基于多组学分析的CaSR-CAV1-EGFR信号轴在肾结石内质网应激中的分子机制研究

结晶-细胞相互作用是影响肾结石形成的关键阶段。申请者先前发现内质网应激是这一相互作用的重要生物学过程，但其上下游分子机制尚不清楚。本项目在肾结石多组学整合分析基础上，聚焦肾结石形成的关键基因CaSR、CAV1和EGFR，拟通过利用CaSR激动剂和抑制剂，揭示结晶诱导内质网应激的发生机制，并通过构建肾脏特异性表达CAV1腺病毒，研究CAV1对CaSR介导的Ca2+释放的调控，进而探讨CaSR-CAV1信号轴对肾结石形成的影响。同时，研究结晶诱导内质网应激情况下，CAV1和EGFR蛋白下调与泛素-蛋白酶体降解途径的关系，揭示CAV1对EGFR及其下游Akt信号通路的调控作用，探讨CAV1-EGFR信号轴对肾结石形成的影响。本项目将阐明CaSR-CAV1-EGFR信号轴在肾结石内质网应激中的分子机制和对肾结石形成的影响，完善肾脏钙化理论，为防治肾结石疾病提供新的靶点和思路。

Crystal-cell interaction is the key step affecting the formation of kidney stone. Applicants have previously found that endoplasmic reticulum stress is an important biological process of the crystal-cell interaction, but its upstream and downstream molecular mechanisms are not clear. Based on the multi-omics analysis of kidney stone, this project focuses on the key genes CaSR, CAV1 and EGFR, of kidney stone. Through the use of CaSR agonists and inhibitors and the construction of renal-specific adenovirus expressing CAV1, we intend to reveal the mechanism of crystals-induced endoplasmic reticulum stress, study the regulation of CAV1 on CaSR-mediated Ca2+ release, and explore the effect of CaSR-CAV1 signal axis on the formation of kidney stone. At the same time, we study the relationship between the down-regulation of CAV1 and EGFR proteins and ubiquitin-proteasome degradation pathway under endoplasmic reticulum stress, to reveal the regulation of CAV1 on EGFR and its downstream Akt signal pathway, and to explore the effect of CAV1-EGFR signal axis on the formation of kidney stone. This project will clarify the role of CaSR-CAV1-EGFR signal axis in endoplasmic reticulum stress and its effect on the formation of kidney stone, improve the theory of nephrocalcinosis, and provide new targets and ideas for the prevention and treatment of kidney stone diseases.