Gene analysis for novel causative mutations in an inherited arrhythmia and functional study for these mutations using zebrafish(Fostering Joint International Research)

• Project/Area Number: 15KK0302
• Research Category: Fund for the Promotion of Joint International Research (Fostering Joint International Research)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine
• Research Institution: Kanazawa University
• Principal Investigator: Hayashi Kenshi 金沢大学, 附属病院, 助教 (00422642)
• Research Collaborator: MacRae Calum ブリガム・アンド・ウィメンズ病院, 循環器内科, 科長兼准教授 ; Mueller Ferenc ; Sik Attila
• Project Period (FY): 2016 – 2018
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥11,830,000 (Direct Cost: ¥9,100,000、Indirect Cost: ¥2,730,000)
• Keywords: 遺伝性不整脈 / 次世代シーケンサー / ゼブラフィッシュ / 網羅的遺伝子解析 / 全エクソーム解析 / 細胞電気生理 / 次世代シークエンサー / 循環器・高血圧 / 遺伝子 / 不整脈

Outline of Final Research Achievements

We performed candidate gene analysis or whole exome sequence for inherited arrhythmia including long QT syndrome and Brugada syndrome using a next generation sequencer. We identified many variants of uncertain significance (VUS) as well as many causative mutations. We performed functional studies for these VUS using zebrafish embryo and could show functional abnormality of them.

Academic Significance and Societal Importance of the Research Achievements

次世代シーケンサ－による網羅的遺伝子解析により、従来の方法で見逃されていた数多くの遺伝子変異を見出すことができた。これまで機能解析が困難だった遺伝子に対し、CRISPR/Cas9システムと呼ばれる新しい手法でゼブラフィッシュの遺伝子改変を行い、ゼブラフィッシュ胚の心臓の光学マッピング、活動電位測定、体表面心電図測定といった画期的な方法で数日間以内に遺伝子変異の病的意義を明らかにすることができた。

