Protein phosphatase SCP4 regulates cartilage development and endochondral osteogenesis via FoxO3a dephosphorylation

Pinger Wang; Kaiao Zou; Jin Cao; Zhengmao Zhang; Wenhua Yuan; Jiali Chen; Jianbo Xu; Zhen Zou; Di Chen; Hongfeng Ruan; Jianying Feng; Xia Lin; Hongting Jin

doi:10.1111/cpr.13691

Cell Proliferation ›› 2024, Vol. 57 ›› Issue (9) : e13691 DOI: 10.1111/cpr.13691

ORIGINAL ARTICLE

Protein phosphatase SCP4 regulates cartilage development and endochondral osteogenesis via FoxO3a dephosphorylation

Pinger Wang ¹^,²
, Kaiao Zou ¹^,²
, Jin Cao ³
, Zhengmao Zhang ⁴
, Wenhua Yuan ¹^,²
, Jiali Chen ¹^,²
, Jianbo Xu ¹^,²
, Zhen Zou ¹^,²
, Di Chen ⁵^,⁶
, Hongfeng Ruan ¹^,²^,^†
, Jianying Feng ⁷^,^†
, Xia Lin ⁸^,^†
, Hongting Jin ¹^,²^,^†

Author information +

¹. Institute of Orthopedics and Traumatology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Chinese Medicine, Hangzhou, Zhejiang, China

². The First College of Clinical Medicine, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China

³. The MOE Key Laboratory of Biosystems Homeostasis and Protection and Zhejiang Provincial Key Laboratory of Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang, China

⁴. Department of Pathology and Laboratory Medicine, Weill Cornell Medicine, New York City, New York, USA

⁵. Research Center for Computer-aided Drug Discovery, Chinese Academy of Sciences, Shenzhen Institute of Advanced Technology, Shenzhen, Guangdong, China

⁶. Faculty of Pharmaceutical Sciences, Chinese Academy of Sciences, Shenzhen Institute of Advanced Technology, Shenzhen, Guangdong, China

⁷. School of Stomatology, Zhejiang Chinese Medical University, Hangzhou, Zhejiang, China

⁸. Department of Hepatobiliary and Pancreatic Surgery and Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

rhf@zcmu.edu.cn

twohorsejy@163.com

linxia79@yahoo.com

hongtingjin@163.com

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Abstract

The regulatory mechanisms involved in embryonic development are complex and yet remain unclear. SCP4 represents a novel nucleus-resident phosphatase identified in our previous study. The primary aim of this study was to elucidate the function of SCP4 in the progress of cartilage development and endochondral osteogenesis. SCP4^–/– and SCP4^Col2ER mice were constructed to assess differences in bone formation using whole skeleton staining. ABH/OG staining was used to compare chondrocyte differentiation and cartilage development. Relevant biological functions were analysed using RNA-sequencing and GO enrichment, further validated by immunohistochemical staining, Co-IP and Western Blot. Global SCP4 knockout led to abnormal embryonic development in SCP4^–/– mice, along with delayed endochondral osteogenesis. In parallel, chondrocyte-specific removal of SCP4 yielded more severe embryonic deformities in SCP4Col2ER mice, including limb shortening, reduced chondrocyte number in the growth plate, disorganisation and cell enlargement. Moreover, RNA-sequencing analysis showed an association between SCP4 and chondrocyte apoptosis. Notably, Tunnel-positive cells were indeed increased in the growth plates of SCP4^Col2ER mice. The deficiency of SCP4 up-regulated the expression levels of pro-apoptotic proteins both in vivo and in vitro. Additionally, phosphorylation of FoxO3a (pFoxO3a), a substrate of SCP4, was heightened in chondrocytes of SCP4^Col2ER mice growth plate, and the direct interaction between SCP4 and pFoxO3a was further validated in chondrocytes. Our findings underscore the critical role of SCP4 in regulating cartilage development and endochondral osteogenesis during embryonic development partially via inhibition of chondrocytes apoptosis regulated by FoxO3a dephosphorylation.

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Pinger Wang, Kaiao Zou, Jin Cao, Zhengmao Zhang, Wenhua Yuan, Jiali Chen, Jianbo Xu, Zhen Zou, Di Chen, Hongfeng Ruan, Jianying Feng, Xia Lin, Hongting Jin. Protein phosphatase SCP4 regulates cartilage development and endochondral osteogenesis via FoxO3a dephosphorylation. Cell Proliferation, 2024, 57(9): e13691 DOI:10.1111/cpr.13691

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