Elucidation of the molecular mechanism and clinical application of increased plasma level of BNP precursor in patients with heart failure

• Project/Area Number: 15K09138
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cardiovascular medicine
• Research Institution: Kyoto University
• Principal Investigator: Nishikimi Toshio 京都大学, 医学研究科, 非常勤講師 (80291946)
• Co-Investigator(Kenkyū-buntansha): 桑原 宏一郎 信州大学, 学術研究院医学系, 教授 (30402887) ; 中川 靖章 京都大学, 医学研究科, 助教 (70452357)
• Co-Investigator(Renkei-kenkyūsha): Minamino Naoto 国立循環器病研究センター, オミックスセンター, センター長 (50124839)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2016: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 心不全 / BNP / poBNP / 前駆体 / O型糖鎖修飾 / プロセシング / 心筋細胞 / macro-proBNP / proBNP / 急性心不全 / microRNA-30 / GALNT

Outline of Final Research Achievements

We introduced human proBNP gene into rat cardiac myocytes and assessed the processing of proBNP by measuring proBNP and total BNP levels using our developed chemiluminescent immunoassay in the culture medium. We also produced proBNP mutant and assessed the effect of each glycosylation site on the proBNP processing. We found that Th71 and Th48 directly inhibit the processing of proBNP, and that other five glycosylation sites were indirectly associated with the inhibition of processing of proBNP. We also showed that the proBNP/total BNP ratio was higher and cGMP/total BNP ratio was lower in severe chronic heart failure compared with mild chronic heart failure. We experienced a case of heart failure whose BNP level was more than 35000pg/mL. We elucidated the mechanism of it and first reported it in the world as a case of macro-proBNP.

Research Products

• [Journal Article] Adrenomedullin as a Biomarker (2018)
  • Author(s): Nishikimi T, Nakagawa Y
  • Journal Title: Heart Fail Clin Volume: 14 Issue: 1 Pages: 49-55
  • DOI: 10.1016/j.hfc.2017.08.006
  • Peer Reviewed
• [Journal Article] Ratio of pro-B-type natriuretic peptide (BNP) to total BNP is decreased in mild, but not severe, acute decompensated heart failure patients: A novel compensatory mechanism for acute heart failure (2018)
  • Author(s): Takahama Hiroyuki、Takashio Seiji、Nishikimi Toshio、Hayashi Tomohiro、Nagai-Okatani Chiaki、Nakagawa Yasuaki、Amaki Makoto、Ohara Takahiro、Hasegawa Takuya、Sugano Yasuo、Kanzaki Hideaki、Yasuda Satoshi、Kangawa Kenji、Minamino Naoto、Anzai Toshihisa
  • Journal Title: International Journal of Cardiology Volume: 258 Pages: 165-171
  • DOI: 10.1016/j.ijcard.2017.12.047
  • Peer Reviewed
• [Journal Article] Endothelium-derived C-type natriuretic peptide contributes to blood pressure regulation by maintaining endothelial integrity. (2017)
  • Author(s): Nakao K, Kuwahara K, Nishikimi T, Nakagawa Y, Kinoshita H, Minami T, Kuwabara Y, Yamada C, Yamada Y, Tokudome T, Nagai-Okatani C, Minamino N, Nakao YM, Yasuno S, Ueshima K, Sone M, Kimura T, Kangawa K, Nakao K
  • Journal Title: Hypertension. Volume: 69 Issue: 2 Pages: 286-296
  • DOI: 10.1161/hypertensionaha.116.08219
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] MiR30-GALNT1/2 Axis-Mediated Glycosylation Contributes to the Increased Secretion of Inactive Human Prohormone for Brain Natriuretic Peptide (proBNP) From Failing Hearts. (2017)
  • Author(s): Nakagawa Y, Nishikimi T, Kuwahara K, Fujishima A, Oka S, Tsutamoto T, Kinoshita H, Nakao K, Cho K, Inazumi H, Okamoto H, Nishida M, Kato T, Fukushima H, Yamashita JK, Wijnen WJ, Creemers EE, Kangawa K, Minamino N, Nakao K, Kimura T.
