021.04.005.

CV

Kawakami Ryoji, Ph.D.
Lab of Experimental Immunology
Program-specific Assistant Professor,
Institute for Life and medical Sciences, Kyoto university.
kawakami.ryoji.4c@kyoto-u.ac.jp[Research Interest]
Immune tolerance, Regulatory T cells, Foxp3, Autoimmunity, Control of autoimmune diseases/allergy/cancer
Epigenetics, Differentiation of T cells, Translation of Dry into Wet[Research History]・Jun, 2020 – Present
Kyoto University
・Apr, 2019 – Present
Invited Researcher, IFReC, Osaka University
・Apr, 2019 - May, 2020 Researcher, INFRONT, Kyoto University[Education]
・Apr, 2015 - Mar, 2019 Majored in Medical Science, Department of Medicine, Osaka University
・Apr, 2013 - Mar, 2015 Department of Pharmaceutical Science, Osaka University

14

14

Ｙ－０６

The role of hypothalamic neurons in sleep, aging and longevity

Akiko Satoh

Department of Integrative Physiology, Institute of Development, Aging, and Cancer, Tohoku University, Japan
Department of Integrative Physiology, National Center for Geriatrics and Gerontology, Japan
It has been reported that the hypothalamus plays an important role in the regulation of aging and longevity in
mammals. However, detailed mechanisms are not fully elucidated. Aging has significant impacts on sleep,
resulting in increases in sleep fragmentation and sleep onset latency, and a decrease in sleep quality in
mammals. We have found that aged mice also display severe sleepiness during sleep deprivation. Intriguingly,
diet restriction (DR), a dietary regimen well known to delay the aging process and to extend lifespan in a wide
variety of organisms including mammals, can dramatically attenuate some of age-associated sleep alterations.
For example, DR significantly reduces the levels of age-associated sleep fragmentation and sleep attemptduring sleep deprivation. Therefore, DR may delay the aging process and extend lifespan by critically altering
sleep control in aged animals. To further elucidate mechanisms by which DR ameliorates age-associated sleep
alterations, we focus on the gene called PR-domain containing factor 13 (Prdm13). Prdm13 is a Sirt1
downstream gene and is exclusively expressed in the dorsomedial hypothalamus (DMH). The DMH has beenshown to control age-associated physiological changes and longevity through Sirt1. We have recently foundthat conditional DMH-specific Prdm13-knockout mice mimic age-associated sleep alterations including sleep
fragmentation and severe sleepiness during sleep deprivation. We are currently testing whether Prdm13 isnecessary to promote DR-induced sleep phenotypes. In this talk, I will discuss about a role of DMH neurons
on age-associated sleep alterations. I will also propose how poor sleep quality promotes the age-associatedpathophysiology in peripheral organs/tissues and ultimately affects longevity.

15

15

Tsuji S, Brace CS, Yao R, Tanie Y, Tokizane K, Tada H, Rensing N, Mizuno S, Almunia J, Kong Y,
Nakamura K, Ogiso N, Toyokuni, S, Takahashi S, Wong M, Imai S, Satoh A#, Prdm13-expressing neurons in
the dorsomedial hypothalamus regulate sleep-wake patterns during aging in mice, under revision
Satoh A#, Central Mechanisms linking age-associated physiological changes to health span through the
hypothalamus, Aging Mechanisms II (Springer), pp289-306, 2022
Satoh A, Imai S, Guarente LP: The brain, ageing, and sirtuins. Nat. Rev. Neurosci. 18:362–374, 2017
Satoh A#, Brace CS, Rensing N, Imai S#: Prdm13, a newly identified dorsomedial hypothalamus-enriched
gene, affects age-associated pathophysiology. Aging Cell 14:209–218, 2015
Satoh A, Brace CS, Rensing N, Clifton P, Wozniak DM, Herzog ED, Yamada KA, Imai S: Sirt1 extends life
span and delays aging in mice through the regulation of Nk2 homeobox 1 in the DMH and LH. Cell Metab.
18:416–430, 2013

Akiko Satoh is an Associate Professor in the Department of Integrative Physiology at Institute of Development,
Aging, and Cancer, Tohoku University, Japan (2021-present) (also an Associate Professor at the National Center
for Geriatrics and Gerontology, Japan). She obtained her PhD in Pharmaceutical Science from the University of
Toyama in 2005. She had her postdoctoral training at Washington University School of Medicine in St. Louis.
Her current research aims to understand the role of hypothalamic sleep regulation in mammalian aging andlongevity control.

1616

Ｓｅｓｓｉｏｎ3

17

18

Ｓ－０５

Novel control mechanisms of lymphocyte trafficking
Suzuki, Kazuhiro1,2,3; Shirai, Taiichiro1; Nakai, Akiko1,2
1

Laboratory of Immune Response Dynamics, WPI Immunology Frontier Research Center (IFReC), Osaka

University, Japan
Department of Immune Response Dynamics, Research Institute for Microbial Diseases (RIMD), Osaka
University, Japan
3
Center for Infectious Disease Education and Research (CiDER), Osaka University, Japan
2

Blood-circulating lymphocytes enter secondary lymphoid organs, including the spleen and lymph nodes(LNs), and migrate to distinct compartments where they survey for antigens. After spending several hours to a
day in a secondary lymphoid organ, lymphocytes exit and traffic to other lymphoid organs where they
continue antigen surveillance. Once lymphocytes encounter cognate antigens, they rapidly change theirmigration programs to stay in the lymphoid organ and induce adaptive immune responses. These lymphocyte
trafficking events are governedby G protein-coupled receptors (GPCRs) that respond to chemotactic

molecules represented by chemokines. Our study showed that inputs from adrenergic neurons inhibit
lymphocyte exit from LNs through the β2-adrenergic receptor (β2AR) expressed on lymphocytes. This ismediated by the crosstalk between β2AR and chemokine receptors, CCR7 and CXCR4, where β2AR
activation enhances LN retention signals mediated by the chemokine receptors. We found that the adrenergic
neuron-mediated control of lymphocyte exit from LNs contributes to diurnal variations of adaptive immune
responses. In investigating molecular mechanisms of the crosstalk between β2AR and chemokine receptors,
we identified a protein complex consisting of copper metabolism MURR1 domain-containing (COMMD) 3
and COMMD8 (COMMD3/8 complex) as an adaptor that recruits GPCR kinase 6 to chemoattractant
receptors and promotes lymphocyte chemotaxis. The COMMD3/8 complex plays essential roles in the controlof B cell migration in vivo and induction of humoral immune responses. Disabling the COMMD3/8 complex
alleviates the pathology in mouse models of autoimmune disease. Based on these findings, we discuss novel
mechanisms for the control of lymphocyte trafficking and immunotherapeutic approaches targeting them.

17

17

References(Related articles)

Nakai, A., Fujimoto, J., Miyata, H., Stumm, R., Narazaki, M., Schulz, S., Baba, Y., Kumanogoh, A. andSuzuki, K. The COMMD3/8 complex determines GRK6 specificity for chemoattractant receptors. J. Exp.
Med. 216 (7): 1630-1647, 2019.
Suzuki, K., Hayano, Y., Nakai, A., Furuta, F. and Noda, M. Adrenergic control of the adaptive immune
response by diurnal lymphocyte recirculation through lymph nodes. J. Exp. Med. 213 (12): 2567-2574, 2016.

Nakai, A., Hayano, Y., Furuta, F., Noda, M. and Suzuki, K. Control of lymphocyte egress from lymph nodes
through β2-adrenergic receptors. J. Exp. Med. 211 (13): 2583-2598, 2014.

CV

04/1994 – 03/1998 School of Science, The University of Tokyo, Tokyo, Japan (BS in Chemistry)

04/1998 – 03/1999 Graduate School of Science, The University of Tokyo, Tokyo, Japan
04/1999 – 03/2003 Medical School, Osaka University, Osaka, Japan (MD)04/2003 – 03/2004 Resident in internal medicine, Osaka University Hospital, Osaka, Japan
04/2004 – 09/2007 Graduate School of Medicine, Osaka University, Osaka, Japan (PhD in Medical Science)
10/2007 – 03/2011 Postdoctoral fellow, University of California, San Francisco, USA
04/2011 – 03/2017 Associate professor, IFReC, Osaka University, Osaka, Japan
04/2017 – present Professor, IFReC, Osaka University, Osaka, Japan
07/2017 – present Professor (concurrent), RIMD, Osaka University, Osaka, Japan
04/2021 – present Professor (concurrent), CiDER, Osaka University, Osaka, Japan

18

18

Ｓ－０６

Roles of the GDF3-ALK7 axis in adiposity

Izumi1, Tetsuro.

1

Laboratory of Molecular Endocrinology and Metabolism, Department of Molecular Medicine Institute for
Molecular and Cellular Regulation, Gunma University, Japan

ALK7, a type I receptor in the transforming growth factor-β superfamily, is predominantly expressed inadipocytes and regulates lipid metabolism by modulating the master adipose transcription factors, C/EBPαand PPARγ. GDF3, a principal candidate of ALK7 ligand, is produced from adipose tissue macrophages in
response to NLRP3 inflammasome activation and/or insulin secreted under nutrient-excess condition. Thissignaling axis within white adipose tissue physiologically plays a major role in storing excess intake energy as
fat. However, the continuous activation enlarges adipocytes and induces a proinflammatory and insulinresistant state. Inactivation of ALK7 or GDF3 protects mice from diet-induced and genetically prone obesity
and insulin resistance. Furthermore, human subjects with ALK7 variants have an association with reduced
waist-to-hip ratio and resistance to development of type 2 diabetes. The blockade of the GDF3-ALK7
signaling pathway, which targets the final outcome of obesity, fat accumulation, may be a safe and robust
treatment for obesity and diabetes.

19

19

References

Hirayama I, Yi Z, Izumi S, Arai I, Suzuki W, Nagamachi Y, Kuwano H, Takeuchi T, and Izumi T (1999).
Genetic analysis of obese diabetes in the TSOD mouse. Diabetes, 48, 1183-1191.
Mizutani S, Gomi H, Hirayama I, and Izumi T (2006). Chromosome 2 locus Nidd5 has a potent effect on
adiposity in the TSOD mouse. Mamm. Genome, 17, 375-384.
Yogosawa S, Mizutani S, Ogawa Y, and Izumi T (2013). Activin receptor-like kinase 7 suppresses lipolysis toaccumulate fat in obesity through downregulation of peroxisome proliferator-activated receptor γ and
C/EBPα. Diabetes, 62, 115-123.

Yogosawa S, and Izumi T (2013). Roles of activin receptor-like kinase 7 signaling and its target, peroxisome
proliferator-activated receptor γ, in lean and obese adipocytes. Adipocyte, 2, 246-250.
Bu Y, Okunishi K, Yogosawa S, Mizuno K, Irudayam MJ, Brown CW, and Izumi T (2018). Insulin regulates
lipolysis and fat mass in adipocytes by upregulating growth/differentiation factor 3 in adipose macrophages.
Diabetes, 67, 1761-1772.

CV
EDUCATION/DEGREES

1982.3: Faculty of Medicine, University of Tokyo, M.D.
1989.4: Faculty of Medicine, University of Tokyo, Ph.D.

POSITIONS/EMPLOYMENT1982 – 1984: Resident (Internal Medicine), University of Tokyo Hospital, Mitsui Memorial Hospital
1984 – 1989: Physician and Research Fellow, Third Department of Internal Medicine, Faculty of Medicine,
University of Tokyo, Institute of Diabetes Care and Research, Asahi Life Foundation
1989 – 1992: Research Associate, Third Department of Internal Medicine, Faculty of Medicine, University ofTokyo
1990 – 1994: Research Associate, Howard Hughes Medical Institute, Department of Pharmacology,
University of Colorado School of Medicine, USA
1994 – 2000: Associate Professor, Department of Molecular Medicine, Institute for Molecular and Cellular
Regulation, Gunma University
2000 – present:
Professor, Institute for Molecular and Cellular Regulation, Gunma University(2015.4-2019.3: Director, Institute for Molecular and Cellular Regulation, Gunma University)

20

20

Ｙ－０７

Premature ovarian insufficiency in mice lacking phosphoinositide-metabolizing
enzymes.
Junko Sasaki1, Shogo Yanai1, Junjie Huang1, Junya Hasegawa1, Takehiko Sasaki1

1

Department of Biochemical Pathophysiology, Medical Research Institute, Tokyo Medical and Dental

University, Tokyo, Japan
Premature ovarian insufficiency (POI) is characterized by the cessation of menstruation before age 40 inhumans. It is estimated that about 1% of women are affected by POI. In POI, ovarian function declines andovulation stops, making pregnancy very difficult. Candidate genes causing follicular dysgenesis have been
discovered mainly by phenotypes obtained from murine knockout models (1). FSH (follicle-stimulating
hormone) receptors and BMP15 (Bone Morphogenetic Protein 15) and GDF9 (Growth Differentiation Factor9), which are oocyte-derived growth factors belonging to the TGF-β (transforming growth factor-β)
superfamily, are found to be genetically abnormal in sporadically occurring human POI. However, more than
50% of POIs are unexplained.
We have analyzed the physiological functions of phosphoinositides (PIPs), which are intracellular signaling
phospholipids (2-5). In the process, we generated conditional

PI(3,4,5)P3 (Phosphatidylinositol 3,4,5-

trisphosphate) phosphatase-deficient mice (cKO mice) using transgenic mice expressing Cre recombinase
under the control of the CD11b promoter and found that males were normal and fertile, while females wereinfertile. When 1-month-old cKO female mice were subjected to superovulation treatment with PMSG
(pregnant mare serum gonadotropin) and hCG (Human chorionic gonadotropin), the cKO female mice did not
ovulate at all, suggesting that they were developing POI. Histological analysis has found that cKO ovaries
contain a large number of follicles with abnormal morphology, which are not found in wild-type ovaries. In
this meeting, I will discuss the pathological analysis of cKO ovaries and report on the molecular mechanismof POI development due to abnormal PI(3,4,5)P3 metabolism.

21

21

References(Related articles)(1) Jiao X, Ke H, Qin Y, Chen ZJ. Molecular Genetics of Premature Ovarian Insufficiency. Trends
Endocrinol Metab, 29, 795-807, 2018
(2) Morioka S, Nakanishi H, Yamamoto T, Hasegawa J, Tokuda E, Hikita T, Sakihara T, Kugii Y, Oneyama
C, Yamazaki M, Suzuki A, Sasaki J, Sasaki T. A mass spectrometric method for in-depth profiling ofphosphoinositide regioisomers and their disease-associated regulation. Nat Commun, 13, 83, 2022
(3) Koizumi A, Narita S, Nakanishi H, Ishikawa M, Eguchi S, Kimura H, Takasuga S, Huang M, Inoue T,
Sasaki J, Yoshioka T, Habuchi T, Sasaki T. Increased fatty acyl saturation of phosphatidylinositol
phosphates in prostate cancer progression. Sci Rep, 9, 13257, 2019(4) Kimura H, Eguchi S, Sasaki J, Kuba K, Nakanishi H, Takasuga S, Yamazaki M, Goto A, Watanabe H,
Itoh H, Imai Y, Suzuki A, Mizushima N, Sasaki T. Vps34 regulates myofibril proteostasis to prevent
hypertrophic cardiomyopathy. JCI Insight, 2, e89462, 2017(5) Kofuji S, Kimura H, Nakanishi H, Nanjo H, Takasuga S, Liu H, Eguchi S, Nakamura R, Itoh R, Ueno N,
Asanuma K, Huang M, Koizumi A, Habuchi T, Yamazaki M, Suzuki A, Sasaki J, Sasaki T. INPP4B is a
PtdIns(3,4,5)P3 phosphatase that can act as a tumor suppressor. Cancer Discov, 5, 730-739, 2015

CV

<Educational carrier>
2002
1995
1993

Ph.D., Graduate School of Pharmaceutical Sciences, University of Tokyo
M.S., Graduate School of Home Economics, Ochanomizu University
B.S., Faculty of Home Economics, Ochanomizu University<Profession carrier>
2018201320082003200019991995-

Associate Professor:
Associate Professor:
Lecturer:
Assistant Professor:
Postdoctoral Fellow:
Technical Staff:
Researcher:

Medical Research Institute Tokyo Medical and Dental University
Akita University Graduate School of Medicine
Akita University Graduate School of Medicine
Akita University Graduate School of Medicine
Tokyo Metropolitan Institute of Medical Science
Ontario Cancer Institute, Canada
Pharmaceutical Research Institute, Kyowa Hakko Kogyo Company

22

22

Ｙ－０８

ACBD3 forms specialized ER-Golgi contact sites to drive the ER exit of
STING
Motani, Kou1; Saito-Tarashima, Noriko2; Nishino, Kohei1,3; Yamauchi, Shunya2;
Minakawa, Noriaki2; Kosako, Hidetaka1.
Division of Cell Signaling, Institute of Advanced Medical Sciences, Tokushima University, Japan.
Graduate School of Pharmaceutical Science, Tokushima University.
3
Kuramoto Division, Technical Support Department, Tokushima University.
1
2

Stimulator of interferon genes (STING) is an endoplasmic reticulum (ER)-resident receptor for cyclic dinucleotides (CDNs) that triggers potent innate immune responses in both microbial infections and sterile
inflammations. Upon CDN binding, STING moves from the ER to the Golgi apparatus, where it activates thedownstream TBK1-IRF3 axis, leading to type-I interferon (IFN) induction. Although ER exit is a critical stepin STING signaling, its mechanism is poorly understood. Here, we visualized the ER exit sites of STING byblocking its transport at low temperature. After ligand binding, STING formed dot-like structures (hereafterreferred to as “STING ER exit foci”) at non-canonical ER exit sites. Using APEX2-based proximity
proteomics and cross-linking immunoprecipitation coupled to mass spectrometry, we identified the Golgiresident protein ACBD3/GCP60 as an essential and specific component of the STING ER exit foci.
Subsequent imaging analysis and in vitro reconstitution using semi-intact cells revealed that ACBD3
recognized ligand-bound STING and concentrated it at specialized ER-Golgi contact sites. Moreover,depletion of ACBD3 diminished STING foci formation and inhibited STING exit from the ER to the Golgi,
resulting in impaired type-I IFN responses. Our results identified the ACBD3-positive specialized ER-Golgi
contact sites that function as platforms to trigger the ER exit of STING.

