Novel functions and regulation of WNK signaling cascade

• Project/Area Number: 16K09642
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Kidney internal medicine
• Research Institution: Tokyo Medical and Dental University
• Principal Investigator: Rai Tatemitsu 東京医科歯科大学, 大学院医歯学総合研究科, 寄附講座教授 (80334431)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Project Status: Completed (Fiscal Year 2018)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2018: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2017: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: KLHL3 / cullin3 / NKCC1 / NCC / FOXO4 / 偽性低アルドステロン症II型 / ユビキチン / 偽性低アルドステロン症II型（PHAII） / ループ利尿薬 / サルコペニア / Cullin3 / PHAII / 高血圧 / 内科 / 生理学 / トランスレーショナルリサーチ / シグナル伝達

Outline of Final Research Achievements

(1) Pathogenesis of defective degradation of WNK kinases by KLHL-Cullin3 complex: By generation and analysis of KLHL3 knockout mice, the dominant-negative effect of mutant KLHL3 was found to be important for the pathogenesis of PHAII.
(2) Function of KLHL3/Cul3/WNK signal in extra-renal organs: By generation and analysis of WNK4 knockout mice, WNK4 was found to be a novel regulating factor of adipocyte differentiation. In addition, NKCC1 was shown to play an important role in muscle differentiation and hypertrophy. Thus, WNK1-FOXO4 signal was identified as a novel signaling system participating in the physiologic and pathogenic association of the muscle and kidney.

Academic Significance and Societal Importance of the Research Achievements

KLHL3とKLHL2によるWNKシグナルの制御メカニズム、慢性腎臓病における塩分感受性高血圧の成立メカニズムについて重要な新知見を得ることができた。この成果は、新たな高血圧治療薬の創薬につながるものである。また腎臓以外の、脂肪組織や筋肉におけるWNKシグナル系の機能についても新規発見を得た。この成果により、WNKシグナル系が肥満やサルコペニアの新しい治療標的となる可能性があることが示された。

Research Products

• [Journal Article] Failure to sense energy depletion may be a novel therapeutic target in chronic kidney disease (2019)
  • Author(s): Kikuchi H, Sasaki E, Nomura N, Mori T, Minamishima YA, Yoshizaki Y, Takahashi N, Furusho T, Arai Y, Mandai S, Yamashita T, Ando F, Maejima Y, Isobe K, Okado T, Rai T, Uchida S, Sohara E
  • Journal Title: Kidney international Volume: 95 Issue: 1 Pages: 123-137
  • DOI: 10.1016/j.kint.2018.08.030
  • Peer Reviewed
• [Journal Article] AKAPs-PKA disruptors increase AQP2 activity independently of vasopressin in a model of nephrogenic diabetes insipidus (2018)
  • Author(s): Ando Fumiaki、Mori Shuichi、Yui Naofumi、Morimoto Tetsuji、Nomura Naohiro、Sohara Eisei、Rai Tatemitsu、Sasaki Sei、Kondo Yoshiaki、Kagechika Hiroyuki、Uchida Shinichi
  • Journal Title: Nature Communications Volume: 9 Issue: 1 Pages: 1411-1411
  • DOI: 10.1038/s41467-018-03771-2
  • Peer Reviewed / Open Access
• [Journal Article] Decreased KLHL3 expression is involved in the pathogenesis of pseudohypoaldosteronism type II caused by cullin 3 mutation in vivo (2018)
  • Author(s): Yoshida S, Araki Y, Mori T, Sasaki E, Kasagi Y, Isobe K, Susa K, Inoue Y, Bomont P, Okado T, Rai T, Uchida S, Sohara E
  • Journal Title: Clinical and experimental nephrology Volume: 22 Issue: 6 Pages: 1251-1257
  • DOI: 10.1007/s10157-018-1593-z
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] WNK1 regulates skeletal muscle cell hypertrophy by modulating the nuclear localization and transcriptional activity of FOXO4 (2018)
