The 11th Nikko
International
Symposium
2014
Inflammation, Cancer and Microenvironment
Opening Remark

Ryozo Nagai
President,Jichi Medical University

Date: Thursday October 30, 2014
Venue: JMU Information and In-service Training Center

Introduction

Yusuke Furukawa
Director, Center for Molecular Medicine, Jichi Medical University

Speakers

Kazuhiro Nakayama, Jichi Medical University
Asuka Sakata, Jichi Medical University
Yasushi Saga, Jichi Medical University
Jiro Kikuchi, Jichi Medical University
Yoshiyuki Morishita, Jichi Medical University
Takanori Komada, Jichi Medical University
Yoshiro Maru, Tokyo Women's Medical University
Kensuke Miyake, Institute of Medical Science, University of Tokyo
Osamu Nureki, University of Tokyo
Orson Moe, University of Texas
James D. Griffin, Harvard Medical School
Closing Remark

Seiji Minota
Vice President, Jichi Medical University

Jichi Medical University

Abstract Book

Tel:0285-58-7550 Fax:0285-40-8303
E-mail:shien@jichi.ac.jp

Sponsored by Center for Molecular Medicine, Jichi Medical University

The 11th Nikko International Symposium 2014
Inflammation, Cancer and Microenvironment
Date: Thursday October 30, 2014
Venue: JMU Information and In-service Training Center
10:00~10:05 Opening Remark Ryozo Nagai, Jichi Medical University
10:05~10:10 Introduction

Yusuke Furukawa, Jichi Medical University

First Session: Therapeutic targeting of microenvironment
Chairpersons: Satoshi Nishimura, Kiyoshi Kawakami
10:10~10:25 Kazuhiro Nakayama, Jichi Medical University
10:25~10:40 Asuka Sakata, Jichi Medical University
Chairpersons: Tatsushi Onaka, Hiroaki Mizukami
10:40~10:55 Yasushi Saga, Jichi Medical University
10:55~11:10 Jiro Kikuchi, Jichi Medical University
Chairpersons: Yutaka Hanazono, Hironori Yamamoto
11:10~11:55 Yoshiro Maru, Tokyo Women's Medical University
12:00~13:00 Lunch Time
Second Session: Inflammation and disease
Chairpersons: Masafumi Takahashi, Makoto Kuro-o
13:00~13:15 Yoshiyuki Morishita, Jichi Medical University
13:15~14:00 Kensuke Miyake, Institute of Medical Science, University of Tokyo
14:00~14:15 Takanori Komada, Jichi Medical University
14:15~15:00 Orson Moe, University of Texas
15:00~15:15 Photo Session and Coffee BreakThird Session: Cancer treatment based on molecular and structural biology
Chairpersons: Naoya Shibayama, Yusuke Furukawa
15:15~16:00 Osamu Nureki, University of Tokyo
16:00~16:45 James D. Griffin, Harvard Medical School
16:45~16:50 Closing Remark
17:00~

Seiji Minota, Jichi Medical University

Buffet-style dinner party

1

LIST OF SPEAKERS
Invited Speakers
Orson Moe
James D. Griffin
Yoshiro Maru
Kensuke Miyake
Osamu Nureki

Speakers of Jichi Medical University
Kazuhiro Nakayama
Asuka Sakata
Yasushi Saga
Jiro Kikuchi
Yoshiyuki Morishita
Takanori Komada

3

CURRICULA VITAE&
ABSTRACTS

5CURRICULUM VITAE
Kazuhiro Nakayama. Ph.D.
Assistant Professor
Division of Human Genetics, Center for Molecular Medicine,
Jichi Medical University
Tel: (+81) 285-58-7341
Fax: (+81)285-44-4902
E-mail: nakayama@jichi.ac.jp
Professional Experience
2004-2005
JSPS Postdoctoral Fellow, The University of Tokyo
21Stt Century COE Postdoctoral Fellow, The University of Tokyo
2005-2006
2006-2013
Research Associate, Jichi Medical University
2013-Present Assistant Professor, Jichi Medical University
Education
1995-1999
1999-2001
2001-2004

B. Eng., Tokai University
M. Sci., Graduate School of Science, The University of Tokyo
Ph.D., Graduate School of Science, The University of Tokyo

Recent Publications
1. Ishizuka Y, NakayamayK, et al. TRIB1 downregulates hepatic lipogenesis and glycolysis via
multiple molecular interactions. J Mol Endocrinol. 52:145-158. 2014
2. NakayamayK et al. Positive natural selection of TRIB2, a novel gene that influences visceralfat accumulation, in East Asia. Hum Genet. 132:201-217. 2013y
K et al. A functional SNP upstream of the beta-2 adrener3. Naka I, Hikami K, Nakayama
gic receptor gene (ADRB2) is associated with obesity in Oceanic populations. Int J Obes 37
1204-1210, 2013
4. NakayamayK et al. Seasonal effects of UCP1 gene polymorphism on visceral fat accumulation in Japanese adults. PLOS ONE 8:e74720, 2013
5. NakayamayK et al. High prevalence of an anti-hypertriglyceridemic variant of the MLXIPL
gene in Central Asia. J Hum Genet. 56:828-833. 20116

