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

2017 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, 保健管理センター, 助教 (50313648) ; 多田 隼人 金沢大学, 大学病院, 助教 (90623653)
• Project Period (FY): 2015-04-01 – 2019-03-31
• 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引き続き試みている。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例である。
②低HDLコレステロール血症患者2人（症例1：28歳女性、HDLコレステロール値2mg/dL、症例2：35歳女性、同6mg/dL）と2人の両親（HDLコレステロール値37mg/dL、41mg/dL）、4人の非血縁正常者（平均HDLコレステロール値107mg/dL）の計8人を対象に全エクソームシーケンスを行った。
305,202個の遺伝子多型が見出され、SnpEffを用い良性変異を除外した結果、21,708個のナンセンス変異、ミスセンス変異、スプライス部変異が同定された。さらに1%以上の頻度を認める変異を除外した結果5,192個の変異に絞られ、劣性遺伝に合致する変異、C-スコア10点未満の変異を除外した結果、ABCA1遺伝子変異c.6230C>A、c.6137G>A、c.2842>A、c.1130C>Tが残り、症例1はc.6230C>A/c.6137G>A、症例2はc.2842>A、c.1130C>Tのそれぞれコンパウンドヘテロ接合体（Tangier病）と診断された（Tada H, Kawashiri M et al. Atherosclerosis 240:324-329;2015）。
①著明な高HDLコレステロール血症の原因「分子X」遺伝子を培養細胞に導入し、コレステロール引き抜き能を解析する予定である。

Current Status of Research Progress

4: Progress in research has been delayed.

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った。
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」の機能解析に着手した。遺伝子改変動物を用いた検討は経済的に困難であり、培養細胞を用いたコレステロール引き抜き研究を優先し着手した。

Strategy for Future Research Activity

遺伝子導入培養細胞を用いたコレステロール引き抜き研究を展開する予定である。本研究は防衛医科大学校の研究グループが過去に行っている。共同研究を視野に研究を進める方針である。

Causes of Carryover

平成29年度は、研究代表者の臨床医としての業務が多忙となった上に、卒業を控えた大学院生の指導や実父の病気入院時の世話などが重なったため研究に十分な時間を割くことが困難であった。
繰り越した研究費は、培養細胞を用いたコレステロール引き抜き実験にあてる予定である。

