RIP1调控分子伴侣介导自噬的机理研究

Chaperone-mediated autophagy is known as an important selective autophagy. Scientists recently demonstrated that chaperone-mediated autophagy is involved in many physiological and pathological processes like metabolism and Parkinson's disease, but little is known about the mechanism and pathway regulating chaperone-mediated autophagy. We previously screened 1434 genes with HTS technology, and found RIP1 involved in regulating chaperone-mediated autophagy. With the help of Mass Spectrometry, we identified a novel phosphorylation modification for RIP1 under chaperone-mediated autophagy induction. Based on the preliminary results, we are going to explore the mechanism how RIP1 regulates chaperone-mediated autophagy. We will confirm that RIP1 is a novel regulator and signal protein for chaperone-mediated autophagy; we will demonstrate that the novel phosphorylation modification for RIP1 is required for the induction of chaperone-mediated autophagy; we will identify the target protein of RIP1 and address their interaction, and we will describe the pathway that RIP1 and its target protein involved regulating chaperone-mediated autophagy. This project will reveal the novel function of RIP1, delineate the novel mechanism regulating chaperone-mediated autophagy，and provide potential target protein for the therapies to major diseases such as Parkinson's disease and cancers.

分子伴侣介导自噬是一种重要的选择性自噬。虽然近年来人们对分子伴侣介导自噬在代谢和帕金森氏症等多种生理和病理过程中的重要作用有了更多的认识，但分子伴侣介导自噬的调控机制尚待进一步研究和明确。在前期的研究中，我们通过高通量基因筛选从1434个基因中发现RIP1参与了分子伴侣介导自噬的调控，运用生物质谱技术我们发现在分子伴侣介导自噬诱导的条件下，RIP1蛋白发生了新的磷酸化修饰。本项目拟在前期结果基础上，进一步研究RIP1调控分子伴侣介导自噬的机理，明确RIP1为新的分子伴侣介导自噬相关蛋白和信号分子；阐明RIP1新的磷酸化修饰在调控分子伴侣介导自噬中的必要性；鉴定RIP1的靶蛋白，并研究其相互作用和调控分子伴侣介导自噬的信号通路。本研究结果将发现RIP1新的功能，对分子伴侣介导自噬调控机制研究提出新的创新性见解，为治疗帕金森氏症、肿瘤等重要疾病提供潜在的靶蛋白。

分子伴侣介导自噬（CMA）是一种溶酶体依赖的降解途径，和多种生理和病理过程密切相关。二甲双胍是一种常用的抗糖尿病药物。我们通过高通量药物筛选鉴定二甲双胍可以诱导分子伴侣介导自噬。二甲双胍通过RIPK1依赖的途径激活IKKα/β，IKKα/β随后调控Hsc70 Ser85位点磷酸化，从而诱导分子伴侣介导自噬。我们进一步证明二甲双胍激活RIPK1-IKKα/β不依赖于AMPK通路。同时，我们筛选新型RIPK1激酶抑制剂，获得了一批具有自主知识产权的新的药物先导化合物，开发了一系列新的有机合成方法。