Elucidation of pathophysiological mechanisms and the development of innovative therapies for lethal arrhythmias caused by calmodulin mutations

• Project/Area Number: 18K15847
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 53020:Cardiology-related
• Research Institution: Kyoto University
• Principal Investigator: Yamamoto Yuta 京都大学, 医学研究科, 研究員 (10802762)
• Project Period (FY): 2018-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2019: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2018: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: カルモジュリン / 不整脈 / iPS細胞 / QT延長症候群 / アンチセンス核酸 / 遺伝子治療 / アンチセンス / 遺伝性心疾患

Outline of Final Research Achievements

Recently, mutations in CALM genes (CALM1-3) encoding calmodulin (CaM) are reported to be associated with severe early-onset arrhythmias known as the calmodulinopathy. However, the pathophysiological mechanisms of calmodulinopathy remains unknown. Moreover, there is no effective treatment for calmodulinopathy.
In the present study, we analyzed KCNQ1 channel function using patch clamp technique and elucidate the KCNQ1 channel dysfunction using human iPS cell model that may explain the clinical phenotype of the LQTS patient carrying the CALM2-N98S mutation. In addition, we demonstrated that antisense oligonucleotides-based therapy might be a promising strategy for the treatment of calmodulinopathy.

Academic Significance and Societal Importance of the Research Achievements

カルモジュリン関連不整脈疾患は若年発症で重篤な症状を引き起こすことが知られているが、その詳細な発症機序は不明な点が多く、有効な治療法も確立されておらず、本疾患の病態を解明し、新規治療法を開発する社会的な意義は大きい。本研究によりCALM2-N98S変異において、KCNQ1チャネルの機能障害を明らかにしたが、これは実際の治療法の選択に関して重要な知見であると考えられる。また、これまでの治療法とは異なりアンチセンス核酸を用いた治療法は変異カルモジュリンをターゲットとした全く新しい治療法であり、本質的な治療となる可能性があり、学術的意義は大きいと考える。