23

23

References(Related articles)

CV
2/2019-Present

Senior Lecturer
Tokushima University

10/2014-2/2019

Assistant Professor
Tokushima University

2/2014-9/2014

Project Assistant Professor
Tokushima University

4/2013-2/2014

Research Fellowship for Young Scientists
Japan Society for the Promotion of Science

4/2011-3/2013

Program Specific Researcher
Kyoto University

3/2011

Ph.D. in Medical Science
Kanazawa University

24

24

Ｙ－０９

Cell type-specific control of reward learning in the brain

Macpherson, Tom1; Hikida, Takatoshi1

1

Laboratory for Advanced Brain Functions, Institute for Protein Research, Osaka University, JapanThe nucleus accumbens (NAc) of the ventral striatum is thought to play a critical role in controlling reward
signaling in the brain, and its dysfunction is associated with several psychiatric conditions associated with
impaired reward learning, including drug addiction, depression, and schizophrenia. Within the NAc, neurons
can be subdivided into two broadly equal populations, medium spiny neurons expressing either dopamine D1
or D2 receptors (D1-/D2-MSNs); however, the contribution of these two neuron types to reward learning has
been unclear. Here in a series of experiments, we used a combination of transgenic and viral technologies toblock neurotransmission from NAc D1- or D2-MSNs in mice performing behavioral tasks that required
various types of reward learning, including Pavlovian and goal-directed learning. Our findings reveal
differential functional roles for NAc cell types, with D1-MSNs necessary for Pavlovian reward learning, andD2-MSNs necessary for goal-directed reward learning as well as adaptive learning when task parameters arechanged. These findings potentially identify new neurocircuit therapeutic targets for reward learning
impairments in psychiatric conditions.

25

25

References(Related articles)
Macpherson T, Kim J, Hikida T. (2022) Nucleus accumbens core dopamine D2 receptor-expressing neurons
control reversal learning but not set-shifting in behavioral flexibility in male mice. Frontiers in Neuroscience.
In Press.
Macpherson T, Matsumoto M, Gomi, H, Morimoto J, Uchibe E, Hikida T. (2021) Parallel andhierarchical neural mechanisms for adaptive and predictive behavioral control. Neural Networks. 144:507521.
Macpherson T, Hikida T. (2019) Role of basal ganglia neurocircuitry in the pathology of psychiatricdisorders. Psychiatry and Clinical Neuroscience.73(6): 289-301.
Macpherson T, Hikida T. (2018) Nucleus accumbens dopamine D1-receptor-expressing neurons control theacquisition of sign-tracking to conditioned cues in mice. Frontiers in Neuroscience. 12: 418.
Macpherson T, Morita M, Wang Y, Sasaoka T, Sawa A, Hikida T. (2016) Nucleus accumbens dopamine D2receptor expressing neurons control behavioral flexibility in a place discrimination task in the IntelliCage.
Learning and Memory. 23 (7): 359-364.

CV
Tom Macpherson completed his PhD in Behavioral & Clinical Neuroscience at the University of Sussex,
UK (2009-2014), studying the role of GABAA receptors in controlling addiction to cocaine in mice. This
work led to new insights into the mechanisms by which GABA receptors in the nucleus accumbens (NAc)
can modulate the influence of local dopamine release following cocaine administration (Maguire et al.,2014).
Upon graduation from his PhD, he was granted a JSPS Postdoctoral Fellowship and moved to Kyoto
University to take up a postodoctoral research position (2014-2016). During this time, his research led toseveral major breakthroughs in our understanding of striatal neurocircuit control of Pavlovian conditioning
and behavioral flexibility and resulted in the award of a second JSPS Postdoctoral Fellowship grant tocontinue his research for a further two years (2016-2018).
Since 2018, Tom Macpherson has been working as an assistant professor at Osaka University’s Institute ofProtein Research. His research incorporates cutting-edge neural imaging, artificial intelligence, genetic,
molecular, and behavioral analyses to elucidate the neural mechanisms underlying motor, cognitive, andlimbic control within the brain.

26

26

Ｓｅｓｓｉｏｎ4

27

28

Ｓ－０７

Molecular mechanisms underlying autophagosome biogenesis

Nobuo N. Noda1,2

Division of Biological Molecular Mechanisms, Institute for Genetic Medicine, Hokkaido University, Japan
Labratory of Structural Biology, Institute of Microbial Chemistry, Japan1
2

Autophagy is an intracellular degradation system conserved among eukaryotes and contributes to cellular
homeostasis through degradation of harmful or superfluous materials. When autophagy is induced, isolation
membrane suddenly appears in the cytosol, which expands and seals into a double-membrane structure termed
an autophagosome, during which cytoplasmic materials are sequestered within it. Autophagosome then fuseswith the lysosome and the sequestered materials are degraded by lysosomal hydrolases. Autophagosome
biogenesis is mediated by dozens of core autophagy-related (Atg) proteins, which are known to gather at the
pre-autophagosomal structure (PAS) upon autophagy induction. However, it remained elusive how Atg
proteins gather and cooperate to execute autophagosome biogenesis.
In order to reveal the molecular mechanisms of autophagosome biogenesis, we performed structural analyses
on all core Atg proteins and established the structural basis of these proteins1,2). During these studies, we
noticed that many of Atg proteins are abundant with intrinsically disordered regions (IDRs), which we could
visualize using high-speed atomic force microscopy (HS-AFM)3,4). Since IDRs are known to be related toliquid-liquid phase separation (LLPS), a common mechanism for constructing membraneless organelles orcondensates, we studied the role of LLPS in autophagy and found that the PAS is a liquid-like condensate
formed by LLPS of the Atg1 complex5). Moreover, we also found that autophagy selectively degrades liquidlike protein condensates formed by LLPS but not solid-like aggregates6,7).
After the PAS is organized, initial isolation membrane is generated from the PAS, which then expands using
phospholipids mainly derived from the endoplasmic reticulum (ER). The mechanism of how bulk
phospholipids are transferred from the ER to the isolation membrane remained elusive. Using in vitro
systems, we found that Atg2 and Atg9, which localize at the ER-isolation membrane contact site, possessintermembrane and interleaflet lipid transfer activities, respectively. Based on in vitro and in vivo data, we
proposed a novel mechanism that Atg2 and Atg9 collaborate to transfer phospholipids from the ER to theisolation membrane for membrane expansion8,9). We also found that Atg8, a ubiquitin-like protein tightly
bound to isolation membranes, possesses membrane perturbation activity in vitro and that Atg8 uses thisactivity to promote autophagosome biogenesis10)

27

27

References(Related articles)1) Suzuki et al., Structural biology of the core autophagy machinery. Curr. Opin. Struct. Biol. 43, 10-17(2017).
2) Matoba & Noda, Structural catalog of core Atg proteins opens new era of autophagy research. J. Biochem.
169, 517-525 (2021).
3) Yamamoto et al., The intrinsically disordered protein Atg13 mediates supramolecular assembly of
autophagy initiation complexes. Dev. Cell 38, 86-99 (2016).
4) Kodera et al., Structural and dynamics analysis of intrinsically disordered proteins by high speed atomic
force microscopy. Nat. Nanotech. 16, 181-189 (2021).
5) Fujioka et al., Phase separation organizes the site of autophagosome formation. Nature 578, 301-305(2020).
6) Yamasaki et al., Liquidity is a critical determinant for selective autophagy of protein condensates. Mol.
Cell 77, 1163-1175 (2020).
7) Fujioka & Noda, Biomolecular condensates in autophagy regulation. Curr. Opin. Cell Biol. 69, 23-29(2021).
8) Osawa et al., Atg2 mediates direct lipid transfer between membranes for autophagosome formation. Nat.
Struct. Mol. Biol. 26, 281-288 (2019).
9) Matoba et al., Atg9 is a lipid scramblase that mediates autophagosomal membrane expansion. Nat. Struct.
Mol. Biol. 27, 1185-1193 (2020).
10) Maruyama et al., Membrane perturbation by lipidated Atg8 underlies autophagosome biogenesis. Nat.
Struct. Mol. Biol. 28, 583-593 (2021).

CV

EDUCATION
2001
1998
1996

Ph.D. from University of Tokyo, Graduate School of Pharmaceutical Sciences
Supervisor: Professor Yoshinori Satow
M.S. in the pharmaceutical sciences from University of Tokyo
B.S. in the pharmaceutical sciences from University of Tokyo

WORK EXPERIENCE
2022 to present
Professor at Institute for Genetic Medicine, Hokkaido University, Sapporo, Japan.
2017 to 2021
Laboratory Head at Institute of Microbial Chemistry, Tokyo, Japan.
2011 to 2017
Chief Researcher at Institute of Microbial Chemistry, Tokyo, Japan.
2008-2011
Lecturer with tenure at Hokkaido University, Sapporo, Japan.
2005-2008
Research Associate at Hokkaido University, Sapporo, Japan.
2001-2005
Postdoctoral fellow at Hokkaido University, Sapporo, Japan.
RESEARCHMy research interest is to understand the molecular mechanisms of autophagy and the roles of liquid-liquid
phase separation in various life phenomena.

28

28

Ｙ－１０

Gut microbiota modulates inflammation in the central nervous system

Miyauchi1,2, Eiji; Desai S3,4., Mahesh; Sasaki1, Nobuo; Ohno2,5,6, Hiroshi
Institute for Molecular and Cellular Regulation, Gunma University, JapanRIKEN Center for Integrative Medical Sciences, Japan3
Department of Infection and Immunity, Luxembourg Institute of Health, Luxembourg
4
Department of Dermatology and Allergy Center, Odense University Hospital, University of Southern
Denmark,
Denmark5
Graduate School of Medical Life Science, Yokohama City University, Japan
6
Graduate School of Medicine, Chiba University, Japan
1
2

While genetic factors are primarily responsible for autoimmune diseases, environmental factors including dietand infections also contribute to their risk. Accumulating evidence has indicated that one of the major
environmental factors, gut microbiota, plays a significant role in the pathogenesis of autoimmune diseases
including multiple sclerosis (MS). MS is an autoimmune disease characterized by chronic inflammation anddemyelination in the central nervous system (CNS), and pro-inflammatory autoreactive T cells, especially T
helper 17 (TH17) cells, are key players in the CNS inflammation. Studies of experimental autoimmune
encephalomyelitis (EAE; an animal model of MS), as well as human studies, have implicated gut microbiota
dysbiosis in the development or severity of MS. However, it remains unclear how gut microbes act on the
inflammation of extra-intestinal tissues such as the CNS.
We found that two distinct gut microbes coordinately activate autoreactive TH17 cells that respondspecifically to myelin oligodendrocyte glycoprotein (MOG) and worsen EAE. After induction of EAE inmice, MOG-specific CD4+ T cells are observed in the small intestine, where a newly isolated strain in thefamily Erysipelotrichaceae acts as an adjuvant to enhance the TH17 responses. Shotgun sequencing of thesmall intestinal microbiota also revealed a strain of Lactobacillus reuteri that possesses peptides that
potentially mimic MOG. Mice that are co-colonized with these two strains showed EAE symptoms that weremore severe than those of germ-free or monocolonized mice. We also performed EAE experiments using
gnotobiotic mice colonized with a synthetic human gut microbiota. Dietary fiber deprivation in these miceleads to the increase in Akkermansia muciniphila and decrease in short chain fatty acids (SCFAs) in theintestine. Although these features have also been observed in MS patients and are thought to contribute to thedisease pathogenesis, removal of A. municiphila and addition of SCFAs did not affect EAE symptoms.
Comprehensive metabolomic analysis of the intestine revealed that metabolite X is increased upon the fiber
deprivation and that the metabolite X promotes the antigen-specific T cell activation.
Our study highlights some potential targets for microbiome-focused therapeutic approaches in MS, althoughfurther studies are needed to validate the their roles in humans.

29

29

References(Related articles)
1.

Miyauchi E, Shimokawa C, Steimle A, Desai MS, Ohno H. The impact of the gut microbiome on extraintestinal autoimmune diseases. Nat Rev Immunol. 2022;10.1038/s41577-022-00727-y.

2.

Miyauchi E, Kim SW, Suda W, et al. Gut microorganisms act together to exacerbate inflammation in
spinal cords. Nature. 2020;585(7823):102-106.

3.

Takewaki D, Suda W, Sato W, et al. Alterations of the gut ecological and functional microenvironment
in different stages of multiple sclerosis. Proc Natl Acad Sci U S A. 2020;117(36):22402-22412.

4.

Duscha A, Gisevius B, Hirschberg S, et al. Propionic Acid Shapes the Multiple Sclerosis Disease
Course by an Immunomodulatory Mechanism. Cell. 2020;180(6):1067-1080.e16.

5.

Desai MS, Seekatz AM, Koropatkin NM, et al. A Dietary Fiber-Deprived Gut Microbiota Degrades theColonic Mucus Barrier and Enhances Pathogen Susceptibility. Cell. 2016;167(5):1339-1353.e21.

6.

Berer K, Gerdes LA, Cekanaviciute E, et al. Gut microbiota from multiple sclerosis patients enables
spontaneous autoimmune encephalomyelitis in mice. Proc Natl Acad Sci U S A. 2017;114(40):1071910724.

CV
2022-current Associate Professors, Laboratory of Mucosal Ecosystem Design, Institute for Molecular andCellular Regulation, Gunma University2021-2022

Senior Researcher, Laboratory for Intestinal Ecosystem, RIKEN Center for Integrative
Medical Sciences (IMS)

2019-2021

Researcher, Laboratory for Intestinal Ecosystem, RIKEN Center for Integrative Medical
Sciences (IMS)

2016-2019

Special Postdoctoral Researcher (SPDR) in RIKEN, Laboratory for Intestinal Ecosystem,
RIKEN Center for Integrative Medical Sciences (IMS)

2013-2015

Research Fellow of Japan Society for the Promotion of Science (PD)2012-2015

Postdoctoral fellow, Laboratory for Intestinal Ecosystem, RIKEN Center for Integrative
Medical Sciences (IMS), RIKEN

2009-2012

Research Fellow of Japan Society for the Promotion of Science (DC1)

2009-2012

Ph.D. (Dr. of Agriculture), Graduate School of Biosphere Science, Hiroshima University

2010-2011

Visiting PhD Student (JSPS Excellent Young Researcher Overseas Visit Program),
Alimentary Pharmabiotic Centre, University College Cork, Ireland

30

30

Ｙ－１１

Immune cell dynamics and role in central nervous system diseases

Minako Ito1

1

Division of Allergy and Immunology, Medical Institute of Bioregulation, Kyushu University, Japan

In recent years, the linkage between the nervous system and the immune system has been the focus of muchattention. In addition to neurodegenerative diseases such as Alzheimer's disease, the involvement of theimmune system has begun to be strongly implicated in the pathogenesis of psychiatric disorders such as
autism spectrum disorder (ASD). It is also becoming clear that immune cells are involved in brain
development and aging. The interaction between various immune cells and nervous system cells is thought tobe important in various events such as pathological conditions, development, and aging.
Using a mouse model of cerebral infarction, we have reported on the regulatory mechanisms of inflammationand neurological symptoms by immune responses after cerebral infarction. In this symposium, we will
introduce immune responses in the brain during various CNS diseases and brain development.