  • Author(s): Mandai S, Mori T, Nomura N, Furusho T, Arai Y, Kikuchi H, Sasaki E, Sohara E, Rai T, Uchida S
  • Journal Title: Scientific reports Volume: 8 Issue: 1 Pages: 9101-9101
  • DOI: 10.1038/s41598-018-27414-0
  • Peer Reviewed / Open Access
• [Journal Article] Loop diuretics are associated with greater risk of sarcopenia in patients with non-dialysis-dependent chronic kidney disease (2018)
  • Author(s): Ishikawa Seiko、Naito Shotaro、Iimori Soichiro、Takahashi Daiei、Zeniya Moko、Sato Hidehiko、Nomura Naohiro、Sohara Eisei、Okado Tomokazu、Uchida Shinichi、Rai Tatemitsu
  • Journal Title: PLOS ONE Volume: 13 Issue: 2 Pages: e0192990-e0192990
  • DOI: 10.1371/journal.pone.0192990
  • Peer Reviewed / Open Access
• [Journal Article] Metformin increases urinary sodium excretion by reducing phosphorylation of the sodium-chloride cotransporter (2018)
  • Author(s): Hashimoto Hiroko、Nomura Naohiro、Shoda Wakana、Isobe Kiyoshi、Kikuchi Hiroaki、Yamamoto Kouhei、Fujimaru Takuya、Ando Fumiaki、Mori Takayasu、Okado Tomokazu、Rai Tatemitsu、Uchida Shinichi、Sohara Eisei
  • Journal Title: Metabolism Volume: 85 Pages: 23-31
  • DOI: 10.1016/j.metabol.2018.02.009
  • Peer Reviewed
• [Journal Article] KLHL3 Knockout Mice Reveal the Physiological Role of KLHL3 and the Pathophysiology of Pseudohypoaldosteronism Type II Caused by Mutant KLHL3. (2017)
  • Author(s): Sasaki E, Susa K, Mori T, Isobe K, Araki Y, Inoue Y, Yoshizaki Y, Ando F, Mori Y, Mandai S, Zeniya M, Takahashi D, Nomura N, Rai T, Uchida S, Sohara E.
  • Journal Title: Mol Cell Biol. Volume: 27 Issue: 7 Pages: 1-13
  • DOI: 10.1128/mcb.00508-16
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] WNK4 is an Adipogenic Factor and Its Deletion Reduces Diet-Induced Obesity in Mice. (2017)
  • Author(s): Takahashi D, Mori T, Sohara E, Tanaka M, Chiga M, Inoue Y, Nomura N, Zeniya M, Ochi H, Takeda S, Suganami T, Rai T, Uchida S.
  • Journal Title: EBioMedicine Volume: 18 Pages: 118-27
  • DOI: 10.1016/j.ebiom.2017.03.011
  • Peer Reviewed / Open Access
• [Journal Article] Impaired degradation of medullary WNK4 in the kidneys of KLHL2 knockout mice. (2017)
  • Author(s): Kasagi Y, Takahashi D, Aida T, Nishida H, Nomura N, Zeniya M, Mori T, Sasaki E, Ando F, Rai T, Uchida S, Sohara E.
  • Journal Title: Biochem Biophys Res Commun. Volume: 487 Issue: 2 Pages: 368-374
  • DOI: 10.1016/j.bbrc.2017.04.068
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Loop diuretics affect skeletal myoblast differentiation and exercise-induced muscle hypertrophy. (2017)
  • Author(s): Mandai S, Furukawa S, Kodaka M, Hata Y, Mori T, Nomura N, Ando F, Mori Y, Takahashi D, Yoshizaki Y, Kasagi Y, Arai Y, Sasaki E, Yoshida S, Furuichi Y, Fujii NL, Sohara E, Rai T, Uchida S.
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 46369-46369
  • DOI: 10.1038/srep46369
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Salt suppresses IFNγ inducible chemokines through the IFNγ-JAK1-STAT1 signaling pathway in proximal tubular cells. (2017)
  • Author(s): Arai Y, Takahashi D, Asano K, Tanaka M, Oda M, Ko SBH, Ko MSH, Mandai S, Nomura N, Rai T, Uchida S, Sohara E.
