腎臓における新規の血圧調節機構ＷＮＫ－ＮＣＣシグナル伝達系の解明

2013 Fiscal Year Annual Research Report

• Project/Area Number: 22390168
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: 頼 建光 東京医科歯科大学, 医学部附属病院, 准教授 (80334431)
• Project Period (FY): 2010-04-01 – 2014-03-31
• Keywords: 偽性低アルドステロン症II型 / KLHL3 / Cullin 3 / ユビキチン / SPAK / Na-K-Cl 共輸送体 / OSR1 / angiotensin II

Research Abstract

(1) 塩分摂取とangiotensin IIによる血管平滑筋におけるWNK-SPAK-NKCC1シグナル伝達系の制御
高塩分負荷により、マウスの大動脈においてSPAKとNKCC1のリン酸化の亢進がみられ、塩分制限下では、SPAKとNKCC1のリン酸化の低下がみられた。また、angiotensin II投与によりSPAKとNKCC1のリン酸化の亢進がみられた。これらの反応はいずれも、WKN3ノックアウトマウスにおいてはみられず、angiotensin IIによる血圧上昇もみられなかった。このことから、angiotensin II血管平滑筋のトーヌスの制御に、WNK3-SPAK-NKCC1シグナル伝達系が関与していることが示された。
(2) 細胞内ユビキチン化によるWNK-NCCシグナル系の制御
最近、kelch-like 3 (KLHL3)とCullin 3の遺伝子変異により偽性低アルドステロン症II型(PHAII)がおこることが明らかになった。細胞内強制発現系において、KLHL3はCullin 3とWNK4と結合してWNK4のユビキチン化を促進し、WNK4の蛋白発現レベルを低下させた。またPHAIIの疾患起因性変異はこれらの結合を阻害して、WNK4のユビキチン化を低下させ、WNK4の蛋白発現量を増加させた。このことから、WNK4はKLHL3によるユビキチン化の基質であり、WNK4のユビキチン化障害が遺伝性高血圧の発症メカニズムであることが示された。

Current Status of Research Progress

Reason

25年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

25年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Dietary salt intake regulates WNK3-SPAK-NKCC1 phosphorylation cascade in mouse aorta through angiotensin II (2013)
  • Author(s): Zeniya M, Sohara E, Kita S, Iwamoto T, Susa K, Mori T, Oi K, Chiga M, Takahashi D, Yang SS, Lin SH, Rai T, Sasaki S, Uchida S
  • Journal Title: Hypertension Volume: 62(5) Pages: 872-8
  • DOI: 10.1161/HYPERTENSIONAHA.113.01543
  • Peer Reviewed
• [Journal Article] Chemical library screening for WNK signalling inhibitors using fluorescence correlation spectroscopy (2013)
  • Author(s): Mori T, Kikuchi E, Watanabe Y, Fujii S, Ishigami-Yuasa M, Kagechika H, Sohara E, Rai T, Sasaki S, Uchida S
  • Journal Title: Biochem J Volume: 455(3) Pages: 339-45
  • DOI: 10.1042/BJ20130597
  • Peer Reviewed
• [Journal Article] Decrease of WNK4 ubiquitination by disease-causing mutations of KLHL3 through different molecular mechanisms (2013)
  • Author(s): Mori Y, Wakabayashi M, Mori T, Araki Y, Sohara E, Rai T, Sasaki S, Uchida S
  • Journal Title: Biochem Biophys Res Commun Volume: 439(1) Pages: 30-4
  • DOI: 10.1016/j.bbrc.2013.08.035
  • Peer Reviewed
• [Journal Article] KLHL2 interacts with and ubiquitinates WNK kinases (2013)
  • Author(s): Takahashi D, Mori T, Wakabayashi M, Mori Y, Susa K, Zeniya M, Sohara E, Rai T, Sasaki S, Uchida S
  • Journal Title: Biochem Biophys Res Commun Volume: 437(3) Pages: 457-62
  • DOI: 10.1016/j.bbrc.2013.06.104
  • Peer Reviewed
• [Journal Article] Impaired KLHL3-mediated ubiquitination of WNK4 causes human hypertension (2013)
  • Author(s): Wakabayashi M, Mori T, Isobe K, Sohara E, Susa K, Araki Y, Chiga M, Kikuchi E, Nomura N, Mori Y, Matsuo H, Murata T, Nomura S, Asano T, Kawaguchi H, Nonoyama S, Rai T, Sasaki S, Uchida S
  • Journal Title: Cell Rep Volume: 3(3) Pages: 858-68
  • DOI: 10.1016/j.celrep.2013.02.024
  • Peer Reviewed
• [Journal Article] Development of enzyme-linked immunosorbent assays for urinary thiazide-sensitive Na-Cl cotransporter measurement (2013)
  • Author(s): Isobe K, Mori T, Asano T, Kawaguchi H, Nonoyama S, Kumagai N, Kamada F, Morimoto T, Hayashi M, Sohara E, Rai T, Sasaki S, Uchida S
  • Journal Title: Am J Physiol Renal Physiol Volume: 305(9) Pages: F1374-81
  • DOI: 10.1152/ajprenal.00208.2013.
  • Peer Reviewed
• [Presentation] Impaired KHLH3-mediated ubiquitination of WNK4 activates OSR1 and SPAK kinases-NaCl cotransporter (NCC) signaling and causes hypertension (2013)
  • Author(s): Wakabayashi M, Mori T, Isobe K, Sohara E, Susa K, Araki Y, Chiga M, Kikuchi E, Nomura N, Mori Y, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-20131110
• [Presentation] WNK3 regulates blood pressure through the regulation of vascular OSR1/SPAK-NKCC1 phosphorylation cascade (2013)
  • Author(s): Zeniya M, Sohara E, Oi K, Chiga M, Susa K, Mori T, Takahashi D, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-20131110
• [Presentation] Clinical significance of urinary thiazide-sensitive Na-Cl cotransporter (NCC) measurement by newly developed enzyme-linked immunosorbent assays (2013)
  • Author(s): Isobe K, Mori T, Sohara E, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-10
• [Presentation] Low salt intake decreased transcription and protein level of KLHL3 in mouse kidney (2013)
  • Author(s): Susa K, Sohara E, Zeniya M, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-10
• [Presentation] Chemical library screening for WNK signaling inhibitors by using fluorescent correlation spectroscopy (2013)
  • Author(s): Mori T, Kikuchi E, Sohara E, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-10
• [Presentation] Analyses of KHLH3 mutants that cause Pseudohypoaldosteronism type II (2013)
  • Author(s): Mori Y, Wakabayashi M, Mori T, Araki Y, Sohara E, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-10
• [Presentation] KHLH2 interacts with and ubiquitinates WNK4 kinases (2013)
  • Author(s): Takahashi D, Mori T, Wakabayashi M, Mori Y, Susa K, Zeniya M, Rai T, Sohara E, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2013
  • Place of Presentation: Atlanta, U.S.A.
  • Year and Date: 20131105-10