Assessing the metabolic impact of Lkb1/Snrk signaling on RGC survival

项目摘要 视网膜神经节细胞(RGC)的退化导致全球数百万人不可逆转的失明。 开发针对视网膜节细胞并防止其退化的治疗方法的努力受到了 对这一过程的理解不全面。我们实验室最近对大约1000种激酶和磷酸酶进行了筛查 确定了视神经损伤后RGC死亡的几个新因素。这些蛋白质中的一组是相关的 对细胞代谢的影响，这是RGC死亡的已知因素。这个项目将探索这些蛋白质是如何贡献的。 损伤的视网膜节细胞内的代谢功能障碍。 我们的筛查确定的最令人兴奋的激酶是肝激酶B1(LKB1)。LKB1是一种主激酶，它可以 在其他类型的细胞中激活各种下游通路，但其在视网膜节细胞中的功能尚未确定。一个非常好的 LKB1可能是应激状态下RGC代谢的主要调节因子之一。 然而，由于其混杂的活动，这一提议是必要的，以破译 RGC中的LKB1信号转导。通过使用我们实验室设计的灵活的、病毒介导的击倒策略，我们将 确定LKB1通路介导的青光眼退行性变的机制及其相关性。通过配对 用单细胞RNA测序和代谢组学方法进行靶向分析，我们将揭示病因 导致RGC死亡的代谢变化，并希望将这些发现转化为可操作的临床 治疗学。在何志刚博士的指导下，这里提出的实验将增长我的技术 我的能力和我的科学思维，为我成为一名独立调查员铺平了道路。

Project Summary The degeneration of retinal ganglion cells (RGCs) causes irreversible blindness in millions of people worldwide. Efforts to develop treatment which target RGCs and prevent their degeneration have been hampered by an incomplete understanding of this process. A recent screen of ~1000 kinases and phosphatases in our lab has identified several novel contributors to RGC death after optic nerve injury. A group of these proteins are related to cellular metabolism, a known contributor to RGC death. This project will explore how these proteins contribute to metabolic dysfunction within injured RGCs. The most exciting kinase identified by our screen is Liver Kinase B1 (LKB1). LKB1 is a master kinase that activates various downstream pathways in other cell types but its function in RGCs is uncharacterized. A very promising possibility is that LKB1 may be one of the principal modulators of RGC metabolism under stress. However, as a result of its promiscuous activity, this proposal is necessary to decipher the exact mechanism of LKB1 signaling in RGCs. Through use of a flexible, viral mediated knockout strategy devised by our lab, we will determine the mechanism of LKB1 pathway mediated degeneration and its relevance in glaucoma. By pairing targeted assays with single cell RNA sequencing and metabolomic approaches, we will reveal the causative metabolic changes that cause RGC death and hope to translate these findings into actionable clinical therapeutics. Under the tutelage of Dr. Zhigang He, the experiments proposed here will grow my technical abilities and my scientific thinking, paving the way for me to become an independent investigator.