CrkL、CrkI/II介导RhoGTPases/Cdc42/Cdc42EP1调控足细胞损伤的作用机制及临床价值研究

足细胞损伤是导致蛋白尿及肾功能进行性恶化的病生理基础，目前维持足细胞结构功能的关键分子及调控机制尚未阐明。CrkL、CrkI/II是一类胞内信号接头蛋白。本研究前期工作发现：CrkL在肾小球疾病患者尿液中表达异常；体外沉默CrkI/II、CrkL可引起足细胞骨架紊乱，黏附、迁移能力下降；差异蛋白质组学筛选发现Crk缺失影响下游RhoGTPases/Cdc42/Cdc42EP1分子表达。因此我们推论：CrkI/II、CrkL缺失可介导RhoGTPases/Cdc42/Cdc42EP1通路并影响足细胞损伤的关联效应分子表达，从而破坏GBM屏障。本项目拟进一步以足细胞、斑马鱼和小鼠模型为研究对象，探讨Crk调控网络及相关差异蛋白在维持足细胞功能及肾脏滤过功能所发挥的作用，通过临床样本检测评价Crk及关联效应分子的临床意义，阐明Crk介导足细胞损伤的作用机制，为延缓肾小球疾病进展提供科学依据。

Podocyte injury is the physiological basis of proteinuria, and progressive deterioration of renal function. At present, the key protein molecules and possible regulatory mechanisms to maintain podocyte structure and function remain incompletely understood. CrkL and Crk I/II are reported to be important intracellular signaling adaptor proteins. Our previous study showed that CrkL was abnormally expressed in the urine of patients with glomerular disease. In vitro experiments, knockdown of Crk I/II and/or CrkL induced podocyte cytoskeleton disorganized, decreased cell adhesion and migration ability. Differential proteomics screening found that knockdown of CrkI/II and/or CrkL affected the RhoGTPases/Cdc42/Cdc42EP1 pathway and related proteins expression. Therefore, we hypothesise that specific knockdown of CrkI/II and/or CrkL mediated RhoGTPases/Cdc42/Cdc42EP1 pathway contributes to related effector molecular expression, which induced podocyte injury and damaged to the glomerular filtration barrier. Next, we intend to use human podocyte and CrkI/II and CrkL knockdown zebrafish and mice models as research subjects. To comprehensively explore the regulation mechanism mediated by Crk, and to investigate the role of related differential proteins in maintaining podocyte and glomerular filtration function. We also further to evaluate the clinical significance of Crk and related differential protein through detecting clinical samples. Together, these will elucidate the molecular mechanism of Crk I/II and CrkL mediating podocyte injury, and to provide a reliable scientific basis for delaying the progression of glomerular diseases.