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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要/摘要这项K99/R00提案旨在表征肥胖的新分子机制和开发用于治疗肥胖症和肥胖相关心血管疾病的非侵入性方法。这提案解决了了解对待的根本差距肥胖。肥胖是多种病理的主要因素，包括2型糖尿病，心血管疾病和癌症。近年来，研究表明，Ca2+信号在脂质中起着至关重要的作用代谢。 Orai蛋白（ORAI1-3）是高度CA2+选择性通道，可无处不在进化配置的受调节Ca2+进入所有细胞的途径，并在无数的一小部分中发挥了重要作用细胞和物理功能，包括脂解。在这里，我建议研究Ca2+的功能通过Orai3（哺乳动物特异性CA2+选择性Orai通道蛋白）进入代谢，神经元神经，神经元激活及其在控制生热和肥胖中的作用。我们有生成的ORAI3全身敲除和用于组织特异性敲除的Orai3flx/Flx小鼠。我的初步数据表明，ORAI3敲除小鼠在15-18°C下容纳，并在常规饮食下迅速肥胖，他们的体重增加来自脂肪的积累。这表明Orai3扮演着重要的角色在脂肪代谢中。这些Orai3 KO小鼠也减少了热量产生，同情减少了对棕色脂肪组织（BAT）的神经支配，表明肥胖是由于蝙蝠减少引起的产生交感神经的去甲肾上腺素（NE）的神经支配或激活改变或激活神经元，调节蝙蝠的热发生。在此应用程序中，我建议解决上述描述了Orai3 KO小鼠肥胖的原因，其以下三个目标和关键实验，1）通过ORAI3进入Ca2+对于蝙蝠线粒体热的产生至关重要，以控制肥胖，这将通过测量线粒体代谢的参数来确定。 2）ORAI3活动对于 Ca2+依赖性的UCP1的基因转录，蝙蝠热发生所需通过研究调节UCP1表达并挽救UCP1表达的转录因子在Orai3 KO小鼠中，并确定ORAI3 KO和UCP1换倒的小鼠的心甲米代谢。 3）ORAI3 在中央和周围神经系统（CNS和PNS）中需要调节蝙蝠活动，这需要将通过使用RNA范围分析CNS和PNS中的ORAI3水平来确定中枢神经系统和测量蝙蝠活性，并在中枢神经系统中删除Orai3，并测量冷脂肪饮食 - 诱发高血压和肥胖症。这项研究将有助于破译Orai3在BAT和IN中的独特作用中央和同情神经元。它将弥合Ca2+稳态之间的差距线粒体功能并揭示了Orai3 Ca2+信号传导在脂质代谢和肥胖中的新作用，将ORAI3作为肥胖和与肥胖相关的心血管疾病的靶标建立。