PHEV具有典型的嗜神经性，侵染机体后导致大脑神经元突起延伸障碍等神经退行性病变，此时Ulk1表达显著下调。Ulk1是神经元形态发生及突触运输等神经活动的关键调节蛋白，其如何介导PHEV致神经损伤的调控过程，目前尚不清楚。为此，本项目拟应用原代培养的大脑皮质神经元建立PHEV感染模型，通过激光共聚焦等技术检测该病毒对神经元形态发生和Ulk1蛋白激酶表达的影响；应用CRISPR/Cas9基因敲除、构建慢病毒过表达载体等方法，进一步验证Ulk1对PHEV致神经损伤的作用；筛选鉴定与Ulk1直接或间接互作的重要蛋白，探讨这些复合物介导的内吞作用、囊泡运输以及轴突运输等下游信号通路在神经损伤中的作用机制，从Ulk1角度系统地阐明该病毒诱发神经退行性病变的分子基础。预期结果不仅为PHEV感染的有效防控及特异性靶向治疗药物的筛选奠定理论基础，也可为深入研究神经退行性疾病的发病机理提供良好的感染模型。

PHEV is a veritable neurotropic coronavirus, invading the central nervous system causes neurodegeneration, such as defects in neurite formation. Unc-51-like kinase 1(Ulk1) is an evolutionarily conserved kinase, plays pivotal role in neuronal morphology and axonal transport, but was significantly down-regulated expression in neurodegeneration caused by PHEV. However, its mechanism attributed to Ulk1 deficit is not clear. Here, we detect the impact of PHEV on neuron morphogenesis and Ulk1 expression in primary cultured cerebral cortical neurons, and establish a model of PHEV-infected neurons. Applications of CRISPR/Cas9 knockout and lentiviral vector-mediated overexpression shall carry to validate the affect of Ulk1 in nerve injury. Furthermore, identifying the essential effectors crossing-talk with Ulk1, basing on endocytosis, vesicular trafficking and axonal transport, to explore the mechanisms in neurological dysfunction of PHEV. Systematically, elucidate the molecular basis of PHEV-induced neurodegenerative disorders. Prospective study represents a theoretical foundation, not only for effective prevention and control of PHEV infection, but also for specific targeted therapy drug screening. Accordingly, comprehensive evidences provide a significative infection model for further research on the pathogenesis of neurodegenerative diseases.