  • Journal Title: Sci Rep. Volume: 7 Issue: 1 Pages: 46580-46580
  • DOI: 10.1038/srep46580
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] WNK4 is indispensable for the pathogenesis of pseudohypoaldosteronism type II caused by mutant KLHL3 (2017)
  • Author(s): Susa Koichiro、Sohara Eisei、Takahashi Daiei、Okado Tomokazu、Rai Tatemitsu、Uchida Shinichi
  • Journal Title: Biochemical and Biophysical Research Communications Volume: 491 Issue: 3 Pages: 727-732
  • DOI: 10.1016/j.bbrc.2017.07.121
  • Peer Reviewed
• [Journal Article] 【腎臓学 この一年の進歩】 尿細管生理 (2017)
  • Author(s): 頼建光、内田信一
  • Journal Title: 日本腎臓学会誌 Volume: 50 Pages: 28-34
• [Journal Article] 【腎臓医が知っておきたい分子遺伝学の進歩】 疾患編 Bartter/Gitelman症候群、Liddle症候群、PHAII (2017)
  • Author(s): 頼建光
  • Journal Title: 腎と透析 Volume: 82 Pages: 431-439
• [Presentation] WNK1-OSR1/SPAK-NCCシグナルの亢進が慢性腎臓病モデルマウスの塩分感受性高血圧に関与する (2018)
  • Author(s): 古荘泰佑、蘇原映誠、萬代新太郎、菊池寛昭、銭谷慕子、野村尚弘、森崇寧、頼建光、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] FMP-API-1はバゾプレシンに匹敵するAQP2水チャネル活性化作用を持つ―新規腎性尿崩症治療薬の開発― (2018)
  • Author(s): 安藤史顕、油井直史、森本哲司、野村尚弘、蘇原映誠、頼建光、佐々木成、根東義明、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] 高カリウム負荷によるナトリウム-クロライド共輸送体の脱リン酸化機構の解明 (2018)
  • Author(s): 正田若菜、野村尚弘、安藤史顕、磯部清志、森崇寧、蘇原映誠、頼建光、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] メトホルミンは腎NCCのリン酸化を低下させ塩分排泄を促進する (2018)
  • Author(s): 橋本博子、蘇原映誠、野村尚宏、正田若菜、磯部清志、菊池寛昭、安藤史顕、森崇寧、頼建光、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] WNK1キナーゼはFOXO4の細胞核内局在，転写活性を調節し，骨格筋肥大を制御する (2018)
  • Author(s): 萬代新太郎、森崇寧、野村尚弘、古荘泰佑、新井洋平、菊池寛昭、佐々木絵美、蘇原映誠、頼建光、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] WNK1はマクロファージにおいてLPS誘導性サイトカイン産生を抑制する (2018)
  • Author(s): 新井洋平、蘇原映誠、浅野謙一、田中正人、頼建光、内田信一
  • Organizer: 第61回日本腎臓学会学術総会
• [Presentation] AKAPs-PKA結合阻害薬はバゾプレシン非依存性に強力にAQP2水チャネルを活性化する (2018)
  • Author(s): 安藤史顕、森修一、油井直史、森本哲司、野村尚弘、蘇原映誠、頼建光、佐々木成、根東義明、影近弘之、内田信一
  • Organizer: 第9回分子腎臓フォーラム
• [Presentation] AKAPs-PKA Disruptors Robustly Increase AQP2 Activity Independently of Vasopressin (2018)
  • Author(s): Ando F, Yui N, Nomura N, Rai T, Sasaki S, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2018
  • Int'l Joint Research
• [Presentation] WNK1-SPAK-NCC Signaling Cascade Is Involved in Salt Sensitive Hypertension Induced by Aristolochic Acid Nephropathy (2018)
  • Author(s): Furusho T, Uchida S, Sohara E, Mandai S, Kikuchi H, Ando F, Zeniya M, Mori T, Nomura N, Rai T
  • Organizer: American Society of Nephrology Kidney Week 2018
  • Int'l Joint Research
• [Presentation] Metformin Has Natriuretic Effects Through Reduction of the Sodium-Chloride Cotransporter Phosphorylation (2018)
  • Author(s): Hashimoto H, Okado T, Rai T, Uchida S, Sohara E, Nomura N, Shoda W, Isobe K, Kikuchi H, Yamamoto K, Fujimaru T, Ando F, Mori T
  • Organizer: American Society of Nephrology Kidney Week 2018
  • Int'l Joint Research
• [Presentation] Molecular Mechanism of High-Potassium Induced Na-Cl Cotransporter Dephosphorylation (2018)
