Glyoxalase 1 and its Role in Metabolic Syndrome

乙二醛酶 1 及其在代谢综合征中的作用

• 批准号: 10656054
• 负责人: James J Galligan
• 金额: $ 41.38万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10656054
• 关键词: 5' -AMP-activated protein kinase; ATAC-seq; Acetyl-CoA Carboxylase; Address; Adipose tissue; Adult; American; Architecture; Area; Binding; Biochemistry; Blood; Bypass; CRISPR/Cas technology; Catabolism; Chromatin; Circulation; Classification; Clinical; Consumption; Data; Diabetes Mellitus; Diet; Dietary Sugars; Disease; Disease Progression; Drug Metabolic Detoxication; Energy Metabolism; Enzymes; Equilibrium; Exhibits; Fatty Acids; Fatty Liver; Fatty acid glycerol esters; Fructose; Genes; Genetic Transcription; Glucose; Glucose-6-Phosphate; Glycolysis; Goals; Health; Hepatic; Hepatocyte; High Fat Diet; Histone H1; Histones; Homeostasis; Hyperglycemia; Incidence; Intestines; Ketones; Knockout Mice; Label; Lactoylglutathione Lyase; Leaky Gut; Link; Lipids; Liver; Measures; Mediating; Metabolic; Metabolic syndrome; Metabolism; Mission; Mitochondria; Modeling; Modification; Mus; Muscle; National Institute of Diabetes and Digestive and Kidney Diseases; Non-Insulin-Dependent Diabetes Mellitus; Nutrient; Obesity; Oral; Output; Overweight; Pancreas; Pathogenesis; Patients; Phosphorylation; Physiological; Post-Translational Protein Processing; Prediabetes syndrome; Prevalence; Process; Proxy; Pyruvaldehyde; Reader; Research; Respiration; Role; Signal Transduction; Site; Stable Isotope Labeling; Sucrose; Testing; Thinness; Tissues; Triglycerides; United States; Weight Gain; Wild Type Mouse; adenylate kinase; carbohydrate metabolism; cohort; comorbidity; design; dietary; epidemiology study; epigenomics; fatty acid oxidation; feeding; fructose-1-phosphate; glucose tolerance; histone modification; in vivo; insulin sensitivity; interdisciplinary approach; lipid biosynthesis; lipid metabolism; liver injury; multiple omics; non-alcoholic fatty liver disease; obesogenic; oxidation; pharmacologic; recruit; response; sensor; sugar; systemic inflammatory response; transcriptome sequencing; uptake

Abstract The overall goal of this proposal is to determine the role of glyoxalase 1 (GLO1) in the pathogenesis of obesity, Type 2 diabetes (T2D), and non-alcoholic fatty liver disease (NAFLD). Greater than 34 million Americans have diabetes, and another 88 million are considered pre-diabetic. This is largely attributed to the prevalence of obesity, with 72% of American adults currently classified as overweight or obese. Among the comorbidities associated with T2D, over 70% of patients have NAFLD. Epidemiological studies have linked high fructose consumption with obesity, T2D, and NAFLD. This proposal identifies GLO1 as pro-NAFLD/obesogenic gene. Using CRISPR-Cas9, we have generated GLO1 knockout mice. When fed a high-fat high sucrose diet, these mice display significantly blunted weight gain, restored glucose tolerance, and reduced hepatic steatosis compared to wild-type counterparts. GLO1 is a ubiquitously expressed enzyme that detoxifies the glycolytic by- product, methylglyoxal (MGO). When GLO1 activity is disrupted, MGO levels increase, resulting in long-lived protein post-translational modifications. We have shown that MGO serves as a metabolic sensor for nutrient flux, regulating glycolytic output and transcriptional responses to sugar. Thus, we hypothesize that GLO1 is a pro- NAFLD/obesogenic gene, reducing MGO and removing the brakes on metabolism. We will test this hypothesis by addressing the following three Specific Aims: In Specific Aim 1 we will quantify the impact of Glo1 on whole- body energetics and hepatic lipid metabolism using a 16-week chow- or high-fat high-sucrose diet. We will quantify lean vs. fat mass, energy expenditure, and total activity. Hepatic fatty acid oxidation and mitochondrial respiration will be quantified in primary hepatocytes. Lastly, lipogeneic genes will be assessed using RNA-seq. In Specific Aim 2 we will determine the impact of Glo1 on intestinal fructose metabolism. The intestine is a primary site of fructose metabolism. Stable isotope labeling via 13C6-fructose oral gavage will be used to quantify intestinal, hepatic, muscle, adipose, and circulating fructose metabolites. This approach will quantitatively determine how Glo1 regulates intestinal carbohydrate metabolism, a previously unexplored area of research. Finally, in Specific Aim 3, we will confirm that MGO-derived histone PTMs regulate transcriptional responses to metabolism in vivo. Site-specific canonical and MGO-derived PTMs will be quantified in each tissue/cohort. Putative reader domains will be identified for MG-H1 histone modifications. Lastly, liver, intestine, muscle, adipose, and pancreas will be subjected to RNA-seq and ATAC-seq. This approach will determine the global epigenomic landscape across multiple tissues in a physiologically relevant model for obesity, T2D and NAFLD. Collectively, this proposal will combine mechanistic biochemistry with a multi-omics approach to determine the mechanisms by which GLO1 propagates disease progression.