Research Products

• [Int'l Joint Research] ブリガム・アンド・ウィメンズ病院(米国) (2016)
  • Year and Date: 2016-09-10
• [Journal Article] Mutant KCNJ3 and KCNJ5 potassium channels as novel molecular targets in bradyarrhythmias and atrial fibrillation. (2019)
  • Author(s): Yamada N, Asano Y, Fujita M, Yamazaki S, Inanobe A, Matsuura N, Kobayashi H, Ohno S, Ebana Y, Tsukamoto O, Ishino S, Takuwa A, Kioka H, Yamashita T, Hashimoto N, Zankov DP, Shimizu A, Asakura M, Asanuma H, Kato H, Nishida Y, Miyashita Y, Shinomiya H, Naiki N, Hayashi K, Makiyama T, Ogita H, et al.
  • Journal Title: Circulation Volume: In press Issue: 18 Pages: 2157-2169
  • DOI: 10.1161/circulationaha.118.036761
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Association of Genetic and Clinical Aspects of Congenital Long QT Syndrome With Life-Threatening Arrhythmias in Japanese Patients (2019)
  • Author(s): Shimizu Wataru、Makimoto Hisaki、Yamagata Kenichiro、Kamakura Tsukasa、Wada Mitsuru、Miyamoto Koji、Inoue-Yamada Yuko、Okamura Hideo、Ishibashi Kohei、Noda Takashi、Nagase Satoshi、Miyazaki Aya、Sakaguchi Heima、Shiraishi Isao、Makiyama Takeru、Ohno Seiko、Itoh Hideki、et al
  • Journal Title: JAMA Cardiology Volume: in press Issue: 3 Pages: 246-254
  • DOI: 10.1001/jamacardio.2018.4925
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Impact of cardiac myosin light chain kinase gene mutation on development of dilated cardiomyopathy (2019)
  • Author(s): Hodatsu A, Fujino N, Uyama Y, Tsukamoto O, Imai-Okazaki A, Yamazaki S, Seguchi O, Konno T, Hayashi K, Kawashiri MA, Asano Y, Kitakaze M, Takashima S, Yamagishi M
  • Journal Title: ESC Heart Fail Volume: 6 Issue: 2 Pages: 06-415
  • DOI: 10.1002/ehf2.12410
  • Peer Reviewed / Open Access
• [Journal Article] Spontaneous Isolated Superior Mesenteric Artery Dissection Requiring Emergent Surgery (2018)
  • Author(s): Tanaka Y, Tada H, Takeda Y, Iino K, Hayashi K, Takemura H, Yamagishi M, Kawashiri MA.
  • Journal Title: Internal Medicine Volume: 57 Issue: 18 Pages: 2681-2684
  • DOI: 10.2169/internalmedicine.0641-17
  • NAID: 130007484205
  • ISSN: 0918-2918, 1349-7235
  • Year and Date: 2018-09-15
  • Peer Reviewed / Open Access
• [Journal Article] Author Correction: Heart Failure Phenotypes Induced by Knockdown of DAPIT in Zebrafish: A New Insight into Mechanism of Dilated Cardiomyopathy (2018)
  • Author(s): Nagata Yoji、Yamagishi Masakazu、Konno Tetsuo、Nakanishi Chiaki、Asano Yoshihiro、Ito Shin、Nakajima Yuri、Seguchi Osamu、Fujino Noboru、Kawashiri Masa-aki、Takashima Seiji、Kitakaze Masafumi、Hayashi Kenshi
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 7768-7768
  • DOI: 10.1038/s41598-018-26012-4
  • Peer Reviewed / Open Access
• [Journal Article] Characteristics of induced pluripotent stem cells from clinically divergent female monozygotic twins with Danon disease. (2018)
  • Author(s): Yoshida S, Nakanishi C, Okada H, Mori M, Yokawa J, Yoshimuta T, Ohta K, Konno T, Fujino N, Kawashiri MA, Yachie A, Yamagishi M, Hayashi K.
  • Journal Title: J Mol Cell Cardiol. Volume: 114 Pages: 234-242
  • DOI: 10.1016/j.yjmcc.2017.11.019
  • Peer Reviewed / Open Access
• [Journal Article] Impact of B-Type Natriuretic Peptide Level on Risk Stratification of Thromboembolism and Death in Patients With Nonvalvular Atrial Fibrillation ― The Hokuriku-Plus AF Registry ― (2018)
  • Author(s): Hayashi Kenshi、Tsuda Toyonobu、Nomura Akihiro、Fujino Noboru、Nohara Atsushi、Sakata Kenji、Konno Tetsuo、Nakanishi Chiaki、Tada Hayato、Nagata Yoji、Teramoto Ryota、Tanaka Yoshihiro、Kawashiri Masa-aki、Yamagishi Masakazu、on behalf of the Hokuriku-Plus AF Registry Investigators
  • Journal Title: Circulation Journal Volume: 82 Issue: 5 Pages: 1271-1278
  • DOI: 10.1253/circj.CJ-17-1085
  • NAID: 130006725972
  • ISSN: 1346-9843, 1347-4820