  • Journal Title: J Am Heart Assoc. Volume: 6 Issue: 2
  • DOI: 10.1161/jaha.116.003601
  • NAID: 120007100125
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Macro-pro-B-type natriuretic peptide (proBNP) and hidden macro-N-terminal proBNP:Case report. (2017)
  • Author(s): Nakagawa Y, Nishikimi T, Sakai H, Ohno S, Kinoshita H, Inazumi H, Moriuchi K, Kuwahara K, Horie M, Kimura T.
  • Journal Title: Clinical Biochemistry Volume: 52 Pages: 148-152
  • DOI: 10.1016/j.clinbiochem.2017.10.019
  • Peer Reviewed
• [Journal Article] The renin-angiotensin system promotes arrhythmogenic substrates andalethal arrhythmias in mice with non-ischemic cardiomyopathy (2016)
  • Author(s): Yamada C, Kuwahara K, Yamazaki M, Nakagawa Y, Nishikimi T, Kinoshita H, Kuwabara Y, Minami T, Yamada Y, Shibata J, Nakao K, , Arai Y, Honjo H, Kamiya K, Nakao K, Kimura T
  • Journal Title: Cardiovascular Research Volume: 109 Issue: 1 Pages: 162-173
  • DOI: 10.1093/cvr/cvv248
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] 心不全における血中BNP分子型の多様性とその意義：proBNP, BNP［1-32］関連分子種, nonglycosylated-NT-BNP,glycosylated-NT-BNPについて. (2016)
  • Author(s): 錦見俊雄,中川靖章
  • Journal Title: 循環器内科 Volume: 79 Pages: 510-516
• [Journal Article] 分子標的薬と高血圧・循環器疾患 (2016)
  • Author(s): 錦見俊雄
  • Journal Title: 腎臓内科・泌尿器科 Volume: 4 Pages: 82-89
• [Journal Article] Longitudinal Changes in Late Systolic Cardiac Load and Serum NT-proBNP Levels in Healthy Middle-Aged Japanese Men. (2015)
  • Author(s): Tomiyama H, Nishikimi T, Matsumoto C, Kimura K, Odaira M, Shiina K, Yamashina A
  • Journal Title: Am J Hypertens Volume: 28 Issue: 4 Pages: 452-458
  • DOI: 10.1093/ajh/hpu174
  • Peer Reviewed
• [Journal Article] Pro-B-type natriuretic peptide is cleaved intracellularly: impact of distance between O-glycosylation and cleavage sites. (2015)
  • Author(s): Nishikimi T, Nakagawa Y, Minamino N, Ikeda M, Tabei K, Fujishima A, Takayama K, Akimoto K, Yamada C, Nakao K, Minami T, Kuwabara Y, Kinoshita H, Tsutamoto T, Ishimitsu T, Kangawa K, Kuwahara K, Nakao K
  • Journal Title: Am J Physiol Regul Integr Comp Physiol. Volume: 309 Issue: 6 Pages: R639-R649
  • DOI: 10.1152/ajpregu.00074.2015
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] O-linked oligosaccharide atachment underestimates N-terminal proBNP(NT-proBNP) levels in heart failure (HF) and chronic renal failure (CRF) on hemodyalysis (HD) (2017)
  • Author(s): Toshio Nishikimi, Yasuaki Nakagawa, Koichiro Kuwahara, Hideyuki Kinoshita, Hideaki Inazumi, Kenji Moriuchi, Kazuwa Nakao, Takeshi Kimura
  • Organizer: The81st Annual Scientific Meeting of the Japanese Circulation Sciety
• [Presentation] Adipocyte-derived adrenomedullin suppresses age-and obesity-related increase in blood pressure (2017)
  • Author(s): Kosai Cho, Takeya Minami, Koichiro Kuwahara, Toshio Nishikimi, Hideaki Inazumi, Kenji Moriuti, Kaoru Koike, Kenji Naoto Minamino, Kenji Kangawa Yasuaki Nakagawa, Hideyuki Kinoshita, Kazuwa Nakao, Takeshi Kimura
  • Organizer: The81st Annual Scientific Meeting of the Japanese Circulation Sciety
• [Presentation] O-linked oligosaccharaide attachment underestimates N-treminal proBNP-76(NT-proBNP) levels in heart failure(HF) and chronic renal failure (CRF) on hemodialysis (HD) (2017)
  • Author(s): Nishikimi T, Nakagawa Y, Kinoshita H, Moriuchi K, Inazumi H, Kuwahara K, Nakao K, Kimura T.
