肾小管坏死不受调控是治疗重症急性肾损伤的难点。我们最近观察到肾小管necroptosis参与急性肾损伤。这是一种可被调控的细胞坏死，RIP3活化和自噬是其标志性特征。TFEB是自噬的关键转录因子，我们发现由它失活所介导的自噬障碍会促进necroptosis。TFEB失活受丝氨酸磷酸化调控，由于RIP3是丝氨酸激酶，我们推测RIP3可能会调控TFEB。我们用CoIP发现它们在necroptosis时相互结合，并观察到RIP3激酶抑制剂可活化TFEB，因此提出“RIP3可能通过激酶作用阻抑TFEB活化”。为了阐明这一推测，课题拟用pull down、液相质谱、磷酸化实验和点突变等技术来研究两者的作用，并利用mRFP-GFP-LC3转基因小鼠来验证抑制RIP3激酶对急性肾损伤的治疗价值。这些工作将为应用和研发RIP3激酶抑制剂提供理论基础，为治疗重症急性肾损伤提供新思路。

Unregulated necrosis in renal tubular epithelial cells is a therapeutic challenge in severe acute kidney injury. We observed recently that necroptosis, a novel proposed regulated necrosis, played a role in renal tubular cells. Activation of RIP3 and autophagy are symbolic characters of necroptosis. TFEB is a key transcription factor of autophagy. We found that TFEB inactivation mediated impaired autophagy promoted necroptosis. Because the TFEB inactivation is modulated by serine phosphorylation and RIP3 is a serine kinase, we speculated that RIP3 might block TFEB through kinase phosphorylation. We confirmed the interaction between RIP3 and TFEB using CoIP, and observed that RIP3 kinase inhibitor was able to activate TFEB. Based on these results above, this program proposes that RIP3 blocks TFEB activation through a kinase pathway. To confirm this hypothesis, we plan to explore the mechanism of interaction between the two molecules, using pull down, HPLC/MS, in vitro phosphorylation assay and point mutant technique. Finally, the therapeutic value of RIP3 kinase inhibitor will be testified in mRFP-GFP-LC3 transgene mouse. These works will facilitate to translate the RIP3 kinase inhibitor into clinical practice and drug development, and provide new ideas for therapy of severe acute kidney injury.