Intraarticular injection of SHP2 inhibitor SHP099 promotes the repair of rabbit full-thickness cartilage defect

Ziying Sun; Xingquan Xu; Zhongyang Lv; Jiawei Li; Tianshu Shi; Heng Sun; Kuoyang Sun; Guihua Tan; Wenqiang Yan; Yannick Xiaofan Yang; Rui Wu; Jia Xu; Hu Guo; Qing Jiang; Dongquan Shi

doi:10.1016/j.jot.2022.01.001

Intraarticular injection of SHP2 inhibitor SHP099 promotes the repair of rabbit full-thickness cartilage defect

J Orthop Translat. 2022 Feb 17:32:112-120. doi: 10.1016/j.jot.2022.01.001. eCollection 2022 Jan.

Authors

Ziying Sun^{1

2}, Xingquan Xu^{1

2}, Zhongyang Lv^{1

2}, Jiawei Li^{1

2}, Tianshu Shi^{1

2}, Heng Sun^{1

2}, Kuoyang Sun^{1

2}, Guihua Tan^{1

2}, Wenqiang Yan^{1

2}, Yannick Xiaofan Yang^{1

3}, Rui Wu^{1

2}, Jia Xu^{1

3}, Hu Guo^{1

2}, Qing Jiang^{1

2}, Dongquan Shi^{1

2}

Affiliations

¹ State Key Laboratory of Pharmaceutical Biotechnology, Department of Sports Medicine and Adult Reconstructive Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, 210008, Jiangsu, PR China.
² Laboratory for Bone and Joint Disease, Model Animal Research Center (MARC), Nanjing University, Nanjing, 210093, Jiangsu, PR China.
³ Drum Tower of Clinical Medicine, Nanjing Medical University, Nanjing, 210008, Jiangsu, PR China.

Abstract

Background: Cartilage repair has been a challenge in the field of orthopaedics for decades, highlighting the significance of investigating potential therapeutic drugs. In this study, we explored the effect of the SHP2 inhibitor SHP099, a small-molecule drug, on cartilage repair.

Methods: Human synovial mesenchymal stem cells (SMSCs) were isolated, and their three-way differentiation potential was examined. After treatment with chondrogenic medium, the chondrogenic effect of SHP099 on SMSCs was examined by western blot, qPCR, and immunofluorescence (IF). Micro-mass culture was also used to detect the effect of SHP099. To explore the chondrogenic effects of SHP099 in vivo, full-thickness cartilage defects with microfractures were constructed in the right femoral trochlea of New Zealand White rabbits. Intraarticular injection of SHP099 or normal saline was performed twice a week for 6 weeks. Cartilage repair was evaluated by haematoxylin and eosin (HE) staining and safranin O/fast green staining. Immunohistochemistry (IHC) for collagen II (COL2) was also conducted to verify the abundance of cartilage extracellular matrix after SHP099 treatment. The mechanism involving yes-associated protein (YAP) and WNT signalling was investigated in vitro.

Results: SMSCs isolated from human synovium have optimal multi-differentiation potential. SHP099 increased chondrogenic marker (SOX9, COL2) expression and decreased hypertrophic marker (COL10, RUNX2) expression in SMSCs. In micro-mass culture, the SHP099-induced cartilage tissues had a better result of Safranin O and Toluidine blue staining and are enriched in cartilage-specific collagen II. Inhibition of YAP and WNT signalling was also observed. Moreover, compared to the normal saline group at 6 weeks, intraarticular injection of SHP099 resulted in better defect filling, forming increased hyaline cartilage-like tissue with higher levels of glycosaminoglycan (GAG) and COL2.

Conclusion: SHP099 promotes the repair of rabbit full-thickness cartilage defects, representing a potential therapeutic drug for cartilage repair.

The translational potential of this article: This study provides evidence that SHP2 inhibition promotes chondrogenesis and the repair of cartilage in defect area, which could be a novel therapeutic approach for cartilage repair.

Keywords: Cartilage repair; Src-homology 2-containing protein tyrosine phosphatase 2; Synovial mesenchymal stem cells.