31

31

References(Related articles)

Ito M*, Komai K, Mise-Omata S, Iizuka-Koga M, Noguchi Y, Kondo T, Sakai R, Matsuo K, Nakayama T,
Yoshie O, Nakatsukasa H, Chikuma S, Shichita T, Yoshimura A*. Brain regulatory T cells suppress
astrogliosis and potentiate neurological recovery. Nature. 2019; 565(7738): 246-250

CV

2020.2-present
Associate Professor
Division of Allergy and Immunology, Medical Institute of Bioregulation, Kyushu University

2019.4-2020.1
Assistant professor
2016.4-2019.3
Postdoctoral research fellow (PD)
Department of Microbiology and Immunology, Keio University School of Medicine.
2016
Ph.D. (Medical Science)
Department of Microbiology and Immunology, Keio University School of Medicine.
2013
M.S. (Medical Science)
Department of Pathological Science, Graduate School of Medical Science, Kyushu University.
2011
B.S. (Biomedical Science)
Department of Biomedical Science, Kyushu University School of Medicine.

32

32

Ｙ－１２

Strategies for the expansion of cerebral cortex in primates

Hatakeyama, Jun1; Sato, Hatuka1; Shimamura, Kenji1;

Department of Brain morphogenesis, Institute of Molecular Embryology and Genetics, Kumamoto
University,

1

Expansion of the cerebral cortex is one of the important factors that archives higher cognitive abilities andsensorimotor functions. Notably, primates including human have markedly expanded cerebral cortex.
However, it is still unclear how species-specific expansion period of neural progenitors is determined andmechanisms underlying massive enlargement of human cerebral cortex. Recent progresses in the field ofneural development have led to the discovery of intrinsic differences of neural progenitors among species.
The differences in proliferative and neurogenic properties of the canonical neural progenitors have been
considered to be responsible for the cortical expansion. On the other hand, the cerebrospinal fluid (CSF),
which is produced from the choroid plexus (ChP), also influences neural progenitors, CSF contains various
secreting molecules including growth factors and supports neural progenitor proliferation. Interestingly,
human embryo has markedly developed ChP, raising a possibility that the differences in CSF contributes tothe expansion of cerebral cortex.
We have identified several secreting factors from the embryonic ChP which are preferentially expressed in
macaque monkey (Macaca fascicularis). We would found that these factors promoted proliferation of primate
neural progenitors, as well as murine ones. We would like to propose that these primate specific CSF factors
play important roles in the expansion of cerebral cortex.

33

33

References(Related articles)
1) Human-specific genomics signatures of neocortical expansion.
Florio M. et al. 2017 42: 33-44, Current Opinion in Neurobiology
2) Cerebral cortex expansion and folding: what have we learned?
Fernandez V. et al. 2016 35(10): 1021-1044, The EMBO journal

CV
Education
2005
Ph.D., Graduate School of Medicine, Kyoto University.
Employment
2021Visiting Assistant Professor, Shiga University of Medical Science
2021JST FOREST scientist
2009Assistant professor, Institute of Molecular Embryology and Genetics,
Kumamoto University
2006-2009 JSPS research fellow (PD), Institute of Molecular Embryology and Genetics,
Kumamoto University
2006
Post-doctoral fellow, Institute of Molecular Embryology and Genetics,
Kumamoto University
2002-2005 JSPS research fellow (DC1), Institute for Virus Research, Kyoto University

34

34

Ｓｅｓｓｉｏｎ5

35

36

Ｓ－０８

Generation of RNA aptamers and their applications in molecular biology

Takahashi Masaki

Project Division of RNA Medical Science, The Institute of Medical Science, The University of Tokyo, JapanRNA has a great potential to specifically bind to various kinds of targets by forming unique three-dimensional
structures. RNA molecules that bind to targets of interest, which widely range from chemicals, recombinant
proteins to viruses and whole living cells, are called as aptamers. Currently, aptamers serve as a new modality
for medical agents and as an experimental tool in molecular biology. Nucleic acid aptamers are isolated fromlarge combinatorial libraries using a unique iterative selection-amplification process known as systematic
evolution of ligands by exponential enrichment (SELEX). By modifying SELEX methods to suit proteins ofinterest, we developed aptamers against P2RY2, a GPCR, and TGF-β1 as a unique GPCR modulator andmedical agent, respectively. As for generating aptamers targeting P2RY2, we established a new selection
strategy named “VLP-SELEX” using virus-like particles (VLPs) for stabilizing GPCRs with their native
structures, and showed unique functions of aptamers as a positive allosteric modulator (PAM) agonist forP2RY2. Regarding anti-TGF-β1 aptamers, we developed the aptamers as medical agents possessing chemical
modifications and showed that the aptamers enhance anti-cancer drug efficacy in xenograft model. Herein, I
would like to introduce and discuss these aptamers and their application in molecular biology.

35

35

References(Related articles)
01. Takahashi M*, Hashimoto Y, Nakamura Y. “Anti-TGF-β1 aptamer enhances therapeutic effect oftyrosine kinase inhibitor, gefitinib, on non-small cell lung cancer in xenograft model.” Mol Ther
Nucleic Acids. 2022 June 29 online.
02. Takahashi M*, Amano R, Ozawa M, Martinez A, Akita K, Nakamura Y*. “Nucleic acid ligands act
as a PAM and agonist depending on the intrinsic ligand binding state of P2RY2.” Proc Natl Acad Sci
U S A.. 2021 May 4 118 (18) e2019497118
03. Takahashi M. “Nucleic acid aptamers emerging as modulators of G-protein coupled receptors:
Challenge to difficult cell-surface proteins" Cells. 2022 June 11(11), 1825 (Review)
04. Takahashi M. “Aptamers targeting cell-surface proteins" Biochimie. 2016 Dec 131, 77-84 (Review)

CV[Education]2008 Ph.D.in Science, Aoyama Gakuin University
2003 B.S. in Science, Aoyama Gakuin University[Professional Career]
2015-present: Project Associate Professor, Project Division of RNA Medical Science, IMSUT
2014-2015: Project Assistant Professor, Project Division of RNA Medical Science, IMSUT
2009-2014: Post-Doctoral Fellow, Department of Molecular Pharmacology, National Institute of
Neuroscience,
NCNP
2007-2009: Research Fellow of Japan Society for the Promotion of Science (JSPS) (DC2, PD)
2005-2007: Junior Research Associate (JRA), RIKEN

36

36

Ｓ－０９

Identification of chromatin instability at the single-cell level fromtranscriptome data
Itoshi NIKAIDO1,2

Medical Research Institute, Tokyo Medical and Dental University (TMDU)
RIKEN Center for Biosystems Dynamics Research

1
2

To understand complex human organs at a single-cell resolution, we need to analyze the function andorganization of each cell type and state and study their interactions. Single-cell transcriptome analysis is adata-driven approach to identifying cell types and functions by acquiring gene expression data on a wholegene scale without prior identification of cell types and states in organs and tissues.
Single-cell RNA-seq methods to obtain reads from the full-length RNA and its data analysis methods areneeded to understand diseases that involve changes in RNA structure, such as cancer. Single-cell RNA
sequencing methods can simultaneously measure thousands to tens of thousands of single-cell transcriptomes(Sasagawa Y. et al. Genome Biol. 2018, Mereu E. et al. Nature Biotech. 2020). However, these highthroughput single-cell RNA sequencing methods determine only 3’-end truncated RNA sequences and cannot
capture changes in RNA structural variation. In addition, in principle, non-poly A RNA, estimated to accountfor 50% of all RNA in typical mammalian cells, cannot be detected. The detection of non-polyA RNAs willlead to the discovery of new disease markers.
Therefore, we developed the world's first Single-cell Full-length Total RNA-seq method RamDA-seq(Hayashi T. et al. Nature Comm. 2018; Nature Digest 2018, Morita R. et al. Nature 2021). We have also
developed a method to identify cell-specific genomic regions expressed from data obtained with this method(Matsumoto H. NARGAB 2020) and its visualization method Millefy (Ozaki H. BMC Genomics, 2020). We
are currently developing robotic experiment automation and artificial intelligence techniques to identify the
genomic instability that leads to abnormal cellular functions. In this talk, I will introduce the progress of thedevelopment of these technologies.

37

37

References(Related articles)
1.

Ritsuko Morita, et al. Tracing the origin of hair follicle stem cells. Nature. 09 June 2021

2.

Hiroshi Ochiai, Tetsutaro Hayashi, Mana Umeda, Mika Yoshimura, Akihito Harada, Yukiko Shimizu,
Kenta Nakano, Noriko Saitoh, Hiroshi Kimura, Zhe Liu, Takashi Yamamoto, Tadashi Okamura,
Yasuyuki Ohkawa, Itoshi Nikaido. Genome-wide analysis of transcriptional bursting-induced noise in
mammalian cells. Science Advances 17 Jun 2020: Vol. 6, no. 25, eaaz6699.

3.

Elisabetta Mereu et al. Benchmarking Single-Cell RNA Sequencing Protocols for Cell Atlas
Projects. Nature Biotechnology. 06 April 2020.

4.

Haruka Ozaki, Tetsutaro Hayashi, Umeda Mana, Itoshi Nikaido. Millefy: visualizing cell-to-cellheterogeneity in read coverage of single-cell RNA sequencing datasets. BMC Genomics volume 21,
Article number: 177, 2020.

5.

Hirotaka Matsumoto, Tetsutaro Hayashi, Haruka Ozaki, Koki Tsuyuzaki, Mana Umeda, Tsuyoshi Iida,
Masaya Nakamura, Hideyuki Okano, Itoshi Nikaido. A NMF-based approach to discover overlooked
differentially expressed gene regions from single-cell RNA-seq data. NAR Genomics andBioinformatics, Volume 2, Issue 1, March 2020.

6.

Tetsutaro Hayashi*, Haruka Ozaki*, Yohei Sasagawa, Mana Umeda, Hiroki Danno, Itoshi
Nikaido. Single-cell full-length total RNA sequencing uncovers dynamics of recursive splicing andenhancer RNAs. Nature Communications. 9, Article number: 619, 2018.

CV
Employment:•
Apr. 2020 - current: Professor, Department of Functional Genome Informatics, Division of Medical
Genomics, Medical Research Institute, Tokyo Medical and Dental University (TMDU)•
Apr. 2019 - current: Team Leader, Laboratory for Bioinformatics Research, RIKEN Center for
Biosystems Dynamics Research•
Apr. 2018 - Mar. 2019: Unit Leader, Laboratory for Bioinformatics Research, RIKEN Center for
Biosystems Dynamics Research•
Apr. 2013 - Mar. 2018: Unit Leader, Bioinformatics Research Unit, Advanced Center for Computing
and Communication, RIKEN, Japan.
•
Apr. 2014 - Mar. 2016: Senior Research Scientist, Preventive medicine and applied genomics unit,
Advanced Center for Computing and Communication, RIKEN, Japan. (concurrent)•
Jun. 2006 – Mar. 2013: Researcher, Functional genomics unit, Center for Developmental Biology,
RIKEN Kobe Institute•Apr. 2004 – May. 2006: Research fellow, Division of Functional Genomics & Systems Medicine,Research Center for Genomic Medicine, Saitama Medical School
Education:•
Apr. 2001 – Mar. 2004 Doctor’s Degree Course Division of Genome Information Resources, Scienceof Biological Supramolecular Systems, Graduate School of Integrated Science, Yokohama City
University

38

38

Ｓ－１０

Chromatin regulation during skeletal muscle regeneration

Yasuyuki Ohkawa

Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, JapanThe selective incorporation of appropriate histone variants into chromatin is critical for the regulation of
genome function. Although many histone variants have been identified, a complete list has not been
compiled. We previously screened the mouse genome in silico, and we identified 14 uncharacterized H3genes, among which13 are similar to H3.3 and one is similar to the human testis-specific H3 variant,
H3T/H3.4. Although some of these genes were previously annotated as pseudogenes, their tissue-specificexpression was confirmed by RNA sequencing. However, many of their functions are still unknown.
Therefore, we have been analyzing the function of these unknown histone H3 genes. In particular, we havebeen analyzing H3mm7, H3mm13, and H3mm18,which are expressed in skeletal muscle. In this talk, I would
like to report on the progress of the analysis and the latest findings, as well as introduce the originaltechnologies that have supported these analyses.

39

39References(Related articles)
1. +Honda M, Oki S, Kimura R, Harada A, Maehara K, Tanaka K, Meno C, *Ohkawa Y.
High-depth spatial transcriptome analysis by photo-isolation chemistry.
Nat Commun. 2021 Jul 20;12(1):4416. doi: 10.1038/s41467-021-24691-8.
2. +Handa T, Harada A, Maehara K, Sato S, Nakao M, Goto N, Kurumizaka H, *Ohkawa Y, *Kimura H.
Chromatin integration labeling for mapping DNA-binding proteins and modifications with low input.
Nat Protoc. 15(10):3334-3360. 2020.
3. *Oka M, Mura S, Otani M, Miyamoto Y, Nogami J, Maehara K, Harada A, Tachibana T, Yoneda Y,
Ohkawa Y.
Chromatin-bound CRM1 recruits SET-Nup214 and NPM1c onto HOX clusters causing aberrant HOX
expression in leukemia cells.
Elife. 8 e46667. 2019.
4. +Harada A, Maehara K, Handa T, Arimura Y, Nogami J, Hayashi-Takanaka Y, Shirahige K,
Kurumizaka H, *Kimura H, *Ohkawa Y.
A chromatin integration labelling method enables epigenomic profiling with lower input.
Nat Cell Biol. Feb;21(2):287-296. 2019.
5. +Harada A, Maehara K, Ono Y, Taguchi H, Yoshioka K, Kitajima Y, Xie Y, Sato Y, Iwasaki T, Nogami
J, Okada S, Komatsu T, Semba Y, Takemoto T, Kimura H, Kurumizaka H, *Ohkawa Y.
Histone H3.3 sub-variant H3mm7 is required for normal skeletal muscle regeneration.
Nat Commun. 9(1):1400. 2018.

CV
2003 Ph.D. Graduate School of Medicine, Osaka University
2003-2006 Postdoctoral Fellow, Department of Cell Biology, University of Massachusetts Medical
School
2006-2011 Project Associate Professor, Graduate School of Medical Sciences, Kyushu University
2011-2015 Associate Professor, Graduate School of Medical Sciences, Kyushu University, Kyushu
University
2016- Professor, Department of Transcriptomics, Medical Institute of Bioregulation, Kyushu University

40

40

Ｓｅｓｓｉｏｎ6

41

42

Ｓ－１１

Regulation on stem cells in myeloid leukemia: a metabolic perspective

Takahiro Ito1

1

Institute for Life and Medical Sciences, Kyoto University, Japan

Cancer cells reprogram and rewire cellular metabolic networks for survival, proliferation, and adaptation to
microenvironment. Such altered metabolism often leads to dependency on specific nutrients, and therefore,targeting cancer-specific metabolism would be an effective therapeutic strategy. Towards this goal it isessential to identify metabolic vulnerabilities in human malignancy. In this talk I would like to share ourrecent findings on the roles of essential role of the branched-chain amino acid (BCAA) metabolism in themaintenance of leukemia stem cells and its functional contribution to disease progression in human cancer.

41

41

References(Related articles)
•

•
•
•

Hattori A, Tsunoda M, Konuma T, Kobayashi M, Nagy T, Glushka J, Tayyari F, McSkimming D,
Kannan N, Tojo A, Edison A, Ito T.
Cancer progression by reprogrammed BCAA metabolism in myeloid leukemia. Nature 545, 500-504,(2017).
Fox RG, Lytle NK, Jaquish DV, Park FD, Ito T, et al.
Image-based detection and targeting of therapy resistance in pancreatic adenocarcinoma. Nature, 534,407-11, (2016).
Zimdahl B*, Ito T*, Blevins A, Bajaj J, Konuma T, Weeks J, et al.
Lis1 regulates asymmetric division in hematopoietic stem cells and in leukemia. Nat. Genet., 46,
245–52, (2014).
Ito T, Kwon HY, Zimdahl B, Congdon KL, Blum J, Lento WE, et al.
Regulation of myeloid leukaemia by the cell-fate determinant Musashi. Nature, 466, 765–8, (2010).