Genetic variation of the Tribbles family and metabolic syndrome
Kazuhiro Nakayama1y, Hiroshi Miyashita2, Yasuo Kagawa3, and Sadahiko Iwamoto1
1Division of Human Genetics, Jichi Medical University, 2Jichi Medical University HealthCare Center, 3Kagawa Nutrition University
Tribbles pseudokinase family (TRIB) consists of three paralogous proteins (TRIB1~3)
that participated in the regulation of various intra-cellular signaling pathways and transcription.
Our large scale genetic association studies showed that single nucleotide polymorphism (SNP)s near the gene encoding TRIB1 strongly influenced plasma triglycerides, low density lipoprotein-cholesterol levels, and susceptibility to non-alcohol fatty liver disease in Japanese. Further
molecular genetic analyses revealed that TRIB1 regulates hepatic lipogenesis and glycogenesis
by multiple molecular interactions. We also discovered that a functional SNP in 3’ untranslated
region of the TRIB2 gene was associated with visceral fat area in Japanese. Further studies of
TRIBs and the interacting partner molecules will shed light on the mechanisms of lipid metabolism dysregulation in multiple organs/tissues.
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7

CURRICULUM VITAE
Asuka Sakata, M.D. , Ph.D.
Research Scientist
Research Division of Cell and Molecular Medicine, Center for Molecular Medicine, Jichi
Medical University
3111-1 Yakushiji, Shimotsuke, Tochigi 329-0498, JAPAN
Tel: +81-285-58-7397
Fax: +81-285-44-7817
E-mail: asusaka@jichi.ac.jp
Professional Experience
2006-2008
Resident in Medicine, Jichi Medical University Hospital
2008-2009
Clinical Fellow in Medicine, Jichi Medical University Hospital
2014-presenet Research Scientist, Jichi Medical University
Education
2000-2006
2009-2014

M.D., Akita University
Ph.D., Jichi Medical University

Recent Publications
1. Sakata A, Ohmori T, Nishimura S, Suzuki H, Madoiwa S, Mimuro J, Kario K, SakataY.
J 2014 Jan
Paxillin is an intrinsic negative regulator of platelet activation in mice. Thromb J.
2;12(1):1. [Epub ahead of print]
2. Koyama K, Madoiwa S, Tanaka S, Koinuma T, Wada M, Sakata A, Ohmori T,Mimuro J,
Nunomiya S, Sakata Y. Evaluation of hemostatic biomarker abnormalities that precede
platelet count decline in critically ill patients with sepsis.
J Crit Care. 2013 Oct;28(5):556-63.
3. Mimuro J, Mizukami H, Hishikawa S, Ikemoto T, Ishiwata A, Sakata A, Ohmori T,
Madoiwa S, Ono F, Ozawa K, Sakata Y. Minimizing the inhibitory effect of neutralizing
antibody for efficient gene expression in the liver with adeno-associated virus 8 vectors. Mol
Ther. 2013 Feb;21(2):318-23.
4. Kashiwakura Y, Mimuro J, Onishi A, Iwamoto M, Madoiwa S, Fuchimoto D, Suzuki S,
Suzuki M, Sembon S, Ishiwata A, Yasumoto A, Sakata A, Ohmori T, Hashimoto M, Yazaki
S, Sakata Y. Porcine model of hemophilia A. PLoS One. 2012;7(11):e49450.
5. Kashiwakura Y, Ohmori T, Mimuro J, Yasumoto A, Ishiwata A, Sakata A, Madoiwa S,
Inoue M, Hasegawa M, Ozawa K, Sakata Y. Intra-articular injection of mesenchymal stem
cells expressing coagulation factor ameliorates hemophilic arthropathy in factor VIII-deficientmice. J Thromb Haemost. 2012 Sep;10(9):1802-13.

8

Establishment of new evaluation methods for treatments of thrombosis with in vivo
animal models.
Asuka Sakata, Tsukasa Ohmori, Satoshi Nishimura
Research Division of Cell and Molecular Medicine, Center for Molecular Medicine; Jichi
Medical University, Japan
Thrombosis is the formation of a blood clot inside a blood vessel, obstructing theflow of blood through the circulatory system. Experimental models of thrombosis that mimic
human vascular disease are essential to unravel the complex factors involved in pathologic
thrombus formation. All reported ones, however, have both advantages and limitations. Using a
confocal laser-scanning microscope, we developed “live thrombus imaging” technique that enables us to observe real time thrombus formation in detail. In our technique, thrombus is triggered by inducing reactive oxygen species production with a laser or by direct laser injury ofendothelial cells. This is the first time thrombi have been visualized with sufficient resolution
to identify individual platelets. We also use venous thrombus models to study factors involvedin thrombus growth, stabilization, lysis, and effectiveness of anticoagulants. Venous thrombus
is triggered by ligation of inferior vena cava. Using this venous thrombus model, we clarify
that efficacy of NOACs for treating venous thrombosis is provided by secondary fibrinolysis.
We now plan to image fibrinolysis using confocal laser-scanning microscope.
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9

