Elucidation of the mechanism of the promoting effect of CNP/NPR-B signaling on endochondral bone growth

• Project/Area Number: 17K09881
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Endocrinology
• Research Institution: Department of Clinical Research, National Hospital Organization Kyoto Medical Center (2019); Kyoto University (2017-2018)
• Principal Investigator: Yasoda Akihiro 独立行政法人国立病院機構(京都医療センター臨床研究センター), その他部局等, 臨床研究センター長 (50378642)
• Project Period (FY): 2017-04-01 – 2020-03-31
• Project Status: Completed (Fiscal Year 2019)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2017: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 骨伸長 / CNP / 器官培養 / 成長板 / シグネチャー / イメージング / 二光子顕微鏡 / 細胞内シグナル / マイクロアレイ / シグナル解析 / 細胞・組織 / 生理活性

Outline of Final Research Achievements

C-type natriuretic peptide (CNP) potently promotes endochondral bone growth as an intrinsic growth factor. Here we aimed to elucidate molecular cascade of the growth promoting effect of CNP in the growth plate by using array analyses of expressing genes, as well as morphological analyses of dynamic imaging of tissue culture using fetal murine long bones. We performed gene set enrichment analysis (GSEA) of two gene sets, i.e., the subtraction of expressing genes in the growth plate of CNP knockout rats treated with vehicle from those with CNP, and that of wild-type rats treated with vehicle from those with an ERK inhibitor, U0126. The resultant two gene sets correlated well, and furthermore, live imaging analyses using two-photon microscope revealed that CNP and U0126 displayed similar growth promoting effect of explanted ulnae from fetal CGA-EGFP mice. These results indicate that ERK pathway is important as an intracellular signaling of the effect of CNP on endochondral bone growth.

Academic Significance and Societal Importance of the Research Achievements

CNP/NPR-Bシグナルの骨伸長促進作用は強力であり、現在、軟骨無形成症をはじめとする骨伸長障害に対する展開研究や臨床応用が進められている。本研究ではCNP/NPR-Bシグナルの分子カスケードについて、最近注目されているシグネチャー解析による解明を試み、さらに二光子蛍光顕微鏡を用いて哺乳類成長板ではじめての4Dイメージング解析をおこなってその成果をフィードバックし、ERKシグナルがCNPの成長板軟骨の伸長促進に重要であることを明らかにした。

Research Products

• [Journal Article] Foramen magnum stenosis and midface hypoplasia in C-type natriuretic peptide-deficient rats and restoration by the administration of human C-type natriuretic peptide with 53 amino acids (2019)
  • Author(s): Yotsumoto Takafumi、Morozumi Naomi、Furuya Mayumi、Fujii Toshihito、Hirota Keisho、Ueda Yohei、Nakao Kazumasa、Yamanaka Shigeki、Yoshikiyo Kazunori、Yoshida Sayaka、Nishimura Tomonari、Abe Yasuyuki、Jindo Toshimasa、Ogasawara Hiroyuki、Yasoda Akihiro
  • Journal Title: PLOS ONE Volume: 14 Issue: 5 Pages: e0216340-e0216340
  • DOI: 10.1371/journal.pone.0216340
  • Peer Reviewed / Open Access
• [Journal Article] オステオクリンとC型ナトリウム利尿ペプチド (2019)
  • Author(s): 八十田 明宏、金井 有吾
  • Journal Title: 腎と骨代謝 Volume: 32 Pages: 158-163
  • Int'l Joint Research
• [Journal Article] Live imaging analysis of the growth plate in a murine long bone explanted culture system (2018)
  • Author(s): Hirota Keisho、Yasoda Akihiro、Kanai Yugo、Ueda Yohei、Yamauchi Ichiro、Yamashita Takafumi、Sakane Yoriko、Fujii Toshihito、Inagaki Nobuya
  • Journal Title: Scientific Reports Volume: 8 Issue: 1 Pages: 1-9
  • DOI: 10.1038/s41598-018-28742-x
  • NAID: 120006502590
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Exogenous C-type natriuretic peptide restores normal growth and prevents early growth plate closure in its deficient rats (2018)
  • Author(s): Hirota Keisho、Furuya Mayumi、… Yasoda Akihiro、Inagaki Nobuya
  • Journal Title: PLOS ONE Volume: 13 Issue: 9 Pages: e0204172-e0204172
  • DOI: 10.1371/journal.pone.0204172
  • Peer Reviewed / Open Access
• [Journal Article] Rats deficient C-type natriuretic peptide suffer from impaired skeletal growth without early death. (2018)
  • Author(s): Fujii T, Hirota K, Yasoda A, Takizawa A, Morozumi N, Nakamura R, Yotsumoto T, Kondo E, Yamashita Y, Sakane Y, Kanai Y, Ueda Y, Yamauchi I, Yamanaka S, Nakao K, Kuwahara K, Jindo T, Furuya M, Mashimo T, Inagaki N, Serikawa T, Nakao K.
  • Journal Title: PROS ONE Volume: 13 Issue: 3 Pages: e0194812-e0194812
  • DOI: 10.1371/journal.pone.0194812
  • NAID: 120006466606
  • Peer Reviewed / Open Access
• [Journal Article] Circulating osteocrin stimulates bone growth by limiting C-type natriuretic peptide clearance. (2017)
  • Author(s): Kanai Y, Yasoda A, Mori KP, Watanabe-Takano H, Nagai-Okatani C, Yamashita Y, Hirota K, Ueda Y, Yamauchi I, Kondo E, Yamanaka S, Sakane Y, Nakao K, Fujii T, Yokoi H, Minamino N, Mukoyama M, Mochizuki N, Inagaki N.
  • Journal Title: Journal of Clinical Investigation Volume: 127 Issue: 11 Pages: 4136-4147
  • DOI: 10.1172/jci94912
  • Peer Reviewed / Open Access