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

基本信息

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
项目类别：
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&apos -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

项目摘要

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.

抽象的该提案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 万美国人糖尿病，另有 8800 万人被认为是糖尿病前期。这很大程度上归因于肥胖，目前 72% 的美国成年人属于超重或肥胖。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与肥胖、T2D 和 NAFLD 的消费。该提案将 GLO1 确定为促 NAFLD/致肥胖基因。使用 CRISPR-Cas9，我们培育出了 GLO1 基因敲除小鼠。当喂食高脂肪高蔗糖饮食时，这些小鼠的体重增加显着减弱，葡萄糖耐量恢复，肝脏脂肪变性减少与野生型对应物相比。 GLO1 是一种普遍表达的酶，可通过以下方式解毒糖酵解：产品，甲基乙二醛（MGO）。当 GLO1 活性受到干扰时，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 可作为营养通量的代谢传感器，调节糖酵解输出和对糖的转录反应。因此，我们假设 GLO1 是亲 NAFLD/致肥胖基因，减少 MGO 并消除新陈代谢的障碍。我们将检验这个假设通过解决以下三个具体目标：在具体目标 1 中，我们将量化 Glo1 对整体的影响：使用 16 周的食物或高脂肪高蔗糖饮食来观察身体能量学和肝脏脂质代谢。我们将量化瘦肉量与脂肪量、能量消耗和总活动量。肝脏脂肪酸氧化和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 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。肠道是一个果糖代谢的主要场所。通过 13C6-果糖口服灌胃的稳定同位素标记将用于定量肠道、肝脏、肌肉、脂肪和循环果糖代谢物。该方法将定量地确定 Glo1 如何调节肠道碳水化合物代谢，这是一个先前未探索的研究领域。最后，在具体目标 3 中，我们将确认 MGO 衍生的组蛋白 PTM 调节转录反应体内代谢。位点特异性规范和 MGO 衍生的 PTM 将在每个组织/队列中进行量化。将确定 MG-H1 组蛋白修饰的假定阅读器域。最后是肝、肠、肌肉、脂肪和胰腺将接受 RNA-seq 和 ATAC-seq。这种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，该提案将机械生物化学与多组学方法相结合，以确定 GLO1 传播疾病进展的机制。