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: 10640181
• 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/10640181
• 关键词: Antioxidants; Autoimmunity; Award; Biological; Biology; CRISPR/Cas technology; Carbon; Cell Line; Cell model; Cells; Cellular Metabolic Process; Chromatin; Cues; Dedications; 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; Post-Translational Protein Processing; Proliferating; 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.

项目概要/摘要 细胞检测和响应代谢信号的能力对于维持体内平衡至关重要，并且 响应代谢通量振荡的传感机制的扰动是许多问题的根本原因 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，通常 疾病中失调。目前，我们对构图的理解存在根本差距， PTM 的丰度和酶促控制以及它们在疾病中如何改变。 我的实验室专注于 PTM 的识别和表征以及它们如何在 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 广泛的生物样品中 PTM 的全球变化。使用这种方法，我们已经确定 一种源自糖酵解副产物的新型赖氨酸 PTM。当乙二醛酶 2 存在时，这些 PTM 会升高 (GLO2) 受到抑制，导致糖酵解输出减少并扰乱一碳代谢。我们的小学 目标是确定 GLO2 抑制策略治疗代谢性疾病的疗效。 我的研究项目致力于理解四个基本问题：1）GLO2 如何控制 一碳代谢和细胞氧化还原？ GLO2 敲除细胞减少了谷胱甘肽并增加了 氧化应激。我们将量化 GLO2 在谷胱甘肽从头合成调节中的作用。此外， GLO2 在抗氧化反应调节中的作用将在氧化细胞模型中进行评估 压力和炎症信号。 2) LactoylLys 修饰是如何调节的？我们将聘用 使用候选蛋白质的 CRISPR-Cas9 敲除细胞系进行定量蛋白质组学鉴定酶 细胞中乳酰赖氨酸修饰的调节因子。 3) GLO2 是治疗糖酵解的可行靶标吗？ 依赖性疾病状态？将使用 GLO2 敲除细胞系的异种移植模型来量化 体内增殖和代谢调节。这将确定靶向 GLO2 的治疗可行性 疾病的治疗。 4) LactoylLys 修饰是否具有功能性组蛋白标记？我们已经确定 组蛋白作为未刺激细胞中 LactoylLys 修饰的修饰目标。这些的存在 PTM 基本上表明在转录调控中具有推定的作用。我们将使用蛋白质组学来识别位点特异性 细胞中 LactoylLys 修饰的修饰和假定的“阅读器”域。 我们的主要目标是确定 GLO2 和 LactoylLys 修饰在细胞代谢和 染色质生物学。该项目将解决我们对细胞代谢如何进行基本理解的根本差距 受到监管。了解这些 PTM 如何调节体内平衡是了解它们的关键的第一步 在疾病中的作用。由于该项目影响深远，治疗应用广泛 对于高度糖酵解疾病状态，该研究项目非常适合 ESI MIRA 奖。

项目成果

A global view of the human post-translational modification landscape.

• DOI: 10.1042/bcj20220251
• 发表时间: 2023-08-30
• 期刊: The Biochemical journal

Chemical Labeling and Enrichment of Histone Glyoxal Adducts.

• DOI: 10.1021/acschembio.1c00864
• 发表时间: 2022-04-15
• 期刊: ACS CHEMICAL BIOLOGY
• 影响因子: 4
• 作者: Ray, Devin M.;Jennings, Erin Q.;Maksimovic, Igor;Chai, Xander;Galligan, James J.;David, Yael;Zheng, Qingfei
• 通讯作者: Zheng, Qingfei

Biochemical Mechanisms of Sirtuin-Directed Protein Acylation in Hepatic Pathologies of Mitochondrial Dysfunction.

• DOI: 10.3390/cells11132045
• 发表时间: 2022-06-28
• 期刊: Cells
• 影响因子: 6

Biochemical genesis of enzymatic and non-enzymatic post-translational modifications.

• DOI: 10.1016/j.mam.2021.101053
• 发表时间: 2022-08
• 期刊: Molecular aspects of medicine
• 影响因子: 10.6
• 作者: Jennings EQ;Fritz KS;Galligan JJ
• 通讯作者: Galligan JJ

Lactoylglutathione promotes inflammatory signaling in macrophages.

乳酰谷胱甘肽促进巨噬细胞中的炎症信号传导。

• DOI: 10.1101/2023.10.10.561739
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Trujillo,MarissaN;Jennings,ErinQ;Hoffman,EmelyA;Zhang,Hao;Phoebe,AidenM;Mastin,GraceE;Kitamura,Naoya;Reisz,JulieA;Megill,Emily;Kantner,Daniel;Marcinkiewicz,MariolaM;Twardy,ShannonM;Lebario,Felicidad;Chapman,Eli;McCullou
• 通讯作者: McCullou