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

2018 Fiscal Year Annual Research Report

• Project/Area Number: 15KK0302
• Research Institution: Kanazawa University
• Principal Investigator: 林 研至 金沢大学, 附属病院, 助教 (00422642)
• Project Period (FY): 2016 – 2018
• Keywords: 遺伝性不整脈 / 網羅的遺伝子解析 / ゼブラフィッシュ

Outline of Annual Research Achievements

遺伝性不整脈102症例に対して次世代シークエンサーによる全エクソーム解析あるいは拡大候補遺伝子解析を行なった。High-Resolution Melting法でLQT1, LQT2, LQT3に異常を認めなかった先天性QT延長症候群65症例に対し、既報の15遺伝子の変異の有無を解析したところ、7症例にKCNQ1変異、6症例にKCNH2変異、1症例にCALM2変異を見出した。また、LQT1およびLQT2についてMLPAを行ったところ、1例に欠失変異を見出した。さらに、1症例にRYR2変異を、2症例にPKP2変異を見出した。Type 1 Brugada症候群25症例に対し、既報の23遺伝子の変異の有無を解析したところ、2症例にSCN5A変異、1症例にGPD1-L変異、1症例にSCN1B変異、1症例にTRPM4変異を見出した。ARVC 12症例に対し、既報の17遺伝子の変異の有無を解析したところ、4症例にDSG2変異、1症例にPKP2変異、1症例にRYR2変異、1症例にLMNA変異、1症例にTTN変異を見出した。
見いだされた希少遺伝子変異の病的意義を明らかにするため、モルフォリノ、Crispr/Cas9システムあるいはENUミュータジェネシスを用いて遺伝子改変ゼブラフィッシュによる機能評価を試みた。若年発症徐脈症例より見出されたLMNA遺伝子変異、LQTSより見いだされたKCNQ1遺伝子変異、KCNH2遺伝子変異について、それぞれの遺伝子のホモログであるゼブラフィッシュlmna遺伝子, kcnh2a遺伝子, kcnq1遺伝子の改変を行った。そのゼブラフィッシュ胚に対し、心拍数計測、心機能評価、光学マッピングによる刺激伝導速度測定、活動電位測定、心電図測定の評価などを行なった。

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 – 2017-02-27
  • Country Name: U.S.A.
  • Counterpart Institution: ブリガム・アンド・ウィメンズ病院
  • Co-investigator Overseas: Calum A MacRae
  • Department: Cardiovascular Division
  • Job Title: Assistant professor
• [Journal Article] Mutant KCNJ3 and KCNJ5 Potassium Channels as Novel Molecular Targets in Bradyarrhythmias and Atrial Fibrillation (2019)
  • Author(s): Yamada Noriaki、Asano Yoshihiro、Hayashi Kenshi, et al.
  • Journal Title: Circulation Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1161/CIRCULATIONAHA.118.036761
  • Peer Reviewed / 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、Hayashi Kenshi、et al
  • Journal Title: JAMA Cardiology Volume: 印刷中 Pages: 印刷中
  • DOI: 10.1001/jamacardio.2018.4925
  • Peer Reviewed
• [Journal Article] Impact of cardiac myosin light chain kinase gene mutation on development of dilated cardiomyopathy (2019)
  • Author(s): Hodatsu Akihiko、Fujino Noboru、Uyama Yuki、Tsukamoto Osamu、Imai‐Okazaki Atsuko、Yamazaki Satoru、Seguchi Osamu、Konno Tetsuo、Hayashi Kenshi、Kawashiri Masa‐aki、Asano Yoshihiro、Kitakaze Masafumi、Takashima Seiji、Yamagishi Masakazu
  • Journal Title: ESC Heart Failure Volume: 6 Pages: 406～415
  • DOI: 10.1002/ehf2.12410
  • Peer Reviewed
• [Journal Article] Spontaneous Isolated Superior Mesenteric Artery Dissection Requiring Emergent Surgery (2018)
  • Author(s): Tanaka Yoshihiro、Tada Hayato、Takeda Yoshimichi、Iino Kenji、Hayashi Kenshi、Takemura Hirofumi、Yamagishi Masakazu、Kawashiri Masa-aki
  • Journal Title: Internal Medicine Volume: 57 Pages: 2681～2684
  • DOI: 10.2169/internalmedicine.0641-17
  • Peer Reviewed
• [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 Pages: -
  • DOI: 10.1038/s41598-018-26012-4
  • Peer Reviewed / Open Access
• [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