CURRICULUM VITAE
Yasushi Saga, M.D., Ph.D.
Associate Professor
Department of Obstetrics and Gynecology; Division of Genetics Therapeutics, Center for
Molecular Medicine, Jichi Medical University
3311-1 Yakushiji, Shimotsuke-shi, Tochigi, 329-0498, Japan
Tel: (+81) 285-58-7376
Fax:(+81) 285-44-8505
E-mail: saga@jichi.ac.jp
Professional Experience
1993-1995 Resident in Medicine, Jichi Medical University Hospital
1995-1997 Clinical Fellow in Medicine, Ani town’s Hospital
2001-2003 Clinical Fellow in Medicine, Public Yonaizawa Hospital
2003-2005 Clinical Associate, Jichi Medical University Hospital
2005-2006 Research Associate, Jichi Medical University
2006-2007 Clinical Fellow in Medicine, International University of Health and Welfare
Hospital
2007-2008 Assistant Professor, International University of Health and Welfare
2008-2013 Assistant Professor, Jichi Medical University
2013-present Associate Professor, Jichi Medical University
Education
1987-1993 M.D., Jichi Medical University
1997-2001 Ph.D., Jichi Medical University
Recent Publications
1. Machida, S., Takei, Y., Yoshida, C., Takahashi, Y., Koyanagi, T., Sato, N., Taneichi,
A., Saga, Y., Fujiwara, H., Suzuki, M.: Radiation therapy for chemotherapy-resistant recurrent
epithelial ovarian cancer. Oncology. 86(4):232-238, 2014
2. Takahashi, K., Mizukami, H., Saga, Y., Takei, Y., Urabe, M., Kume, A., Machida, S.,
Fujiwara, H., Suzuki, M., Ozawa, K.: Suppression of lymph node and lung metastases ofendometrial cancer by muscle-mediated expression of soluble vascular endothelial growth
factor receptor-3. Cancer Sci. 104(8):1107-1111, 2013.
3. Fujiwara, H., Takei, Y., Ishikawa, Y., Saga, Y., Machida, S., Taneichi, A., Suzuki, M.:
Community-based interventions to improve HPV vaccination coverage among 13- to 15-yearold females: measures implemented by local governments in Japan. PLoS One. 8(12):e84126,
2013.
4. Koyanagi, T., Suzuki, Y., Saga, Y., Machida, S., Takei, Y., Fujiwara, H., Suzuki, M., Sato,Y.: In vivo delivery of siRNA targeting vasohibin-2 decreases tumor angiogenesis andsuppresses tumor growth in ovarian cancer. Cancer Sci. 104(12):1705-1710, 2013.
5. Uchibori, R., Tsukahara, T., Mizuguchi, H., Saga, Y., Urabe, M., Mizukami, H., Kume, A.,
Ozawa, K.: NF-κB activity regulates mesenchymal stem cell accumulation at tumor sites.
Cancer Res. 73(1):364-372, 2013.

10

Robust treatment for cervical cancer targeting human papillomavirus E6/E7
Yasushi Saga
g 1,2, Naoto Sato1,2, Hiroaki Mizukami2, Ryosuke Uchibori2, Tomonori Tsukahara2, Masashi Urabe2, Akihiro Kume2, Mitsuaki Suzuki1, Keiya Ozawa2,3
1Department of Obstetrics and Gynecology; 2Division of Genetics Therapeutics, Center for
Molecular Medicine, Jichi Medical University, Shimotsuke-shi, Tochigi, Japan
3Present address: Director, IMSUT Hospital, The Institute of Medical Science, The Universityof Tokyo, Minato-ku, Tokyo
Human papillomavirus (HPV) is the major causative agent of cervical cancer. The
HPV oncoproteins E6 and E7 induce carcinogenesis by inactivating host tumor suppressor
genes. Therefore, stable expression of specific inhibitors of E6 and E7 in cancer cells could
provide effective treatment without affecting normal tissue. Here, we propose a novel therapeutic approach that uses an adeno-associated virus (AAV) vector encoding a short-hairpin (sh)RNA to target E6 and E7 (shE6E7) of HPV type 16 (HPV-16). Three different HPV-16-positive
cervical cancer cell lines (BOKU, SiHa, and SKG-IIIa) were tested for gene transfer efficiency
using AAV vectors of different serotypes. In all three cervical cancer cell lines, the highest
gene transfer efficiency was obtained using AAV2 vectors. The proportions of GFP-positive
cells at the dose of 1 × 105 vg/cell were 87.3%, 98.3%, and 87.9%, respectively. Transductionof cervical cancer cells by AAV2-shE6E7, an shRNA-encoding AAV vector that targets both
E6 and E7, resulted in apoptosis, G1 arrest, and cell growth inhibition. In transduced cells, E6,
E7, and p16 expression was reduced, whereas p53, p21, and pRb expression was enhanced.
Next, AAV2-shE6E7 was directly injected into cervical cancer cell-derived subcutaneous tumors in mice. Tumor growth was markedly inhibited by a single AAV2-shE6E7 administration,and most tumors showed complete regression without any adverse effects. We are currently extending this strategy to target other HPVs, especially, type 18. These results suggest the utilityof AAV2-shE6E7 as a novel therapeutic approach for cervical cancer.
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11

