The role of Hsp70 in quality control of mutant hERG protein associated LQT2 and its therapeutic application.

• Project/Area Number: 25461109
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cardiovascular medicine
• Research Institution: Tottori University
• Principal Investigator: Li Peili 鳥取大学, 医学(系)研究科(研究院), 助教 (40464292)
• Co-Investigator(Kenkyū-buntansha): Shirayoshi Yasuaki 鳥取大学, 医学系研究科, 准教授 (90249946) ; 池田 信人 鳥取大学, 医学(系)研究科(研究院), 助教 (50620316)
• Co-Investigator(Renkei-kenkyūsha): Ikeda Nobuhito 鳥取大学, 医学系研究科, 助教 (50620316)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2015: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
• Keywords: LQT2 / hERG / histone deacetylase 6 / acetylation / ubiquitination / lysine / 変異hERG / アセチル化 / ユビキチン化 / HDAC6 / HDAC阻害剤 / histone deacetylase / mutation / mutant hERG / histine deacetylase / Histone deacetylase 6 / histone deacetylase 10 / chaperone / protein quality control

Outline of Final Research Achievements

LQT2 results from the mutations in hERG causing reduced hERG protein expression on cell membrane and hERG currents. We found a novel role of histone deacetylase (HDAC) 6 in hERG. Three HDAC pan-inhibitors, a HDAC6 selective inhibitor (TBA) or knockdown of HDAC6 increased wild-type (WT) protein expression and induced two mutant hERGs’ maturation with enhanced the acetylation of hERG protein, whereas, decreased the ubiquitin with prolonged half-life. Co-expression of HDAC6 posed opposite effects on hERG. Immunochemistry and electrophysiological studies confirmed the results. Substitutions of the three lysine residues (116, 495 and 757) with arginine in hERG erased the effects of HDAC6 on hERG protein level, acetylation and ubiquitintation levels, respectively. TBA treatment enhanced the expression of ERG in mouse cardiomyocytes HL-1, increased IKr and shortened action potential duration. The results indicate HDAC6 regulates hERG by altering acetylation/ubiquitination of hERG proteins.