  • Author(s): Shoda W, Nomura N, Ando F, Isobe K, Mori T, Sohara E, Rai T, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2018
  • Int'l Joint Research
• [Presentation] WNK1 Regulates Skeletal Muscle Hypertrophy by Modulating Phosphorylation, Nuclear Localization, and Transcriptional Activity of FoxO4 (2018)
  • Author(s): Mandai S, Uchida S, Mori T, Nomura N, Furusho T, Arai Y, Kikuchi H, Sasaki E, Sohara E, Rai T
  • Organizer: American Society of Nephrology Kidney Week 2018
  • Int'l Joint Research
• [Presentation] 塩は近位尿細管細胞においてJAK/STAT系を介してIFNgamma関連ケモカインを抑制する (2017)
  • Author(s): 新井洋平、蘇原映誠、高橋大栄、浅野謙一、田中正人、小田真由美、洪繁、洪実、萬代新太郎、野村尚弘、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] ループ利尿薬はマウス骨格筋細胞分化および運動による筋肥大効果を抑制する (2017)
  • Author(s): 萬代新太郎、古川晋、小高愛未、畑裕、森崇寧、野村尚弘、蘇原映誠、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] WNK4は脂肪細胞分化の制御因子である (2017)
  • Author(s): 高橋大栄、森崇寧、蘇原映誠、田中都、千賀宗子、井上佑一、野村尚弘、銭谷慕子、越智広樹、竹田秀、菅波孝祥、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] 変異CUL3によるPHAIIの発症にはKLHL3の発現低下が関与する (2017)
  • Author(s): 吉田紗矢香、蘇原映誠、荒木雄也、笠木祐里、佐々木絵美、須佐紘一郎、森崇寧、井上佑一、磯部清志、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] KLHL2は腎臓でWNK4蛋白の分解を行う (2017)
  • Author(s): 笠木祐里、蘇原映誠、相田知海、高橋大栄、西田秀範、野村尚弘、銭谷慕子、森崇寧、佐々木絵美、安藤史顕、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] WNK1はマクロファージにおけるLPS誘導性サイトカイン産生を抑制する (2017)
  • Author(s): 新井洋平、蘇原映誠、浅野謙一、田中正人、頼建光、内田信一
  • Organizer: 第60回日本腎臓学会学術総会
• [Presentation] WNK4は脂肪細胞分化の制御因子である (2017)
  • Author(s): 高橋大栄、森崇寧、蘇原映誠、田中都、千賀宗子、井上佑一、野村尚弘、銭谷慕子、越智広樹、竹田秀、菅波孝洋、頼建光、内田信一
  • Organizer: 第8回分子腎臓フォーラム
• [Presentation] WNK4 deletion inhibits adipogenesis in vitro and in vivo (2017)
  • Author(s): Takahashi D, Mori T, Sohara E, Miyako Tanaka M, Inoue Y, Nomura N, Chiga M, Zeniya M, Suganami T, Rai T, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2017
  • Int'l Joint Research
• [Presentation] Decreased protein expression of KLHL3 is involved in the pathogenesis of PHAII caused by CUL3 mutation in vivo (2017)
  • Author(s): Yoshida S, Araki Y, Mori T, Sasaki E, Kasagi Y, Isobe K, Susa K, Inoue Y, Rai T, Uchida S, Sohara E
  • Organizer: American Society of Nephrology Kidney Week 2017
  • Int'l Joint Research
• [Presentation] Generation and analysis of KLHL2 knockout mice (2017)
  • Author(s): Kasagi Y, Takahashi D, Aida T, Nishida H, Nomura N, Zeniya M, Mori T, Sasaki E, Ando F, Rai T, Uchida S, Sohara E
  • Organizer: American Society of Nephrology Kidney Week 2017
  • Int'l Joint Research
• [Presentation] A high salt diet suppressed CXCL9 and CXCL10 in proximal tubules through the IFNγ-JAK1-STAT1 signaling pathway (2017)
  • Author(s): Arai Y, Takahashi D, Asano K, Tanaka M, Oda M, Ko SB, Ko MS, Mandai S, Nomura N, Rai T, Uchida S, Sohara E
  • Organizer: American Society of Nephrology Kidney Week 2017
  • Int'l Joint Research
• [Presentation] Generation of p62/SQSTM1 (p62) KO cell lines using CRISPR/Cas9 -The role of p62 on WNK1 regulation by changes of extracellular potassium concentration-. (2016)