CV

Bio:
Dr. Ito received his Ph.D. in Pharmaceutical Sciences from the University of Tokyo in 2006 for his work ontranscriptional regulation by eukaryotic RNA polymerase II. He then shifted his academic focus to stem celland cancer biology and joined the group of Tannishtha Reya at Duke University Medical Center and later UC
San Diego. After the postdoctoral training, Dr. Ito was appointed as an assistant professor in 2013 in the
Department of Biochemistry and Molecular Biology at the University of Georgia and then promoted to an
Associate Professor. In 2019, Dr. Ito joined the Institute of Life and Medical Sciences at Kyoto University as
a professor. His research group studies the molecular mechanisms and regulation of cell fates in stem cells
and cancer with a particular focus on the role of RNA binding proteins and metabolic reprogramming incancer progression and drug resistance.

42

42

Ｓ－１２

Modeling hematopoietic stem cell development in a dishOgawa1, Minetaro

1

Department of Cell Differentiation, Division of Stem Cell Research, Institute of Molecular Embryology andGenetics, Kumamoto University, Kumamoto, Japan
Inducing transplantable hematopoietic stem cells (HSCs) from pluripotent stem cells (PSCs) in culture has
been a long outstanding challenge in developmental biology and regenerative medicine. During the ontogenyof vertebrate species, HSCs derive from a specialized population of endothelial cells known as hemogenic
endothelial cells (HECs). HECs are not merely the developmental origin of HSCs: they also generate various
types of blood cells independently of the HSC development. In the case of mouse embryos, HECs in theextraembryonic yolk sac give rise to erythro-myeloid progenitors and lymphoid-myeloid progenitors.
However, the yolk sac HECs do not generate HSCs. In the intraembryonic region, HECs of the dorsal aorta
generate diverse blood precursors, including multipotent hematopoietic progenitors, before the emergence of
HSCs. HSCs are eventually formed from the late HECs in the dorsal aorta as the last wave of the definitive
hematopoiesis. The HSC-independent hematopoietic lineages with different developmental origins andpathways have been an obstacle to the induction of HSCs from PSCs in culture. The successful derivation of
HSCs from PSCs will require the culture method to correctly follow the developmental pathway toward thedefinitive HSC without deviating to the easy-to-make HSC-independent lineages. To this end, it is necessary
to precisely identify intermediate precursors in various developmental stages between the pluripotent
epiblasts and the definitive HSCs. The signaling molecules driving the proliferation and differentiation ofeach intermediate precursor should also be identified. Single-cell RNA-sequencing of mouse embryos
provides valuable insights into the process of HSC development. This technology helps identify candidate cell
populations of intermediate precursors and deduce the signaling pathways activated in the precursors. For
verification, the candidate precursors are isolated from mouse embryos and cultured in the presence ofdeduced signaling molecules to induce their differentiation into transplantable HSCs. Once the culture
conditions are determined by tracing the developmental stages step-by-step, these conditions can be directly
applied to mouse embryonic stem cells to induce HSCs in culture. An example of our ongoing approach inline with this strategy will be discussed.

43

43References(Related articles)
Tsuruda, M., S. Morino-Koga, and M. Ogawa. Hematopoietic stem cell-independent differentiation of mast
cells from mouse intraembryonic VE-cadherin+ cells. Stem Cells 40: 332-345, 2022. DOI:
10.1093/stmcls/sxac001
Tsuruda, M., S. Morino-Koga, and M. Ogawa. Bone morphogenetic protein 4 differently promotes distinct
VE-cadherin+ precursor stages during the definitive hematopoietic development from embryonic stem cellderived mesodermal cells. Exp. Hematol. 103: 40-51.e7, 2021. DOI: 10.1016/j.exphem.2021.08.008
Tsuji-Tamura, K., S. Morino-Koga, S. Suzuki, and M. Ogawa. The canonical smooth muscle cell marker
TAGLN is present in endothelial cells and is involved in angiogenesis. J. Cell Sci. 134 (15): jcs254920, 2021.
DOI: 10.1242/jcs.254920
Ahmed, T., K. Tsuji-Tamura, and M. Ogawa. CXCR4 signaling negatively modulates the bipotential state ofhemogenic endothelial cells derived from embryonic stem cells by attenuating the endothelial potential. StemCells 34:2814-2824, 2016. DOI: 10.1002/stem.2441

CV
Name (Surname, First name): Ogawa, Minetaro
Education:
1980–1984
1984–1986
1986–1987
1992

Faculty of Pharmaceutical Science, Kanazawa University, Kanazawa, Japan
Bachelor of Pharmacy
The Master’s Course in Pharmaceutical Science, Kanazawa University
Master of Pharmacy
The Doctoral Course in Pharmaceutical Science, Kanazawa University
Ph.D. (Doctor of Pharmacy, Kanazawa University)

Work Experience:
1987–1992
Assistant Professor, Institute for Medical Immunology,
Kumamoto University School of Medicine, Kumamoto, Japan
1992–1993
Assistant Professor, Institute of Molecular Embryology and Genetics,
Kumamoto University School of Medicine
1993–1995
Assistant Professor, Department of Molecular Genetics,
Faculty of Medicine, Kyoto University, Kyoto, Japan
1995–1997
Member Scientist, Basel Institute for Immunology, Basel, Switzerland
1997–2000
Assistant Professor, Department of Molecular Genetics,
Graduate School of Medicine, Kyoto University
2000–2002
Associate Professor, Department of Molecular Genetics,
Graduate School of Medicine, Kyoto University
2002 to present
Professor, Institute of Molecular Embryology and Genetics, Kumamoto University

44

44

Poster Session

45

46

Ｐ－０１

A cross-talk between microtubules and focal adhesions regulates dynamics ofactin cytoskeleton
Nishimura, Yukako1,2; Nisha Bte Mohd Rafiq2 ;Sergey V. Plotnikov3; Visalatchi
Thiagarajan2; Zhen Zhang2; Meenubharathi Natarajan2; Shidong Shi2; Virgile Viasnoff2,4,5;
Gareth E. Jones6; Pakorn Kanchanawong2,7 and Alexander D. Bershadsky2,81
Division of Developmental Physiology, IGM, Hokkaido University, Japan; 2Mechanobiology Institute,
National University of Singapore, Singapore; 3Department of Cell and Systems Biology, University ofToronto, Canada; 4CNRS UMI 3639, Singapore; 5Department of Biological Sciences, National university ofSingapore, Singapore; 6Randall Centre for Cell & Molecular Biophysics, King's College London, UK;
7
Deparment of Biomedical Engineering, National University of Singapore, Singapore; 8Department of
Molecular Cell Biology, Weizmann Institute of Science, Israel

The interrelationship between microtubules and the actin cytoskeleton in mechanoregulation of integrinmediated adhesions is poorly understood. Here, we show that the effects of microtubules on a major type ofcell-matrix adhesion, focal adhesions, are mediated by KANK family proteins connecting the adhesion
protein talin with microtubule tips. Both total microtubule disruption and microtubule uncoupling fromadhesions by manipulations with KANKs trigger a massive assembly of myosin-IIA filaments. Myosin-IIA
filaments, augmenting the focal adhesions, are indispensable effectors in the microtubule-dependent
regulation of integrin-mediated adhesions. Myosin-IIA filament assembly depends on Rho activation by the
RhoGEF, GEF-H1, which is trapped by microtubules when they are connected with integrin-mediatedadhesions via KANK proteins but released after their disconnection. Thus, microtubule capturing by integrinmediated adhesions modulates the GEF-H1-dependent effect of microtubules on the myosin-IIA filaments.
Subsequent actomyosin reorganization then remodels the focal adhesions, closing the regulatory loop.

Ｐ－０２

Extracellular Domains I and II of CD44 mediate its trans-homophilic
dimerization and tumor cluster aggregation
Madoka Kawaguchi1, Nurmaa Dashzeveg2, Yue Cao3, Yuzhi Jia2, Xia Liu4, Yang Shen3*,
Huiping Liu2,5,6*.
Department of Gene Expression Regulation, Institute of Department, Aging and Cancer, Tohoku University,
Japan; 2Department of Pharmacology, Finberg School of Medicine, Northwestern University, USA;3
Department of Electrical and Computer Engineering, TEES-AgriLife Center for Bioinformatics andGenomic Systems Engineering, Texas A&M University, USA; 4Department of Toxicology and Cancer
Biology, College of Medicine, University of Kentucky, USA; 5Department of Medicine,
Hematology/Oncology Division, Finberg School of Medicine, Northwestern University, USA; 6Robert H.
Lurie Comprehensive Cancer Center, Finberg School of Medicine, Northwestern University, USA1

CD44 molecule (CD44) is a surface glycoprotein on cancer stem cells. However, its utility as a therapeutictarget for managing metastases remains to be fully evaluated. We examined the biochemical properties ofCD44 in homotypic tumor cell aggregation. The standard CD44 form (CD44s) mainly assembled asintercellular homodimers (trans-dimers) in tumor clusters rather than intracellular dimers (cis-dimers) present
in single cells. Based on machine learning-based computational modeling combined with experimental
mutagenesis tests, substituted 10 these residues in Domain I or 5 residues in Domain II abolished CD44
dimerization and reduced tumor cell aggregation in vitro. Importantly, the substitutions in Domain II
dramatically inhibited lung colonization in mice. The CD44 dimer-disrupting substitutions decreased
downstream P21-activated kinase 2 (PAK2) activation, suggesting that PAK2 activation in tumor cell clusters
is CD44 trans-dimer-dependent. These results shed light on the biochemical mechanisms of CD44-mediatedtumor cell cluster formation and may help inform the development of therapeutic strategies to prevent tumor
cluster formation and block cluster-mediated metastases.

45
45

Ｐ－０３

Epigenetic dysregulations in the lysine methyltransferase (KMT2) deficient
endometrial cancer cells
Ryosuke Kobayashi1, Reika-Kawabata-Iwakawa2, Makoto Sugiyama3, Takehiro Yokobori2,
Sumiyo Morita1, Takuro Horii1, Masahiko Nisiyama2 and Izuho Hatada1
1 Laboratory of Genome Science, Biosignal Genome Resource Center, Institute for Molecular and Cellular

Regulation, Gunma University, Gunma, 371-8512, Japan. 2Division of Integrated Oncology Research,
Gunma University Initiative for Advanced Research (GIAR), Gunma, 371-8511, Japan. 3Faculty of
Veterinary Medicine, Kitasato University School of Veterinary Medicine, Aomori, 034-8628, Japan

Endometrial cancer (EMC) is the most common gynecological malignancy in Japan. The lysine
methyltransferase 2 (KMT2) family genes are frequently mutated in patients of EMC. Specially, KMT2C/D,
which are involved in H3K4me1 deposition in enhancer regions, were mutated in 28.7% and 21.0%,
respectively in EMC. However, the biological consequences of KMT2 mutations for the endometrial cancer
development has not been shown. To challenge these issues, we established KMT2-deficient EMC cell lines
using CRISPR/Cas9 genome editing. Ablation of KMT2 showed repression of cell growth and migration invitro. On the other hand, three-dimensional culture system revealed that spheroids formed from KMT2deficient cells had structural abnormality with spike-like protrusion compared to those from intact cells.
Morphological analysis by transmission electron microscope revealed that KMT2-deficient cells showeddisruption of cellular polarity. RNA-seq and ChIP-seq analysis demonstrated that genes involved in epithelial
functions such as adhesion or extracellular matrix organization were largely repressed in KMT2-deficient
cells, and active enhancer marks accompanied with these genes were also disappeared. These data suggestthat KMT2 ablation disrupt epithelial functions through epigenetic dysregulations in EMC cells.

Ｐ－０４

A novel E3 ligase is involved in Unfolded Protein Response throughUbiquitination of eS7A and up-regulation of Hac1 production in yeast
Sato1, Nichika; Nakano2, Yu; Matsuki2, Yasuko; Matsuo1, Yoshitaka; Inada1, Toshifumi
Division of RNA and Gene Regulation, The University of Tokyo, Japan; 2Lab of Gene Regulation, Tohoku

1

University, JapanThe endoplasmic reticulum (ER) is the major intracellular calcium reservoir in the cell and is arranged in a
dynamic tubular network involved in gluconeogenesis and lipid synthesis. Accumulation of unfolded or
misfolded proteins in ER due to various stresses induces ER stress. Unfolded stress response (UPR) is aconserved cellular pathway involved in protein quality control to maintain homeostasis under ER stress. In
the yeast UPR signaling pathway, the ER-located transmembrane protein Ire1 is crucial for splicing of the
HAC1 pre mRNA, HAC1(u) to produce HAC1i mRNA that encodes the Hac1, a transcriptional factor
essential for UPR.
We recently reported that E3 ligase Not4 was involved in UPR in yeast, Saccharomyces cerevisiae. Not4mediated mono-ubiquitination of eS7A is crucial for increasing the translation efficiency of HAC1i mRNA.
More recently, we identified GRR1 as an essential E3 ligase for resistance to Tunicamycin (Tm), an ER
stress-inducing drug. Grr1p is involved in eS7A ubiquitination and translation of HAC1i mRNA, however,
the role of eS7 ubiquitination by Grr1p in the translation of HAC1i mRNA remains unknown.
In this study, we have investigated the relationship between Grr1p-mediated ubiquitination of eS7A and upregulation of Hac1 expression to reveal the function of GRR1 under ER stress conditions. We found that
Grr1p forms a polyubiquitin chain on the K83 residue of eS7A under ER stress conditions. In addition, Hel2,
another E3 ligase that polyubiquitinated eS7A via K63 linkage, was not involved in the translation of HAC1i
mRNA. These results suggest that non-K63-linked polyubiquitination by Grr1p may be required for thetranslation of HAC1i mRNA. In summary, we propose that polyubiquitination of eS7A by Grr1p is crucial forthe translation of HAC1i and UPR.

46
46

Ｐ－０５

Molecular mechanism of YAP-dependent hepatocyte elimination
Okamoto Yoshimi 1; Endo Yoshiki 1; Nishina Hiroshi1*
Department of Developmental and Regenerative Biology, Medical Research Institute, Tokyo Medical and
Dental University (TMDU), Tokyo, Japan

1

Background and Purpose: The oncogene product Yes-associated protein (YAP) is a transcriptional cofactor
that serves in the nucleus to enhance cell proliferation. It has been reported that YAP is highly expressed in
the nucleus in cancer tissues of various organs including liver, and that the nuclear level of YAP correlateswith poor prognosis of patients. Activation of YAP in mouse liver also induces hepatocarcinogenesis. On theother hand, we found that when YAP activated forms were induced in a mosaic fashion with liver injury,
YAP-expressing hepatocytes were eliminated within one week. However, the molecular mechanism of how
activated YAP-expressing precancerous hepatocytes are eliminated is unknown. The aim of this study was toelucidate this hepatocyte elimination mechanism.
Methods: Using the hydrodynamic tail vein injection (HTVi) method, we introduced a plasmid expressing
YAP (2SA), an activated form of YAP, into mouse liver. Hepatocytes expressing activated YAP wereisolated by FACS and subjected to comprehensive gene expression analysis using RNA-seq.
Results: Several genes were altered in an activated YAP-dependent manner. Ontology analysis of theidentified genes showed that transcription of stress-responsive genes was upregulated by YAP. These genes
included Rho-guanine nucleotide exchange factors, which functions in actin cytoskeletal remodeling, andseveral chemokines involved in macrophage activation. On the other hand, transcription of metabolismrelated genes was suppressed by YAP.
Discussion: These results suggest that cell morphology changes mediated by actin remodeling, immune cell
recruitment mediated by chemokines, and changes in intracellular metabolism are involved in the elimination
of YAP-activated cells. In this way, the liver maintains homeostasis by eliminating YAP-activatedprecancerous hepatocytes, and disruption of this elimination mechanism is thought to increase the risk ofhepatocarcinogenesis. The results of this study are also expected to lead to the development of therapeuticand preventive methods for hepatic cancer targeting the mechanism of abnormal cell elimination.

