Store-operated Orai3 calcium channels in metabolism and obesity

商店操纵的 Orai3 钙通道在代谢和肥胖中的作用

• 批准号: 10427729
• 负责人: Trayambak Pathak
• 金额: $ 10.45万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427729
• 关键词: Address; Adipocytes; American; Area; Attenuated; Biological Process; Blood Pressure; Blood Vessels; Brain; Brown Fat; Calcium; Calcium Channel; Cardiovascular Diseases; Cell Nucleus; Cell membrane; Cell physiology; Cells; Complex; Data; Deacetylation; Development; Diet; Disease; Dyslipidemias; Electron Microscopy; Epidemic; Fatty acid glycerol esters; Gene Expression; Generations; Genetic Transcription; Glucose tolerance test; Goals; Growth; Heart; Heart Diseases; High Fat Diet; Homeostasis; House mice; Hyperglycemia; Hypertension; Insulin Resistance; Knock-out; Knockout Mice; Knowledge; Label; Life; Light; Lipolysis; Malignant Neoplasms; Mammals; Measures; Mediating; Medical; Membrane Potentials; Metabolic; Metabolic Diseases; Metabolism; Methods; Mitochondria; Modality; Molecular; Mus; Nerve; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Norepinephrine; Obese Mice; Obesity; Obesity associated cardiovascular disease; Obesity associated disease; Organelles; Overweight; Oxidative Phosphorylation; Oxidoreductase; PPAR gamma; Pathology; Pathway interactions; Peripheral Nervous System; Persons; Phenotype; Physiological; Physiology; Play; Population; Predisposition; Protein Isoforms; Proteins; RNA; Rabies virus; Regulation; Research; Risk Factors; Role; Route; SLC17A8 gene; Second Messenger Systems; Signal Pathway; Signal Transduction; Signaling Molecule; Spinal; Structure; Sympathetic Ganglia; Telemetry; Testing; Thermogenesis; Tissues; Transgenic Mice; Weight Gain; Work; cancer type; cardiometabolism; cell type; experimental study; heart function; insulin tolerance; lipid metabolism; mitochondrial metabolism; nerve supply; neuronal excitability; novel; obesity treatment; receptor; receptor coupling; spatiotemporal; transcription factor; uncoupling protein 1

Project Summary/Abstract This K99/R00 proposal aims to characterize novel molecular mechanisms of obesity and develop non-invasive methods for treating obesity and obesity-related cardiovascular diseases. The proposal addresses a fundamental gap in knowledge with a significant impact on the treatment of obesity. Obesity is a major contributor to several pathologies, including type 2 diabetes, cardiovascular disease, and cancer. In recent years, studies have shown that Ca2+ signals play a crucial role in lipid metabolism. Orai proteins (Orai1-3) are highly Ca2+ selective channels that contribute ubiquitous and evolutionary conserved routes of regulated Ca2+ entry into all cells and play a major role in a myriad of cellular and physiological functions, including lipolysis. Here, I propose to study the function of Ca2+ entry through Orai3, a mammalian specific Ca2+ selective Orai channel protein, in metabolism, neuronal innervation, neuronal activation, and its role in regulating thermogenesis and obesity. We have generated Orai3 whole-body knockout and Orai3flx/flx mice for tissue-specific knockouts. My preliminary data show that Orai3 knockout mice housed at 15-18°C and on regular diets quickly become obese, and their weight gain is derived from fat accumulation. This suggests that Orai3 plays an essential role in fat metabolism. These Orai3 KO mice also have reduced heat generation, reduced sympathetic innervation to brown adipose tissue (BAT), suggesting that adiposity is due to reduced BAT thermogenesis and/or altered innervation or activation of norepinephrine (NE) producing sympathetic neurons, which regulate BAT thermogenesis. In this application, I propose to address the above- described causes of adiposity in Orai3 KO mice with the following three aims and key experiments, 1) Ca2+ entry through Orai3 is crucial for BAT mitochondrial heat generation to control adiposity, which will be determined by measuring parameters of mitochondrial metabolism. 2) Orai3 activity is critical for Ca2+-dependent gene transcription of UCP1, required for BAT thermogenesis, which will be assessed by studying the transcription factors that regulate UCP1 expression and by rescuing UCP1 expression in Orai3 KO mice and determining cardio-metabolism on Orai3 KO and UCP1-rescued mice. 3) Orai3 is required in the central and peripheral nervous system (CNS and PNS) to regulate BAT activity, which will be determined by analyzing Orai3 levels in both CNS and PNS using RNA scope, deleting Orai3 in CNS and measuring BAT activity, and deleting Orai3 in CNS and measuring cold and high fat diet- induced hypertension and obesity. This study will help decipher the unique role of Orai3 in BAT and in both central and sympathetic neurons. It will bridge the gap between Ca2+ homeostasis and mitochondrial function and unveil a novel role for Orai3 Ca2+ signaling in lipid metabolism and obesity, establishing Orai3 as a target in obesity and obesity-related cardiovascular disease.