  • Organizer: ESC Congress 2017
  • Int'l Joint Research
• [Presentation] Adipocyte-derived adrenomedullin suppresses age-and obesity-related increase in blood pressure (2017)
  • Author(s): Cho Kozai, Takeya Minami, Toshio Nishikimi, Yasuaki Nakagawa, Kazuwa Nakao, Koichiro Kuwahara, Kaoru Koike, Takeshi Kimura
  • Organizer: ESC Congress 2017
  • Int'l Joint Research
• [Presentation] Impact of continuous reduction in B-type natriuretic peptide bioactivity during hospitalization in patients with advanced heart failure (2017)
  • Author(s): Takahama H, Takashio S, Hayashi T, Okatani-Nagai C, Nishikimi T, Nakagawa Y, Amaki M, Ohara T, Hasegawa T, Sugano Y, Kanzaki H, Yaduda S, Kangawa K, Minamino N, Anzai T.
  • Organizer: AHA Scientific Session 2017
  • Int'l Joint Research
• [Presentation] Molar ratio of N-terminal fragment of pro-B-type natriuretic peptide(BNP) to proBNP predicts occurrence of acute kiney injury in patients with acute heart failure (2017)
  • Author(s): Takahama H, Takashio S, Hayashi T, Nishikimi T, Nakagawa Y, Amaki M, Hasegawa T, Sugano Y, Kanzaki H, Yaduda S, Kangawa K, Minamino N, Anzai T.
  • Organizer: AHA Scientific Session 2017
  • Int'l Joint Research
• [Presentation] マクロproBNP血症の一例 (2017)
  • Author(s): 中川靖章、錦見俊雄、酒井宏、大野聖子、木下秀之、稲住英明、森内健史、柳澤洋、桑原宏一郎、堀江稔、木村剛
  • Organizer: 第21回心血管内分泌代謝学会
• [Presentation] 病的心筋におけるmiR30-GALNT経路による糖鎖修飾を介したproBNP分泌制御 (2016)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、木下秀之、稲住英明、森内健史、南野直人、中尾一和、木村剛
  • Organizer: 第20回日本心血管内分泌代謝学会総会(CVMW2016)
  • Place of Presentation: 東京
  • Year and Date: 2016-12-16
• [Presentation] Secretion of proBNP is regulated by MiR30-GALNTs axis dependent O-linked glycosylation in the pathological myocardium. (2016)
  • Author(s): Yasuaki Nakagawa, Toshio Nishikimi, Koichiro Kuwahara, Hideyuki Kinoshita, Hideaki Inazumi, Kenji Moriuchi, Takao Kato, Naoto Minamino, Kazuwa Nakao, Takeshi Kimura
  • Organizer: AHA Scientific Session 2016
  • Place of Presentation: New Orleans, Louisiana, USA
  • Year and Date: 2016-11-13
  • Int'l Joint Research
• [Presentation] Endogenous plasma NT-BNP is glycosylated and undetectable with the current NT-BNP assay system (2016)
  • Author(s): Toshio Nishikimi, Yasuaki Nakagawa, Koichiro Kuwahara, Hideyuki Kinoshita, Chinatsu Yamada, Takeshi Kimura, Kazuwa Nakao
  • Organizer: The 26th Scientific meeting of the International Society of Hypertension
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2016-09-24