  • Peer Reviewed / Open Access
• [Journal Article] Emerin plays a crucial role in nuclear invagination and in the nuclear calcium transient. (2017)
  • Author(s): Shimojima M, Yuasa S, Motoda C, Yozu G, Nagai T, Ito S, Lachmann M, Kashimura S, Takei M, Kusumoto D, Kunitomi A, Hayashiji N, Seki T, Tohyama S, Hashimoto H, Kodaira M, Egashira T, Hayashi K, Nakanishi C, Sakata K, Yamagishi M, Fukuda K.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 44312-44312
  • DOI: 10.1038/srep44312
  • NAID: 120006325351
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Genotype-phenotype correlation of SCN5A mutation for clinical and electrocardiographic characteristics of probands with Brugada syndrome: Japanese Multicenter Registry. (2017)
  • Author(s): Yamagata K, Horie M, et al.
  • Journal Title: Circulation Volume: 印刷中 Issue: 23 Pages: 0-0
  • DOI: 10.1161/circulationaha.117.027983
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ventricular Fibrillation Associated With Dynamic Changes in J-Point Elevation in a Patient With Silent Thyroiditis. (2017)
  • Author(s): Karashima S, Tsuda T, Wakabayashi Y, Kometani M, Demura M, Ichise T, Kawashiri MA, Takeda Y, Hayashi K, Yoneda T.
  • Journal Title: J Endocr Soc. Volume: 2 Issue: 2 Pages: 135-139
  • DOI: 10.1210/js.2017-00276
  • Peer Reviewed
• [Journal Article] Heat Failure Phenotypes Induced by Knockdown of DAPIT in Zebrafish: A New Insight into Mechanism of Dilated Cardiomyopathy. (2017)
  • Author(s): Nagata Y, Yamagishi M, Konno T, Nakanishi C, Asano Y, Ito S, Nakajima Y, Seguchi O, Fujino N, Kawashiri MA, Takashima S, Kitakaze M, Hayashi K.
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 17417-17417
  • DOI: 10.1038/s41598-017-17572-y
  • Peer Reviewed / Open Access
• [Journal Article] Dynamical mechanisms of phase-2 early afterdepolarizations in human ventricular myocytes: insights from bifurcation analyses of two mathematical models (2017)
  • Author(s): Kurata Y, Tsumoto K, Hayashi K, Hisatome I, Tanida M, Kuda Y, Shibamoto T
  • Journal Title: Am J Physiol Heart Circ Physiol Volume: 312 Issue: 1 Pages: H106-H127
  • DOI: 10.1152/ajpheart.00115.2016
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] The genetics of atrial fibrillation. (2017)
  • Author(s): Hayashi K, Tada H, Yamagishi M.
  • Journal Title: Curr Opin Cardiol Volume: 32 Pages: 10-16
  • Peer Reviewed
• [Journal Article] The genetics underlying acquired long QT syndrome: impact for genetic screening. (2016)
  • Author(s): Itoh H, Crotti L, Aiba T, Spazzolini C, Denjoy I, Fressart V, Hayashi K, Nakajima T, Ohno S, Makiyama T, Wu J, Hasegawa K, Mastantuono E, Dagradi F, Pedrazzini M, Yamagishi M, Berthet M, Murakami Y, Shimizu W, Guicheney P, Schwartz PJ, Horie M.
  • Journal Title: European Heart Journal Volume: 37 (18) Issue: 18 Pages: 1456-1464
  • DOI: 10.1093/eurheartj/ehv695
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Impact of Updated Diagnostic Criteria for Long QT Syndrome on Clinical Detection of Diseased Patients: Results from Study of Patients Carrying Gene Mutations. (2016)
  • Author(s): Hayashi K, Konno T, Fujino N, Itoh H, Fujii Y, Imi-Hashida Y, Tada H, Tsuda T, Tanaka Y, Saito T, Ino H, Kawashiri M, Ohta K, Horie M, Yamagishi.M.
  • Journal Title: JACC Clin Electrophysiol Volume: 2 Issue: 3 Pages: 279-287
  • DOI: 10.1016/j.jacep.2016.01.003
  • Peer Reviewed
• [Presentation] Cardiac Events during Competitive Sports and Daily Activities in Children and Adolescents with Long QT Syndrome (2019)
  • Author(s): Kenshi Hayashi, Kunio Ohta, Takekatsu Saito, Noboru Fujino, Yasuhiro Todo, Kenji Sakata, Hayato Tada, Akihiro Nomura, Toyonobu Tsuda, Yoshihiro Tanaka, Masakazu Yamagishi, Masa-aki Kawashiri
  • Organizer: JCS2019
