猪血凝性脑脊髓炎病毒（PHEV）是一种嗜神经性冠状病毒，主要侵害仔猪中枢神经系统诱发神经细胞损伤。本团队前期研究发现，PHEV感染神经细胞可诱导自噬体形成，但自噬启动激酶ULK1表达却显著下调，提示PHEV诱导神经细胞自噬的机制可能较经典的ULK1依赖性途径更为复杂。为解析这一科学问题，本项目拟以ULK1及其自噬相关同源物ULK2激酶为研究对象，分别建立其基因敲除细胞系，通过检测自噬体形成、自噬溶酶体成熟和降解等，明确ULK1/2在神经细胞基础自噬及PHEV诱导自噬中的作用；利用营养/能量缺失条件性培养、自噬信号抑制剂/激活剂诱导、脂质体激酶测定等方法，系统阐明ULK1/2与自噬通路重要分子之间的广泛交叉和反馈环在PHEV诱导神经细胞自噬中的调控作用。预期成果不仅可阐明PHEV诱导神经细胞自噬的分子机制，有利于解释PHEV致神经功能障碍的机理，并为新型抗病毒药物研发提供重要的参考策略。

Porcine hemagglutinating encephalomyelitis virus (PHEV) is a neurotropic coronavirus that primarily invades the central nervous system in piglets and causes nerve cells damage. Our previous study found that PHEV infection induces autophagosome formation in nerve cells, but the expression of the key autophagy-initiating kinase ULK1 is significantly down-regulated, suggesting that the mechanism of PHEV-induced autophagy may be more complicated than the classical ULK1-dependent pathway. In our project, to gain insight into the ULK1 and its autophagy-related homologue ULK2 kinase, its knockout cell line is established, respectively. By detecting autophagosome formation, autophagy lysosome maturation and degradation in these cells, we first clarify the role of ULK1/2 in neuronal basal autophagy and PHEV-induced autophagy. To elucidate the role of the crossover and feedback loop between ULK1/2 and other important autophagy-related regulators in the PHEV-induced autophagic signal, we subsequently perform nutrient/energy deficient conditioning, autophagy signaling inhibitor/activator induction, liposome kinase assay, etc. The expected results not only clarify the autophagy-core molecular machinery in PHEV-infected nerve cells, but also explain the mechanism of PHEV-induced neurological dysfunction, and also provide an important reference strategy for the development of new antiviral drugs.