CURRICULUM VITAE
Jiro Kikuchi, Ph.D.
Associate Professor
Division of Stem Cell Regulation, Center for Molecular Medicine
Jichi Medical University
3311-1 Yakushiji, Shimotsuke-City, Tochigi 329-0498, JAPAN
Tel: (+81) 285-58-7400
Fax: (+81) 285-44-7500
E-mail: kiku-j@jichi.ac.jp
Professional Experience
2001-2002
Research Fellow, Division of Cell and Molecular Medicine, Center for
Molecular Medicine, Jichi Medical University, Japan
2002-2005
Research Associate, Cell Regulation Analysis Team, Research Center for
Glycoscience, National Institute of Advanced Industrial Science and Technology(AIST), Japan
2005-2007
Research Associate, Division of Stem Cell Regulation, Center for
Molecular Medicine, Jichi Medical University, Japan
2007-2011
Lecturer, Division of Stem Cell Regulation, Center for
Molecular Medicine, Jichi Medical University, Japan
2011-present Associate Professor, Division of Stem Cell Regulation, Center for
Molecular Medicine, Jichi Medical University, Japan
Education
1985-1989
1993-2001

BSc., Science University of Tokyo
Ph.D., Jichi Medical School

Recent Publications
1. Koyama D, Kikuchi J, Hiraoka N, Wada T, Kurosawa H, Chiba S, and Furukawa Y.
Proteasome inhibitors exert cytotoxicity and increase chemosensitivity via transcriptional
repression of Notch1 in T-cell acute lymphoblastic leukemia. Leukemia. 28, 1216-1226,
2014.
2. Kikuchi J, Koyama D, Mukai H, and Furukawa Y. Suitable drug combination withbortezomib for multiple myeloma under stroma-free conditions and in contact withfibronectin or bone marrow stromal cells.
Int J Hematol. 99, 726-736, 2014.
3. Kikuchi J, Yamada S, Koyama D, Wada T, Nobuyoshi M, Izumi T, Akutsu M, Kano Y,
and Furukawa Y. The novel orally active proteasome inhibitor K-7174 exerts anti-myeloma
activity in vitro and in vivo by down-regulating the expression of class I histone deacetylases.
J Biol Chem, 288, 25593-25602, 2013
4. Kikuchi J, Wada T, Shimizu R, Izumi T, Akutsu M, Mitsunaga K, Noborio-Hatano K,
Nobuyoshi M, Ozawa K, Kano Y, and Furukawa Y. Histone deacetylase is a critical target ofbortezomib-induced cytotoxicity in multiple myeloma.
Blood, 116, 406-417, 2010.
5. Noborio-Hatano K, Kikuchi J, Takatoku M, Shimizu R, Wada T, Ueda M, Nobuyoshi M, Oh
I, Sato K, Suzuki T, Ozaki K, Mori M, Nagai T, Muroi K, Kano Y, Furukawa Y, and Ozawa
K. Bortezomib overcomes cell adhesion-mediated drug resistance via down-regulation of
VLA-4 expression in multiple myeloma.
Oncogene, 28, 231-242, 2009.

12

Phosphorylation-mediated EZH2 Inactivation as a Principal Mechanism of Drug
Resistance in Multiple Myeloma
Jiro Kikuchi and Yusuke Furukawa
Division of Stem Cell Regulation, Center for Molecular Medicine,
Jichi Medical University, Japan
Multiple myeloma (MM) is one of the most intractable malignancies characterized
by the infiltration and growth of malignant plasma cells in the bone marrow (BM). MM cells
are much less sensitive to chemotherapeutic agents in the BM microenvironment depending on
the interaction with bone marrow stromal cells . This type of drug resistance, termed cell adhesion-mediated drug resistance (CAM-DR), is a major obstacle in achieving cure in MM patients. Although epigenetics has critical roles in various aspects of MM biology, little is known
about their specific roles in CAM-DR.In this study, we identified trimethylation of histone H3
at lysine-27 (H3K27me3) as a critical histone mark for CAM-DR in MM cells. Cell adhesion
counteracted drug-induced hypermethylation of H3K27 via inactivating phosphorylation of
EZH2, leading to sustained expression of anti-apoptotic genes including IGF-1, Bcl-2 and HIF1a. Inhibition of the IGF-1R/PI3K/Akt pathway was able to reverse CAM-DR by promoting
EZH2 dephosphorylation and H3K27 hypermethylation. Among them, the IGF-1R inhibitor
OSI-906 appears to be particularly effective to overcome CAM-DR in vitro and in refractorymurine MM models.These results suggest that phosphorylation-mediated EZH2 inactivationand subsequent H3K27 hypermethylation are crucial for CAM-DR. This is the first report ofthe epigenetic mechanism of CAM-DR involving a regulatory circuit from the membrane to
the nucleus and provides a rationale for the inclusion of kinase inhibitors counteracting EZH2
phosphorylation. The IGF-1R inhibitor OSI-906 may be translated to the clinic to improve thetreatment outcome of myeloma patients in combination with conventional anti-MM agents.
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13

