Gene analysis for novel causative mutations in an inherited arrhythmia and determination of the underlying mechanism of arrhythmia

2020 Fiscal Year Annual Research Report

• Project/Area Number: 18K08100
• Research Institution: Kanazawa University
• Principal Investigator: 林 研至 金沢大学, 保健学系, 准教授 (00422642)
• Co-Investigator(Kenkyū-buntansha): 野村 章洋 金沢大学, 附属病院, 特任准教授 (30707542) ; 藤野 陽 金沢大学, 保健学系, 准教授 (40361993)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Keywords: 遺伝性不整脈 / ゼブラフィッシュ / 機能解析

Outline of Annual Research Achievements

遺伝性不整脈であるQT延長症候群（LQTS）で見いだされたKCNQ1遺伝子バリアントに対し、ゼブラフィッシュ胚を用いた機能解析がその病的意義決定に有用かどうか検討した。ヒトKCNQ1遺伝子のホモログであるゼブラフィッシュkcnq1遺伝子配列を認識するcrisprRNAを設計し、受精卵にcrisprRNA、trans-activating crRNA、CAS9蛋白を注入した。受精72時間後に遺伝子解析を行い、ゲノム編集時にさまざまな挿入欠失変異が生じていることを確認した。F0世代のゼブラフィッシュを飼育し、野生型ゼブラフィッシュと交配させた。産まれたF1世代ゼブラフィッシュ胚の遺伝子解析を行ったところkcnq1遺伝子のin-frame mutationをもつゼブラフィッシュを多数確認した。このin-frame mutationをもつゼブラフィッシュを飼育し、変異体どおしを交配させ、産まれたF2世代においてホモ接合性の変異をもつゼブラフィッシュを確認した。このホモ接合性変異体を飼育し、変異体どおしを交配させ、F3世代のゼブラフィッシュ胚の心電図、活動電位測定を行った。ホモ接合性変異体の心電図で有意なQT延長を認め、活動電位測定で活動電位持続時間の有意な延長を認めた。次に、ホモ接合性変異体の受精卵に野生型ヒトKCNQ1遺伝子mRNAを注入し、その72時間後に活動電位を測定したところ、活動電位持続時間の有意な短縮を認めた。一方、変異型KCNQ1 mRNAを注入したゼブラフィッシュ胚の活動電位持続時間は延長したままで短縮を認めなかった。このようにゼブラフィッシュ胚を用いた機能解析はLQTSで見いだされるKCNQ1遺伝子バリアントの病的意義の評価に有用であると考えられた。

