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.

抽象的 该提案的总体目标是确定糖酶1（GLO1）在肥胖症的发病机理中的作用， 2型糖尿病（T2D）和非酒精性脂肪肝病（NAFLD）。超过3400万美国人 糖尿病和另外8800万糖尿病被认为是糖尿病前期的。这主要归因于 肥胖症，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。流行病学研究已将高果糖联系起来 肥胖，T2D和NAFLD的消费。该提案将GLO1识别为促nafld/rolig opoogent基因。 使用CRISPR-CAS9，我们生成了GLO1敲除小鼠。喂高脂高蔗糖饮食时 小鼠表现出显着钝化的体重增加，恢复的葡萄糖耐受性和肝脂肪变性降低 与野生型对手相比。 GLO1是一种普遍表达的酶，可解毒 产物，甲基甘氨酸（MGO）。当glo1活性破坏时，MGO水平增加，导致长寿 蛋白质后翻译。我们已经证明MGO用作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 NAFLD/肥胖基因，减少MGO并消除新陈代谢的制动器。我们将检验这个假设 通过解决以下三个特定目标：在特定目标1中，我们将量化GLO1对整个的影响 人体能量和肝脂质代谢，使用16周的Chow或高脂高核饮食。我们将 量化瘦肉与脂肪质量，能量消耗和总活动。肝脂肪酸氧化和线粒体 将在原发性肝细胞中量化呼吸。最后，将使用RNA-Seq评估脂肪酸基因。 在特定目标2中，我们将确定GLO1对肠道果糖代谢的影响。肠是一个 果糖代谢的主要部位。通过13C6-果糖口服块的稳定同位素标记将用于量化 肠，肝，肌肉，脂肪和循环果糖代谢产物。这种方法将定量 确定GLO1如何调节肠道碳水化合物代谢，这是以前未开发的研究领域。 最后，在特定的目标3中，我们将确认MGO衍生的组蛋白PTM调节转录反应 体内代谢。位点特异性的规范和MGO衍生的PTM将在每个组织/队列中进行量化。 将针对MG-H1组蛋白修饰确定推定的读取器域。最后，肝脏，肠，肌肉， 脂肪和胰腺将受到RNA-SEQ和ATAC-SEQ的约束。这种方法将决定全球 肥胖，T2D和NAFLD的生理相关模型中多个组织跨多个组织的表观基因组景观。 总的来说，该提案将结合机械生物化学与多词的方法来确定 GLO1传播疾病进展的机制。