Research Products

• [Journal Article] Prominent Tendon Xanthomas and Abdominal Aortic Aneurysm Associated with Cerebrotendinous Xanthomatosis Identified Using Whole Exome Sequencing. (2018)
  • Author(s): Tada H, Inaba S, Pozharitckaia D, Kawashiri MA.
  • Journal Title: Intern Med. Volume: 57 Pages: 1119-1122
  • DOI: 10.2169/internalmedicine.9687-17.
  • Peer Reviewed
• [Journal Article] Clinical characteristics of spontaneous isolated visceral artery dissection. (2018)
  • Author(s): Tanaka Y, Yoshimuta T, Kimura K, Iino K, Tamura Y, Sakata K, Hayashi K, Takemura H, Yamagishi M, Kawashiri MA.
  • Journal Title: J Vasc Surg. Volume: 67 Pages: 1127-1133
  • DOI: 10.1016/j.jvs.2017.08.054.
  • Peer Reviewed
• [Journal Article] Traumatic Esophageal Injury Detected by Transthoracic Echocardiography. (2018)
  • Author(s): Oe K, Araki T, Hayashi K, Kawashiri MA.
  • Journal Title: Intern Med. Volume: 57 Pages: 441-442
  • DOI: 10.2169/internalmedicine.9494-17.
  • Peer Reviewed
• [Journal Article] Molecular and functional characterization of familial chylomicronemia syndrome. (2018)
  • Author(s): Teramoto R, Tada H, Kawashiri MA, Nohara A, Nakahashi T, Konno T, Inazu A, Mabuchi H, Yamagishi M, Hayashi K.
  • Journal Title: Atherosclerosis. Volume: 269 Pages: 272-278
  • DOI: 10.1016/j.atherosclerosis.2017.11.006.
  • Peer Reviewed
• [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
• [Journal Article] Assessment of arterial stiffness in patients with familial hypercholesterolemia. (2018)
  • Author(s): Tada H, Kawashiri MA, Nohara A, Inazu A, Mabuchi H, Yamagishi M.
  • Journal Title: J Clin Lipidol. Volume: 12 Pages: 397-402
  • DOI: 10.1016/j.jacl.2017.12.002.
  • Peer Reviewed
• [Journal Article] Duration of cardiopulmonary resuscitation in patients without prehospital return of spontaneous circulation after out-of-hospital cardiac arrest: Results from a severity stratification analysis. (2018)
  • Author(s): Funada A, Goto Y, Tada H, Teramoto R, Shimojima M, Hayashi K, Kawashiri MA, Yamagishi M.
  • Journal Title: Resuscitation. Volume: 124 Pages: 69-75
  • DOI: 10.1016/j.resuscitation.2018.01.008.
  • Peer Reviewed
• [Journal Article] Unsolved Antiatherogenic Mechanism of n-3 Polyunsaturated Fatty Acids. (2018)
  • Author(s): Kawashiri MA, Tada H, Yamagishi M.
  • Journal Title: Circ J. Volume: 82 Pages: 332-333
  • DOI: 10.1253/circj.CJ-17-1252.
• [Journal Article] Late Gadolinium Enhancement for Prediction of Mutation-Positive Hypertrophic Cardiomyopathy on the Basis of Panel-Wide Sequencing. (2018)
  • Author(s): Teramoto R, Fujino N, Konno T, Nomura A, Nagata Y, Tsuda T, Tada H, Sakata K, Yamagishi M, Hayashi K, Kawashiri MA.
  • Journal Title: Circ J. Volume: 82 Pages: 1139-1148
  • DOI: 10.1253/circj.CJ-17-1012.
  • Peer Reviewed
• [Journal Article] Development and external validation of a post-discharge bleeding risk score in patients with acute coronary syndrome: The BleeMACS score. (2018)
  • Author(s): Raposeiras-Roubin S, Kawashiri MA, Nakahayshi T, Sakata K, Yamagishi M, Abu-Assi E, et al.
  • Journal Title: Int J Cardiol. Volume: 254 Pages: 10-15
  • DOI: 10.1016/j.ijcard.2017.10.103.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] How to implement clinical guidelines to optimise familial hypercholesterolaemia diagnosis and treatment. (2017)
  • Author(s): Farnier M, Civeira F, Descamps O, Kawashiri MA, Grigore L, Vohnout B
  • Journal Title: Atheroscler Suppl. Volume: 26 Pages: 25-35
  • DOI: 10.1016/S1567-5688(17)30022-3.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Protein-Truncating Variants at the Cholesteryl Ester Transfer Protein Gene and Risk for Coronary Heart Disease. (2017)
  • Author(s): Nomura A, Tada H, Kawashiri MA, Inazu A, Yamagishi M, Rader DJ, Kathiresan S, et al.
  • Journal Title: Circ Res. Volume: 121 Pages: 81-88
  • DOI: 10.1161/CIRCRESAHA.117.311145.
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Effect of Reverse Vessel Remodeling on Regression of Coronary Atherosclerosis in Patients Treated With Aggressive Lipid- and Blood Pressure-Lowering Therapy - Insight From MILLION Study. (2017)
  • Author(s): Gamou T, Sakata K, Tada H, Konno T, Hayashi K, Ino H, Yamagishi M, Kawashiri MA; MILLION Study Group.
  • Journal Title: Circ J. Volume: 81 Pages: 1490-1495
  • DOI: 10.1253/circj.CJ-16-1253.
  • Peer Reviewed
• [Journal Article] Early endothelialization associated with a biolimus A9 bioresorbable polymer stent in a porcine coronary model. (2017)
  • Author(s): Mori M, Sakata K, Nakanishi C, Nakahashi T, Kawashiri MA, Yoshioka K, Takuwa Y, Okada H, Yokawa JI, Shimojima M, Yoshimuta T, Yoshida S, Yamagishi M, Hayashi K.
  • Journal Title: Heart Vessels. Volume: 32 Pages: 1244-1252
  • DOI: 10.1007/s00380-017-0992-5.
  • Peer Reviewed
