自噬调节蛋白LRPPRC介导的纤连蛋白降解在角膜损伤修复中的作用及机制研究

Cornea wound healing is a normal physiological response after corneal injury to maintain cornea function. Most of glycoprotein extracellular matrix including fibronectin have been shown to modulate cornea wound healing, and the absence of glycoprotein extracellular matrix could lead to cornea wound healing delay and visual impairment. Recently, a new study in our group found fibronectin could be degraded in common pathway involving autophagy and endolysosomal pathway by forming amphisome, and LRPPRC could affect the level of autophagy in ocular fibroblasts. Therefore, we speculate the autophagy induced by LRPPRC may influence the degradation of fibronectin in cornea and LRPPRC could be as an important regulatory factor in the process of cornea wound healing. In our study, we plan to detect the autophagy level and degradation of fibronectin when the level of LRPPRC change in human ocular fibroblasts; compare the degradation of FN-FITC in LRPPRC knockout mice and wide type mice ocular fibroblasts; try to explain the effect of LRPPRC on autophagy and multiple biological processes involved in cornea wound healing by establishing cornea wound model in these two different mice separately. This study will find a new research way about the molecular mechanism in the process of cornea wound healing, and provide a potential target for clinical treatment of corneal injury.

角膜损伤后机体将产生正常的生理反应即角膜损伤修复来维持角膜的生理功能。角膜中纤连蛋白等糖蛋白类细胞外基质的缺失将造成角膜损伤修复延迟，并导致视力的严重损害。本课题组最新研究发现纤连蛋白能通过自噬与内吞溶酶体途径的交叉通路发生降解，并且自噬调节蛋白LRPPRC可以调控角膜成纤维细胞内的自噬水平。因此，我们推测自噬调节蛋白LRPPRC可能通过介导自噬影响角膜中纤连蛋白的降解，并成为调控角膜损伤修复的重要因子。本课题拟通过检测人角膜成纤维细胞中LRPPRC蛋白水平的改变对自噬以及纤连蛋白降解的影响；培育野生型小鼠与LRPPRC基因敲除小鼠，比较两种小鼠角膜成纤维细胞中外源性FN-FITC的降解情况；并在两种小鼠中建立角膜损伤模型，以阐述LRPPRC蛋白水平对自噬及角膜损伤修复的多个生物学过程的影响。本研究将为角膜损伤修复过程中的分子机制开辟新的研究思路，为角膜损伤的临床治疗提供潜在的靶标。