Establishing Glyoxalase 2 as a Viable Target for the Treatment of Disease

将乙二醛酶 2 确立为治疗疾病的可行靶点

• 批准号: 10415990
• 负责人: James J Galligan
• 金额: $ 37.74万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-10 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10415990
• 关键词: Address; Antioxidants; Autoimmunity; Award; Biological; Biology; CRISPR/Cas technology; Carbon; Cell Line; Cell model; Cells; Cellular Metabolic Process; Chromatin; Cues; Diabetes Mellitus; Disease; Global Change; Glutathione; Glycolysis; Goals; Health; Histones; Homeostasis; Inflammatory; Knock-out; Laboratories; Lactoylglutathione Lyase; Lysine; Malignant Neoplasms; Metabolic; Metabolic Diseases; Metabolism; Methods; Modification; Output; Oxidation-Reduction; Oxidative Stress; Plant Roots; Post-Translational Protein Processing; Proteins; Proteomics; Reader; Reduced Glutathione; Regulation; Research; Role; Sampling; Sepsis; Signal Transduction; Site; Site-Directed Mutagenesis; Therapeutic; Transcriptional Regulation; Treatment Efficacy; Xenograft Model; in vivo; novel; programs; response; sensor; treatment strategy

PROJECT SUMMARY/ABSTRACT The ability for cells to detect and respond to metabolic cues is critical to maintaining homeostasis, and perturbations in the sensing mechanisms that respond to oscillations in metabolic flux are the root cause of many diseases, including sepsis, autoimmunity, cancer, and diabetes. There is mounting evidence that protein post- translational modifications (PTMs) are the critical sensors for these metabolic fluctuations and are often dysregulated in disease. Currently, we have a fundamental gap in our understanding of the composition, abundance, and enzymatic control of PTMs and how they are altered in disease. My laboratory focuses on the identification and characterization of PTMs and how they are regulated in both health and disease. To accomplish this goal, we have developed sensitive methods to identify and quantify global changes in PTMs across a broad spectrum of biological samples. Using this approach, we have identified a novel lysine PTM that is derived from a glycolytic by-product. These PTMs are elevated when glyoxalase 2 (GLO2) is inhibited, resulting in reduced glycolytic output and disrupted one-carbon metabolism. Our primary goal is to establish the therapeutic efficacy of a GLO2 inhibition strategy for the treatment of metabolic disorders. My research program is dedicated to understanding four fundamental questions: 1) How does GLO2 control one-carbon metabolism and cellular redox? GLO2 knockout cells have reduced glutathione and increased oxidative stress. We will quantify the role of GLO2 in the regulation of de novo glutathione synthesis. In addition, the role of GLO2 in the regulation of antioxidant responses will be evaluated in a cellular model for oxidative stress and inflammatory signaling. 2) How are LactoylLys modifications regulated? We will employ quantitative proteomics using CRISPR-Cas9 knockout cell lines of candidate proteins to identify enzymatic regulators of LactoylLys modifications in cells. 3) Is GLO2 a viable target for the treatment of glycolysis- dependent disease states? A xenograft model will be employed using GLO2 knockout cell lines to quantify proliferation and metabolic regulation in vivo. This will determine the therapeutic feasibility of targeting GLO2 for the treatment of disease. 4) Are LactoylLys modifications functional histone marks? We have identified histones as targets for modification by LactoylLys modifications in unstimulated cells. The presence of these PTMs basally suggests a putative role in transcriptional regulation. We will use proteomics to identify site-specific modifications and putative ‘reader’ domains for LactoylLys modifications in cells. Our primary goal is to establish the role of GLO2 and LactoylLys modifications in cell metabolism and chromatin biology. This project will address a fundamental gap in our basic understanding of how cell metabolism is regulated. Understanding how these PTMs regulate homeostasis is a critical first step to understanding their role in disease. Due to the far-reaching implications of this project and the broad applications for the treatment of highly glycolytic disease states, this research program is an ideal fit for the ESI MIRA Award.

项目总结/摘要 细胞检测和响应代谢信号的能力对维持体内平衡至关重要， 对代谢通量振荡作出响应的传感机制中的扰动是许多疾病的根本原因。 疾病，包括败血症、自身免疫、癌症和糖尿病。越来越多的证据表明蛋白质后- 翻译修饰（PTM）是这些代谢波动的关键传感器， 在疾病中失调。目前，我们对组成的理解存在根本性的差距， 丰度和PTM的酶控制以及它们在疾病中如何改变。 我的实验室专注于PTMs的识别和表征以及它们在细胞中的调节方式。 健康和疾病。为了实现这一目标，我们开发了敏感的方法来识别和量化 跨广谱生物样品的PTM的全球变化。通过这种方法，我们确定了 衍生自糖酵解副产物的新型赖氨酸PTM。当乙二醛酶2时，这些PTM升高 （GLO 2）被抑制，导致糖酵解输出减少和一碳代谢中断。我们的首要 目的是建立GLO 2抑制策略用于治疗代谢紊乱的治疗功效。 我的研究计划致力于了解四个基本问题：1）GLO 2如何控制 一碳代谢和细胞氧化还原GLO 2敲除细胞具有减少的谷胱甘肽和增加的 氧化应激我们将量化GLO 2在从头谷胱甘肽合成的调节中的作用。此外，本发明还提供了一种方法， GLO 2在调节抗氧化反应中的作用将在细胞模型中进行评价， 应激和炎症信号。2)LactoylLys修饰如何调控？我们会委聘 使用候选蛋白的CRISPR-Cas9敲除细胞系的定量蛋白质组学来鉴定酶促 LactoylLys修饰的调节剂。3)GLO 2是治疗糖酵解的可行靶点吗？ 依赖性疾病状态？将采用使用GLO 2敲除细胞系的异种移植模型来定量 增殖和体内代谢调节。这将确定靶向GLO 2以治疗肿瘤的治疗可行性。 疾病的治疗4)LactoylLys修饰是功能性组蛋白标记吗？我们已经确定 在未刺激的细胞中，组蛋白作为LactoylLys修饰的靶点。存在这些 PTMs基本上表明在转录调控中的假定作用。我们将使用蛋白质组学来识别特定位点 修饰和细胞中乳酰赖氨酸修饰的推定“阅读器”结构域。 我们的主要目标是确定GLO 2和LactoylLys修饰在细胞代谢中的作用， 染色质生物学这个项目将解决一个根本的差距，在我们的基本理解如何细胞代谢 是受管制的。了解这些PTM如何调节体内平衡是了解它们的关键第一步。 疾病中的作用。由于该项目的深远影响和治疗的广泛应用， 高度糖酵解疾病状态，这项研究计划是一个理想的适合ESI MIRA奖。