エクソームシーケンスを用いた新規高比重リポ蛋白代謝制御因子の発見とその機能解析

2015 Fiscal Year Research-status Report

• Project/Area Number: 15K09118
• Research Institution: Kanazawa University
• Principal Investigator: 川尻 剛照 金沢大学, 医学系, 准教授 (90345637)
• Co-Investigator(Kenkyū-buntansha): 野原 淳 金沢大学, 医薬保健学総合研究科, 特任准教授 (50313648) ; 多田 隼人 金沢大学, 大学病院, 助教 (90623653)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 高比重リポ蛋白 / 動脈硬化 / 全エクソームシーケンス

Outline of Annual Research Achievements

動脈硬化症予防を目的に、症例に立脚した高比重リポ蛋白（HDL）代謝異常症の病態解明を目指した研究を試みた。発端者は①著明な高HDLコレステロール血症（170mg/dL）と②著明な低HDLコレステロール血症（2、6mg/dL）症例である。
低HDLコレステロールコレステロール血症の遺伝子DNAを用い、②2人の低HDLコレステロール血症患者と2人の家系内正常者、4人の非血縁正常者を対象に全エクソームシーケンスを行った。SnpEffを用い両性変異、1%以上の頻度を認める変異、劣性遺伝に合致しない変異、C-スコア<10を除外した結果、ABCA1遺伝子にコンパウンドヘテロ変異（c.6230C>A/c.6137G>A、c.2842G>A/c.1130C>T）を認め、いずれの症例もTangier病と診断した（Tada H, Kawashiri M et al. Atherosclerosis 240:324-329;2015）。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

全エクソームシーケンス法で著明な低HDLコレステロール血症（2、6mg/dL）2例の遺伝子解析を終了した。新規遺伝子ではなかったため、機能解析は見送ることにした。

Strategy for Future Research Activity

著明な高HDLコレステロール血症（170mg/dL）症例に見出された新規候補「分子X」に関し、機能解析を行う予定である。具体的には、遺伝子過剰発現マウス・遺伝子欠損マウスを樹立し、脂質値を検討し、アポE欠損マウスと交配し、遺伝子改変マウスの動脈硬化病理組織を検討する予定である。

Causes of Carryover

低HDLコレステロール血症の原因遺伝子がABCA1（Tangier病）で新規でなかったため、機能解析を行う必要性がなくなったため。

Expenditure Plan for Carryover Budget

高HDLコレステロール血症の新規候補「分子X」の機能解析が現在のところ進捗していない。遺伝子欠損マウス、遺伝子導入マウスの樹立にはかなりの出費が想定されており、これらに充てる予定である。