• [Presentation] EXOME SEQUENCING AND FUNCTIONAL ANALYSIS FOR PATIENTS WITH EARLY ONSET CARDIAC CONDUCTION-SYSTEM DISEASES (2018)
  • Author(s): Kenshi Hayashi, Akihiro Nomura, Yoshihiro Asano, Manu Beerens, Yasutaka Kurata, Noboru Fujino, Hayato Tada, Calum A MacRae, Seiji Takashima, Masakazu Yamagishi
  • Organizer: INTERNATIONAL ACADEMY OF CARDIOLOGY ANNUAL SCIENTIFIC SESSIONS 2018 23rd WORLD CONGRESS ON HEART DISEASE
  • Int'l Joint Research
• [Presentation] J Wave for Predicting Cardiac Events in Hypertrophic Cardiomyopathy (2018)
  • Author(s): Kenshi Hayashi, Toyonobu Tsuda, Akihiro Nomura, Noboru Fujino, Masakazu Yamagishi
  • Organizer: 11th Asia Pacific Heart Rhythm Society Scientific Session
  • Int'l Joint Research
• [Presentation] Predictors for Exacerbating Kidney Function in Patients with Nonvalvular Atrial Fibrillation: The Hokuriku-plus AF Registry (2018)
  • Author(s): Kenshi Hayashi, Toyonobu Tsuda, Akihiro Nomura, Noboru Fujino, Atsushi Nohara, Kenji Sakata, Chiaki Nakanishi, Hayato Tada, Yoshihiro Tanaka, Masa-aki Kawashiri, and Masakazu Yamagishi
  • Organizer: AHA Scientific sessions 2018
  • Int'l Joint Research
• [Presentation] Exome Sequencing for Patients with Early-onset Cardiac Conduction Diseases (2017)
  • Author(s): Kenshi Hayashi, Akihiro Nomura, Yoshihiro Asano, Noboru Fujino, Kenji Sakata, Tetsuo Konno, Hayato Tada, Toyonobu Tsuda, Ryota Teramoto, Yoji Nagata, Yoshihiro Tanaka, Masa-aki Kawashiri, Calum MacRae, Seiji Takashima, Masakazu Yamagishi
  • Organizer: The 81st Annual Scientific Meeting of the Japanese Circulation Society
  • Place of Presentation: Kanazawa
  • Year and Date: 2017-03-17
• [Presentation] Exome Sequencing for Patients with Early-onset Cardiac Conduction Diseases (2017)
  • Author(s): Kenshi Hayashi
  • Organizer: The 81st Annual Scientific Meeting of the Japanese Circulation Society
• [Presentation] CRISPR-mediated insertions or deletions of the human LMNA homolog in zebrafish as a model of early-onset cardiac conduction disease (2017)
  • Author(s): Kenshi Hayashi
  • Organizer: The 64ｔｈ Annual Meeting of the Japanese Heart Rhythm Society
• [Presentation] CRISPR-mediated insertions or deletions of the human LMNA homolog in zebrafish as a model of early-onset cardiac conduction disease (2017)
  • Author(s): Kenshi Hayashi
  • Organizer: AHA 2017
  • Int'l Joint Research
• [Presentation] Mechanisms of Fever-induced QT Prolongation in Patients with KCNH2 Mutations in the S5-pore Region: Evidence from Genotypic and Functional Analyses (2016)
  • Author(s): Kenshi Hayashi, Tadashi Nakajima, Yasutaka Kurata, Shoichi Tange, Noboru Fujino, Kenji Sakata, Tetsuo Konno, Toyonobu Tsuda, Yoji Nagata, Ryota Teramoto, Yoshihiro Tanaka, Takekatsu Saito, Masa-aki Kawashiri, Kunio Ohta, Yoshiaki Kaneko, Masahiko Kurabayash, Masakazu Yamagishi
  • Organizer: AHA2016
  • Place of Presentation: New Orleans, LA
  • Year and Date: 2016-11-12
  • Int'l Joint Research
• [Presentation] Updated Diagnostic Criteria for Long QT Syndrome (2016)
  • Author(s): Kenshi Hayashi, Tetsuo Konno, Noboru Fujino, Hideki Itoh, Yusuke Fujii, Yoko Imi-Hashida, Hayato Tada, Toyonobu Tsuda, Yoshihiro Tanaka, Takekatsu Saito, Hidekazu Ino, Masa-aki Kawashiri, Kunio Ohta, Minoru Horie, Masakazu Yamagishi.
  • Organizer: APHRS 2016
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2016-10-12
  • Int'l Joint Research
• [Presentation] The Role of Common and Rare Genetic Variants Implicated in Susceptibility to Atrial Fibrillation (2016)
  • Author(s): Kenshi Hayashi, Hayato Tada, and Masakazu Yamagishi
  • Organizer: APHRS 2016
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2016-10-12
  • Int'l Joint Research
• [Book] 循環器ジャーナル (2017)
  • Author(s): 林 研至 山岸正和
  • Total Pages: 216
  • Publisher: 医学書院