CURRICULUM VITAE
Yoshiro Maru, M.D., Ph.D.
Professor and chairman
Department of Pharmacology, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan.
Tel:(+81) 3-5269-7417
Fax:(+81) 3-5269-7417
E-mail:ymaru@research.twmu.ac.jp
Professional Experience
1983-1985
Resident in Internal Medicine, Toranomon Hospital
1985-1989
Medical Staff, Third Department of Internal Medicine, The University of Tokyo
Hospital
1989-1993
Associate, Howard Hughes Medical Institute, University of California, Los Angeles
1993-1996
Assistant Professor, Department of Genetics, Institute of Medical Science, University of Tokyo
1996-2002
Associate Professor, Department of Genetics, Institute of Medical Science, University of Tokyo
2002-present Professor and Chairman, Department of Pharmacology,
Tokyo Women’s Medical University
Education
1977-1983
M.D., The University of Tokyo
1985-1989
Ph.D., The University of Tokyo
Recent Publications
1. Tomita T., Ieguchi K., Coin F., Kato Y., Kikuchi H., Oshima Y., Kurata S., and Maru Y.
ZFC3H1, a zinc finger protein, modulates IL-8 transcription by binding with Celastramycin A,
a potential immune suppressor. PLOS One, 2014, in press.
2. Ieguchi K, Tomita T, Omori T, Komatsu A, Deguchi A, Masuda J, Duffy S, Coulthard M,
Boyd A, Maru Y. ADAM12-cleaved ephrin-A1 contributes to lung metastasis. Oncogene.
33,2179-2190, 2014
3. Hiratsuka S, Ishibashi S, Tomita T, Watanabe A, Tamura SA, Murakami M, Kijima H, Miyake K, Aburatani H, Maru Y. Primary tumours modulate innate immune signalling to create
pre-metastatic vascular hyperpermeability foci. Nature communications. 4, 1853, 2013.
4. Deguchi A, Tomita T, Omori T, Komatsu A, Ohto U, Takahashi S, Tanimura N, AkashiTakamura S, Miyake K, Maru Y. Serum amyloid A3 binds MD-2 to activate p38 and NFkappaB pathways in a MyD88-dependent manner. Journal of immunology 191,1856-1864,2013.
5. Maru Y. Molecular biology of chronic myeloid leukemia. Cancer Sci. 103,1601-1610, 2012.

14

How pre-metastatic milieu is establishedYoshiro Maru Department of Pharmacology, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan
Circulating tumor cells in advanced cancer are never guaranteed for their growthin organs distinct from the original site of carcinogenesis. Innate immune responses to tumor
are both anti-metastatic and pro-metastatic in conditions where tumor cells exist in the beginning. However, primary tumor tissues can alter microenvironments, whether physically andfunctionally, in the organs that are distant from the primary site. This remote control cultivates
so-called soil before the actual arrival of tumor cells as seed from the primary site. The disseminated tumor cells may die in circulation or after reaching tissues separate from the primarysite or stay alive in a quiescent state before acquisition of signals for re-growth. Irrespectiveof where the primary tumor cells remain alive in the body, establishment of metastatic soil isrequired for metastatic progression. Information on substances produced from the tumor cells
are accumulating, which include actively secreted molecules upon stimulation by growth factors or upon senescence-associated secretory phenotype, and passively released molecules bycell death caused by chemotherapy and irradiation. Those substances can stimulate immune
cells directly or indirectly via non-immune cells such as endothelial cells and resident epithelial cells. The major biological roles are played by pattern recognition receptors (PRRs) that
include TLR and inflammasomes. Here I discuss on molecular mechanisms of pre-metastatic
lungs in which TLR4 is clearly engaged.
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15

CURRICULUM VITAE
Yoshiyuki Morishita, M.D., Ph. D.
LectureDivision of Nephrology, Department of Internal Medicine, Jichi Medical University
3311-1 Yakushiji, Shimotsuke city, Tochigi 329-0498, Japan
Tel: (+81) 285-58-7346
Fax: (+81) 285-44-4869
E-mail: ymori@jichi.ac.jp
Professional Experience
1996-1998
Resident in Medicine, Okayama Red Cross General Hospital
2002-2004
Research Fellow in Nephrology, Jichi Medical University
2007-2009
Research Fellow in Immune Disease Institute, Harvard Medical School, USA
2009-2013
Assistant professor, Division of Nephrology, Department of Internal Medicine,
Jichi Medical University
2013-present Associate Professor, Division of Nephrology, Department of Internal Medicine,
Jichi Medical University
Education
1990-1996 M.D., Jichi Medical University
2004-2008 Ph.D., Okayama University
Recent Publications
1. Morishita, Y., Yoshizawa, H., Watanabe, M., Ishibashi, K., Muto, S., Kusano, E., Nagata, D.:
siRNAs targeted to Smad4 prevent renal fibrosis in vivo. Sci Rep, 2014 (in press)
2. Morishita, Y., Watanabe, M., Nakazawa, E., Ishibashi, K., Kusano, E.: The interaction of
LFA-1 on mononuclear cells and ICAM-1 on tubular epithelial cells accelerates TGF-β1induced renal epithelial-mesenchymal transition. PLoS One. 6: e23267, 2011
3. Morishita, Y., Ohnishi, A., Watanabe, M., Ishibashi, K., Kusano, E.: Establishment of acutekidney injury mouse model by 0.75% adenine ingestion. Ren Fail. 33: 1013-1018, 2011
4. Peer, D., Park, EJ., Morishita, Y., Carman, CV., Shimaoka, M.: Systemic leukocyte-directed
siRNA delivery revealing cyclin D1 as an anti-inflammatory target. Science. 319: 627-630,
2008
5. Morishita, Y., Uenaka, A., Kaya, S., Sato, S., Aji, T., Nakayama, E: HLA-DRB1*0410restricted recognition of XAGE-1b37-48 peptide by CD4 T cells. Microbiol Immunol. 51:
755-62, 2007