Research Products

• [Journal Article] Common and Rare Variant Association Study for Plasma Lipids and Coronary Artery Disease. (2016)
  • Author(s): Tada H, Kawashiri MA, Konno T, Yamagishi M, Hayashi K.
  • Journal Title: J Atheroscler Thromb. Volume: 23 Pages: 241-256
  • DOI: 10.5551/jat.31393.
  • Peer Reviewed / Open Access
• [Journal Article] Lipoprotein(a) in Familial Hypercholesterolemia With Proprotein Convertase Subtilisin/Kexin Type 9 (PCSK9) Gain-of-Function Mutations. (2016)
  • Author(s): Tada H, Kawashiri MA, Yoshida T, Teramoto R, Nohara A, Konno T, Inazu A, Mabuchi H, Yamagishi M, Hayashi K.
  • Journal Title: Circ J. Volume: 80 Pages: 512-518
  • DOI: 10.1253/circj.CJ-15-0999.
  • Peer Reviewed / Open Access
• [Journal Article] Usefulness of Electrocardiographic Voltage to Determine Myocardial Fibrosis in Hypertrophic Cardiomyopathy. (2016)
  • Author(s): Konno T, Nagata Y, Teramoto R, Fujino N, Nomura A, Tada H, Sakata K, Furusho H, Takamura M, Nakamura H, Kawashiri MA, Yamagishi M, Hayashi K.
  • Journal Title: Am J Cardiol. Volume: 117 Pages: 443-449
  • DOI: 10.1016/j.amjcard.2015.11.015.
  • Peer Reviewed / Open Access
• [Journal Article] A de novo mutation of the LDL receptor gene as the cause of familial hypercholesterolemia identified using whole exome sequencing. (2016)
  • Author(s): Tada H, Hosomichi K, Okada H, Kawashiri MA, Nohara A, Inazu A, Tomizawa S, Tajima A, Mabuchi H, Hayashi K.
  • Journal Title: Clin Chim Acta. Volume: 453 Pages: 194-196
  • DOI: 10.1016/j.cca.2015.12.028.
  • Peer Reviewed / Open Access
• [Journal Article] Exome sequencing in suspected monogenic dyslipidemias. (2015)
  • Author(s): Stitziel NO, Kawashiri MA, Nohara A, Yamagishi M, Kathiresan S et al.
  • Journal Title: Circ Cardiovasc Genet. Volume: 8 Pages: 343-350
  • DOI: 10.1161/CIRCGENETICS.114.000776.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Infantile Cases of Sitosterolaemia with Novel Mutations in the ABCG5 Gene: Extreme Hypercholesterolaemia is Exacerbated by Breastfeeding. (2015)
  • Author(s): Tada H, Kawashiri MA, Takata M, Matsunami K, Imamura A, Matsuyama M, Sawada H, Nunoi H, Konno T, Hayashi K, Nohara A, Inazu A, Kobayashi J, Mabuchi H, Yamagishi M.
  • Journal Title: JIMD Rep. Volume: 21 Pages: 115-122
  • DOI: 10.1007/8904_2014_404.
  • Peer Reviewed
• [Journal Article] Impact of Enhanced Production of Endogenous Heme Oxygenase-1 by Pitavastatin on Survival and Functional Activities of Bone Marrow-derived Mesenchymal Stem Cells. (2015)
  • Author(s): Kawashiri MA, Nakanishi C, Tsubokawa T, Shimojima M, Yoshida S, Yoshimuta T, Konno T, Yamagishi M, Hayashi K.
  • Journal Title: J Cardiovasc Pharmacol. Volume: 65 Pages: 601-606
  • DOI: 10.1097/FJC.0000000000000231.
  • Peer Reviewed / Open Access
• [Journal Article] Expansion of the clinical application of optical coherence tomography to percutaneous coronary intervention and assessment of the instability of coronary atherosclerosis. (2015)
  • Author(s): Sakata K, Kawashiri MA, Hayashi K.
  • Journal Title: Circ J. Volume: 79 Pages: 513-514
  • DOI: 10.1253/circj.CJ-15-0103.
  • Peer Reviewed / Open Access
• [Journal Article] Extreme Contrast of Postprandial Remnant-Like Particles Formed in Abetalipoproteinemia and Homozygous Familial Hypobetalipoproteinemia. (2015)
  • Author(s): Kawashiri MA, Tada H, Hashimoto M, Taniyama M, Nakano T, Nakajima K, Inoue T, Mori M, Nakanishi C, Konno T, Hayashi K, Nohara A, Inazu A, Koizumi J, Ishihara H, Kobayashi J, Hirano T, Mabuchi H, Yamagishi M.
  • Journal Title: JIMD Rep. Volume: 22 Pages: 85-94
  • DOI: 10.1007/8904_2015_415.
  • Peer Reviewed / Open Access