项目总结/摘要 该K99/R 00提案旨在表征肥胖的新分子机制， 开发治疗肥胖症和肥胖相关心血管疾病的非侵入性方法。的 建议解决了一个根本的知识差距，对治疗有重大影响， 肥胖肥胖是几种病理的主要贡献者，包括2型糖尿病、心血管疾病、糖尿病性糖尿病和糖尿病性糖尿病。 疾病和癌症。近年来的研究表明，钙离子信号在脂质代谢中起着至关重要的作用， 新陈代谢.奥赖蛋白（Orai 1 -3）是高度选择性的Ca 2+通道，其贡献了普遍存在的并且 调节Ca 2+进入所有细胞的进化保守途径，并在无数的 细胞和生理功能，包括脂肪分解。在此，我建议研究Ca 2+的功能 Orai 3是一种哺乳动物特异性的钙离子选择性奥赖通道蛋白，在代谢、神经元 神经支配、神经元活化及其在调节产热和肥胖中的作用。我们有 产生Orai 3全身敲除和Orai 3flx/flx小鼠用于组织特异性敲除。我的初步 数据显示，在15-18 ℃和常规饮食下饲养的Orai 3敲除小鼠迅速变得肥胖， 而他们的体重增加来自于脂肪的积累。这表明Orai 3在 在脂肪代谢中。这些Orai 3 KO小鼠还具有减少的产热、减少的交感神经传导、减少的神经传导和减少的神经传导。 棕色脂肪组织（BAT）的神经支配，表明肥胖是由于减少BAT 产热和/或改变的神经支配或去甲肾上腺素（NE）产生交感神经的激活 神经元，调节BAT产热。在本申请书中，本人建议处理上述事宜─ 描述了Orai 3 KO小鼠肥胖的原因，具有以下三个目的和关键实验，1） Ca 2+通过Orai 3进入对于BAT线粒体产热控制肥胖至关重要， 将通过测量线粒体代谢的参数来确定。2)Orai 3活性对于 UCP 1的Ca 2+依赖性基因转录，BAT产热所需，将对其进行评估 通过研究调节UCP 1表达的转录因子， 并测定Orai 3 KO和UCP 1挽救的小鼠的心脏代谢。3)Orai3 是中枢和外周神经系统（CNS和PNS）调节BAT活性所必需的， 将通过使用RNA scope分析CNS和PNS中的Orai 3水平来确定， CNS和测量BAT活性，以及在CNS中缺失Orai 3和测量冷和高脂肪饮食- 诱发高血压和肥胖。这项研究将有助于破译Orai 3在BAT中的独特作用， 中枢神经元和交感神经元。它将弥合Ca 2+稳态和Ca 2+代谢之间的差距。 线粒体功能，并揭示了Ora 13 Ca 2+信号在脂质代谢和肥胖中的新作用， 建立Orai 3作为肥胖和肥胖相关心血管疾病的靶点。