Research Products

• [Journal Article] Phenotype-Based High-Throughput Classification of Long QT Syndrome Subtypes Using Human Induced Pluripotent Stem Cells (2019)
  • Author(s): Yoshinaga Daisuke、Baba Shiro、Makiyama Takeru、Shibata Hirofumi、Hirata Takuya、Akagi Kentaro、Matsuda Koichi、Kohjitani Hirohiko、Wuriyanghai Yimin、Umeda Katsutsugu、Yamamoto Yuta、Conklin Bruce R.、Horie Minoru、Takita Junko、Heike Toshio
  • Journal Title: Stem Cell Reports Volume: 13 Issue: 2 Pages: 394-404
  • DOI: 10.1016/j.stemcr.2019.06.007
  • NAID: 120006786948
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Calmodulinopathy(カルモジュリン遺伝子関連不整脈疾患) iPS細胞技術を用いた疾患モデルの作製と遺伝子治療へのアプローチ (2019)
  • Author(s): 山本 雄大,牧山 武
  • Journal Title: 心電図 Volume: 39 Pages: 273-282
  • NAID: 130007903608
• [Journal Article] Clinical Manifestations and Long-Term Mortality in <i>Lamin A/C</i> Mutation Carriers From a Japanese Multicenter Registry (2018)
  • Author(s): 14.Nakajima K, Aiba T, Makiyama T, Nishiuchi S, Ohno S, Kato K, Yamamoto Y, Doi T, Shizuta S, Onoue K, Yagihara N, Ishikawa T, Watanabe I, Kawakami H, Oginosawa Y, Murakoshi N, Nogami A, Aonuma K, Saito Y, Kimura T, Yasuda S, Makita N, Shimizu W, Horie M, Kusano K.
  • Journal Title: Circulation Journal Volume: 82 Issue: 11 Pages: 2707-2714
  • DOI: 10.1253/circj.CJ-18-0339
  • NAID: 40021697176
  • ISSN: 1346-9843, 1347-4820
  • Year and Date: 2018-10-25
  • Peer Reviewed / Open Access
• [Journal Article] Complex aberrant splicing in the induced pluripotent stem cell?derived cardiomyocytes from a patient with long QT syndrome carrying KCNQ1-A344Aspl mutation (2018)
  • Author(s): Wuriyanghai Yimin、Makiyama Takeru、Sasaki Kenichi、Kamakura Tsukasa、Yamamoto Yuta、Hayano Mamoru、Harita Takeshi、Nishiuchi Suguru、Chen Jiarong、Kohjitani Hirohiko、Hirose Sayako、Yokoi Fumika、Gao Jingshan、Chonabayashi Kazuhisa、Watanabe Ken、Ohno Seiko、Yoshida Yoshinori、Kimura Takeshi、Horie Minoru
  • Journal Title: Heart Rhythm Volume: 15 Issue: 10 Pages: 1566-1574
  • DOI: 10.1016/j.hrthm.2018.05.028
  • Peer Reviewed / Open Access
• [Presentation] Modeling Overlap Syndrome Of Long QT And Brugada Syndrome Associated With CACNA1C-E1115K Which Alters Ion Selectivity With Induced Pluripotent Stem Cells (2019)
  • Author(s): Asami Kashiwa, Takeru Makiyama, Hirohiko Kohjitani, Yimin Wuriyanghai, Yuta Yamamoto, Gao Jungshan, Hai Huang, Taisuke Ishikawa, Seiko Ohno, Futoshi Toyoda, , Seiichi Sato, Minoru Horie , Naomasa Makita, Takeshi Kimura.
  • Organizer: 40th Heart Rhythm Scientific Sessions
  • Int'l Joint Research
• [Presentation] Automatic Computational Action Potential Analyzing System Enables Accurate Drug Response Interpretation and Prediction in Human Induced Pluripotent Stem Cell-derived Cardiomyocytes (2019)
  • Author(s): Hirohiko Kohjitani, Shigeya Kouda, Yukiko Himeno, Takeru Makiyama, Asami Kashiwa, Yimin Wuriyanghai, Yuta Yamamoto, Minoru Horie, Takeshi Kimura, Akinori Noma, Akira Amano.
  • Organizer: 40th Heart Rhythm Scientific Sessions
  • Int'l Joint Research
• [Presentation] Structural Insights into Catecholaminergic Polymorphic Ventricular Tachycardia-associated RyR2 Mutant Channels using a Three-dimensional In Silico Model. (2019)
  • Author(s): J. Gao, T. Makiyama, S. Ohno, Y. Yamamoto, Y. Wuriyanghai, H. Kohjitani, A. Kashiwa, H. Huang, K. Katou, M. Horie, T. Kimura.
  • Organizer: European Society of Cardiology Congress 2019
  • Int'l Joint Research
• [Presentation] KCNQ1 Channel Dysfunction in human induced pluripotent stem cell-derived cardiomyocytes carrying a CALM2-N98S mutation associated with long QT syndrome (2019)
  • Author(s): Yuta Yamamoto, Takeru Makiyama, Yimin Wuriyanghai, Hirohiko Kohjitani, Sayako Hirose, Jingshan Gao, Asami Kashiwa, Huang Hai, Takanori Aizawa, Tomohiko Imamura, Taisuke Ishikawa, Seiko Ohno, Kazuhisa Chonabayashi, Hideki Motomura, Yoshinori Yoshida, Minoru Horie, Naomasa Makita, Takeshi Kimura
  • Organizer: American Heart Association Scientific Sessions 2019
  • Int'l Joint Research
• [Presentation] Morphological and electrophysiological characterization of homozygous lamin A/C knockout human induced-pluripotent stem cell-derived cardiomyocytes. American Heart Association Scientific Sessions 2019 (2019)
