人椎间盘退变过程中lncRNA-C15orf2-10促进髓核细胞MMP-2表达和ECM降解的机制研究

Intervertebral disc degeneration (IVDD) has important clinical significance, but its mechanism has not been clarified. The degradation of extracellular matrix (ECM) resulting from over-secretion of matrix metalloproteinases (MMPs) in nucleus pulposus cell is the key process. However the mechanism underneath the abnormal expression of MMPs in IVDD is unclear. In the early stage of the project, we found miR-874 has the ability to regulate MMP2 expression in nucleus pulposus cells; further screening indicated that the expression level of LncRNA C15orf2-10 increased significantly in the nucleus pulposus cells of IVDD. The preliminary result and predicted results suggest that C15orf2-10 might promote the expression of MMP2 by inhibiting miR-874 through competitive endogenous RNA mechanism (ceRNA), leading to excessive degradation of ECM. This study plan to investigate the role of C15orf2-10 in IVDD and the associated ceRNA mechanism by using the technology of gene-expression chip, bioinformatics analysis, qRT-PCR, RIP，Gene -reporter system, and the CRISPR-Cas9 gene editing tool. This project aims to provide new theoretical reference for understanding and eventually curing of IVDD.

椎间盘退变(Intervertebral disc degeneration，IVDD)具有重要的临床意义，但其发生机制尚未阐明，髓核细胞过度分泌基质金属蛋白酶（MMPs）导致的细胞外基质（ECM）过度降解可能是其中关键环节，然而MMPs异常表达的机制尚不明确。项目组前期发现髓核细胞中miR-874可以调节MMP-2的表达，进一步芯片筛选发现LncRNA C15orf2-10在人退变椎间盘髓核细胞中显著增高，预实验及预测结果提示，其可能通过内源性竞争性RNA(ceRNA)机制抑制miR-874并促进MMP-2表达，导致ECM过度降解。本研究拟综合运用基因芯片、生物信息学分析、CRISPR-Cas9技术、qRT-PCR、RIP及报告基因系统等研究技术，分析C15orf2-10参与椎间盘退变调控网络中的病理意义及其ceRNA相关机制，为认识IVDD发生机制和新的诊疗策略研究提供新的参考。

椎间盘退变(Intervertebral disc degeneration，IVDD)具有重要的临床意义，但其发生机制尚未阐明，髓核细胞过度分泌基质金属蛋白酶（MMPs）导致的细胞外基质（ECM）过度降解可能是其中关键环节，然而MMPs异常表达的机制尚不明确。本项目组研究发现miR-874在人IVDD中表达下调，直接导致MMP-2、MMP-3的转录后上调，并最终促进髓核细胞凋亡及细胞外基质的过度降解而引发椎间盘退变，而上调miR-874可显著下调MMPs的转录水平，抑制髓核细胞凋亡和细胞外基质降解，从而对椎间盘退变起保护作用。同时通过构建一款新型水凝胶纤维支架，在为髓核细胞提供稳定三维微环境，引导髓核细胞取向性生长和加快髓核再生的同时，具备 “疾病触发释放”功能，按时按需将金属蛋白酶抑制剂（AgomiRNA）作用于病变部位，抑制MMPs的表达，促进椎间盘修复。在此基础上，深入诠释相关抗凋亡及胶原合成代谢细胞信号通路，为防治IVDD提供理论依据。