过表达基因FADD抑制视网膜神经节细胞程序性坏死的机制研究

青光眼是一类以视网膜神经节细胞（RGCs）程序性死亡及轴突损伤为特征的疾病，已有研究表明，程序性坏死与RGCs的死亡及轴突损伤密切相关，基因FADD参与细胞的程序性坏死过程。我们最近的研究发现，在体内和体外模型中，RGCs和视网膜神经细胞的死亡方式可表现为凋亡和程序性坏死两种形式，且过表达基因FADD可通过抑制RIPK1和RIPK3的相互作用来显著减轻细胞程序性坏死的发生比例。本项目将在该发现的基础上进一步探讨程序性坏死与RGCs存活状态及轴突生长的关系，阐明基因FADD影响RGCs程序性坏死发生的相关机制，明确基因FADD表达水平及其上下游信号通路与原发性青光眼的相关性。我们将运用分子生物学、细胞生物学和视觉电生理等技术，从AKT和Sarm1-MAPK两个信号通路入手，从动物、细胞和患者三个方面全面探讨过表达基因FADD抑制RGCs程序性坏死发生的机制，为今后RGCs的保护提供科学依据。

Glaucoma is a blinding disease that is characterized by programmed cell death of retinal ganglion cells (RGCs) and axons damage. As reported, necroptosis is tightly related to RGCs and their axons damage and gene FADD is involved in the regulation of necroptosis. Our recent study found that RGCs death could be classified as apoptosis and necroptosis in vivo and in vitro and overexpression of FADD could significantly ameliorate the RGCs necroptosis through inhibiting the interaction of RIPK1 and RIPK3. Based on above, this project will further explore the relationship among necroptosis, RGCs survival and their axons regeneration, the mechanism of FADD expression on RGCs necroptosis, and the correlation between FADD signal pathway and glaucoma occurrence. Moreover, to study the regulatory elements which influence gene FADD expression, we will focus on the upstream microRNA of this target gene. To elucidate the molecular mechanism of FADD anti-necroptosis, we would focus on two signal pathway: AKT and Sarm1-MAPK, through molecular biology, cell biology and visual electrophysiology techniques in animals, cells and patients. To observe the effect of gene FADD on RGCs necroptosis，gene FADD will be overexpressed, and the expression of RIPK1, RIPK3, MLKL and so on, will be detected. We will observe the effect when necroptosis was inhibited and provide a new therapeutic target for the prevention of RGCs damage.