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

评估 Lkb1/Snrk 信号对 RGC 存活的代谢影响

• 批准号: 10537481
• 负责人: Ryan J Donahue
• 金额: $ 6.72万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10537481
• 关键词: Address; Adult; Apoptosis; Binding; Biological Assay; Blindness; Cell Death; Cell Survival; Cellular Metabolic Process; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Complex; Data; Dependence; Disease; Distress; Drug Targeting; Exhibits; Family; Gene Deletion; Genes; Glaucoma; Hour; Immunohistochemistry; Injury; Knock-out; Knockout Mice; Lead; Link; Mediating; Metabolic; Metabolic dysfunction; Metabolic stress; Metabolism; Modeling; Mus; Nerve Crush; Nerve Degeneration; Optic Nerve; Optic Nerve Injuries; Pathway interactions; Patients; Persons; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphotransferases; Physiologic Intraocular Pressure; Procedures; Process; Proteins; Research Personnel; Retina; Retinal Ganglion Cells; Risk Factors; STK11 gene; Signal Transduction; Stress; Testing; Therapeutic; Thinking; Time; Translating; Treatment Efficacy; Viral; axon regeneration; cell type; clinically actionable; clinically relevant; comparative; conditional knockout; experimental study; flexibility; injured; insight; metabolomics; modifiable risk; novel; optic nerve disorder; prevent; programs; protective effect; response; response to injury; retinal ganglion cell degeneration; single-cell RNA sequencing

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.

项目摘要 视网膜神经节细胞（RGC）的变性导致全世界数百万人不可逆转的失明。 开发靶向RGC并防止其退化的治疗方法的努力受到了阻碍， 对这个过程的不完全理解。我们实验室最近筛选了约1000种激酶和磷酸酶， 确定了几个新的贡献者RGC死亡后视神经损伤。其中一组蛋白质与 细胞代谢，已知的RGC死亡的贡献者。该项目将探索这些蛋白质如何有助于 受损的视网膜节细胞内的代谢功能障碍。 我们筛选出的最令人兴奋的激酶是肝激酶B1（LKB 1）。LKB 1是一种主激酶， 在其他细胞类型中激活各种下游途径，但其在RGC中的功能尚不清楚。一个非常 有希望的可能性是LKB 1可能是应激下RGC代谢的主要调节剂之一。 然而，由于其混杂的活动，这个建议是必要的，以破译的确切机制， RGC中的LKB 1信号传导。通过使用我们实验室设计的灵活的病毒介导的敲除策略， 确定LKB 1通路介导的变性机制及其在青光眼中的相关性。通过配对 通过单细胞RNA测序和代谢组学方法的靶向测定，我们将揭示 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，这里提出的实验将提高我的技术水平。 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。