• [Journal Article] Whole exome sequencing combined with integrated variant annotation prediction identifies asymptomatic Tangier disease with compound heterozygous mutations in ABCA1 gene. (2015)
  • Author(s): Tada H, Kawashiri MA, Nohara A, Saito R, Tanaka Y, Nomura A, Konno T, Sakata K, Fujino N, Takamura T, Inazu A, Mabuchi H, Yamagishi M, Hayashi K.
  • Journal Title: Atherosclerosis. Volume: 240 Pages: 324-329
  • DOI: 10.1016/j.atherosclerosis.2015.04.003.
  • Peer Reviewed / Open Access
• [Journal Article] Whole Exome Sequencing in Monogenic Dyslipidemias. (2015)
  • Author(s): Tada H, Kawashiri MA, Yamagishi M, Hayashi K.
  • Journal Title: J Atheroscler Thromb. Volume: 22 Pages: 881-885
  • DOI: 10.5551/jat.ED016.
  • Peer Reviewed / Open Access
• [Journal Article] Right ventricular hypertrophy is associated with cardiovascular events in hypertrophic cardiomyopathy: evidence from study with magnetic resonance imaging. (2015)
  • Author(s): Nagata Y, Konno T, Fujino N, Hodatsu A, Nomura A, Hayashi K, Nakamura H, Kawashiri MA, Yamagishi M.
  • Journal Title: Can J Cardiol. Volume: 31 Pages: 702-708
  • DOI: 10.1016/j.cjca.2014.12.036.
  • Peer Reviewed / Open Access
• [Journal Article] Impact of Elevated D-Dimer on Diagnosis of Acute Aortic Dissection With Isolated Neurological Symptoms in Ischemic Stroke. (2015)
  • Author(s): Yoshimuta T, Yokoyama H, Okajima T, Tanaka H, Toyoda K, Nagatsuka K, Higashi M, Hayashi K, Kawashiri MA, Yasuda S, Yamagishi M.
  • Journal Title: Circ J. Volume: 79 Pages: 1841-1845
  • DOI: 10.1253/circj.CJ-15-0050.
  • Peer Reviewed / Open Access
• [Journal Article] Atrial fibrillation: an inherited cardiovascular disease-a commentary on genetics of atrial fibrillation: from families to genomes. (2015)
  • Author(s): Tada H, Kawashiri MA, Yamagishi M, Hayashi K.
  • Journal Title: J Hum Genet. Volume: 61 Pages: 3-4
  • DOI: 10.1038/jhg.2015.63.
  • Peer Reviewed / Open Access
• [Journal Article] Xiphodynia Mimicking Acute Coronary Syndrome. (2015)
  • Author(s): Tanaka Y, Sakata K, Waseda Y, Fujimura T, Yamada K, Oyama T, Kawashiri MA, Yamagishi M.
  • Journal Title: Intern Med. Volume: 54 Pages: 1563-1566
  • DOI: 10.2169/internalmedicine.54.3449.
  • Peer Reviewed / Open Access
• [Journal Article] Electrocardiographic QRS Fragmentation as a Marker for Myocardial Fibrosis in Hypertrophic Cardiomyopathy. (2015)
  • Author(s): Konno T, Hayashi K, Fujino N, Oka R, Nomura A, Nagata Y, Hodatsu A, Sakata K, Furusho H, Takamura M, Nakamura H, Kawashiri MA, Yamagishi M.
  • Journal Title: J Cardiovasc Electrophysiol. Volume: 26 Pages: 1081-1087
  • DOI: 10.1111/jce.12742.
  • Peer Reviewed / Open Access
• [Journal Article] Functional Characterization of Rare Variants Implicated in Susceptibility to Lone Atrial Fibrillation. (2015)
  • Author(s): Hayashi K, Konno T, Tada H, Tani S, Liu L, Fujino N, Nohara A, Hodatsu A, Tsuda T, Tanaka Y, Kawashiri MA, Ino H, Makita N, Yamagishi M.
  • Journal Title: Circ Arrhythm Electrophysiol. Volume: 8 Pages: 1095-1104
  • DOI: 10.1161/CIRCEP.114.002519.
  • Peer Reviewed / Open Access
• [Journal Article] Impact of real-time three-dimensional transesophageal echocardiography on procedural success for mitral valve repair. (2015)
  • Author(s): Mori M, Yoshimuta T, Ohira M, Yagi M, Sakata K, Konno T, Kawashiri MA, Tomita S, Watanabe G, Iino K, Takemura H, Yamagishi M, Hayashi K.
  • Journal Title: J Echocardiogr. Volume: 13 Pages: 100-106
  • DOI: 10.1007/s12574-015-0255-3.
  • Peer Reviewed / Open Access
• [Journal Article] Clinical characteristics of Japanese patients with severe hypertriglyceridemia. (2015)
