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）的退化会导致全球数百万人的不可逆失明。 为针对RGC和防止其变性的治疗的努力受到了阻碍 对这个过程的理解不完全。在我们实验室中，最近〜1000个激酶和磷酸酶的屏幕具有 确定了视神经损伤后RGC死亡的几个新贡献者。这些蛋白质一组是相关的 对细胞代谢，这是RGC死亡的已知贡献者。该项目将探讨这些蛋白质如何贡献 在受伤的RGC中代谢功能障碍。 我们屏幕鉴定的最令人兴奋的激酶是肝激酶B1（LKB1）。 LKB1是一种大师激酶 激活其他细胞类型的各种下游途径，但其在RGC中的功能未经表征。非常 有希望的可能性是LKB1可能是压力下RGC代谢的主要调节剂之一。 但是，由于其滥交活动，该建议是破译的确切机制所必需的 RGC中的LKB1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，我们将 确定LKB1途径介导的变性的机理及其在青光眼中的相关性。通过配对 具有单细胞RNA测序和代谢组方法的有针对性测定，我们将揭示因果关系 代谢变化会导致RGC死亡并希望将这些发现转化为可行的临床 疗法。在Zhigang博士的指导下，这里提出的实验将增长我的技术 能力和我的科学思维，为我铺平了成为独立调查员的道路。