Discovery and characterization of a novel acylcarnitine transporter in brown adipocytes

棕色脂肪细胞中新型酰基肉碱转运蛋白的发现和表征

基本信息

批准号：
10681400
负责人：
Judith Anne Simcox
金额：
$ 31.1万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-15 至 2027-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10681400
关键词：
Address Adipocytes Adipose tissue Adult Affect American Biological Assay Biological Markers Blood Brown Fat CRISPR library CRISPR screen Cardiovascular Diseases Cardiovascular system Carrier Proteins Cell Culture Techniques Cell membrane Cell physiology Cells Cessation of life ChIP-seq Development Diabetes Mellitus Disease Energy Metabolism Gene Expression Genomics Glucose Health Heart Homeostasis In Vitro Individual Insulin Resistance Knock-out Knowledge Link Lipids Liver Luciferases Mass Spectrum Analysis Mediating Membrane Transport Proteins Metabolic Metabolic Diseases Metabolism Modeling Molecular Mus Non-Insulin-Dependent Diabetes Mellitus Obesity Obesity associated disease Overweight PPAR gamma Pathway interactions Pharmacological Treatment Physiologic Thermoregulation Physiological Plasma Population Primary carcinoma of the liver cells Process Production Regulation Reporter Respiration Signal Transduction Skeletal Muscle Source Supercritical Fluid Chromatography Thermogenesis Time Tissue Sample Tissues Transcript United States Work acylcarnitine aged blood glucose regulation chromatin immunoprecipitation experimental study fatty acid oxidation glucose tolerance glucose uptake improved in vivo in vivo Model insight insulin regulation insulin sensitivity insulin signaling insulin tolerance mouse model novel obesity treatment overexpression prevent promoter protein expression resistance mechanism response solute targeted treatment theories therapeutic target tissue/cell culture uptake

项目摘要

Project Summary/Abstract In the United States, 42% of the population is obese, and this obesity predisposes individuals for several diseases including type 2 diabetes which affects 29 million Americans. One established mechanism for the causal relationship between obesity and type 2 diabetes is the development of lipotoxicity, a state where excess lipids build-up to cause increased ectopic lipid droplets, elevated plasma lipids, and subsequently, decreased cellular insulin signaling. These plasma lipids that are increased with lipotoxicity include acylcarnitines, a key intermediate in fatty acid oxidation. Acylcarnitines are an established biomarker of type 2 diabetes, hepatocellular carcinoma, and cardiovascular disease, and are known to cause insulin resistance. Despite the known importance of plasma acylcarnitines in the development of insulin resistance, we do not know how they are imported into cells, how this uptake is regulated, or how imported acylcarnitines are metabolized once they enter the cell from the circulatory system. In this proposal we examine the regulation of acylcarnitine uptake by transporters we have identified through a CRISPR/Cas9 screen. Through targeted knockout of these transporters, we will conduct cell culture experiments to highlight the precise molecular pathways through which acylcarnitines enter the cell and signal for insulin sensitivity. We will also explore tissue specific loss of acylcarnitines in mouse models to determine the contribution of plasma acylcarnitine uptake on whole-body energy expenditure, glucose regulation, and plasma lipid pools. Finally, we will determine how these transporters for plasma acylcarnitines are regulated in cell culture and tissue samples from mice to establish potential therapeutic targets for the treatment of metabolic disease. The proposed work will address long-standing questions on the molecular regulation of plasma acylcarnitine import, metabolism of acylcarnitines after entry into the cell, and the regulation of insulin sensitivity by acylcarnitines. In terms of translatability, this work will generate a mechanistic understanding of how plasma acylcarnitine abundance is controlled and how plasma acylcarnitines impact disease vulnerability to enable the identification of new targets for the pharmacological treatment of obesity and diabetes.

项目概要/摘要在美国，42% 的人口肥胖，这种肥胖使个人容易患上多种疾病包括 2 型糖尿病在内的疾病影响着 2900 万美国人。一项既定机制肥胖与 2 型糖尿病之间的因果关系是脂毒性的发展，脂毒性是一种过量摄入脂肪的状态。脂质积聚导致异位脂滴增加、血浆脂质升高，随后降低细胞胰岛素信号传导。这些随脂毒性而增加的血浆脂质包括酰基肉碱，这是一种关键的脂肪酸氧化的中间体。酰基肉碱是 2 型糖尿病、肝细胞癌的既定生物标志物癌症和心血管疾病，并且已知会导致胰岛素抵抗。尽管已知血浆酰基肉碱在胰岛素抵抗发展中的重要性，我们不知道它们是如何发生的导入细胞，如何调节这种摄取，或者导入的酰基肉碱进入细胞后如何代谢来自循环系统的细胞。在本提案中，我们研究了我们通过以下方法确定的转运蛋白对酰基肉碱摄取的调节： CRISPR/Cas9 筛选。通过靶向敲除这些转运蛋白，我们将进行细胞培养实验强调酰基肉碱进入细胞并发出胰岛素信号的精确分子途径敏感性。我们还将探索小鼠模型中酰基肉碱的组织特异性损失，以确定血浆酰基肉碱摄取对全身能量消耗、葡萄糖调节和血浆的贡献脂质池。最后，我们将确定这些血浆酰基肉碱转运蛋白如何在细胞中受到调节小鼠的培养物和组织样本，以建立治疗代谢性疾病的潜在治疗靶点疾病。拟议的工作将解决血浆酰基肉碱分子调节的长期存在的问题酰基肉碱进入细胞后的输入、代谢以及胰岛素敏感性的调节酰基肉碱。就可翻译性而言，这项工作将对等离子体如何产生机械性的理解控制酰基肉碱丰度以及血浆酰基肉碱如何影响疾病易感性，从而使确定肥胖和糖尿病药物治疗的新靶点。