Depalmitoylation regulates hepatic glucose metabolism

去棕榈酰化调节肝脏葡萄糖代谢

基本信息

批准号：
10387053
负责人：
Sarah Lilly Speck
金额：
$ 3.2万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10387053
关键词：
Acylation Adipose tissue American Bacteria Biotin Biotinylation Blood Glucose Body Composition Body measure procedure C-terminal Cardiovascular Diseases Cell membrane Cell surface Cells Chronic Chronic Kidney Failure Cysteine Cytosol Data Development Diabetes Mellitus Diagnosis Drug Targeting Environment Enzymes Fatty Acids Fatty acid glycerol esters Fellowship Female Fluorescent Probes Gluconeogenesis Glucose Glucose Transporter Glycogen Goals HepG2 Hepatic High Fat Diet Human Hyperglycemia Immunofluorescence Immunologic In Vitro Incubated Insulin Insulin Resistance Investigation Knock-out Knockout Mice Label Ligase Link Lipids Liver LoxP-flanked allele Mass Spectrum Analysis Mediating Metabolic Metabolic Diseases Metabolic Pathway Metabolism Methods Modification Molecular Biology Molecular Biology Techniques Molecular and Cellular Biology Mus N-terminal Non-Insulin-Dependent Diabetes Mellitus Obesity Palmitates Physicians Physiology Plant Resins Plasma Plasmids Post-Translational Protein Processing Primary carcinoma of the liver cells Process Protein Isoforms Proteins Proteome Pyruvate Recording of previous events Regulatory Pathway Research Research Personnel Risk Role Saturated Fatty Acids Scientist Streptavidin Testing Training Universities Vertebrates Vesicle Washington Work blood glucose regulation career clay cohort experience fasting plasma glucose fatty acid metabolism glucose metabolism glucose tolerance in vivo inhibitor/antagonist insulin sensitivity insulin signaling intermolecular interaction lipid metabolism medical schools novel palmitoylation response skills

项目摘要

PROJECT SUMMARY/ABSTRACT Hundreds of thousands of Americans are diagnosed with type 2 diabetes every year, increasing their risk of developing cardiovascular disease and chronic kidney disease. While insulin signaling in the liver typically modulates blood glucose levels by suppressing the synthesis of new glucose and inducing glucose storage, hepatic insulin resistance can exacerbate hyperglycemia. Insulin resistance is associated with dysregulated fatty acid metabolism. Fatty acids can directly modify proteins through a reversible process known as palmitoylation. Depalmitoylating acyl protein thioesterases (APTs), such as APT1 and APT2, can remove these posttranslational modifications. Although multiple proteins involved in glucose homeostasis have been shown to be palmitoylated, the role of depalmitoylases in metabolism is still under investigation. Preliminary data suggest that APT1 liver knockout (APT1LKO) female mice have insulin resistance, while APT2LKO female mice demonstrate increased fasting plasma glucose levels. However, the redundancy of APT1 and APT2 in the glucose metabolic pathway is unclear. The objective of this proposal is to elucidate the mechanisms and consequences of functional redundancy of APT1 and APT2 in the context of glucose homeostasis. The overall hypothesis is that depalmitoylation by both APT1 and APT2 regulates hepatic glucose metabolism. Aim 1 of this proposal will determine the substrate redundancy of APT1 and APT2 using in vitro proximity labeling. This aim will generate APT-biotin ligase fusion constructs to label proteins proximal to APT1 and APT2. Mass spectrometry of biotinylated proteins will facilitate identification of shared APT-protein interactions. Aim 2 of this proposal will investigate the functional redundancy of APT1 and APT2 in murine hepatic glucose metabolism. Chow- and high fat-fed mice with dual deletion of hepatic APT1 and APT2 will be tested for glucose, insulin, and pyruvate tolerance. Hepatic insulin signaling will also be evaluated in single and double liver-KO mice. The long-term goal of the proposed research is to implicate reversible lipid modifications in the development of metabolic disease. During the fellowship, the applicant will develop important skills for becoming an independent investigator of metabolism, emphasizing cellular and molecular biology techniques. The sponsor of this work, Dr. Clay Semenkovich, has vast experience studying the relationship between fatty acid and glucose metabolism, and the institutional environment provides supportive, collaborative experts in liver physiology and molecular biology. Washington University School of Medicine has a long history of helping physician-scientists build successful careers. The proposed training plan will facilitate the applicant’s transition into becoming an independent physician-scientist, using research to discover novel targets for treating chronic metabolic diseases.

项目摘要/摘要每年有成千上万的美国人被诊断出患有2型糖尿病，增加了他们的风险发展心血管疾病和慢性肾脏疾病。而通常在肝脏中发出胰岛素信号通过抑制新葡萄糖和诱导葡萄糖储存的合成来调节血糖水平肝胰岛素抵抗会加剧高血糖。胰岛素抵抗与失调有关脂肪酸代谢。脂肪酸可以通过称为可逆过程直接修饰蛋白质棕榈酰化。去氨木酰基酰基蛋白硫酯酶（APTS）（例如APT1和APT2）可以去除这些翻译后修饰。尽管已经显示了参与葡萄糖稳态的多种蛋白质为了被棕榈油，deplmitoylases在代谢中的作用仍在研究中。初步数据建议APT1肝敲除（APT1LKO）雌性小鼠具有胰岛素抵抗，而APT2LKO雌性小鼠表现出升高的空腹血浆葡萄糖水平。但是，APT1和APT2的冗余葡萄糖代谢途径尚不清楚。该提议的目的是阐明机制和在葡萄糖稳态的背景下，APT1和APT2功能冗余的后果。总体假设是APT1和APT2的deplimitoyl化调节肝葡萄糖代谢。目标1 该建议将使用体外接近标记确定APT1和APT2的底物冗余。这 AIM将产生APT-BIOTIN连接酶融合构建体，以标记靠近APT1和APT2的蛋白质。大量的生物素化蛋白的光谱法将促进鉴定共同的Apt-蛋白质相互作用。目标2 提案将研究鼠肝葡萄糖代谢中APT1和APT2的功能冗余。用双重缺失的肝apt1和APT2的Chow和高脂肪喂养的小鼠将测试葡萄糖，胰岛素，和丙酮酸耐受性。肝胰岛素信号传导也将在单肝脏和双肝脏小鼠中进行评估。这拟议的研究的长期目标是在开发中实施可逆的脂质修饰代谢疾病。在奖学金期间，申请人将发展重要技能，成为成为独立研究者代谢，强调细胞和分子生物学技术。这项工作的赞助商，克莱博士 Semenkovich，在研究脂肪酸与葡萄糖代谢之间的关系以及机构环境提供了肝生理和分子的支持性协作专家生物学。华盛顿大学医学学院有悠久的历史，帮助身体科学家建立成功的职业。拟议的培训计划将促进申请人的过渡到成为独立的物理科学家，使用研究发现治疗慢性代谢的新靶标疾病。