Ｐ－０６

Application of CDK4/6 inhibition therapies to PDACs
Yuanyuan Zhang1, Santosh Kumar Gothwal1, Susumu Kohno1, Jindan Sheng2, Chiaki Takahashi1
1.
Cancer Research Institute,Kanazawa University, Japan.2.Medical Structural Biological Shanghai Cancer
Institute, Renji Hospital affiliated to Shanghai Jiao Tong University School of Medicine, China.

The CDK4/6-dependent RB1 mono-phosphorylation triggers cells to pass through the G1-S checkpoint incell cycle following RB1 hyper-phosphorylation. Thus, CDK4/6 inhibitors (CDK4/6i) are emerging cancer
therapeutic tools. However, many intrinsic or extrinsic factors and mutational landscapes often halt theeffectiveness of CDK4/6i therapies. Pancreatic ductal adenocarcinomas (PDAC) are depicted by veryfrequent incidence of Kras mutation and rare RB1 mutation. Here we investigated the effects of a CDK4/6i,
Palbociclib, on PDAC lines harboring KrasG12D or KrasG12C mutation, and assessed the mechanisms
leading to therapy resistance. RB7LP is a constitutively active form mutant of RB1, lacking N-terminal end
with all serines substituted to unphosphorylatable forms. RB7LP overexpression in cells phenocopies the
CDK4/6i effects including senescence-like phenotypes and partial apoptosis. Therefore, RB7LPoverexpressing cells may serve as a good model to parallelly examine the mechanism of intrinsic resistanceto CDK4/6i. We screened chemical libraries to seek molecules that enhance the cytotoxic action of RB7LP in
PDACs, which identified several kinase inhibitors. One of them appeared to target a signaling molecule X.
We further observed that CDK4/6 inhibition in PDACs increased the activity of X and moreover that of a
receptor kinase Y upstream of X. Importantly, inhibition of either X or Y dramatically synergized with
Palbociclib treatment in PDAC lines. We are currently investigating the mechanism whereby CDK4/6i
treatment increases activity of X and Y, and gathering proof of concepts that may support future applicationof CDK4/6i to PDAC patients in combination with a rationally selected kinase inhibitor.

47
47

Ｐ－０７

Structural basis of Marburg virus helical nucleoprotein-RNA complex
formation
Fujita-Fujiharu1,2,3, Yoko; Sugita1,3,4, Yukihiko; Takamatsu1, Yuki; Houri1,2,3, Kazuya;
Igarashi5, Manabu; Muramoto1,2,3, Yukiko; Nakano1,2,3, Masahiro; Tsunoda1,2,3, Yugo;
Taniguchi6, Ichiro; Becker7,8, Stephan; Noda1,2,3, Takeshi*Laboratory of Ultrastructural Virology, Institute for Frontier Life and Medical Sciences, Kyoto University,
Japan; 2Laboratory of Ultrastructural Virology, Graduate School of Biostudies, Kyoto University, Japan;
3
CREST, Japan Science and Technology Agency, Japan; 4Hakubi Center for Advanced Research, Kyoto
University, Japan; 5Division of Global Epidemiology, International Institute for Zoonosis Control, Hokkaido
University, Japan; 6Laboratory of RNA system, Institute for Frontier Life and Medical Sciences, Kyoto
University, Japan; 7Institute of Virology, University of Marburg, Germany; 8German Center for Infection
Research (DZIF), Marburg-Gießen-Langen Site, University of Marburg, Germany
1

Marburg virus (MARV), which belongs to the family Filoviridae in the order Mononegavirales, causes lethal
hemorrhagic fever in humans and non-human primates. MARV poses a threat to public health; however, novaccine or antiviral drugs have yet been licensed.
The nucleoprotein (NP) binds the non-segmented, single-stranded, negative-sense viral genomic RNA(vRNA) to form the helical NP – RNA complex. The helical NP-RNA complex acts as a scaffold fornucleocapsid formation and as a template for viral genome transcription and replication. Proper interactions
between NPs and vRNA are required for the helical assembly and their function. However, the structural
basis for the helical assembly remains largely elusive.
Here, to elucidate the structural basis for the nucleocapsid formation, we determined the structure of the
MARV NP-RNA complex using cryo-electron microscopy. Structure-based mutational analysis of MARVand Ebola virus (EBOV) NPs identified residues critical for helix assembly and subsequent viral RNA
synthesis. Most of the residues were conserved in filoviruses, suggesting that filoviruses share common
molecular mechanisms for nucleocapsid formation. The findings advance our understanding of filovirus
morphogenesis and contribute to the development of novel antivirals against MARV and EBOV.

Ｐ－０８

Abnormal pH environment interrupts cell competition-mediateddevelopmental robustness
Akieda, Yuki1; Ishitani, Tohru1.
Department of Homeostatic Regulation, Division of Cellular and Molecular Biology, Research Institute forMicrobial Diseases, Osaka University, Japan1

Our bodies are formed through repeated cell division from a single fertilized egg. During this process ofembryonic development, defective cells are suddenly generated due to genomic errors. We found that these
defective cells are eliminated by cell competition in zebrafish embryos. The defective cells are recognized by
the neighboring normal cells and are eliminated via cell death. This cell competition supports precise
embryogenesis and confers developmental robustness. Interestingly, recent NGS analysis has revealed thatcell population originating from mutant cell at early embryo are involved in various diseases such as
Alzheimer's disease and diabetes. These mutant cells occurring during embryogenesis may have evaded
elimination due to disruption of cell competition. However, the factors that disrupt the elimination by cellcompetition are not well understood. In this study, we focused on the environmental perturbations. As aresult, we found that acidic pH environment disrupts cell competition. As does alkaline pH environment. The
mechanism of suppression of cell competition by the abnormal pH environment is becoming clear that theabnormal pH environment acts on the sensing mechanism of abnormal cells by neighboring normal cells.
Thus, a new factor for disrupting cell competition has been uncovered. Interestingly, it has been reported thatthe extracellular pH environment changes to acidic due to various situations such as cancer
microenvironment, hypoxia, and inflammation, suggesting that cell competition may also be disrupted inthese situations.

48
48

Ｐ－０９

Altered behavior and increased neuroinflammation in Importin α4/KPNA4
KO mice
Sakurai, Koki1,2; Miyamoto, Yoichi3; Makiko, Morita1; Macpherson, Tom1,2; Ozawa,
Takaaki1,2; Masahiro, Oka3; Takatoshi, Hikida1,2*
Lab. for Advanced Brain Functions, Inst. for Protein Research, Osaka Univ., Japan; 2Deaprtment of
Biological Sciences, Graduate School of Science, Osaka Univ., Japan; 3Lab. for Nuclear Transport Dynamics,
National inst. for Biomedical Innovation
1

Importins are a family of proteins that assist newly synthesized polypeptide molecules to be transported fromthe cytoplasm into the nucleus. Importin αs (Imp αs/KPNAs), are a type of Importin that bind to classicalnuclear localization signals contained in the sequences of cargo proteins and form a cargo-Impα-Importin β
trimer, which allow the cargo to move through nuclear pore complexes into the nucleus. The characteristics of
Imp αs related to nucleocytoplasmic transport have been extensively studied, but the physiologicalimplications of such cellular functions are still under investigation. 3 Impα subtypes are expressed highly inthe mammalian brain and have been associated to psychiatric disease and/or alterations in mouse behavior,bringing up the importance of Impαs in brain function and behavioral regulation.
In particular, the Importin α4 protein (KPNA4) has been known to transport NF-κB transcription factors to
the nucleus, and decreased expression has been associated to psychiatric symptoms of schizophrenia anddecreases in NF-κB signaling. To examine the behavioral phenotypes of Kpna4 KO mice, we conducted a
behavioral task battery. A KPNA4 KO mouse line showed behavioral abnormalities including increased
anxiety and reduced sensorimotor gating, providing evidence that KPNA4 is associated with brain function
and behavior. To further characterize the alterations in Kpna4 KO mice, we measured inflammatory cytokines
from several different brain regions and primary cultured cells and found increases in levels of inflammatory
cytokines in KO. We aim to elucidate the molecular background underlying the behavioral abnormalities
through characterization of cellular gene expression patterns.

Ｐ－１０

RHBDL2 has essential roles for glutaminolysis and chemoresistance in triple
negative breast cancer
Matsushita Yosuke1, Okumura Kazumasa1, Komatsu Masato1, Yoshimaru Tetsuro1, Ono
Masaya 2, Tangoku Akira3, Miyoshi Yasuo4, Sasa Mitsunori5, Katagiri Toyomasa1
Div. Genome Med., Inst. Advanced Med. Sci., Tokushima Univ, Japan 2 Div. Chemother. Clin. Res., Natl.
Cancer Ctr. Res. Inst. Japan, 3 Dept. Thoracic, Endocrine Surg. Oncology, Inst. Biomed. Sci., Tokushima
Univ. Grad. Sch. Japan, 4 Dept. Breast and Endocrine Surg., Hyogo College Med. Japan, 5 Dept. Surg.,
Tokushima Breast Care Clinic. Japan1

The current therapeutic modalities for triple-negative breast cancer (TNBC) are limited, and the developmentof new molecular targeting therapeutic approaches is required. We here report the pivotal role of cancerspecific intramembrane serine protease RHBDL2 (Rhomboid-like 2) in regulating glutaminolysis in
chemoresistance of TNBC. RHBDL2 expression is significantly associated with poor prognosis in TNBC.
Furthermore, RNA interference (RNAi)-mediated attenuation of RHBDL2 in TNBC cells significantly
suppressed cell proliferation and progression, whereas ectopic RHBDL2 overexpression in HCC1806 cells
drove their proliferation. Notably, RHBDL2 interacts with ASCT2 (SLC1A5: solute carrier family one
member 5), a significant glutamine transporter, thereby inducing glutamine uptake, promoting glutaminolysis
and oxidative phosphorylation, resulting in chemoresistance to docetaxel and taxanes in TNBC cells. Thesefindings suggest that the RHBDL2-ASCT2 axis plays a critical role in the regulation of glutaminolysis andmay be a promising therapeutic target for TNBC treatment.

49
49

Ｐ－１１

Physiological functions of novel proteins encoded by hidden ORFs
Matsumoto, Akinobu; Ichihara, Kazuya; Nita, Akihiro; Mise, Shintaro; Nakayama, Keiichi
I.
Department of Molecular Cellular Biology, Medical Institute of Bioregulation, Kyushu University, Japan

1

Although significant progress has been made in genome, epigenome, and transcriptome analyses in recent
years, but the progress of translatome analysis remains behind compared to them. In translatome analysis, weare particularly focusing on "extension of known open reading frames (ORFs)" and "identification of novel
ORFs".
Translatome analysis relies on ribosome profiling (Ribo-seq), but Ribo-seq data contain a lot of experimental
noise. We have therefore established a novel method termed TISCA to precisely identify translation initiationsites, and have identified many proteins that are translated from Non-AUG initiation codons (Ichihara K.,
Matsumoto A*. et al., Nucleic Acids Res., 2021). Such Non-AUG initiation codons are often present
upstream of the canonical AUG initiation codon, producing N-extended proteins (extension of known ORFs).
Some of the extended N-terminal regions contain localization signals, which result in different subcellular
localizations. We are currently analyzing mice that specifically lack N-extended proteins.
Furthermore, although long non-coding RNAs (lncRNAs) are defined as RNAs that do not translate proteins,
we found that some lncRNAs translate polypeptides of less than 100 amino acids (identification of novel
ORFs) (Matsumoto A. et al., Nature, 2017). These novel polypeptides regulate the functions of VDAC andHDAC3, and knockout mice showed diverse phenotypes such as abnormal social behavior, delayed skinregeneration and infertility (Nita A., Matsumoto A*. et al., PLoS Genet., 2021, Mise S., Matsumoto A*. et al.,Nat. Commun., 2022).
Thus, we have succeeded in "extension of known ORFs" and "identification of novel ORFs" and revealed thatthese hidden proteins contribute to various physiological functions.

Ｐ－１２

The female-specific regulation of meiotic cell cycle in murine germ cells
Ryuki Shimada1, Ishiguro Kei-ichiro*
1

IMEG, Kumamoto University, JapanThe meiosis is specialized cell cycle to produce haploid cells that is triggered by the activation of meiotic
genes by STRA8-MEIOSIN. As mitosis-meiosis transition occurred in S phase, meiotic gene activation
should coincide with S phase. In the mammalian gametogenesis, meiotic initiation is occurred in sex-specifictiming. In testis, meiosis continuously initiated throughout their life after puberty. In contrast, in the ovary, allgerm cells enter meiosis synchronously in embryonic stage and keep cells in meiotic prophase for long term.
Although such sexual difference is well known, the underlying mechanism to ensure sexually different modeof meiosis remain elusive. Here, we identified retinoblastoma family protein, RB1 and p107 (RB-like 1) as
STRA8 interacting protein. RB is well known as a repressor of S phase entry during cell cycle. To elucidate
the physiological relevance of STRA8-RB interaction, we generated mutant mice that express STRA8 lacking
LXCXE motif (Stra8ΔRB), in which STRA8 cannot bind to RB but preserves intact interaction withMEIOSIN. In Stra8ΔRB mutant mice, whereas male mice produce functional sperm, female prematurely lose
oocytes before puberty. To address the role of RB-STRA8 interaction in meiotic initiation, we conducted
single cell-RNA-sequencing analyses using STRA8-expressing cells from, control, Stra8ΔRB and STRA8null ovaries. Transcriptome analysis revealed that STRA8-null cells highly expressed pluripotent genes,suggesting STRA8 functions to induce differentiation prior to meiosis. We also revealed that without RBSTRA8 interaction, S phase entry and expression of the Meiosin were compromised. As a result, meiosis
progression was delayed in Stra8∆RB germ cells. Our present data suggests that STRA8 have central role toinduce cell differentiation, S phase entry and meiotic initiation to ensure production of functional oocytes.

50
50

Ｐ－１３

Formation of pericentromeric heterochromatin via ZNF518s that link satellite
DNA to heterochromatin
Shinya Ohta*, Ken-Ichi Noma
Institute for Genetic Medicine, Hokkaido University, Kita-15, Nishi-7, Kita-Ku, Sapporo, 060-0815, Japan
Aneuploidy is caused by chromosomal missegregation and frequently observed in cancers and hematologicaldiseases, making it important to understand the molecular mechanisms underlying chromosomal segregation.
The intricate structure formed at the centromere/kinetochore plays an integral role in the faithful segregationof chromosomes. For instance, heterochromatin present at pericentromeric alpha-satellites is essential for
proper chromosomal segregation, although how heterochromatin is targeted to pericentromeres is not fully
understood. Here, we demonstrate a novel mechanism by which two homologous zinc-finger proteins of
ZNF518s (ZNF518A and ZNF518B) play roles in heterochromatin formation at pericentromeres. We
investigate this mechanism by determining the cellular localization of ZNF518s, centromeric proteins, suchas CENP-A and CENP-B, and heterochromatin factors, including HP1 and histone H3K9 tri-methylation.
The results indicate that the multiple segments within ZNF518s contribute to their interactions with CENP-B,
HP1, and G9a histone H3K9 methyltransferase. This study identifies the novel mechanism underlying
pericentromeric heterochromatin formation, which is mediated by the central components, ZNF518s.

Ｐ－１４

Aurora A-dependent polyubiquitination of OLA1 regulates centrosome number
through controlling the recruitment of PCM proteins
Fang1, Zhenzhou; Li1, Xingming; Yoshino1, Yuki; Murooka1, Hinari; Watababe1, Minori;Mori2, Takahiro; Chiba1, Natsuko
Departmentof Cancer Biology, IDAC, Tohoku University, Japan; 2Genome Medical Science Project,
National Center for Global Health and Medicine, Japan1

BRCA1/BARD1 functions in a variety of cellular processes, including DNA repair and centrosome
regulation. We have identified Obg-like ATPase 1 (OLA1) as a BARD1-interacting protein. OLA1 binds to
BRCA1 and BARD1 and localizes to centrosome throughout the cell cycle, whereas its localization decreases
in G2 phase. Knockdown or overexpression of OLA1 causes centrosome amplification, suggesting that
adequate level of OLA1 expression is important for the regulation of centrosome number. We have found that
OLA1 is polyubiquitinated by Aurora A, which is known as a mitotic kinase, following degradation via theproteasome in G2 phase.
In this study, we found that the expression level of OLA1 negatively regulates the recruitment of g-tubulin
and pericentrin, a conserved pericentriolar materials (PCM) scaffold protein, to centrosome in G2 phase.
Furthermore, the E3 ubiquitin ligase activity of Aurora A was involved in the recruitment of these PCM
proteins to centrosome in G2 phase and in the regulation of centrosome number. Interestingly, we also foundthat the phosphomimetic variant, OLA1-T124E, enhanced the polyubiquitination of OLA1, but not the nonphosphorylated variant, OLA1-T124A. NIMA-related kinase 2 (NEK2), which is a potential kinase for T124
residue of OLA1, promoted OLA1 polyubiquitination by Aurora A.
These suggest that Aurora A-dependent ubiquitination of OLA1 regulates centrosome number throughcontrolling the recruitment of PCM proteins to G2 phase.