摘要 这项提议的总体目标是确定乙二醛酶1(GLO1)在肥胖发病机制中的作用。 2型糖尿病(T2D)和非酒精性脂肪肝(NAFLD)。超过3400万美国人 另有8800万人被认为是糖尿病前期。这在很大程度上归因于 肥胖，目前有72%的美国成年人被归类为超重或肥胖。在合并症中 与T2D相关，超过70%的患者患有NAFLD。流行病学研究表明，高果糖 肥胖、T2D和NAFLD的消费。该建议将GLO1确定为NAFLD前/肥胖基因。 使用CRISPR-Cas9，我们已经产生了GLO1基因敲除小鼠。当喂食高脂肪高蔗糖饮食时，这些 小鼠体重增加明显减慢，糖耐量恢复，肝脏脂肪变性减轻。 与野生型的同类相比。GLO1是一种普遍表达的酶，通过以下方式解毒糖酵解- 产品甲基乙二醛(MGO)。当GLO1活动中断时，MGO水平会增加，从而导致长寿 蛋白质翻译后修饰。我们已经证明，MGO是营养流量的新陈代谢传感器， 调节糖酵解的输出和对糖的转录反应。因此，我们假设GLO1是一个专业- NAFLD/肥胖基因，减少MGO，消除代谢刹车。我们将检验这一假设 通过处理以下三个具体目标：在具体目标1中，我们将量化全球一体化对整个世界的影响-- 身体能量和肝脏脂肪代谢，使用16周的食物或高脂肪高蔗糖饮食。我们会 量化精瘦与脂肪的质量、能量消耗和总活动量。肝脂肪酸氧化与线粒体 呼吸作用将在原代肝细胞中进行量化。最后，将使用RNA-seq来评估脂肪生成基因。 在具体目标2中，我们将确定Glo1对肠道果糖代谢的影响。肠道是一种 果糖代谢的主要部位。通过13C6-果糖口服稳定同位素标记将用于定量 肠道、肝脏、肌肉、脂肪和循环中的果糖代谢物。这一方法将在数量上 确定Glo1如何调节肠道碳水化合物代谢，这是一个以前从未探索过的研究领域。 最后，在特定的目标3中，我们将确认MGO衍生的组蛋白PTMS调节转录反应 体内代谢。将在每个组织/队列中量化特定部位的经典性和MGO来源的PTM。 将确定MG-H1组蛋白修饰的假定阅读器结构域。最后是肝、肠、肌肉， 脂肪，胰腺将受到RNA-SEQ和ATAC-SEQ的影响。这一方法将决定全球 在肥胖、T2D和NAFLD的生理相关模型中，跨多个组织的表观基因组景观。 总的来说，这项提案将结合机械性生物化学和多组学方法来确定 GLO1传播疾病进展的机制。