The clarification of aldosterone production machinery and the identification of therapeutic factors based on the posttranscriptional regulation

• Project/Area Number: 21K08557
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 54040:Metabolism and endocrinology-related
• Research Institution: Hiroshima University
• Principal Investigator: 沖 健司 広島大学, 病院(医), 講師 (30638995)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Project Status: Discontinued (Fiscal Year 2022)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2023: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2022: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2021: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 原発性アルドステロン症 / アルドステロン / アルドステロン合成酵素 / 転写 / 転写後調節 / CYP11B2 / アンギオテンシンII / 遺伝子変異 / 蛋白発現調節

Outline of Research at the Start

CYP11B2 mRNA結合蛋白から転写後調節因子を同定し，その因子を調節した細胞株からリボソームプロファイリングの手法を導入し，CYP11B2 mRNA結合因子がCYP11B2の翻訳制御に直接関与することを明らかにする．さらにこれらの因子がCYP11B2の転写後調節に直接かかわっていることを明らかにする．これらの候補因子の絞り込みをAPAモデル細胞株およびPAモデルマウスにおいて行い，絞り込んだ因子の阻害薬投与またはノックアウトマウスを作成し，その表現型の解析によってCYP11B2のmRNAに結合する因子を介したアルドステロン合成制御機序を明らかにする．

Outline of Annual Research Achievements

試験１. CYP11B2 RNA結合蛋白の機能解析
前年度に同定したCYP11B2 RNA結合因子によるアルドステロン合成機構を明らかにするため，同因子の過剰発現およびknock downを行った．原発性アルドステロン症モデル細胞株でのshRNAによるknock downを行ったところ，アルドステロン合成は抑制され、HAC15細胞株での過剰発現解析では，CYP11B2 mRNAは高発現し，アルドステロン分泌は抑制された．
さらに，RNA結合因子のRNA結合部位のアミノ酸変異を加えたplasmidを作成し、同様の解析を行ったところ，アルドステロン分泌に変化はみられなかった．
試験２. CYP11B2 RNA結合蛋白を用いたChiP-seq解析
CYP11B2 RNA結合蛋白がCYP11B2との結合を明らかにするため，同タンパクの抗体を用いたChiP-seq解析を行った．本解析で得られたmRNAにCYP11B2の配列が確認され，その定量解析で原発性アルドステロン症モデル細胞株では対照細胞株に比較し有意に高値であることが示された．