51
51

Ｐ－１５

The role of small GTPase protein Rab35 in brain development
Ikuko Maejima1 Taichi Hara2, Satoshi Tsukamoto3, Hiroyuki Koizumi4,5, Takeshi
Kawauchi6, Tomoko Akuzawa1, Rika Hirai1, Hisae Kobayashi1, Inoya Isobe1, Kazuo
Emoto4, Hidetaka Kosako7, and Ken Sato1,*
Laboratory of Molecular Traffic, Institute for Molecular and Cellular Regulation, Gunma University, Japan;
Laboratory of Food and Life Science, Faculty of Human Sciences, Waseda University, Japan, 3 Laboratory
Animal and Genome Sciences Section, National Institutes for Quantum and Radiological Science andTechnology, Japan, 4 Department of Biological Sciences, School of Science, The University of Tokyo, Japan,
5
Department of Molecular and Cellular Biology, School of Pharmaceutical Sciences, Ohu University, Japan,
6
Laboratory of Molecular Life Science, Institute of Biomedical Research and Innovation, Foundation for
Biomedical Research and Innovation at Kobe (FBRI), Japan, 7 Division of Cell Signaling, Fujii Memorial
Institute of Medical Sciences, Tokushima University, Japan.
1
2

The largest family of small GTPases, Rab proteins function as molecular switches that regulate intracellular
membrane trafficking. Rab35, a highly conserved Rab in metazoans, is a multifunctional small GTPase that
regulates endocytic recycling of protein cargos to plasma membrane, cytoskeletal rearrangement, cytokinesis.
However, its physiological importance in mammalian development remains unclear. Here, we generated
systemic Rab35-knockout mice and found that Rab35 is essential for early embryogenesis. Interestingly,
brain-specific Rab35-knockout mice exhibited defects in brain functions such as anxiety-related behaviours.
Moreover, they showed hippocampal malformation due to the impaired migration of pyramidal neurons,
although evident cerebral cortex formation defects were not observed. Quantitative proteomics indicated thatthe loss of Rab35 significantly affected the levels of several proteins associated with endocytic trafficking, as
well as some neural cell adhesion molecules. Altogether, our findings reveal that Rab35 plays a crucial rolein the developing hippocampus by regulating the expression of cell adhesion molecules.

Ｐ－１６

Identification and validation of multi-specific interactions of a tumor
suppressor protein PRELP
Kosuge1,2, Hirofumi; Nakakido1,2, Makoto; Nagatoishi1, Satoru; Fukuda3, Tetsuya; Bando3,
Yasuhiko; Ohnuma4, Shin-ichi; Tsumoto1,2, KouheiInst. of Med. Sci., Univ. of Tokyo, Japan; 2Sch. of Eng., Univ. of Tokyo, Japan; 3Biosys Technologies, Japan;
4Inst. of Ophthalmology, Univ. College London, UK
1

The small leucine-rich repeat proteoglycans (SLRPs) are a family of secreted proteins, many of that regulate
various cellular functions through the interactions with different proteins in the extracellular environment.
Proline and arginine-rich end leucine-rich repeat protein (PRELP) is a SLRP, known to function as a tumor
suppressor and that mRNA of PRELP is down-regulated in many types of cancer, although the details of themolecular mechanism have yet to be fully clarified. Since we hypothesized that PRELP regulates tumor cellproliferation through some interactions with different proteins as other SLRPs, we investigated the newinteractions of PRELP to characterize the molecular mechanism of PRELP-mediated tumor suppression.
We first assessed the interaction of PRELP with transforming growth factor-β1 (TGFβ1), a growth factor
known to interact with several SLRPs. A pull-down assay showed that PRELP interacted with TGFβ1 asother SLRPs. Then, we also carried out co-immunoprecipitation coupled with mass spectrometry (CoIP-MS)
to comprehensively identify novel interactions between PRELP and membrane proteins. The proteomic
analysis identified various membrane proteins that would interact with PRELP, including two growth factor
receptors, insulin-like growth factor I receptor (IGFI-R) and low-affinity nerve growth factor receptor(p75NTR). Surface plasmon resonance (SPR) analysis using recombinant proteins validated the directbinding of PRELP to TGFβ1, IGFI-R, and p75NTR with around micromolar affinities. Furthermore, cellbased analysis using recombinant PRELP protein showed that PRELP suppressed cell growth and affected
cell morphology of A549 lung carcinoma cells, also at micromolar concentration, which is consistent with themicromolar affinities measured by SPR analysis. Our results indicate that PRELP regulates the tumor cellproliferation triggered by the weak-affinity multi-specific interactions (H. Kosuge et al., J. Biol. Chem. 2021;
296: 100278).

52

52

Ｐ－１７

Establishment of a high-content polymer array screening system
to explore niche mimicries for cancerous cells
Murota1, Yoshitaka; Tabu1, Kouichi; Taga1, Tetsuya
Department of Stem Cell Regulation, Medical Research Institute, Tokyo Medical and Dental University(TMDU), Japan
1

Chemical-based approaches are one of the gold standard therapies for cancers. However, fundamentally newmodalities still need to be developed due to the resistant properties of refractory cancers against conventional
small molecular compounds that act inside cells. Thus, macromolecules controlling cells from outside (i.e.,
microenvironment) are considered possible key options to tackle this issue. Although some synthetic polymers
with bio-functionalities have been reported, the presence of polymer scaffolds that affect cancer nature andtheir detailed mechanisms of action remains to be demonstrated. Here, the groundworks for a high-content
polymer screening system have been set up to explore functional polymers that specifically regulate cancer
cells. To fabricate the microwell plates for such screening, sixty-nine acrylate heteropolymers were designedwith two from 14 kinds of biocompatible monomers, photo-polymerized at the defined condition of 254 nm
UV illumination and immobilized on the acrylate-functionalized glass bottom surface. Residual monomers
were washed out by ethanol and water, and the immobilization of polymers was confirmed by using
fluorescein-O-acrylate. Finally, adult p53-/- mouse-derived astrocytes (AS) and their transformed ones by
RasV12 oncogenic mutant (AS-RasV12) were mixed and seeded onto polymer arrays. The numbers of cells
were evaluated at 24 and 96 hours after seeding, and two hit polymers with different growth-supporting
preferences for AS and AS-RasV12 cells were identified. One cancer-friendly polymer grew AS-RasV12 by
22.7-fold and AS by 14.2-fold, and another polymer grew AS-RasV12 3.2-fold and AS 5.8-fold. Further
investigation to reveal the mechanisms of action how these hit polymers specifically support cancerous cells
will help us uncover unknown cancer characteristics, leading to the development of new modalities and theidentification of new therapeutic targets.

Ｐ－１８

Lysosomes regulate oncogenic signals in acute myelogenous leukemia
Masaya Ueno1,2, Loc Thi Pham1, Kenta Kurayoshi1, Atsushi Hirao1,2
1

Division of Molecular Genetics, Cancer Research Institute, Kanazawa University, Japan

2

WPI Nano Life Science Institute (WPI-Nano LSI), Kanazawa University, Japan

Even though a majority of leukemia cells are killed by chemotherapy, persistent leukemic stem cells (LSCs)
are still not eradicated with current treatments, due to various drug-resistance mechanisms. Recent studies
have revealed that nutrients, such as amino acid, sugar, and lipid, are critical determinants of cell proliferationand differentiation of LSCs, indicating metabolic pathways have critical roles in regulation of "stemness" in
LSCs. In this study, to understand hallmarks of metabolic regulation of stemness in acute myelogenous
leukemia (AML), we screened metabolic pathways that critical for maintenance of undifferentiated status of
AML cells. For this purpose, we establisted sgRNA libraries comprehensively targeting molecules in humanmetabolisms. Cas9-expressing HL-60 cells were transduced with these sgRNA libraries, and then forceddifferentiated cells by CRISPR/Cas9-mediated gene-deletion were sorted. We found that sgRNA in the
differentiated cells enriched targeted-genes that contribute particular metabolic pathways: lysosomal
acidification, iron-sulfur clusters, and electron transport in mitochondrial. We confirmed that pharmacologicalinhibition of lysosome acidification efficiently induced cell differentation and apoptosis in AML cell lines.
RNA-seq indicated that inhibition of lysosome acidification suppress transcription of oncogenes. We also
found that H3K27ac occupancy around an oncogene was significantly reduced by inhibition of lysosome
acidification. Furthermore, combination therapy with another anti-cancer drug and inhibitor of lysosome
acidification, synergistically suppressed tumor formation in vivo. These data indicated that lysosome regulates
expression of oncogenes, and lysosome acidification is a promissing therapetic target for AML.

53

Ｐ－１９

Identification of factors essential for CXCR4 signaling in leukemic cells using
CRISPR dropout screens
Aoki1, Kazunari; Yusa1, Kosuke
1

Stem cell genetics, LiMe Kyoto University, Japan

Blood cells, including immune cells, are generated from hematopoietic stem and progenitor cells (HSPCs) inthe bone marrow. HSPCs are in contact with and maintained by special microenvironments, known as niches,in the bone marrow. CXCL12 as well as stem cell factor, which are produced by HSPC niches, are essential
for HSPC maintenance.
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive malignant neoplasm of the bone marrow, andthe outcome of treatment-refractory and/or relapsed T-ALL is poor. CXCL12 is essential for the migration ofleukemic cells to the bone marrow and the retention and maintenance of leukemic cells in the bone marrow;
however, factors essential for CXCL12-induced chemotaxis of leukemic cells are not fully described.
To identify genes essential for the regulation of CXCL12-induced chemotaxis of leukemic cells, we carriedout a genome-wide CRISPR/Cas9 screen in a chemotaxis assay with a human T-ALL cell line. Cas9expressing leukemic cells were transduced with the human CRISPR library, cultured for 10 days, and used fora chemotaxis assay toward CXCL12. Input cells and migrated cells were harvested to determine their gRNA
content. The genome-wide screen identified 1,072 genes which were significantly depleted from the migrated
cells, suggesting that these genes are essential for the regulation of CXCL12-indeced chemotaxis of theleukemic cells. CXCR4 (encoding CXCR4, the main physiological receptor for CXCL12) and RHOA
(encoding RHOA, a guanosine triphosphatase essential for CXCL12-induced chemotaxis) were highly
depleted from the migrated cells, confirming the robustness of our screen. Some genes which had not beenreported to regulate CXCL12-induced chemotaxis were highly depleted from the migrated cells.
Thus, we develop a new screening system which effectively identify genes essential for a CXCL12-induced
chemotaxis by combining the genome-wide CRISPR-Cas9 dropout screening system and the chemotaxis
assay system. Our screening strategy can be used to explore factors essential for another chemokine-induced
chemotaxis of other types of cells.

Ｐ－２０

An infectivity-enhancing site on the SARS-CoV-2 spike protein targeted by
antibodies
Liu Yafei1,2, Arase Hisashi1,2,*
1
2

Department of Immunochemistry, Research Institute for Microbial Diseases, Osaka University, Japan;
Laboratory of Immunochemistry, WPI Immunology Frontier Research Center, Osaka University, Japan;

Antibodies against the receptor-binding-domain of the SARS-CoV-2 spike protein prevent SARS-CoV-2
infection. However, the effects of antibodies against other spike protein domains are largely unknown. Here,we screened a series of anti-spike monoclonal antibodies from COVID-19 patients, and found that some ofantibodies against the N-terminal-domain (NTD) induced the open conformation of receptor binding domain(RBD) and thus enhanced the binding capacity of the spike protein to ACE2 and infectivity of SARS-CoV-2.
Mutational analysis revealed that all the infectivity-enhancing antibodies recognized a specific site on the
NTD. Structural analysis demonstrated that all the infectivity-enhancing antibodies bound to NTD in asimilar manner. The antibodies against this infectivity-enhancing site were detected at high levels in severe
patients. Moreover, we identified antibodies against the infectivity-enhancing site in uninfected donors, albeit
at a lower frequency. These findings demonstrate that not only neutralizing antibodies but also enhancing
antibodies are produced during SARS-CoV-2 infection.

54
54

Ｐ－２１

Induction of cellular senescence by sustained NFκB activation
Tabata, Sho1; Izumi, Yoshihiro2; Takahashi, Masatomo2; Bamba, Takeshi2; Okada, Mariko1

IPR, Osaka Univ., Japan; 2 Div. of Metabolomics, MIB, Kyushu Univ., Japan

1

Nuclear Factor-kappa B (NFκB) is an important transcription factor implicated in various biologicalphenomena such as cancer, inflammation, aging, and immune response. The canonical pathway of NFκB has
multiple feedbacks, and activation of NFκB is known to exhibit oscillation behavior. NFκB inhibitor alpha(IκBα) is one of target genes of NFκB, but also mediates a powerful negative feedback loop that may remove
NFκB from DNA and result in oscillatory NFκB activity. Knockout of the IκBα gene induces sustained NFκB
activation in cells stimulated with tumor necrosis factor α (TNFα), leading to gene expression distinct fromoscillatory NFκB activation (Cheng et al., Science, 2021). However, whether and how this change in the
dynamics of NFκB activity link to cell properties or fate is not fully understood.
In breast cancer MCF7 cells, knockdown of IκBα and stimulation of TNFα induced sustained activation of
NFκB, which promoted cellular senescence phenotypes, including increased cell size, cell cycle arrest,
elevated SA-βgal activity, and upregulated senescence-associated secretory phenotype (SASP) gene
expression. Furthermore, metabolomic and transcriptomic analyses revealed that the sustained activation of
NFκB induced some characteristic metabolic alterations associated with cellular senescence. In thispresentation, we would like to discuss the molecular mechanisms of cellular senescence caused by the altereddynamics of NFκB activity due to IκBα knockdown.

Ｐ－２２

Endoplasmic Reticulum Stress Sensor IRE1 Directly Detects Misfolded
Insulin by Oligomerization
Motonori, Matsusaki1,2,3; Takeshi, Yokoyama4; Atsushi, Tsugita4; Shingo, Kanemura5;
Michiko, Tajiri6; Satoko, Akashi6; Tomohide, Saio1; Kenji, Inaba3; Masaki, Okumura2
Inst. of Adv. Med. Sci., Tokushima Univ., Japan; 2FRIS, Tohoku Univ., Japan; 3IMRAM, Tohoku Univ.,
Japan; 4Grad. Sch. of Life Sci., Tohoku Univ., Japan; 5Grad. Sch. of Sci., Kwansei Gakuin Univ., Japan;
6Grad. Sch. of Med. Life Sci., Yokohama City Univ., Japan
1

The mammalian endoplasmic reticulum (ER) is an organelle that produces secretory proteins such as insulin,collagen, and IgG. Sometimes, the accumulation of misfolded proteins (ER stress) is caused by external
environmental changes or the production of secretory proteins in excess of acceptable levels. Since ER stresscan cause various diseases, cells have an ER stress response mechanism (UPR) that regulates the amount ofmolecular chaperones. The ER stress sensor, IRE1, initiates the UPR by oligomer formation at a microscopic
scale under stress conditions. Interestingly, cells regulate UPR depending on the quantity and quality ofstress, but the molecular mechanism of regulation is unknown. In particular, it is completely unclear how
IRE1 recognizes the "stress level". In this study, we investigated the hypothesis that the more misfolded
proteins there are, the larger IRE1 oligomers are formed. In the presence of misfolded insulin, the IRE1
oligomeric state was investigated by electrophoresis, mass spectrometry, and so on. The results showed that
IRE1 changes its oligomeric state depending on the amount of misfolded insulin. This change in theoligomeric state suggests that IRE1 senses the accumulation level of misfolded proteins as the level of ER
stress.

