Post-doc The Chinese University of Hong Kong (CUHK), Hong Kong
Abstract: As a skeletal neurotrophin, nerve growth factor (NGF) participates in the maintenance of skeletal pain, and NGF signal pathway involves in bone formation and fracture healing. Bone loss occurs as a result of traumas and injuries, and poor bone healing leads to functionally debilitating conditions of patients. Human bone marrow mesenchymal stem cells (BMSC) contribute to the homeostasis of the skeletal system under the regulation of different cytokine profiles. NGF can promote BMSC survival during BMSC transplantation therapy. The NGF-derived peptides were investigated here because of the advantages of low immunogenicity, high bioactivity, and specific binding to tropomyosin receptor kinase A (TrkA, high-affinity receptor for NGF) in our earlier study. In this study, we used artificial intelligence-based approaches to generate NGF-derived peptides. Two peptide candidates were screened by molecular docking simulation and surface plasmon resonance (SPR) experiments. Human primary BMSCs were isolated from hip femoral heads obtained from patients who underwent total hip arthroplasty. To study the biological effects of the peptide candidates, hBMSCs were cultured in an osteogenic medium with NGF or NGF-derived peptides for 4 weeks. Alizarin red and von Kossa staining intensity revealed that NGF-derived peptides significantly enhanced the calcification of hBMSC compared to the control group. We then established an immortalized hBMSC-derived single-cell clone. RNA sequence analysis in this cell line demonstrated that inflammation- and angiogenesis-related signalings were induced by NGF-derived peptides. At last, the peptide candidates were delivered into the osseous cavities in rat models with bone defects to verify their function on bone formation in vivo, and fast bone healing was observed in the treatment groups at week 1 and week 4.
Funding Source: 1. National Key R&D Program, the Ministry of Science and Technology (MOST, China), (2019YFA0111900 to Yangzi Jiang) 2. Center for Neuromusculoskeletal Restorative Medicine (CNRM, Health@InnoHK to Yangzi Jiang)
