自噬是一种高度保守的降解胞内蛋白及受损细胞器的主要途径，对维持细胞稳态、调控多种疾病具有重要作用。近年发现一些化合物可以在某些自噬基因缺失的情况下诱导非经典自噬，但具体调节机制并不明确。我们近期研究发现，新化合物AMDE1可以不依赖ULK1和Beclin1诱导非经典自噬的发生，泛素化水平和定位与自噬作用密切相关。特异性地抑制V-ATPase或沉默其亚基可以阻断非经典自噬的发生，且该作用具有普遍性。由此提出V-ATPase介导非经典自噬这一新观点。本项目拟在前期基础上，确定化合物AMDE1诱导非经典自噬发生的具体通路；阐明V-ATPase所在细胞器对自噬体形成的影响；明确V-ATPase、泛素蛋白和Atg16之间的作用关系及在自噬调控中的作用。本项目的开展有助于在细胞和分子水平揭示非经典自噬的发生平台和调节机制，拓宽对不同种类自噬作用方式的理解，为自噬相关疾病的干预提供新的见解。

Autophagy is a highly evolutionarily conserved pathway to degrade intracellular proteins and damaged organelles. Autophagy plays important physiological roles in maintaining cell homeostasis and regulating various diseases. Recently, studies showed that chemicals could bypass some core autophagic genes to induce autophagy, but the regulatory mechanisms are still unclear. In our recent work, one novel chemical AMDE1 was found to induce ULK1- and Beclin1-independent non-canonical autophagy. The ubiquitin level is tightly realated to the function of AMDE1-induced autophagy. We also found that specific inhibition of V-ATPase could generally suppress non-canonical autophagy. So a novel hypothesis was put forward that V-ATPase could mediate the induction of non-canonical autophagy. Based on our previous works, we try to determine the signal pathways of AMDE1 treatment in detail, clarify the effect of V-ATPase positive structure on the formation of autophagosome, understand the interaction among V-ATPase, ubiquitin, and Atg16, and their roles in the process of non-canonical autophagy. The implementation of this work will be helpful to elucidate the regulatory mechanism of non-canonical autophagy in the cellular and molecular level, broaden the understanding of different working pattern of autophagy, and provide the new ideas for regulating autophay-related pathophysiological processes.