16

The delivery of miR-146 with polyethylenimine nanoparticle inhibits renal fibrosis in vivo
Yoshiyuki
yMorishita, Hiromichi Yoshizawa, Toshimi Imai, Minami Watanabe, Shigeaki Muto
and Daisuke Nagata
Division of Nephrology, Department of Internal Medicine, Jichi Medical University, Tochigi,
Japan
Renal fibrosis is the final common pathway leading to decreased renal function. No
therapy has been established to prevent it. Recently, the roles of micro RNAs (miR) for the development of renal fibrosis have been reported; however, few studies have focused the effects
of exogenous miR delivery in an in vivo therapeutic setting for the real fibrosis. In the present study, we investigated the effects of the delivery of miR-146 mimic with polyethylamine
nanoparticle (PEI-NPs) for renal fibrosis in vivo. miR-146 mimic oligo / PEI-NPs complex (N/P ratio: 6) were injected into each renal fibrosis mice induced by unilateral ureteral obstruction
by systemic tail vein injection three times. Non-targeted oligo (Control oliogo) / PEI-NPs were
injected in the same way for a control group. miR-146 mimic-PEI-NPs significantly increased
miR-146 expression (>20 fold) in the kidney compared with those of control group. They
inhibited kidney fibrosis area and the expression of α-SMA, collagen type1 and fibroblastic
specific protein 1. They also inhibited Samd4 expression. Control oligo-PEI-NPs did not showthese effects. The results of this study suggest that the delivery of miR-146 mimic with PEINPs attenuated renal fibrosis and it may be one of the therapeutic options for the prevention ofrenal fibrosis in vivo.
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17

CURRICULUM VITAE
Kensuke Miyake, M.D. , Ph.D.
Professor
Division of Innate Immunity,
Department of Microbiology and Immunology,
The Institute of Medical Science, The University of Tokyo
4-6-1, Shirokanedai, Minatoku, Tokyo 108-8639, Japan
Phone: +81-3-5449-5294
FAX: +81-3-5449-5410
E-mail: kmiyake@ims.u-tokyo.ac.jp
Professional Experience
1988-1990
Research associate, Oklahoma Medical Research Foundation, USA
1990-1993
Assistant professor, Saga Medical School
1993-2001
Associate professor, Saga Medical School
2001-current
Professor, The Institute of Medical Science, The University of Tokyo
Education
Undergraduate
Graduate

1978-1984 Okayama University Medical School
1984-1988 Osaka University Medical School,
Ph. D. awarded 3/88

Recent Publications
1. Miyake K, Kaisho T. Homeostatic inflammation in innate immunity. Curr Opin Immunol.
2014 doi: 10.1016/j.coi.2014.08.003.
2. Huh JW, Shibata T, Hwang M, Kwon EH, Jang MS, Fukui R, Kanno K, Jung DJ, Jang
MH, *Miyake K, and *Kim YM (*: co-corresponding author) UNC93B1 is essential for theplasma membrane localization and signaling of Toll-like receptor 5. Proc. Natl. Acad. Sci.
USA 2014, 111:7072-7077. doi:10.1073/pnas.1322838111.
3. Onji M, Kanno A, Saitoh S, Fukui R, Motoi Y, Shibata T, Matsumoto F, Lamichhane A,
Sato S, Kiyono H, Yamamoto K, Miyake K. An essential role for the N-terminal fragment ofToll-like receptor 9 in DNA sensing. Nat. Commun. 2013, 4: 1949, doi:10.1038/ncomms2949
4. Fukui R, Saitoh S-I, Kanno A, Onji M, Shibata T, Ito A, Onji M, Matsumoto M, Akira
S, Yoshida N, Miyake K. Unc93B1 restricts systemic lethal inflammation by orchestrating
TLR7- and TLR9-trafficking. Immunity, 2011, 35:69-81.
5. Fukui R, Saitoh S-I, Matsumoto F, Kozuka-Hara H, Oyama M, Tabeta K, Beutler B, andMiyake K. Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells
towards DNA- but against RNA-sensing. J. Exp. Med., 2009, 206:1339-1350

