BRE1A在骨关节炎软骨退变中的调控机制研究

骨关节炎软骨退变是目前国际研究热点，对指导临床治疗具有重要意义。我们前期研究发现新的抑制骨关节炎软骨退变的核心分子且能作用于基质金属蛋白酶ADAMTS的蛋白，即泛素化E3连接酶BRE1A。本研究拟对BRE1A在骨关节炎软骨退变中的调控机制进行研究，通过利用荧光素酶报告基因系统、ChIP和re-ChIP等技术，确定BRE1A对ADAMTS的调节机制；利用转录因子芯片、RNA-seq和ChIP-seq等确定BRE1A对上游调节因子和下游靶基因的影响；通过检测细胞外基质成分、细胞生长和细胞代谢等体外实验检测BRE1A对骨关节炎软骨细胞的影响；通过对BRE1A软骨特异性基因敲除小鼠进行组织病理学、μCT、MRI和功能学评定等体内实验检测BRE1A在骨关节炎软骨退变中的生物学功能；利用临床标本确定BRE1A与ADAMTS和下游靶基因表达的相关性及其临床意义，为阐明骨关节炎软骨退变机制提供新的思路。

Osteoarthritis articular cartilage degeneration has currently been a hot topic of international academic research, which is of great significance to guide clinical treatment. Our previous study found that BRE1A, a novel molecule and E3 ubiquitin ligase, could inhibit the osteoarthritis cartilage degeneration and regulate the ADAMTS expression. This study aims to investigate the function and regulatory mechanism of BRE1A in osteoarthritis cartilage degeneration. Luciferase reporter system, ChIP and re-ChIP assay will be performed to study the mechanism of BRE1A in regulating ADAMTS. Transcription factor chip, RNA-seq and ChIP-seq will be applied to determine the effect of BRE1A on upstream regulatory factors and downstream target genes. The effect of BRE1A on chondrocyte phenotype in osteoarthritis will be investigated by detecting extracellular matrix, cell growth and cell metabolism in vitro. Histopathology, μCT, MRI and functional evaluation in vivo in cartilage-specific BRE1A knockout mice will be examined to reveal the biological function of BRE1A in cartilage degeneration of osteoarthritis. The clinical specimens will be used to determine the correlation and clinical significance among the expression of BRE1A, ADAMTS and the downstream target genes, contributing to provide new avenues for elucidating the mechanism of osteoarthritis cartilage degeneration.