  • Author(s): Yimin Wuriyanghai, Takeru Makiyama, Yuta Yamamoto, Hirohiko Kohjitani, Sayako Hirose, Jingshan Gao, Asami Kashiwa, Hai Huang, Takanori Aizawa, Tomohiko Imamura, Seiko Ohno, Minoru Horie, Takeshi Kimura
  • Organizer: American Heart Association Scientific Sessions 2019
  • Int'l Joint Research
• [Presentation] Response of KCNQ1 to Adrenergic Stimulation was Impaired in Human Induced Pluripotent Stem Cell-derived Cardiomyocytes Carrying a CALM2-N98S Mutation Associated with Long QT Syndrome. (2019)
  • Author(s): Yuta Yamamoto, Takeru Makiyama, Yimin Wuriyanghai, Hirohiko Kohjitani, Sayako Hirose, Jingshan Gao, Asami Kashiwa, Huang Hai, Takanori Aizawa, Tomohiko Imamura, Taisuke Ishikawa, Seiko Ohno, Kazuhisa Chonabayashi, Hideki Motomura, Yoshinori Yoshida, Minoru Horie, Naomasa Makita, Takeshi Kimura
  • Organizer: 12th Asia Pacific Heart Rhythm Society Scientific Session (APHRS2019)
  • Int'l Joint Research
• [Presentation] Algorithmic Auto-Recreation System of hiPSC-CMs Simulation and Prediction of Drug Testing. (2019)
  • Author(s): Hirohiko Kohjitani, Shigeya Kouda, Yukiko Himeno, Takeru Makiyama, Asami Kashiwa, Yimin Wuriyanghai, Yuta Yamamoto, Minoru Horie, Takeshi Kimura, Akinori Noma, Akira Amano
  • Organizer: 12th Asia Pacific Heart Rhythm Society Scientific Session (APHRS2019)
  • Int'l Joint Research
• [Presentation] Patients characteristics of idiopathic ventricular fibrillation carrying SCN5A mutations without Brugada or long QT syndrome. (2019)
  • Author(s): Takanori Aizawa, Takeru Makiyama, Hai Huang, Tomohiko Imamura, JingShan Gao, Yimin Wuriyanghai, Takeru Makiyama, Yuta Yamamoto, Hirohiko Kohjitani, Sayako Hirose, Jingshan Gao, Asami Kashiwa, Hai Huang, Wuriyanghai Yimin, Hirohiko Kohjitani, Yuta Yamamoto, Seiko Ohno, Minoru Horie
  • Organizer: 12th Asia Pacific Heart Rhythm Society Scientific Session (APHRS2019)
  • Int'l Joint Research
• [Presentation] Algorithmic Auto-Recreation System of hiPSC-CMs Simulation and Prediction of Drug Testing (2019)
  • Author(s): Hirohiko Kohjitani, Shigeya Kouda, Yukiko Himeno, Takeru Makiyama, Hai Huang, Jingshan Gao, Asami Kashiwa, Yimin Wuriyanghai, Daisuke Yoshinaga, Takeshi Kimura, Akira Amano.
  • Organizer: 第83回日本循環器学会学術集会
• [Presentation] Overlapping Sick Sinus Syndrome Associated with SCN5A Mutations in Patients with Brugada Syndrome (2019)
  • Author(s): Asami Kashiwa, Takeru Makiyama, Seiko Ohno, Hisaaki Aoki, Hirohiko Kohjitani Yimin Wuriyanghai, Yuta Yamamoto, Jingshan Gao, Huang Hai, Minoru Horie, Takeshi Kimura.
  • Organizer: 第83回日本循環器学会学術集会
• [Presentation] Three-Dimensional Structural Analysis of Mutant Ryanodine Receptor Type 2 Channels Associated with Catecholaminergic Polymorphic Ventricular Tachycardia. (2019)
  • Author(s): Jingshan Gao, Takeru Makiyama, Seiko Ohno, Yuta Yamamoto, Yimin Wuriyanghai, Hirohiko Kohjitani, Asami Kashiwa, Minoru Horie, Takeshi Kimura.
  • Organizer: 第83回日本循環器学会学術集会
• [Presentation] Modeling Overlapping Phenotypes of Long-QT and Brugada Syndrome with CACNA1C-E1115K that Alters Ion Selectivity of the Cardiac L-Type Calcium Channel Using Induced Pluripotent Stem Cells. (2019)
  • Author(s): Asami Kashiwa, Takeru Makiyama, Hirohiko Kohjitani, Yimin Wuriyanghai, Yuta Yamamoto, Jingshan Gao, Hai Huang, Taisuke Ishikawa, Seiko Ohno, Futoshi Toyoda, Seiichi Sato, Minoru Horie, Naomasa Makita, Takeshi Kimura
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Algorithmic Auto-Recreation System of hiPSC-CMs Simulation and Prediction of Drug Testing. (2019)
  • Author(s): Hirohiko Kohjitani, Shigeya Kouda, Yukiko Himeno, Takeru Makiyama, Hai Huang, Jingshan Gao, Asami Kashiwa, Yimin Wuriyanghai, Daisuke Yoshinaga, Takeshi Kimura, Akira Amano.
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Single Cell Electrophysiological Analysis of iPS Cell-derived Cardiomyocytes Generated from Long-QT Syndrome Patients Carrying a CALM2 Mutation Using a Membrane Potential Imaging System. (2019)
  • Author(s): Yuta Yamamoto, Takeru Makiyama(代発表), Yimin Wuriyanghai, Hirohiko Kohjitani, Sayako Hirose, Jingshan Gao, Asami Kashiwa, Huang Hai, Taisuke Ishikawa, Seiko Ohno, Kazuhisa Chonabayashi, Kenji Suda, Yoshinori Yoshida, Minoru Horie, Naomasa Makita, Takeshi Kimura,