18

Mechanisms regulating nucleic acid-sensing Toll-like receptors
Kensuke MiyakeyInstitute of Medical Science, University of Tokyo
The Toll family of receptors has critical roles in microbial recognition and activation of defense responses. Cell surface Toll-like receptors (TLRs) including TLR4/MD-2,
TLR1/TLR2, TLR2/TLR6, or TLR5 recognize microbial membrane lipids or flagellin, whereas
TLR3, 7, 8, and 9 reside in intracellular organelle and sense nucleic acids. Recent progresses
have revealed that self-pathogen discrimination by TLRs is error prone and TLRs have been
implicated in a variety of autoimmune diseases. Although the ligand specificity and downstream signaling pathways of each TLR have been extensively studied, much less is known as
to how innate immune responses to self-products are controlled. Nucleic acid-sensing TLRs,
TLR7, 8 and 9, have a risk of responding to self-derived nucleic acids. To prevent autoimmune
responses, these TLRs are thought to be controlled by restricting nucleic acid sensing in endolysosomes, not the cell surface. Extracellular self-nucleic acids are instantly degraded anddo not get to endolysosomes, whereas microbial nucleic acids are protected by microbial membranes and get to the endolysosomes. To limit nucleic acid sensing in endolysosomes, trafficking of nucleic acid-sensing TLRs from ER to endolysosomes is tightly controlled. The subcellular distribution of TLR7 and 9 are dependent on the TLR transporter Unc93B1. Unc93B1 is
associated with TLR7 and 9 and transports them from ER to the endolysosomes. Interestingly,
Unc93B1 is not just the TLR transporter but also has a role in controlling the balance between
TLR7 and TLR9. Unc93B1-dependent control of TLR7 and 9 balance prevents TLR7-dependent autoimmunity. TLR7 has a unique risk of autoimmunity. TLR7 is a promising target fortherapeutic intervention in autoimmune diseases.
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CURRICULUM VITAE
Takanori Komada, M.D.
Graduate Student
Division of Inflammation Research, Center for Molecular Medicine,
Department of Nephrology, Jichi Medical University
3311-1, Yakushiji, Shimotsuke-shi, Tochigi, 329-0431,Japan
Tel: +81-28-558-7346 (Office)Fax: +81-28-544-4869 (Office)E-mail: t_komada@jichi.ac.jp
Professional Experience
2002~2004: Junior Resident, Mie Prefectural Hospital
2004~2006: Medical Staff in Internal Medicine, Kinan Hospital
2006~2007: Senior Resident in Department of Nephrology, Jichi Medical University Saitama
Medical Center
2007~2008: Medical Staff in Internal Medicine, Minamiise Municipal Hospital
2008~2010: Medical Staff in Department of Nephrology, Fujisawa City Hospital
2010~2011: Senior Resident in Department of Nephrology,
Jichi Medical University Saitama Medical Center
2011~Present :Graduate School of Medicine, Jichi Medical University
Education
2002: M.D., Jichi Medical University
Recent Publications
1. Komada T, Usui F, Shirasuna K, Kawashima A, Kimura H, Karasawa T, Nishimura S,
Sagara J, Noda T, Taniguchi S, Muto S, Nagata D, Kusano E, Takahashi M: ASC in Renal
Collecting Duct Epithelial Cells Contributes to Inflammation and Injury after Unilateral
Ureteral Obstruction. Am. J. Pathol. 2014
2. Komada T, Morishita Y, Kitamura M, Iwazu K, Numata A, Kobayashi T, Yamamoto H,
Akimoto T, Saito O, Ando Y, Takemoto F, Muto S, Yumura W, Kusano E: Acute kidney injury
in a patient with nephrotic syndrome due to focal segmental glomerular nephritis induced
d 52:
by a single oral administration of high-dose bisphosphonate (minodronate). Intern Med
1383–1387, 2013
3. Suzuki J, Komada T, Hirai K, Tsuruoka H, Mori H, Yoshida I, Tabei K: An adult case ofd 61:
fulminant Epstein-Barr virus infection with acute tubulointerstitial nephritis. Intern Med
629–634, 2012
4. Komada T, Yoshida I, Sakuma Y, Mori H, Amemiya M, Nakazato Y, Tabei K: Comparison
between long-acting and short-acting loop diuretics in patients with chronic kidney disease:evaluation of efficacy and safety. Jichi Medical University Journall 33: 79–89, 201020

