SPARC抑制VEGF/VEGFR信号通路调控星形胶质细胞CD59表达参与NMO中枢特异性病灶形成的机制研究

视神经脊髓炎(NMO)是一种严重的中枢神经系统(CNS)炎性脱髓鞘疾病。上一课题我们发现星形胶质细胞CD59的表达缺失导致了NMO中枢特异性病灶的形成，但其表达调控机制不清。预研发现神经血管内皮细胞(NECs)与星胶共培养或其分泌的SPARC蛋白可抑制星胶CD59的表达，而星胶中CD59可能与细胞周期检查点基因互为ceRNA，但二者如何关联并影响CD59的表达尚需探索。本研究拟分析不同组织标本及不同处理条件下星胶中CD59的表达规律，利用比较蛋白质组学及RNA-seq分析NECs特征性作用于星胶的分泌物以及星胶对此响应的信号通路，并结合“knock in”、“knock down”、信号通路抑制剂等方法揭示影响星胶CD59表达的信号通路及其调控机制。研究结果对进一步明确NMO中枢特异性病灶形成机制及为以阻断NECs分泌物调节星胶CD59表达的信号通路为基础的药物新靶点的发现及干预奠定基础。

Neuromyelitis optica (NMO) is a serious inflammatory demyelinating disease in central nervous system (CNS). With support of the last NSFC grant, we have found that lost of the expression of CD59 in astrocytes contributed to the CNS-restricted lesion development in NMO, but the detailed mechanisms of CD59 regulation remained unknown. Our preliminary data showed that the co-culture of astrocytes with neurovascular endothelial cells (NECs) or adding the SPARC protein secreted by NECs decreased the expression of CD59 in astrocytes, and also found that the CD59 and some cell cycle check-point genes worked as ceRNA with each other, however, how these two phenomenon are related to regulate the expression of CD59 remains to be explored. This project will: 1) Analyze the expression profile of CD59 in brain and primary astrocytes with different stimulation by immunohistochemistry and real-time PCR methods; 2) Use comparative proteomics and RNA-Seq methods to identify the secretes from NECs to regulate the expression of CD59 and the responders which expressed on astrocytes membrane to the secretes; 3) Identify and confirm the regulation mechanisms for CD59 expression by the “knock in”, “knock down” methods and signaling pathway inhibitors. The results will help us better understand the mechanisms of CNS-restricted lesion development of NMO, and it will also help us to identify the novel therapeutic target by blocking the secretes-responder communications.