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Structural Insights into Catecholaminergic Polymorphic Ventricular Tachycardia-associated RyR2 Mutant Channels using a Three-dimensional in Silico Model. (2019)
  • Author(s): Jingshan Gao, Takeru Makiyama, Seiko Ohno, Yuta Yamamoto, Yimin Wuriyanghai, Hirohiko Kohjitani, Asami Kashiwa, Hai Huang, Kouichi Katou, Minoru Horie, Takeshi Kimura.
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Phenotype-based High-throughput Classification of Long-QT Syndrome Subtypes Using Human Induced Pluripotent Stem Cells. (2019)
  • Author(s): Daisuke Yoshinaga, Shiro Baba, Takeru Makiyama, Hirofumi Shibata, Takuya Hirata, Kentaro Akagi, Koichi Matsuda, Hirohiko Kohjitani, Yimin Wuriyanghai, Katsutsugu Umeda, Yuta Yamamoto, Bruce R. Conklin, Minoru Horie, Junko Takita, Toshio Heike.
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Complex aberrant splicing in the induce pluripotent stem cell-derived cardiomyocytes from a patient with long-QT syndrome carrying KCNQ1-A344Aspl mutation. (2019)
  • Author(s): Wuriyanghai Y, Makiyama T, Sasaki K, Kamakura T, Yamamoto Y, Hayano M, Harita T, Nishiuchi S, Chen J, Kohjitani H, Hirose S, Yokoi F, Gao J, Chonabayashi K, Watanabe K, Ohno S, Yoshida Y, Kimura T, Horie M
  • Organizer: 第66回日本不整脈心電学会学術大会
• [Presentation] Cell-specific Mathematical Modeling of hiPSC-CMs and Its Potential for Prediction of Drug Testing. (2018)
  • Author(s): 糀谷泰彦、山本雄大、牧山武
  • Organizer: 23rd Annual Scientific Meeting of the International Society of Cardiovascular Pharmacotherapy
  • Int'l Joint Research
• [Presentation] Development of Homozygous Lamin A/C Deficient Human Induced Pluripotent Stem Cell Model for Analyzing the Pathophysiological Mechanism of Lamin A/C-related Cardiomyopathy. (2018)
  • Author(s): Wuriyanghai Yimin、山本雄大、牧山武
  • Organizer: 23rd Annual Scientific Meeting of the International Society of Cardiovascular Pharmacotherapy
  • Int'l Joint Research
• [Presentation] Significance of cell-specific precise computer simulation using new mathematical models of human induced pluripotent stem cell derived cardiomyocyte in drug testing. (2018)
  • Author(s): 糀谷泰彦、山本雄大、牧山武
  • Organizer: ESC Congress 2018
  • Int'l Joint Research
• [Presentation] Overlapping Brugada Syndrome Associated with SCN5A Mutations in Patients with Sick Sinus Syndrome Treated with Permanent Pacemakers. (2018)
  • Author(s): 柏麻美、山本雄大、牧山武
  • Organizer: The 11th Asia Pacific Heart Rhythm Society Scientific Session
  • Int'l Joint Research
• [Presentation] Single Cell Electrophysiological Analysis of iPS Cell-derived Cardiomyocytes generated from Long-QT syndrome patients carrying a CALM2 mutation using membrane voltage imaging system. (2018)
  • Author(s): 山本雄大、牧山武
  • Organizer: The 11th Asia Pacific Heart Rhythm Society Scientific Session
  • Int'l Joint Research
• [Presentation] Analysis of Disease-Causing Mechanism of Lamin A/C-Related Cardiomyopathy Using Homozygous Lamin A/C Knock out Human Induced Pluripotent Stem Cell Model. (2018)
  • Author(s): Wuriyanghai Yimin、山本雄大、牧山武
  • Organizer: The 11th Asia Pacific Heart Rhythm Society Scientific Session
  • Int'l Joint Research
• [Presentation] Three-dimensional Structural Analysis of Mutant Ryanodine Receptor Type 2 Channels Associated with Catecolaminergic Polymorphic Ventricular Tachycardia. (2018)
  • Author(s): 高景山、山本雄大、牧山武
  • Organizer: The 11th Asia Pacific Heart Rhythm Society Scientific Session
  • Int'l Joint Research
• [Presentation] The Inactivation of L-type Ca2+ Cannel was Impaired in Human iPS Cell Model of Long-QT Syndrome with CALM2-D134H Mutation. (2018)
  • Author(s): 山本雄大、牧山武
  • Organizer: 第 82 回 日本循環器学会学術集会
  • Int'l Joint Research
• [Presentation] Single cell analysis of CALM-related iPS cell model using membrane voltage imaging system. (2018)
  • Author(s): 山本雄大、牧山武
  • Organizer: 第 2 回 日本循環器学会基礎研究フォーラム
• [Patent(Industrial Property Rights)] アンチセンスオリゴヌクレオチド医薬のスクリーニング方法 (2020)
  • Inventor(s): 牧山武、山本雄大、入山友輔、近藤嘉紀、日高夕貴、石川龍太郎
  • Industrial Property Rights Holder: 牧山武、山本雄大、入山友輔、近藤嘉紀、日高夕貴、石川龍太郎
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
• [Patent(Industrial Property Rights)] ＣＡＬＭ２のアンチセンスオリゴヌクレオチド (2020)
  • Inventor(s): 入山友輔、近藤嘉紀、日高夕貴、石川龍太郎、牧山武、山本雄大
  • Industrial Property Rights Holder: 入山友輔、近藤嘉紀、日高夕貴、石川龍太郎、牧山武、山本雄大
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020