  • Author(s): Tada H, Kawashiri MA, Nakahashi T, Yagi K, Chujo D, Ohbatake A, Mori Y, Mori S, Kometani M, Fujii H, Nohara A, Inazu A, Mabuchi H, Yamagishi M, Hayashi K.
  • Journal Title: J Clin Lipidol. Volume: 9 Pages: 519-524
  • DOI: 10.1016/j.jacl.2015.05.004.
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of Autosomal Dominant Hypercholesterolemia Caused by PCSK9 Gain of Function Mutations and Its Specific Treatment With Alirocumab, a PCSK9 Monoclonal Antibody. (2015)
  • Author(s): Hopkins PN, , Mabuchi H, Kawashiri MA, Nohara A, Tada H, Swergold GD et al.
  • Journal Title: Circ Cardiovasc Genet. Volume: 8 Pages: 823-831
  • DOI: 10.1161/CIRCGENETICS.115.001129.
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Determination of Early and Late Endothelial Progenitor Cells in Peripheral Circulation and Their Clinical Association with Coronary Artery Disease. (2015)
  • Author(s): Tagawa S, Nakanishi C, Mori M, Yoshimuta T, Yoshida S, Shimojima M, Yokawa J, Kawashiri MA, Yamagishi M, Hayashi K.
  • Journal Title: Int J Vasc Med. Volume: 2015 Pages: 674213
  • DOI: 10.1155/2015/674213.
  • Peer Reviewed / Open Access
• [Journal Article] Abrupt progression of ventricular septal perforation after primary angioplasty for acute myocardial infarction. (2015)
  • Author(s): Nakahashi T, Sakata K, Tsuda T, Tanaka Y, Ohira M, Mori M, Takata M, Kawashiri MA, Takemura H, Yamagishi M, Hayashi K.
  • Journal Title: Cardiovasc Diagn Ther. Volume: 5 Pages: 479-483
  • DOI: 10.3978/j.issn.2223-3652.2015.06.06.
  • Peer Reviewed / Open Access
• [Presentation] PCSK9の発見と新たなコレステロール低下剤の開発 (2015)
  • Author(s): 川尻剛照
  • Organizer: 日本人類遺伝学会第60回大会
  • Place of Presentation: 東京
  • Year and Date: 2015-10-14 – 2015-10-17
  • Invited
• [Presentation] 家族性高コレステロール血症の診断と治療の重要性 冠動脈疾患発症抑制のために (2015)
  • Author(s): 川尻剛照
  • Organizer: 第24回日本心血管インターベンション治療学会
  • Place of Presentation: 福岡
  • Year and Date: 2015-07-30 – 2015-08-01
• [Presentation] A patient with compound heterozygous familial hypercholesterolemia accompanied with supravalvular aortic stenosis detected by Whole Exome Sequencing (2015)
  • Author(s): 川尻剛照、多田隼人、井上健、中西千明、森三佳、野原淳、稲津明広、馬渕宏、山岸正和
  • Organizer: 第47回日本動脈硬化学会総会・学術集会
  • Place of Presentation: 仙台
  • Year and Date: 2015-07-09 – 2015-07-10
• [Presentation] 家族性高コレステロール血症を見逃さない (2015)
  • Author(s): 川尻剛照
  • Organizer: 第1回国際心血管薬物療法学会日本部会学術集会
  • Place of Presentation: 京都
  • Year and Date: 2015-06-20 – 2015-06-21
  • Invited
• [Presentation] Vascular Complications in Japanese FH Patients (2015)
  • Author(s): Kawashiri M
  • Organizer: The Satellite Symposium of the ISA 2015 in Tokyo
  • Place of Presentation: Tokyo
  • Year and Date: 2015-05-21
  • Int'l Joint Research / Invited
• [Presentation] Myocardial ischemia treated with PCI and plaque regression by lipid lowering & blood pressure controlling assessed by intravascular ultrasonography (2015)
  • Author(s): Kawashiri M, Yamagishi M, MILLION study group
  • Organizer: The Society for Cardiovascular Angiography and Intervention
  • Place of Presentation: San Diego
  • Year and Date: 2015-05-06 – 2015-05-09
  • Int'l Joint Research
• [Book] 診断と治療のABC 高LDL-C血症・低HDL-C血症 (2015)
  • Author(s): 川尻剛照
  • Total Pages: 8
  • Publisher: 最新医学社
• [Book] PCSK9阻害剤 (2015)
  • Author(s): 川尻剛照
  • Total Pages: 9
  • Publisher: メディカルビュー社 血管医学
• [Book] PCSK9をターゲットとした新しい治療を識る (2015)
  • Author(s): 川尻剛照
  • Total Pages: 6
  • Publisher: メディカルビュー社 Heart View
• [Book] LDLアフェレシス (2015)
  • Author(s): 川尻剛照、多田隼人、山岸正和
  • Total Pages: 7
  • Publisher: メディカルレビュー社 The Lipid
• [Book] 頚動脈エコーで石灰化していない低輝度～等輝度プラークに効果的な薬剤は？ (2015)
  • Author(s): 川尻剛照
  • Total Pages: 2
  • Publisher: 日本医事新報社 週刊日本医事新報