• [Journal Article] Do We Expect Any Pleiotropic Effect of Proprotein Convertase Subtilisin/Kexin Type 9 Inhibition for Reducing Cardiovascular Events Beyond Low-Density Lipoprotein Cholesterol Reduction? (2017)
  • Author(s): Tada H, Kawashiri MA, Yamagishi M.
  • Journal Title: Circ J. Volume: 81 Pages: 1098-1099
  • DOI: 10.1253/circj.CJ-17-0586.
• [Journal Article] Prognostic superiority of coronary artery bypass grafting to percutaneous coronary intervention in non-diabetic patients with anatomically complex multivessel coronary artery disease. (2017)
  • Author(s): Sakata K, Nakahashi T, Kawashiri MA.
  • Journal Title: Cardiovasc Diagn Ther. Volume: 7 Pages: S77-S81
  • DOI: 10.21037/cdt.2017.01.14.
• [Journal Article] Impact of Aging on High-sensitivity Cardiac Troponin T in Patients Suspected of Acute Myocardial Infarction. (2017)
  • Author(s): Ichise T, Tada H, Sakata K, Kawashiri MA, Yamagishi M, Hayashi K.
  • Journal Title: Intern Med. Volume: 56 Pages: 2097-2102
  • DOI: 10.2169/internalmedicine.8510-16.
  • Peer Reviewed
• [Journal Article] Application of an enhanced device to transluminal retrieval of malappositioned coronary stents: An experimental study. (2017)
  • Author(s): Mori M, Sakata K, Yokawa J, Nakanishi C, Okada H, Shimojima M, Yoshida S, Yoshimuta T, Hayashi K, Yamagishi M, Kawashiri MA.
  • Journal Title: J Interv Cardiol. Volume: 30 Pages: 537-543
  • DOI: 10.1111/joic.12429.
  • Peer Reviewed
• [Journal Article] Impact of evolocumab treatment on low-density lipoprotein cholesterol levels in heterozygous familial hypercholesterolemic patients withdrawing from regular apheresis. (2017)
  • Author(s): Kawashiri MA, Nohara A, Higashikata T, Tada H, Nakanishi C, Okada H, Konno T, Sakata K, Hayashi K, Inazu A, Mabuchi H, Yamagishi M.
  • Journal Title: Atherosclerosis. Volume: 265 Pages: 225-230
  • DOI: 10.1016/j.atherosclerosis.2017.09.011.
  • Peer Reviewed
• [Journal Article] Assessments of Carotid Artery Plaque Burden in Patients With Familial Hypercholesterolemia. (2017)
  • Author(s): Tada H, Kawashiri MA, Okada H, Nakahashi T, Sakata K, Nohara A, Inazu A, Mabuchi H, Yamagishi M, Hayashi K.
  • Journal Title: Am J Cardiol. Volume: 120 Pages: 1955-1960
  • DOI: 10.1016/j.amjcard.2017.08.012.
  • Peer Reviewed
• [Journal Article] Huge right ventricular mass lesion associated with genital malignant tumor: a case report. (2017)
  • Author(s): Higashi M, Hodatsu A, Uchiyama K, Tada H, Mori M, Ohira M, Hayashi K, Kawashiri MA.
  • Journal Title: J Med Case Rep. Volume: 11 Pages: 282
  • DOI: 10.1186/s13256-017-1439-4.
  • 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 Pages: 17417
  • DOI: 10.1038/s41598-017-17572-y.
  • Peer Reviewed / Open Access
• [Journal Article] Impact of triple antithrombotic therapy in patients with acute coronary syndrome undergoing percutaneous coronary intervention in real-world practice. (2017)
  • Author(s): Yan Y, Kawashiri MA, Kedev S, et al.
  • Journal Title: J Geriatr Cardiol. Volume: 14 Pages: 679-687
  • DOI: 10.11909/j.issn.1671-5411.2017.11.003.
  • Peer Reviewed / 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 Pages: 135-139
  • DOI: 10.1210/js.2017-00276.
  • Peer Reviewed
• [Presentation] PCSK9を標的とした新たな冠動脈疾患治療薬の誕生：疫学研究からの新薬開発 (2018)
  • Author(s): 川尻剛照
  • Organizer: 日本慢性疾患重症化予防学会 第4回年次学術集会
  • Invited
• [Presentation] 冠動脈、脳血管疾患予防におけるコレステロール低下療法の意義 (2017)
  • Author(s): 川尻剛照、多田隼人、野原淳、山岸正和
  • Organizer: 第4回日本心血管脳卒中学会学術集会
  • Invited
• [Presentation] 家族性高コレステロール血症の診断と治療 (2017)
  • Author(s): 川尻剛照
  • Organizer: 日本内科学会第71回北陸支部生涯教育講演会
  • Invited
• [Presentation] 家族性高コレステロール血症の診断 (2017)
  • Author(s): 川尻剛照、多田隼人、野原淳、馬渕宏、山岸正和
  • Organizer: 第49回日本動脈硬化学会総会・学術集会
  • Invited
• [Presentation] PCSK9阻害剤による超積極的LDLコレステロール低下療法の意義 (2017)
  • Author(s): 川尻剛照
  • Organizer: 第49回日本動脈硬化学会総会・学術集会
  • Invited
• [Presentation] Impact of evolocumab, a PCSK9 inhibitor, withdrawing from regular LDL apheresis therapy in patients with heterozygous familial hypercholesterolemia (2017)
  • Author(s): Kawashiri M, Higashikata T, Nohara A, Nakanishi C, Tada H, Okada H, Yamagishi M
  • Organizer: European Society of Cardiology 2017
  • Int'l Joint Research
• [Book] 医療と検査機器・試薬 (2017)
  • Author(s): 川尻剛照、多田隼人、山岸正和
  • Total Pages: 545(299-306)
  • Publisher: 宇宙堂八木書店
• [Book] 動脈硬化予防 (2017)
  • Author(s): 川尻剛照
  • Total Pages: 364(95-97)
  • Publisher: メジカルビュー社
• [Book] 循環器専門医 (2017)
  • Author(s): 川尻剛照、山岸正和
  • Total Pages: 318(145-149)
  • Publisher: 日本循環器学会
• [Book] 診断と治療 (2017)
  • Author(s): 川尻剛照、多田隼人、山岸正和
  • Total Pages: 1607(1191-1196)
  • Publisher: 診断と治療社
• [Book] 循環器内科 (2017)
  • Author(s): 川尻剛照、多田隼人、山岸正和
  • Total Pages: 592(196-202)
  • Publisher: 科学評論社
• [Book] 総合検診 (2017)
  • Author(s): 川尻剛照
  • Total Pages: 876(100-107)
  • Publisher: 日本総合健診医学会