  • Int'l Joint Research
• [Presentation] miR30-GALNT経路によるO型糖鎖修飾調節を介した病的心からのproBNP分泌亢進機序 (2016)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、保野慎治、木下秀之、桒原佳宏、稲住英明、森内健史、上嶋健治、南野直人、中尾一和、木村剛
  • Organizer: 第２回 国際心血管薬物療法学会日本部会(J-ICPS)
  • Place of Presentation: 徳島
  • Year and Date: 2016-06-25
• [Presentation] O型糖転移酵素GALNTのBNPプロセシング調節への関与 (2016)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、木下秀之、山田千夏、稲住英明、南野直人、中尾一和、木村剛
  • Organizer: 第89回 日本内分泌学会学術総会
  • Place of Presentation: 京都
  • Year and Date: 2016-04-21
• [Presentation] miR30-GALNT経路によるO型糖鎖修飾を介したBNPプロセシング調節が病的心筋からのproBNP分泌亢進に寄与する (2016)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、保野慎治、木下秀之、桒原佳宏、山田千夏、稲住英明、上嶋健治、南野直人、中尾一和、木村剛
  • Organizer: 第53回 日本臨床分子医学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2016-04-15
• [Presentation] MiR30-GALNTs axis-mediated O-glycosylation plays an important role in the increased secretion of unprocessed pro-brain natriuretic peptide by failing heart (2016)
  • Author(s): Yasuaki Nakagawa, Toshio Nishikimi, Koichiro Kuwahara, Kazuwa Nakao and Takeshi Kimura
  • Organizer: Keystone symposia
  • Place of Presentation: Snowbird, Utah, USA
  • Year and Date: 2016-04-03
  • Int'l Joint Research
• [Presentation] miR-30-GALNT1,2経路による糖鎖修飾調節が病的心筋からのproBNP分泌に関与する (2016)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、木下秀之、稲住英明、森内健史、南野直人、中尾一和、木村剛
  • Organizer: 第26回 日本循環薬理学会
  • Place of Presentation: 松本
• [Presentation] Secretion and metabolism of proBNP in patients with heart failure (2015)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、木下秀之、山田千夏、南野直人、中尾一和、木村剛
  • Organizer: 第19回日本心血管内分泌代謝学会学術総会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-10
• [Presentation] O型糖鎖修飾を介したヒトBNPプロセシング調節に関与する糖転移酵素についての検討 (2015)
  • Author(s): 中川靖章、錦見俊雄、桑原宏一郎、木下秀之、山田千夏、南野直人、中尾一和、木村剛
  • Organizer: 第19回日本心血管内分泌代謝学会学術総会
  • Place of Presentation: 神戸国際会議場（兵庫県、神戸市）
  • Year and Date: 2015-12-10
• [Presentation] BNP up to date (2015)
  • Author(s): 錦見俊雄、中川靖章。桑原宏一郎
  • Organizer: 日本循環器学会総会
  • Place of Presentation: 大阪国際会議場（大阪府、大阪市）
  • Year and Date: 2015-04-24
  • Invited
• [Book] Endocrinology of the Heart in Health and Disease. Chapter 2 Adrenomedullin. (2016)
  • Author(s): T. Nishikimi, K. Kuwahara, Y. Nakagawa, K Kangawa, and K. Nakao
  • Publisher: Elsevier Scinence Imprint:Academic Press
• [Remarks] 京都大学医学部付属病院循環器内科 基礎研究グループの活動 心血管内分泌代謝グループ
  • URL: http://kyoto-u-cardio.jp/kisokenkyu/cat236/