Research Products

• [Journal Article] Association of proteinuria with incident atrial fibrillation in the general Japanese population (2021)
  • Author(s): Tanaka Yoshihiro、Tada Hayato、Hara Satoshi、Hayashi Kenshi、Patel Ravi B.、Nishikawa Tetsuo、Hashiba Atsushi、Takamura Masayuki、Greenland Philip、Kawashiri Masa-aki
  • Journal Title: Journal of Cardiology Volume: 77 Pages: 100～105
  • DOI: 10.1016/j.jjcc.2020.08.001
  • Peer Reviewed
• [Journal Article] Myocyte-specific enhancer factor 2c triggers transdifferentiation of adipose tissue-derived stromal cells into spontaneously beating cardiomyocyte-like cells (2021)
  • Author(s): Takashima Shinichiro、Usui Soichiro、Inoue Oto、Goten Chiaki、Yamaguchi Kosei、Takeda Yusuke、Cui Shihe、Sakai Yoshio、Hayashi Kenshi、Sakata Kenji、Kawashiri Masa-aki、Takamura Masayuki
  • Journal Title: Scientific Reports Volume: 11 Pages: 1520~1520
  • DOI: 10.1038/s41598-020-80848-3
  • Peer Reviewed
• [Journal Article] Impact of functional studies on exome sequence variant interpretation in early-onset cardiac conduction system diseases (2020)
  • Author(s): Hayashi Kenshi、et al.
  • Journal Title: Cardiovascular Research Volume: 116 Pages: 2116～2130
  • DOI: 10.1093/cvr/cvaa010
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Risk Factors Associated With Ischemic Stroke in Japanese Patients With Nonvalvular Atrial Fibrillation (2020)
  • Author(s): Hayashi Kenshi、et al.
  • Journal Title: JAMA Network Open Volume: 3 Pages: e202881～e202881
  • DOI: 10.1001/jamanetworkopen.2020.2881
  • Peer Reviewed / Open Access
• [Journal Article] Serum sitosterol level predicting ABCG5 or ABCG8 genetic mutations (2020)
  • Author(s): Kojima Nobuko、Tada Hayato、Usui Soichiro、Sakata Kenji、Hayashi Kenshi、Nohara Atsushi、Inazu Akihiro、Takamura Masayuki、Kawashiri Masa-aki
  • Journal Title: Clinica Chimica Acta Volume: 507 Pages: 11～16
  • DOI: 10.1016/j.cca.2020.04.003
  • Peer Reviewed
• [Journal Article] Fulminant myocarditis and pulmonary cavity lesion induced by disseminated mucormycosis in a chronic hemodialysis patient: Report of an autopsied case (2020)
  • Author(s): Fujisawa Yuhei、Hara Satoshi、Zoshima Takeshi、Maekawa Naoto、Inoue Dai、Sasaki Motoko、Gamou Tadatsugu、Nagata Yoji、Hayashi Kenshi、Takeji Akari、Ito Kiyoaki、Mizushima Ichiro、Fujii Hiroshi、Kawano Mitsuhiro
  • Journal Title: Pathology International Volume: 70 Pages: 557～562
  • DOI: 10.1111/pin.12943
  • Peer Reviewed
• [Journal Article] Transient swallowing‐induced atrial tachycardia in a patient with genotyped hypertrophic cardiomyopathy (2020)
  • Author(s): Fujino Noboru、Hayashi Kenshi、Sakata Kenji、Tada Hayato、Nakanishi Chiaki、Tsuda Toyonobu、Nomura Akihiro、Yoshida Shohei、Sakamoto Yuichiro、Kato Takeshi、Furusho Hiroshi、Kawashiri Masa‐aki、Takamura Masayuki
  • Journal Title: Clinical Case Reports Volume: 8 Pages: 1809～1813
  • DOI: 10.1002/ccr3.3015
  • Peer Reviewed
• [Journal Article] Prolongation of QT interval after pulmonary vein isolation for paroxysmal atrial fibrillation (2020)
  • Author(s): Chikata Akio、Kato Takeshi、Usuda Kazuo、Fujita Shuhei、Maruyama Michiro、Otowa Kan‐ichi、Usuda Keisuke、Niwa Satoru、Tsuda Toyonobu、Hayashi Kenshi、Takamura Masayuki
  • Journal Title: Journal of Cardiovascular Electrophysiology Volume: 31 Pages: 2371～2379
  • DOI: 10.1111/jce.14625
  • Peer Reviewed
• [Journal Article] Personalized medicine for cardiovascular diseases (2020)
  • Author(s): Tada Hayato、Fujino Noboru、Nomura Akihiro、Nakanishi Chiaki、Hayashi Kenshi、Takamura Masayuki、Kawashiri Masa-aki
  • Journal Title: Journal of Human Genetics Volume: 66 Pages: 67～74
  • DOI: 10.1038/s10038-020-0818-7
  • Peer Reviewed
• [Journal Article] Enhancing rare variant interpretation in inherited arrhythmias through quantitative analysis of consortium disease cohorts and population controls (2020)
  • Author(s): Hayashi Kenshi, et al.
  • Journal Title: Genetics in Medicine Volume: 23 Pages: 47～58
  • DOI: 10.1038/s41436-020-00946-5
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hokuriku-plus familial hypercholesterolaemia registry study: rationale and study design (2020)
  • Author(s): Hayashi Kenshi, et al.
  • Journal Title: BMJ Open Volume: 10 Pages: e038623～e038623
  • DOI: 10.1136/bmjopen-2020-038623
  • Peer Reviewed
• [Presentation] 遺伝情報を考慮した不整脈診療 (2021)
  • Author(s): 林 研至
  • Organizer: 不整脈心電学会東海北陸地方会
  • Invited
• [Presentation] Rare variant of the Glycerol-3-Phosphate Dehydrogenase-1 Like (GPD1-L) Gene in Japanese Patients with Brugada Syndrome (2021)
  • Author(s): Keisuke Usuda, Kenshi Hayashi, et al.
  • Organizer: JCS2021
• [Presentation] Clinical Characteristics and Outcomes in Elderly Japanese Patients With Nonvalvular Atrial Fibrillation -Insights From the Results of Japanese AF Registry- (2021)
  • Author(s): Toyonobu Tsuda, Kenshi Hayashi, et al.
  • Organizer: JCS2021
• [Presentation] Atrial fibrillation as an independent predictor for developing renal dysfunction (2021)
  • Author(s): Yoshihiro Tanaka, Kenshi Hayashi, et al.
  • Organizer: JCS2021
• [Presentation] Rare SCN10A variants associated with cardiac conduction system diseases (2020)
  • Author(s): Kenshi Hayashi, Noboru Fujino, Hiroshi Furusho, Soichiro Usui, Kenji Sakata, Takeshi Kato, Toyonobu Tsuda, Satoru Niwa, Keisuke Usuda, Kazutaka Takeuchi, Masa-aki Kawahiri, and Masayuki Takamura.
  • Organizer: ESC2020
  • Int'l Joint Research
• [Presentation] Lamin A Deficiency Leads to Electrophysiological and Nuclear Abnormalities Reminiscent of Atrial Cardiomyopathy in Zebrafish (2020)
  • Author(s): Teramoto, Hayashi K, Beerens M, Chiang D, Kiviniemi T, Satoh M, KC A, Burch M, Zhu W , Dai W, Liu J, Mahadevan A, Buys E, and MacRae C
  • Organizer: AHA2020
  • Int'l Joint Research
• [Book] 不整脈診療ロジック×プラクティス (2020)
  • Author(s): 加藤 武史、松尾 征一郎
  • Total Pages: 392
  • Publisher: 南江堂
  • ISBN: 978-4524248940