Research Products

• [Journal Article] Genetic Risk of Primary Aldosteronism and Its Contribution to Hypertension: A Cross-Ancestry Meta-Analysis of Genome-Wide Association Studies (2023)
  • Author(s): Naito Tatsuhiko、Inoue Kosuke、Sonehara Kyuto、Baba Ryuta、Kodama Takaya、Otagaki Yu、Okada Akira、Itcho Kiyotaka、Kobuke Kazuhiro、Kishimoto Shinji、Yamamoto Kenichi、Morisaki Takayuki、Higashi Yukihito、Hinata Nobuyuki、Arihiro Koji、Hattori Noboru、Okada Yukinori、Oki Kenji、BioBank Japan
  • Journal Title: Circulation Volume: 147 Issue: 14 Pages: 1097-1109
  • DOI: 10.1161/circulationaha.122.062349
  • Peer Reviewed / Open Access
• [Journal Article] A hyperaldosteronism subtypes predictive model using ensemble learning (2023)
  • Author(s): Karashima S, Kawakami M, Nambo H, Kometani M, Kurihara I, Ichijo T, Katabami T, Tsuiki M, Wada N, Oki K, Ogawa Y, Okamoto R, Tamura K, Inagaki N, Yoshimoto T, Kobayashi H, Kakutani M, Fujita M, Izawa S, Suwa T, Kamemura K, Yamada M, Tanabe A, Naruse M, Yoneda T; JPAS/JRAS Study Group.
  • Journal Title: Scientific Reports Volume: 13 Issue: 1 Pages: 3043-3043
  • DOI: 10.1038/s41598-023-29653-2
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Effect of Intraprocedural Cortisol Measurement on ACTH-stimulated Adrenal Vein Sampling in Primary Aldosteronism (2022)
  • Author(s): Kometani Mitsuhiro、Yoneda Takashi、Karashima Shigehiroら
  • Journal Title: Journal of the Endocrine Society Volume: 6 Issue: 9
  • DOI: 10.1210/jendso/bvac104
  • Peer Reviewed / Open Access
• [Journal Article] NFIA determines the cis-effect of genetic variation on Ucp1 expression in murine thermogenic adipocytes (2022)
  • Author(s): Hiraike Y, Tsutsumi S, Wada T, Oguchi M, Saito K, Nakamura M, Ota S, Koebis M, Nakao H, Aiba A, Nagano G, Ohno H, Oki K, Yoneda M, Kadowaki T, Aburatani H, Waki H, Yamauchi T.
  • Journal Title: iScience Volume: 25 Issue: 8 Pages: 104729-104729
  • DOI: 10.1016/j.isci.2022.104729
  • Peer Reviewed / Open Access
• [Journal Article] Association of cardiovascular disease risk and changes in renin levels by mineralocorticoid receptor antagonists in patients with primary aldosteronism (2022)
  • Author(s): Nomura Motoko、Kurihara Isao、Itoh Hiroshi、Ichijo Takamasa、Katabami Takuyuki、Tsuiki Mika、Wada Norio、Yoneda Takashi、Sone Masakatsu、Oki Kenji、et al
  • Journal Title: Hypertension Research Volume: 45 Issue: 9 Pages: 1476-1485
  • DOI: 10.1038/s41440-022-00960-x
  • Peer Reviewed / Open Access
• [Journal Article] Association of DNA methylation with steroidogenic enzymes in Cushing’s adenoma (2022)
  • Author(s): Kodama Takaya、Oki Kenji、Otagaki Yu、Baba Ryuta、Okada Akira、Itcho Kiyotaka、Kobuke Kazuhiro、Nagano Gaku、Ohno Haruya、Hinata Nobuyuki、Arihiro Koji、Gomez-Sanchez Celso E、Yoneda Masayasu、Hattori Noboru
  • Journal Title: Endocrine-Related Cancer Volume: 29 Issue: 8 Pages: 495-502
  • DOI: 10.1530/erc-22-0115
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Hypomethylation associated vitamin D receptor expression in ATP1A1 mutant aldosterone-producing adenoma (2022)
  • Author(s): Nanao Yuta、Oki Kenji、Kobuke Kazuhiro、Itcho Kiyotaka、Baba Ryuta、Kodama Takaya、Otagaki Yu、Okada Akira、Yoshii Yoko、Nagano Gaku、Ohno Haruya、Arihiro Koji、Gomez-Sanchez Celso E.、Hattori Noboru、Yoneda Masayasu
  • Journal Title: Molecular and Cellular Endocrinology Volume: 548 Pages: 111613-111613
  • DOI: 10.1016/j.mce.2022.111613
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Subtype-specific trends in the clinical picture of primary aldosteronism over a 13-year period (2021)
  • Author(s): Saito Kohei、Yamada Tetsuya、Kobayashi Hiroki、Tamura Kouichi、Ogawa Yoshihiro、Kawashima Junji、Inagaki Nobuya、Yamamoto Koichi、Yamada Masanobu、Kamemura Kohei、Fujii Yuichi、Suzuki Tomoko、Yasoda Akihiro、Tanabe Akiyo、Naruse Mitsuhide et. al.
  • Journal Title: Journal of Hypertension Volume: 39 Issue: 11 Pages: 2325-2332
  • DOI: 10.1097/hjh.0000000000002924
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Renal Injuries in Primary Aldosteronism: Quantitative Histopathological Analysis of 19 Patients With Primary Adosteronism (2021)
  • Author(s): Ogata Hiroko、Yamazaki Yuto、Tezuka Yuta、Gao Xin、Omata Kei、Ono Yoshikiyoら
  • Journal Title: Hypertension Volume: 78 Issue: 2 Pages: 411-421
  • DOI: 10.1161/hypertensionaha.121.17436
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] ATP1A1 Mutant in Aldosterone-Producing Adenoma Leads to Cell Proliferation (2021)
  • Author(s): Kobuke Kazuhiro、Oki Kenji、Gomez-Sanchez Celso E.、Gomez-Sanchez Elise P.、Itcho Kiyotaka、Ohno Haruya、Nagano Gaku、Yoshii Yoko、Baba Ryuta、Kodama Takaya、Arihiro Koji、Hattori Noboru、Yoneda Masayasu
  • Journal Title: International Journal of Molecular Sciences Volume: 22 Issue: 20 Pages: 10981-10981
  • DOI: 10.3390/ijms222010981
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Update on Genetics of Primary Aldosteronism (2021)
  • Author(s): Itcho Kiyotaka、Oki Kenji、Ohno Haruya、Yoneda Masayasu
  • Journal Title: Biomedicines Volume: 9 Issue: 4 Pages: 409-409
  • DOI: 10.3390/biomedicines9040409
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Should Adrenal Venous Sampling Be Performed in PA Patients Without Apparent Adrenal Tumors? (2021)
  • Author(s): Okamoto Kentaro、Ohno Youichi、Sone Masakatsu、Inagaki Nobuya、Ichijo Takamasa、Yoneda Takashi、Tsuiki Mika、Wada Norio、Oki Kenji、Tamura Kouichi、Kobayashi Hiroki、Izawa Shoichiro、Tanabe Akiyo、Naruse Mitsuhide
  • Journal Title: Frontiers in Endocrinology Volume: 12 Pages: 645395-645395
  • DOI: 10.3389/fendo.2021.645395
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Association of aldosterone and blood pressure with the risk for cardiovascular events after treatments in primary aldosteronism. (2021)
  • Author(s): 1.Haze T, Hirawa N, Yano Y, Tamura K, Kurihara I, Kobayashi H, Tsuiki M, Ichijo T, Wada N, Katabami T, Yamamoto K, Oki K, Inagaki N, Okamura S, Kai T, Izawa S, Yamada M, Chiba Y, Tanabe A, Naruse M.
  • Journal Title: Atherosclerosis. Volume: 324 Pages: 84-90
  • DOI: 10.1016/j.atherosclerosis.2021.03.033
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Presentation] 遺伝子変異・発現解析による副腎性クッシング症候群の細胞内分子機構の解析 (2021)
  • Author(s): 馬場 隆太, 沖 健司, 岡田 晃, 大田垣 裕, 児玉 尭也, 小武家 和博, 一町 澄宜, 長野 学, 大野 晴也, 米田 真康
  • Organizer: 第25回日本臨床内分泌病理学会学術総会
• [Presentation] 両側副腎腫瘍にPRKACA変異を認めたクッシング症候群の解析 (2021)
  • Author(s): 大田垣 裕, 沖 健司, 馬場 隆太, 小武家 和博, 一町 澄宜, 長野 学, 大野 晴也, 米田 真康
  • Organizer: 第22回日本内分泌学会中国支部学術集会