  • Author(s): Mori Y, Mandai S, Mori T, Sohara E, Rai T, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2016
  • Place of Presentation: Chicago USA
  • Year and Date: 2016-11-17
  • Int'l Joint Research
• [Presentation] Drug repositioning screening for the discovery of inhibitors of Keap1-Nrf2 interaction using fluorescent correlation spectroscopy. (2016)
  • Author(s): Yoshizaki Y, Sohara E, Mori T, Kikuchi E, Takahashi D, Zeniya M, Araki Y, Mori Y, Ando F, Nomura N, Rai T, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2016
  • Place of Presentation: Chicago USA
  • Year and Date: 2016-11-17
  • Int'l Joint Research
• [Presentation] The pseudohypoaldosteronism type II-causing mutant Cul3 protein forms dimer with KLHL3 and inhibits the degradation of WNK4 with a dominant-negative effect. (2016)
  • Author(s): Araki Y, Tatemitsu Rai T, Sohara E, Mori T, Inoue Y, Mori Y, Uchida S
  • Organizer: American Society of Nephrology Kidney Week 2016
  • Place of Presentation: Chicago USA
  • Year and Date: 2016-11-17
  • Int'l Joint Research
• [Presentation] Generation and Analysis of KLHL3 Knockout Mice. (2016)
  • Author(s): Sasaki E, Susa K, Mori T, Isobe K, Araki Y, Inoue Y, Rai T, Uchida S, Sohara E.
  • Organizer: American Society of Nephrology Kidney Week 2016
  • Place of Presentation: Chicago USA
  • Year and Date: 2016-11-17
  • Int'l Joint Research
• [Presentation] CRISPR/Cas9システムを用いた高血圧関連遺伝子多型STK39 rs3754777ノックイン細胞の樹立と解析 (2016)
  • Author(s): 萬代新太郎、森 崇寧、蘇原 映誠、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Presentation] CRISPR/Cas9システムを用いたp62ノックアウト細胞の作成とその解析-カリウムによるWNK1制御におけるp62の役割- (2016)
  • Author(s): 森雄太郎、小川 祥、萬代 新太郎、森 崇寧、蘇原 映誠、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Presentation] CaMK1gはWNK4 S1190をリン酸化する (2016)
  • Author(s): 新井洋平、蘇原 映誠、高橋 大栄、小田 真由美、洪 繁、洪 実、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Presentation] 血管平滑筋のカリウムによるKLHL2を介したWNK3シグナル制御機構解明 (2016)
  • Author(s): Wang Yuanlong、蘇原 映誠、銭谷 慕子、森 雄太郎、内藤 省太郎、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Presentation] KLHL3ノックアウトマウスの作製と解析 (2016)
  • Author(s): 佐々木絵美、蘇原 映誠、須佐 紘一郎、森 崇寧、磯部 清志、荒木 雄也、井上 佑一、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Presentation] 変異KLHL3による偽性低アルドステロン症II型の発症にはWNK4が必要である (2016)
  • Author(s): 須佐紘一郎、蘇原 映誠、高橋 大栄1、頼 建光、内田 信一
  • Organizer: 第59回日本腎臓学会学術総会
  • Place of Presentation: パシフィコ横浜会議センター 神奈川県横浜市
  • Year and Date: 2016-06-17
• [Remarks] FMP-API-1が腎臓において尿濃縮に重要な水チャネルを強力に活性化する
  • URL: http://tmd-kid.jp/wp-content/uploads/2019/04/20180413_1-Ando.pdf
• [Remarks] 慢性腎臓病の新たな治療標的としてエネルギー不全の感知障害を発見
  • URL: http://tmd-kid.jp/wp-content/uploads/2019/04/20181116_1-Kikuchi.pdf
• [Remarks] 糖尿病治療薬メトホルミンが、腎臓からの塩分排泄を増加させる
  • URL: http://www.tmd.ac.jp/archive-tmdu/kouhou/20180309_1.pdf
• [Remarks] 保存期腎不全患者におけるループ利尿薬使用はサルコペニア合併と関連する可能性がある
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