Research Products

• [Journal Article] Electrophysiological properties of iPS cell-derived cardiomyocytes from a patient with long QT syndrome type 1 harboring the novel mutation M437V of KCNQ1 (2016)
  • Author(s): Sogo T, Morikawa K, Kurata Y, Li P, Ichinose T, Yuasa S, Nozaki D, Miake J, Ninomiya H, Shimizu W, Fukuda K, Yamamoto K, Shirayoshi Y, Hisatome I
  • Journal Title: Regenerative Therapy Volume: 4 Pages: 9-17
  • DOI: 10.1016/j.reth.2015.12.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] E3 ligase CHIP and Hsc70 regulate Kv1.5 protein expression and function in mammalian cells. (2015)
  • Author(s): Li P, Kurata Y, Maharani N, Mahati E, Higaki K, Hasegawa A, Shirayoshi Y, Yoshida A, Kondo T, Kurozawa Y, Yamamoto K, Ninomiya H, Hisatome I.
  • Journal Title: J Mol Cell Cardiol. Volume: 86 Pages: 138-146
  • DOI: 10.1016/j.yjmcc.2015.07.018
  • Peer Reviewed / Open Access
• [Journal Article] Molecular Mechanisms Underlying Urate-Induced Enhancement of Kv1.5 Channel Expression in HL-1 Atrial Myocytes (2015)
  • Author(s): Maharani N, Ting YK, Cheng J, Hasegawa A, Kurata Y, Li P, Nakayama Y, Ninomiya H, Ikeda N, Morikawa K, Yamamoto K, Makita N, Yamashita T, Shirayoshi Y, Hisatome I.
  • Journal Title: Circulation Journal Volume: 79 Issue: 12 Pages: 2659-2668
  • DOI: 10.1253/circj.CJ-15-0416
  • NAID: 130005110934
  • ISSN: 1346-9843, 1347-4820
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Stabilization of Kv1.5 channel protein by the inotropic agent olprinone. (2015)
  • Author(s): Endo R, Kurata Y, Notsu T, Li P, Morikawa K, Kondo T, Ogura K, Miake J, Yoshida A, Shirayoshi Y, Ninomiya H, Higaki K, Kuwabara M, Yamamoto K, Inagaki Y, Hisatome I.
  • Journal Title: Eur J Pharmacol. Volume: 765 Pages: 488-494
  • DOI: 10.1016/j.ejphar.2015.09.013
  • Peer Reviewed / Open Access
• [Journal Article] Apoptosis induced by an uromodulin mutant C112Y and its suppression by topiroxostat. (2014)
  • Author(s): Utami SB, Mahati E, Li P,
  • Journal Title: Clin Exp Nephrol. Volume: Epub ahead of print
  • Peer Reviewed / Open Access
• [Journal Article] Instability of KCNE1-D85N that Causes Long QT Syndrome: Stabilization by Verapamil (2014)
  • Author(s): Sakata S, Kurata Y, Li P, Notsu T, Morikawa K, Miake J, Higaki K, Yamamoto Y, Yoshida A, Shirayoshi Y, Yamamoto K, Horie M, Ninomiya H, Kanzaki S, Hisatome I.
  • Journal Title: Pacing Clinical Electrophysiology Volume: Epub ahead of print Issue: 7 Pages: 1-11
  • DOI: 10.1111/pace.12360
  • Peer Reviewed / Open Access
• [Journal Article] AMP deaminase 3 plays a critical role in remote reperfusion lung injury (2013)
  • Author(s): Li P, Ogino K, Morikawa K, Shirayoshi Y, Hisatome I, et al.
  • Journal Title: Biochem Biophys Res Commun. Volume: 431(2) Issue: 1 Pages: 131-6
  • DOI: 10.1016/j.bbrc.2013.03.056
  • Peer Reviewed
• [Journal Article] Hsp90 prevents interaction between CHIP and HERG proteins to facilitate maturation of wild-type and mutant HERG proteins (2013)
  • Author(s): Iwai C, P Li,
  • Journal Title: Cardiovasc Res Volume: ３ Issue: 3 Pages: 520-528
  • DOI: 10.1093/cvr/cvt200
  • Peer Reviewed
• [Journal Article] Simultaneous Treatment with Azelnidipine and Olmesartan Inhibits Apoptosis of Hl-1 Cardiac Myocytes Expressing E334k cMyBPC (2013)
  • Author(s): Bahrudin U, Ikeda N, Utami SB, Maharani N, Morikawa K, Li P
  • Journal Title: Drug Res (Stuttg). Volume: １０ Issue: 10 Pages: 515-520
  • DOI: 10.1055/s-0033-1347188
  • Peer Reviewed
• [Journal Article] Bepridil Suppresses Apoptosis in HL-1 Cardiac Atrial Myocytes Expressing Mutant E334K cMyBPC. (2013)
  • Author(s): Endo R, Notsu T, Mishima M, Morikawa K, Li P