55
55

Ｐ－２３

Essential role of ER membrane complex subunit 1 (EMC1) in B cell
development
Kazuhiko Kawata1, Yoshihiro Baba1
1
Division of Immunology and Genome Biology, Medical Institute of genome bioregulation, Kyushu
University, Japan

Introduction
Endoplasmic reticulum (ER) is a multifunctional organelle which contributes to quality control of proteinsand cellular calcium homeostasis. It has been suggested that the ER homeostasis is closely involved in B celldifferentiation and antibody secretion, but its molecular basis remains unclear. Store-operated Ca2+ entry(SOCE) mediated by STIM1, which can induce a sustained influx of extracellular Ca2+, is chief source ofCa2+ signals and extremely important for maintaining ER homeostasis. Recently, we have identified the ER
membrane complex subunit 1 (EMC1) as a novel STIM1-binding partner to regulate SOCE. In addition,
EMC1 has been reported to be a member of EMC family which consist of ten subunits and function as“insertase” which promotes protein insertion into plasma membrane, suggesting the important role of EMC1
on ER homeostasis by regulating Ca2+ and protein quality control. In this study, we aimed to understand thefunction of EMC1 in B cell development.
Results and Discussion
We generated mice with B cell-specific deletion of EMC1 (EMC1 BKO mice) and found that SOCE and B
cell development was impaired in the absence of EMC1. Given that B cell differentiation in STIM BKO miceis normal, it is considered that the abnormal B cell differentiation in EMC1 BKO mice is due to reasons otherthan SOCE disorders. To address the cause of defect in B cell development, we performed proteome analysis.
The results showed that chemokine receptors (CXCR4, CCR6, EBI2) are decreased in EMC1-deficient B
cells. CXCR4 is particularly associated with maintaining peripheral B cell compartment. Therefore, ourfindings suggest that EMC1 support insertion of CXCR4 into plasma membrane and that regulates B cell
development.

Ｐ－２４

RNA phase transition disrupts α-Synuclein proteostasis
Yabuki1,2, Yasushi; Matsuo1, Kazuya; Shioda1,2, Norifumi
Institute of Molecular Embryology and Genetics, Kumamoto University, Japan; 2Graduate School of
Pharmaceutical Sciences, Kumamoto University, Japan

1

The mechanism underling dysfunction of cellular proteostasis on prion-like proteins (such as α-synuclein (αSyn), tau and FUS) leading to pathogenesis of neurodegeneration remains unclear. Recently, we reported thatthe binding of an RNA secondary structure G-quadruplex (G4RNA) to a prion-like protein FMRpolyG causes
its liquid-to-solid phase transition, leading to neurodegeneration in a hereditary neurodegenerative disease,
Fragile X-associated Tremor/Ataxia Syndrome (FXTAS) (Sci Adv. 2021). Here, we introduce the possibility
that G4RNA is a key pathogen on the phase transition of α-Syn that cause sporadic synucleinopathy. Purified
α-Syn protein preferentially bound to guanine-rich RNA sequences by RNA Bind-n-Seq using a pool ofrandom RNA oligonucleotides in vitro. We confirmed that purified α-Syn protein binds to G4 structure
formed RNA specifically, but not other structures in EMSA. In addition, purified α-Syn protein underwent
liquid-liquid phase separation, and the addition of G4RNA promoted the liquid-to-solid phase transition
under molecular crowding. In mouse primary neurons, G4RNA aggregation was immediately observed undercellular stress conditions, thereafter co-aggregation of α-Syn with G4RNA was occurred. We confirmed that
artificial assembly of G4RNA using an optogenetic approach initiated α-Syn aggregation, thereby elicits
neuronal dysfunction in mouse primary neurons. These results suggest that G4RNA assembly evoked byvarious cellular stress trigger to develop aggregation of α-Syn, which may be a cellular mechanism
underlying onset of sporadic synucleinopathy.

56

56

International Symposium

57

58

Ｉ－０１

The potential role of multi-drug resistance protein ABCB1 in tumor
suppression in bats
Itahana, Koji

Division, Cancer & Stem Cell Biology Program, Duke-NUS Medical School, Singapore

Most cancer research utilizes cancer-prone, short-lived models such as mice and rats. However, emerging
evidence suggests that long-lived mammals such as elephants, whales, and naked mole rats have uniqueanticancer strategies that could potentially inform new treatment strategies in humans. Although longevity is
positively correlated with body size in general, bats are unique since bats live much longer (up to 40 years)
than other mammals of equivalent size. Bats are also known to have extremely low cancer incidence.
However, the underlying unique anti-cancer mechanisms of bats remain elusive. We discovered that batderived cells exhibit enhanced DNA damage resistance to toxic chemical exposure compared to human andmouse cells. We found that bat cells accumulate less chemicals than human and mouse cells and that efficient
drug efflux mediated by the ABC transporter, ABCB1, underlies this improved response to genotoxic
reagents. Increased drug efflux and high expression of ABCB1 are conserved across multiple bat species,
while ABCB1 is expressed in only limited tissues in humans and mice. As the accumulation of DNA damage
is one of the primary causes of cancer and senescence, we propose that genotoxic efflux by ABCB1 is apotential mechanism that contributes to the low cancer incidence and longevity in bats. In this talk, I would
like to introduce the novel role of ABCB1 in tumor suppression in bats and our current research on
developing potential novel treatment strategies in humans using bat ABCB1 as a tool.

57

57

References(Related articles)

Koh J, Itahana Y, Mendenhall I, Low D, Soh EXY, Guo AK, Chionh YT, Wang LF, Itahana K (2019) ABCB1
protects bat cells from DNA damage induced by genotoxic compounds. Nat Commun. 0:1:2820.

(Others)
1.

Lee Y, Itahana Y, Ong CC, Itahana K (2022) Redox-dependent AMPK inactivation disrupts metabolic
adaptation to glucose starvation in xCT-overexpressing cancer cells J Cell Sci. jcs.259090.
2. Guo AK, Itahana Y, Seshachalam VP, Chow HY, Ghosh S, Itahana K (2021) Mutant TP53 interacts with
BCAR1 to contribute to cancer cell invasion Br J Cancer. 124:1:299-312.
3. Alagu J, Itahana Y, Sim F, Chao SH, Bi X, Itahana K (2018) Tumor suppressor p14ARF enhances IFNγactivated immune response by inhibiting PIAS1 via SUMOylation. J Immunol. 201:2:451-464.
4. Lee HY, Itahana Y, Schuechner S, Ogris E, Virshup DM, Itahana K (2018) Ca2+-dependent demethylationof phosphatase PP2Ac promotes glucose deprivation-induced cell death independently of inhibiting
glycolysis. Sci Signal. 11:512: pii: eaam7893.
5. Itahana Y, Zhang J, Göke J, Vardy LA, Han R, Iwamoto K, Engin C, Robson P, Pouladi MA, Colman A,
Itahana K (2016) Histone modifications and p53 binding poise the p21 promoter for activation in humanembryonic stem cells. Sci Rep. 6: 28112.
6. Yeo SY, Itahana Y, Guo AK, Han R, Iwamoto K, Nguyen TH, Bao Y, Kleiber K, Wu YJ, Bay BH,
Voorhoeve PM, Itahana K (2016) Transglutaminase 2 contributes to a TP53-induced autophagy program
to prevent oncogenic transformation. eLife 5:e07101.
7. Itahana Y, Han R, Barbier S, Lei Z, Rozen S, Itahana K (2015) The uric acid transporter SLC2A9 is adirect target gene of the tumor suppressor p53 contributing to antioxidant defense. Oncogene 34:14:1799810.
8. Neo SH, Itahana Y, Alagu J, Kitagawa M, Guo A, Lee SH, Tang K, Itahana K (2015) TRIM28 is an E3
ligase for ARF-mediated NPM1/B23 SUMOylation that represses centrosome amplification. Mol Cell
Biol. 35:16:2851-63.
9. Itahana K and Zhang Y (2008) Mitochondrial p32 is a critical mediator of ARF-induced apoptosis. Cancer
Cell 13:6:542-553 (featured article).
10. Itahana K, Mao H, Jin A, Itahana Y, Clegg HV, Lindström MS, Bhat K, Godfrey VL, Evan GI, Zhang Y(2007) Targeted inactivation of Mdm2 RING finger E3 ubiquitin ligase activity in the mouse reveals
mechanistic insights into p53 regulation. Cancer Cell 12:4:355-366 (cover article).
11. Itahana K, Bhat KP, Jin A, Itahana Y, Hawke D, Kobayashi R, Zhang Y (2003) Tumor suppressor ARF
degrades B23, a nucleolar protein involved in ribosome biogenesis and cell proliferation. Mol Cell.
12:5:1151-1164.

CV

Dr. Itahana obtained B.S. and Ph.D. degrees at the Faculty of Science, Kyoto University in Japan and received
postdoctoral training at the laboratory of Dr. Judith Campisi, Lawrence Berkeley National Laboratory to study
the role of p53 in cellular senescence (Itahana et al., JBC 2002 and MCB 2003). Subsequently, he joined thelaboratory of Dr. Yanping Zhang, University of North Carolina at Chapel Hill to study the regulation of
MDM2 degradation in vivo (Itahana et al., Cancer Cell 2007) and the novel functions of ARF (Itahana et al.,Mol Cell 2003 and Cancer Cell 2008). After he started his own laboratory at Cancer & Stem Cell Biology
Program, Duke-NUS Medical School Singapore in 2009, he continued to focus on tumor suppression and the
ARF-MDM2-p53 pathway. Recent mouse models challenged the long-held view of canonical functions of the
ARF-MDM2-p53 pathway such as cell cycle arrest, apoptosis, and senescence and highlighted the importanceof non-canonical, diverse roles of this pathway. Therefore, his laboratory has focused on uncovering novel
non-canonical functions of this pathway by searching for new downstream targets of p53 and new binding
proteins of ARF and p53. He demonstrated novel roles of ARF in preventing genomic instability caused bycentrosome amplification (MCB 2015) and activating immune response by inhibiting PIAS1 via
SUMOylation (J Immunol 2018). He also demonstrated the first beneficial link between p53 and uric acid foran antioxidant function to prevent cancer (Oncogene 2015) and identified that the p53-TGM2 pathway is a
critical barrier to prevent oncogenic transformation by promoting autophagy (eLife 2016). He revealed theunique epigenetic strategies in human embryonic stem cells (hESCs) that control p53 target gene expression
for maintaining homeostasis and genomic stability in ESCs (Sci Rep 2016). He also revealed new roles for
p53 in response to histone methylation inhibitor (Clin Cancer Res 2012) and to irradiation in quiescent cells(BBRC 2015) and the role of BCAR1 in mutant p53-mediated invasion (Br J Cancer 2020). His other ongoing
research projects include the role of PP2A (Sci Signal 2018) and AMPK (J Cell Sci 2022) in cancer
metabolism and the role of ABCB1 in tumor suppression and longevity in bats (Nat Commun 2019). He iscurrently an Associate Professor at Duke-NUS Medical School, and he continues to work on the noncanonical functions of ARF and p53, tumor suppression in bats, cancer metabolism, and aging.

58

58

Ｙ－０４

The membrane-linked adaptor FRS2b fashions a cytokine-richmicroenvironment that promotes breast cancer carcinogenesis
Yasuto Takeuchi, Tatsunori Nishimura, Mengjiao Li, Yuming Wang, Noriko Gotoh

Cancer Research Institute of Kanazawa University, Division of Cancer Cell Biology, Japan

Although it is held that pro-inflammatory changes precede the onset of breast cancer, the underlying
mechanisms remain obscure. Here, we demonstrate that FRS2β, an adaptor protein expressed in a small
subset of mammary epithelial cells, triggers the pro-inflammatory changes that induce stroma in precancerous
mammary tissues and is responsible for the disease onset. FRS2β-deficiency in a mouse mammary tumor
virus (MMTV)-ErbB2 genetic background markedly attenuated mammary tumorigenesis. FRS2β (+/+)
tumors contained ample stroma, on the other hand, very little stroma was observed in FRS2β (-/-) tumors,suggesting that FRS2β is required for the formation of tumor stroma. Importantly, tumor cells derived from
MMTV-ErbB2 mice failed to form tumors when they were grafted in the FRS2β-deficient precancerous
mammary tissues, indicating that FRS2β plays essential role in mammary tumorigenesis.
We also demonstrated that co-localization of FRS2β and the NEMO subunit of the IkB kinase (IKK) complex
in early endosomes led to activation of nuclear factor-kB (NF-kB), a master regulator of inflammation.
Moreover, inhibition of the activities of the NF-kB-induced cytokines, CXC chemokine ligand (CXCL)12
and insulin-like growth factor (IGF)1, abrogated tumorigenesis, indicating that the production of IGF1 andCXCL12 in premalignant mammary tissues creates a cytokine-enriched microenvironment that is necessary
for mammary tumorigenesis.
Human breast cancer tissues that express higher levels of FRS2β contain more stroma. Furthermore, patientswith higher expression levels of FRS2β in breast cancer tissues had poorer prognosis. In conclusion, theelucidation of the FRS2β-NF-kB axis uncovers the unknown molecular link between the pro-inflammatorychanges and the onset of breast cancer.

59

59

References(Related articles)

Y. Takeuchi†, N. Kimura†, T. Murayama†, Y. Machida, D. Iejima, T. Nishimura, M. Terashima, Y. Wang, M.
Li, R. Sakamoto, M. Yamamoto, N. Itano, Y. Inoue, M. Ito, N. Yoshida, J. Inoue, K. Akashi, H. Saya, K.
Fujita, M. Kuroda, I. Kitabayashi, D. Voon, T. Suzuki, A. Tojo and N. Gotoh. The membrane-linked adaptor
FRS2b fashions a cytokine-rich inflammatory microenvironment that promotes breast cancer carcinogenesis.
†
Equally contribution. Proc Natl Acad Sci USA., 2021 Oct 26;118(43):e2103658118.

CV

Yasuto Takeuchi, D.D.S, Ph.D.
Dec. 2019 – Present: Assistant Professor, Division of Cancer Cell Biology, Cancer Research Institute ofKanazawa University

60

60

Ｉ－０２

Predicting primary resistance to cancer targeted therapies at diagnosis: lessons
from chronic myeloid leukemia
S. Tiong Ong
Cancer & Stem Cell Biology Signature Research Programme,Duke-NUS Medical School, Singapore

Primary resistance to targeted therapies is a significant barrier to achieving optimal responses in a broad rangeof human cancers with druggable oncogenes. This includes chronic myeloid leukemia, (CML) where
pharmacologic targeting of the BCR-ABL1 kinase with tyrosine kinase inhibitors (TKI) has proved immensely
successful. However, even in CML, the promise of targeted therapies has not been fully realized sinceclinical responses to first-line TKIs remain highly heterogeneous with 10−15% of patients at either end of the
response spectrum enjoying either major molecular responses (MMR) or progressing to blast crisis (BC).1, 2
To date, the factors contributing to CML response heterogeneity and primary TKI resistance remain largely
unknown, and cannot be fully accounted for by somatic mutations in the BCR-ABL1 kinase domain that are acommon cause of secondary TKI resistance, i.e. following an initial clinical response.3 In order to identify
bone marrow (BM) cell types contributing to response heterogeneity, we performed single cell RNAsequencing of pretreatment chronic phase (CP) BM mononuclear cells (MNC) samples from patients with:
optimal responses (MMR) to first-line imatinib, sub-optimal responses to imatinib but favorable responses tosecond- or third-line TKIs; and pan-TKI resistance with eventual BC progression. We generated a single cellatlas from 163,146 BM MNCs, and comparing across prognostic groups, obtained a transcriptional and cellular
landscape of progressive TKI resistance. Using our atlas, and employing machine-learning, we identified
leukemia stem cell (LSC) and NK cell gene expression profiles predicting imatinib response with >80%
accuracy, as well as eight statistically significant features in pretreatment BM MNCs which correlated witheither sensitivity (MMR) or extreme resistance to imatinib (eventual BC transformation). We will discuss: 1.
Ongoing work to understand how differential transcriptional programs within CML LSCs and NK cells
functionally contribute to primary TKI response heterogeneity; 2. Development of antibody-based panels to
predict TKI responses at the time of diagnosis; and 3. Novel hypotheses arising from our observations.