Inflammasome Activation in Renal Collecting Duct Epithelial Cells: The Potential Role of
Non-Immune cells in Renal Inflammation and Fibrosis
Takanori Komada1,2, Fumitake Usui1, Koumei Shirasuna1, Akira Kawashima, 1 Hiroaki
Kimura1, Tadayoshi Karasawa1, Shigeaki Muto2, Daisuke Nagata2, Eiji Kusano2, and Masafumi Takahashi1.
1Division of Inflammation Research, Center for Molecular Medicine, 2Department of Nephrology, Jichi Medical University, Tochigi; Japan
Inflammation plays a crucial role in the pathophysiological characteristics of chronic
kidney disease; however, the inflammatory mechanisms underlying the chronic kidney disease
process remain unclear. Recent evidence indicates that sterile inflammation triggered by tissue
injury is mediated through a multiprotein complex called the inflammasome. Therefore, weinvestigated the role of the inflammasome in the development of chronic kidney disease using
a murine unilateral ureteral obstruction (UUO) model. Inflammasome-related molecules were
up-regulated in the kidney after UUO. Apoptosis- associated speck-like protein containing a
caspase recruitment domain deficiency significantly reduced inflammatory responses, such as
inflammatory cell infiltration and cytokine expression, and improved subsequent renal injury
and fibrosis. Furthermore, apoptosis-associated speck-like protein containing a caspase recruitment domain was specifically up-regulated in collecting duct (CD) epithelial cells of the UUOtreated kidney. In vitro experiments showed that extracellular adenosine triphosphate (ATP)induced inflammasome activation in CD epithelial cells through P2X7-potassium efflux and reactive oxygen species-dependent pathways. These results demonstrate the molecular basis forthe inflammatory response in the process of chronic kidney disease and suggest the CD inflammasome as a potential therapeutic target for preventing chronic kidney disease progression.
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CURRICULUM VITAE
Osamu Nureki, PhD
Professor
Department of Biological Sciences, Graduate School of Science
The University of Tokyo
2-11-16 Yayoi, Bunkyo-ku, Tokyo 113-0032
Tel: (+81)3-5841-4392
FAX: (+81)3-5841-8057
E-mail: nureki@bs.s.u-tokyo.ac.jp
Education and Employment
1993.3
Graduation from Department of Biophysics and Biochemistry, Graduate Schoolof Science, the University of Tokyo (Doctor of Science)
1994.4-1995.3 Post Doctor at Protein Engineering Research Institute (PERI)
1994.4-1995.4 Special Researcher at Crystallography Laboratory in RIKEN
1995.4-2003.4 Assistant Professor at the University of Tokyo
2002.4-2003.4 Associate Professor at the University of Tokyo
2003.5Professor at Department of Biological Information, Graduate School of
Biosciences and Biotechnology, Tokyo Institute of Technology
2008.4Professor at Department of Basic Medical Sciences, The Institute of Medical
Science, The University of Tokyo
2010.4Professor at Department of Biophysics and Biochemistry, Graduate School of
Science, The University of Tokyo
Awards
JSPS PRIZE in 2008 by JSPS, Mochida Memorial Research Award in 2009, Kihara Memorial
Research Award in 2010, Inoue Prize for Science in 2011, Uehara Award in 2014, Takeda Prize
for Medical Science in 2014
Recent publications.
1. “Structural basis for Sec-independent membrane protein insertion by YidC” K. Kumazaki,
S. Chiba, M. Takemoto, A. Furukawa, K. Nishiyama, Y. Sugano, T. Mori, N. Dohmae, K.
Hirata, Y. Nakada-Nakura, A. D. Maturana, Y. Tanaka, H. Mori, Y. Sugita, F. Arisaka, K. Ito,
R. Ishitani, T. Tsukazaki and O. Nureki
Nature 509, 516-520 (2014).
2. “Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA” H. Nishimasu,
F. A. Ran, P. D. Hsu, S. Konermann, S. I. Shehata, N. Dohmae, R. Ishitani, F. Zhang F and O.
Nureki.
Cell 156, 935-949 (2014).
3. “Structural basis for the counter-transport mechanism of a H+/Ca2+ exchanger” T. Nishizawa,
S. Kita, A. D. Maturana, N. Furuya, K. Hirata, G. Kasuya, S. Ogasawara, N. Dohmae, T.
Iwamoto, R. Ishitani, O. Nureki Science 341, 168-172 (2013).
4. “Structural basis for the novel mechanism of drug extrusion by a MATE multidrug
transporter.” Y. Tanaka, C. J. Hipolito, A. D. Maturana, K. Ito, T. Kuroda, T. Higuchi, T.
Katoh, H. E. Kato, M. Hattori, K. Kumazaki, T. Tsukazaki, R. Ishitani, H. Suga and O. Nureki
Nature 496, 247–251 (2013)
5. “Crystal structure of the channelrhodopsin light-gated cation channel” H. E. Kato, F. Zhang,
O. Yizhar, C. Ramakrishnan, T. Nishizawa, K. Hirata, J. Ito, Y. Aita, T. Tsukazaki, S. Hayashi,
P. Hegemann, A. D. Maturana, R. Ishitani, K. Deisseroth and O. Nureki
Nature 428, 369-374 (2012).

22

Crystal structure of Cas9 complexed with guide RNA and target DNA
Hiroshi Nishimasu1, Ryuichiro Ishitani1, Feng Zhang2 and Osamu Nureki1
1The University of Tokyo, 2MIT
The CRISPR-associated endonuclease Cas9 can be targeted to specific genomic lociby single guide RNAs (sgRNAs). Here, we solved the crystal structure of Streptococcus pyrogenes Cas9 in complex with sgRNA and its target DNA at 2.5 A resolution. The structure revealed a bilobed architecture consisting of target recognition and nuclease lobes (Rec and Nuclobes, respectively), accommodating the sgRNA:DNA heteroduplex in a positively-charged
groove at their interface. While Rec lobe is essential for binding sgRNA and DNA, Nuc lobe
contains the HNH and RuvC nuclease domains, which are properly located for cleavage of thecomplementary and noncomplemantary strands of the target DNA, respectively. Nuc lobe also
contains a C-terminal domain responsible for the recognition of the protospacer adjacent motif(PAM). This high-resolution structure combined with functional analyses have revealed themolecular mechanism of RNA-guided DNA targeting by Cas9, thus paving the way for rational design of new, versatile genome-editing technologies.
<Reference>“Crystal Structure of Cas9 in Complex with Guide RNA and Target DNA” H. Nishimasu, F. A.
Ran, P. D. Hsu, S. Konermann, S. I. Shehata, N. Dohmae, R. Ishitani, F. Zhang F and O. Nureki.
Cell 156, 935-949 (2014).
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MEMO

MEMO

MEMO

MEMO

The 11th Nikko International Symposium 2014
Jichi Medical University
Tel:0285-58-7550 Fax:0285-40-8303 E-mail:shien@jichi.ac.jp