  • Journal Title: Yonago Acta Med Volume: Epub
• [Presentation] Epigenetic and post-translational modification of hERG to control the channel protein expression and function (2016)
  • Author(s): Peili Li, et al
  • Organizer: 第80回日本循環器学会学術集会
  • Place of Presentation: 宮城県仙台市仙台国際センター
  • Year and Date: 2016-03-18
• [Presentation] Hyperuricemia-induced Stabilization of Kv1.5 (2016)
  • Author(s): Maharani Nani, Li Peili, et al.
  • Organizer: 第49回 日本痛風・核酸代謝学会
  • Place of Presentation: 大阪府大阪市大阪千里ライフサイエンスセンター
  • Year and Date: 2016-02-18
• [Presentation] ビルジカイニドの慢性投与によるHERGチャネル増加作用に関する検討 (2015)
  • Author(s): 大野原岳史、久留一郎、李 佩俐、et al.
  • Organizer: 第１０７回日本循環器学会中国地方会
  • Place of Presentation: 広島県広島市広島アステールプラザ
  • Year and Date: 2015-11-28
• [Presentation] Heat shock factor-1 は変異型A78THERGチャネル蛋白の成熟化に関与する (2015)
  • Author(s): 近藤健人、李 佩俐、et al
  • Organizer: 第１０７回日本循環器学会中国地方会
  • Place of Presentation: 広島県広島市広島アステールプラザ
  • Year and Date: 2015-11-28
• [Presentation] Histone deacetylase plays a pivotal role in expression and function of wild-type and mutant hERGs (2015)
  • Author(s): Peili Li, et al.
  • Organizer: 第30回日本不整脈学会学術大会 第32回日本心電学会学術集会
  • Place of Presentation: 京都府京都市国立京都国際会館
  • Year and Date: 2015-07-28
• [Presentation] Stabilization of Kv1.5 channel protein by Olprinone (2015)
  • Author(s): Ryo Endo, Peili Li, et al.
  • Organizer: 第30回日本不整脈学会学術大会 第32回日本心電学会学術集会
  • Place of Presentation: 京都府京都市国立京都国際会館
  • Year and Date: 2015-07-28
• [Presentation] A novel Double Mutation of hERG protein in LQT2 syndrome: Identification and rescue. (2015)
  • Author(s): Endang Mahati, Peili Li, et al
  • Organizer: 第30回日本不整脈学会学術大会 第32回日本心電学会学術集会
  • Place of Presentation: 京都府京都市国立京都国際会館
  • Year and Date: 2015-07-28
• [Presentation] Electrophysiological properties of iPS cell-derived cardiomyocytes from a patient with long QT syndrome type 1 harboring a novel mutation of KCNQ1 (2015)
  • Author(s): Yasutaka Kurata, peili Li, et al
  • Organizer: 第30回日本不整脈学会学術大会 第32回日本心電学会学術集会
  • Place of Presentation: 京都府京都市国立京都国際会館
  • Year and Date: 2015-07-28
• [Presentation] Hsp90 Promotes Maturation of Wild-type and Mutant HERG Proteins (2014)
  • Author(s): Peili Li
  • Organizer: 第29回日本不整脈学会・第31回日本心電学会合同学術大会
  • Place of Presentation: 東京都港区ザ・プリンスパークタワー東京,プリンスホテル
  • Year and Date: 2014-07-22 – 2014-07-25
• [Presentation] The Activity of Urate to Increases Kv1.5 Channel Expression in Atrial Myocytes Depends on Uric acid Transporters, Xanthine Oxidase and ERKs (2014)
  • Author(s): Nani Maharani, Peili li,
  • Organizer: 第29回日本不整脈学会・第31回日本心電学会合同学術大会
  • Place of Presentation: 東京都港区ザ・プリンスパークタワー東京,プリンスホテル
  • Year and Date: 2014-07-22 – 2014-07-25
• [Presentation] Topiroxiostat improved the apoptpsis of cells expressing Uromodulin Mutation C112Y Causing Familial Juvenile Hyperuricemic Nephropathy (FJHN) (2014)
  • Author(s): Peili Li , Sulistiyati Bayu Utami,
  • Organizer: 第47回日本痛風・核酸代謝学会総会
  • Place of Presentation: 神戸国際会議場
• [Presentation] Hsp90 Prevents Interaction between CHIP and HERG Proteins to Facilitate Maturation of Wild-type and Mutant HERG Proteins (2014)
  • Author(s): Peili Li, Chisato Iwai
  • Organizer: 第78回日本循環器学会学術集会
  • Place of Presentation: 東京国際フォーラム
• [Presentation] E3 ligase CHIP interacts with Hsc70 to regulate Kv1.5 expression and repolarization in HL-1 cells (2013)
  • Author(s): Peili Li, Masaru Kato
  • Organizer: 第30回日本心電学会学術集会
  • Place of Presentation: 青森市文化会館 ホテル青森
• [Presentation] ユビキチンリガーゼCHIPに介してHsp90は変異hERG蛋白発現とチャネル活性を改善する (2013)
  • Author(s): 李佩俐、岩井ちさと
  • Organizer: 第103日本循環器学会中国地方会
  • Place of Presentation: 米子コンベンションセンター