61

References
1.
Braun TP, Eide CA, Druker BJ. Response and Resistance to BCR-ABL1-Targeted Therapies. Cancer
Cell 2020 Apr 13; 37(4): 530-542. PMC7722523.
2.
Holyoake TL, Vetrie D. The chronic myeloid leukemia stem cell: stemming the tide of persistence.
Blood 2017 Mar 23; 129(12): 1595-1606.
3.
Krishnan V, Kim DDH, Hughes TP, Branford S, Ong ST. Integrating genetic and epigenetic factors in
CML risk assessment: toward gene expression-based biomarkers. Haematologica 2021 Oct 7; 107(2): 358370.
4.
Takeuchi S, Hase T, Shimizu S, et al. Phase I study of vorinostat with gefitinib in BIM deletion
polymorphism/epidermal growth factor receptor mutation double-positive lung cancer. Cancer Sci 2020 Feb;
111(2): 561-570. PMC7004511.
5.
Tanimoto A, Takeuchi S, Arai S, Fukuda K, Yamada T, Roca X, Ong ST, Yano S. Histone Deacetylase
3 Inhibition Overcomes BIM Deletion Polymorphism-Mediated Osimertinib Resistance in EGFR-Mutant
Lung Cancer. Clin Cancer Res 2017 Jun 15; 23(12): 3139-3149.
6.
Ng KP, Hillmer AM, Chuah CT, et al. A common BIM deletion polymorphism mediates intrinsic
resistance and inferior responses to tyrosine kinase inhibitors in cancer. Nat Med 2012 Apr; 18(4): 521-528.
7.
Sinnakannu JR, Lee KL, Cheng S, Li J, Yu M, Tan SP, Ong CCH, Li H, Than H, Anczukow-Camarda
O, Krainer AR, Roca X, Rozen SG, Iqbal J, Yang H, Chuah C, Ong ST. SRSF1 mediates cytokine-induced
impaired imatinib sensitivity in chronic myeloid leukemia. Leukemia 2020 Jul; 34(7): 1787-1798.
PMC7682023.
8.
Ng KP, Manjeri A, Lee KL, Huang W, Tan SY, Chuah CT, Poellinger L, Ong ST. Physiologic hypoxia
promotes maintenance of CML stem cells despite effective BCR-ABL1 inhibition. Blood 2014 May 22;123(21): 3316-3326.
9.
Ko TK, Javed A, Lee KL, et al. An integrative model of pathway convergence in geneticallyheterogeneous blast crisis chronic myeloid leukemia. Blood 2020 Jun 25; 135(26): 2337-2353.
10.
Lim S, Saw TY, Zhang M, Janes MR, Nacro K, Hill J, Lim AQ, Chang CT, Fruman DA, Rizzieri DA,
Tan SY, Fan H, Chuah CT, Ong ST. Targeting of the MNK-eIF4E axis in blast crisis chronic myeloid
leukemia inhibits leukemia stem cell function. Proc Natl Acad Sci U S A 2013 Jun 18; 110(25): E2298-2307.
PMC3690864.

CV
I completed my undergraduate and medical training at Cambridge University in the UK, andsubsequently trained as a hematologist/oncologist at the University of Chicago, where I also began mylaboratory research with Michelle Le Beau. My independent research career started in 2001 when I
began my studies in cancer drug resistance. While our studies focus on chronic myeloid leukemia(CML), the implications of our work have been extended to other human cancers, including EGFRmutated non-small cell lung cancer,4-6 and more recently, glioblastoma. In 2007, I joined Duke-NUS, a
collaboration between Duke University and the National University of Singapore to help build the first
American-style post-baccalaureate medical school in Asia.
My research career began with foundational work on the role of dysregulated mRNA translation inresistance to imatinib (Gleevec). In recent years my team has made significant contributions to theunderstanding of BCR-ABL1-independent factors in CML patient response heterogeneity, including
microenvironmental factors (extracellular cytokine signaling via SRSF1, physiologic hypoxia),7, 8
germline polymorphisms (the BIM deletion polymorphism6), and more recently, BCR-ABL1independent epigenetic reprogramming of leukemia stem cells.9 Several of our discoveries have led toearly phase clinical trials conducted by myself or international collaborators, including leading Japanese
researchers,4, 5 and drug development efforts uncovered by our discovery-driven projects.10 A common
thread that runs through our work is the use of cutting-edge technologies to interrogate primary patient
samples, efforts undertaken in close collaboration with leading computational biologists, followed bybench-based experiments to dissect, confirm, and therapeutically overcome novel mechanisms of drug
resistance. Our work has been published in leading scientific journals including Nature Medicine,
Proceedings of the National Academy of Sciences (USA), Blood, Cancer Research, Leukemia, andOncogene. We have also received current and past funding from the US NIH, National Medical
Research Council Singapore, and the Leukemia & Lymphoma Society. Our past and current service
activities include membership of the Singapore Medical Council, NIH and ASH Global Research Awardreview panels, and Directorship of the Duke-NUS MD-PhD Programme. Our research teams have also
received international awards and recognition, including career development awards from the American
Society of Clinical Oncology (STO), European Society of Hematology International John Goldman
Award (Vaidehi Krishnan), and election to the American Society of Clinical Investigation (STO).

62

62

Ｓ－０３

Identification and functional analysis of senescent cells in tumor
microenvironment
Johmura1, Yoshikazu; Nakanishi2, Makoto

1

Division of Cancer and Senescence Biology, Cancer Research Institute Kanazawa University, Japan

2

Division of Cancer Cell Biology, Institute of Medical Science University of Tokyo, Japan

Cellular senescence is triggered by diverse genotoxic stimuli. An important hallmark of senescent cells is its
inability to proliferate in response to physiological mitotic stimuli, which can prevent tumorigenesis. Anotheris the appearance of Senescence-associated secretory phenotypes (SASP), such as robust secretion ofnumerous cytokines, which may cause deleterious effects on tissue microenvironment. Recent studies indicatethat the elimination of senescent cells accumulated in the body during aging ameliorates age-related diseases
including cancer, thus promoting the healthy lifespan (references 1 and 2). Therefore, cellular senescencelikely plays both positive and negative roles in cancer. Recently, we generated a p16Ink4a-CreERT2tdTomato mouse to analyze the in vivo characteristics of senescent cells (reference 3). Our new mouse modeland single-cell analyses suggest that in vivo senescent cells exhibited heterogeneous senescence-associatedphenotypes during aging. Currently, we are trying to apply this approach to some cancer models, and we
found that cancer-related senescent cells also are composed of several cell types and have various
characteristics. Further analysis will provide a molecular basis for context-dependent effects of cellularsenescence in cancer.

63

63

References
1. Baker DJ, Childs BG, Durik M, Wijers ME, Sieben CJ, Zhong J, Saltness RA, Jeganathan KB,
Verzosa GC, Pezeshki A, Khazaie K, Miller JD, van Deursen JM. Naturally occurring p16(Ink4a)positive cells shorten healthy lifespan.Nature. 2016 11;530(7589):184-9. doi: 10.1038/nature16932
2. Johmura Y, Yamanaka T, Omori S, Wang TW, Sugiura Y, Matsumoto M, Suzuki N, Kumamoto S,
Yamaguchi K, Hatakeyama S, Takami T, Yamaguchi R, Shimizu E, Ikeda K, Okahashi N, Mikawa R,
Suematsu M, Arita M, Sugimoto M, Nakayama KI, Furukawa Y, Imoto S, Nakanishi M. Senolysis byglutaminolysis inhibition ameliorates various age-associated disorders. Science. 2021 15;371(6526):265270. doi: 10.1126/science.abb5916.
3. Omori S, Wang TW, Johmura Y, Kanai T, Nakano Y, Kido T, Susaki EA, Nakajima T, Shichino S,
Ueha S, Ozawa M, Yokote K, Kumamoto S, Nishiyama A, Sakamoto T, Yamaguchi K, Hatakeyama S,
Shimizu E, Katayama K, Yamada Y, Yamazaki S, Iwasaki K, Miyoshi C, Funato H, Yanagisawa M,
Ueno H, Imoto S, Furukawa Y, Yoshida N, Matsushima K, Ueda HR, Miyajima A, Nakanishi M.
Generation of a p16 Reporter Mouse and Its Use to Characterize and Target p16 high Cells In Vivo. Cell
Metab. 2020 3;32(5):814-828.e6. doi: 10.1016/j.cmet.2020.09.006.

CV[Education]
2003 - B.A.
2005 - M.A.
2008 - Ph.D.

Pharmaceutical Sciences, Nagoya City University, Japan
Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan
Graduate School of Pharmaceutical Sciences, Nagoya City University, Japan[Career]
2008-2008
2008-20112011-20162016-2022
2022-

Postdoctoral Researcher, Pharmaceutical Sciences, Nagoya City University, Japan
Postdoctoral Researcher, National Cancer Institute, NIH, USA
Assistant Professor, Department of Medical Sciences, Nagoya City University, Japan
Assistant Professor, Institute of Medical Science, University of Tokyo, Japan
Professor, Cancer Research Institute, Kanazawa University, Japan[Honors]
1. Award at 71th Meeting of The Japanese Biochemical Society, Chubu Branch, 2007
2. Fellows Award for Research Excellence in NIH, USA, 2010
3. Award for best faculty member in Nagoya City University, Japan, 2015
4. Award for best presentation at 2019 meeting for Advanced Animal Model support, Japan, 2020
[Email Address]
johmuray@staff.kanazawa-u.ac.jp

64

64

Ｉ－０３

Systems Genetics identifies WWP2 as a new target for fibrotic disease infibroblasts and macrophages
Petretto Enrico
Cardiovascular and Metabolic Disorder (CVMD) Signature Programme and Centre for Computational Biology(CCB) of Duke-National University of Singapore (NUS) Medical School, Singapore
2
Institute of Clinical Sciences of Imperial College London, London, UK
3
Institute of Molecular and Translational Cardiology of IRCCS Policlinico San Donato, Italy
4
Institute for Big Data and Artificial Intelligence in Medicine, School of Science, China Pharmaceutical
University, Nanjing, China
1

Pathological cardiac fibrosis is a final common pathology in inherited and acquired heart diseases that causes
cardiac electrical and pump failure. Here, we use systems genetics to identify a pro-fibrotic gene network in
the diseased heart and show that this network is regulated by the E3 ubiquitin ligase WWP2, specifically by
the WWP2-N terminal isoform. Importantly, the WWP2-regulated pro-fibrotic gene network is conserved
across different cardiac diseases characterized by fibrosis: human and murine dilated cardiomyopathy andrepaired tetralogy of Fallot. Transgenic mice lacking the N-terminal region of the WWP2 protein show
improved cardiac function and reduced myocardial fibrosis in response to pressure overload or myocardial
infarction. In primary cardiac fibroblasts, WWP2 positively regulates the expression of pro-fibrotic markers
and extracellular matrix genes. TGFβ1 stimulation promotes nuclear translocation of the WWP2 isoforms
containing the N-terminal region and their interaction with SMAD2. WWP2 mediates the TGFβ1-induced
nucleocytoplasmic shuttling and transcriptional activity of SMAD2.
Moreover, we show that its myeloid-specific deletion reduces cardiac fibrosis. Using the same model, weestablish the functional heterogeneity of macrophages and define an early pro-fibrogenic phase driven by
Ccl5-expressing Ly6chigh monocytes. Among other cardiac macrophage subtypes, WWP2 dysfunction
primarily affects the Ccl5-dependent infiltration and activation of Ly6chigh monocytes, which causes reduced
myofibroblast trans-differentiation. WWP2 interacts with IRF7, promoting its non-degradative
monoubiquitination, nuclear translocation and transcriptional activity, including upstream Ccl5. Thus, WWP2
plays also a role as a key regulator of IRF7-mediated Ccl5/Ly6chigh monocyte axis in heart fibrosis.

65

References(Related articles)

Schafer, S. et al. IL-11 is a crucial determinant of cardiovascular fibrosis. Nature 552, 110–115 (2017)
Moreno-Moral, A., Pesce, F., Behmoaras, J. & Petretto, E. Systems genetics as a tool to identify mastergenetic regulators in complex disease. Methods Mol. Biol. 1488, 337–362 (2017)
Petretto, E. et al. Integrated genomic approaches implicate osteoglycin (Ogn) in the regulation of left
ventricular mass. Nat. Genet. 40, 546–552 (2008)
Moreno-Moral, A., Mancini, M., D’Amati, G., Camici, P. & Petretto, E. Transcriptional network analysis forthe regulation of left ventricular hypertrophy and microvascular remodeling. J. Cardiovasc. Transl. Res. 6,
931–944 (2013)
Rotival, M. & Petretto, E. Leveraging gene co-expression networks to pinpoint the regulation of complex
traits and disease, with a focus on cardiovascular traits. Brief. Funct. Genomics 13, 66–78 (2014)
Heinig, M. et al. A trans-acting locus regulates an anti-viral expression network and type 1 diabetes risk.
Nature 467, 460–464 (2010)
Kang, H. et al. Kcnn4 is a regulator of macrophage multinucleation in bone homeostasis and inflammatory
disease. Cell Rep. 8, 1210–1224 (2014)

CV
Associate Professor Enrico Petretto is Director of the Centre for Computational Biology (CCB) and leads the
Systems Genetics Group at Duke-NUS Medical School in Singapore, which was stablished as a landmark
collaboration between two world-ranking institutions of higher education: Duke University (USA) and the
National University of Singapore (NUS). He is also Theme Leader for Bioinformatics and Systems Genetics
at the Institute of Molecular and Translational Cardiology (IMTC) for the study of Brugada Syndrome andSudden Death, Policlinico San Donato University and Research Hospital in Milan (Italy), Adjunct Professor
at the Institute of Big Data and Artificial Intelligence, China Pharmaceutical University (CPU) in Nanjing(China) and Senior Lecturer at Imperial College London.
Enrico Petretto has developed a research programme in Systems Genetics to study the genetic regulation ofcellular pathways and gene networks that drive human disease. His lab uses Systems Biology and integrative
genomics approaches to identify new drug targets for various complex disease, including cardiometabolic(e.g., cardiomyopathies, Brugada Syndrome) and fibrotic disease, diabetic nephropathy, cancer and important
brain disorders such as epilepsy and Alzheimer’s disease. The identified drug targets are then followed up
experimentally in Petretto’s lab using several cellular and pre-clinical disease models. Enrico Petretto has
published more than 130 papers in top tier journals such as Nature, Nature Genetics, Nature
Communications, Nature Neuroscience, Nature Immunology, Cell, Circulation, European Heart Journal, etc.,with more than 9,000 citations to date (see Google Scholar).
Selected papers include: Integrated genomic approaches implicate osteoglycin (Ogn) in the regulation of left
ventricular mass (Nature Genetics 2008); A trans-acting locus regulates an anti-viral expression network andtype 1 diabetes risk (Nature 2010); Systems-genetics identifies Sestrin 3 as a regulator of a proconvulsant
gene network in human epileptic hippocampus (Nature Communications 2015); Systems genetics identifies a
convergent gene network for cognition and neurodevelopmental disease (Nature Neuroscience 2016); A
predictive computational framework for direct reprogramming between human cell types (Nature Genetics
2016); IL11 is a critical determinant of cardiovascular fibrosis (Nature 2017); A systems-level framework for
drug discovery identifies Csf1R as an anti-epileptic drug target (Nature Communications 2018); Changes inmacrophage transcriptome associate with systemic sclerosis and mediate GSDMA contribution to disease risk
(Annals of the Rheumatic Diseases 2018); WWP2 regulates pathological cardiac fibrosis by modulating
SMAD2 signaling (Nature Communications 2019); A single-cell atlas of entorhinal cortex from individuals
with Alzheimer's disease reveals cell-type-specific gene expression regulation (Nature Neuroscience 2019);
Whole-genome sequencing of Finnish type 1 diabetic siblings discordant for kidney disease reveals DNA
variants associated with diabetic nephropathy (Journal of the American Society of Nephrology 2020); Wntregulated lncRNA discovery enhanced by in vivo identification and CRISPRi functional validation (Genome
Medicine 2020); Brugada Syndrome Genetics is Associated with Phenotype Severity (European Heart
Journal 2020); EpiMogrify models H3K4me3 data to identify signaling molecules that improve cell fatecontrol and maintenance (Cell Systems 2020); Transcriptional signature in microglia associated with Aβplaque phagocytosis (Nature Communications 2021).
For more details on Enrico Petretto’s group and research activities please visit enricopetretto.com.

66

66

The 17th International Symposium of the Institute Network for
Biomedical Sciences International Symposium on Tumor Biology in Kanazawa 2022
2022 CRI & DUKE-NUS Joint Symposium

Fundamental
Biological
Principles and Cancer
Cancer Research Institute of Kanazawa University

October 13-14, 2022
Kanazawa University, Japan