Follistatin promotes browning and influences energy metabolism

卵泡抑素促进褐变并影响能量代谢

• 批准号: 8740378
• 负责人: RAJAN SINGH
• 金额: $ 28.7万
• 依托单位: CHARLES R. DREW UNIVERSITY OF MED & SCI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8740378
• 关键词: Activins; Adipocytes; Adipose tissue; Adrenergic Receptor; Affinity; Age; Agonist; Area; BMP7 gene; Bioenergetics; Biogenesis; Biological Assay; Body Composition; Brown Fat; Burn injury; Calories; Cardiovascular Diseases; Cells; Consultations; Data; Development; Diabetes Mellitus; Diet; Disease; Doctor of Philosophy; Drug Design; Embryo; Energy Intake; Energy Metabolism; Enzyme-Linked Immunosorbent Assay; Epidemic; Exercise; Expenditure; Extracellular Protein; FGF21 gene; Fatty acid glycerol esters; Fibroblasts; Follistatin; Gene Expression; Gene Expression Profiling; Genus Hippocampus; Gifts; Glucose; Glucose tolerance test; Grant; Health; In Vitro; Inflammatory; Insulin Receptor; Insulin Resistance; Knock-out; Knowledge; Link; Lipids; Mediator of activation protein; Metabolic; Metabolic Diseases; Minority; Mission; Mitochondria; Molecular; Molecular Profiling; Mus; Muscle; Non-Insulin-Dependent Diabetes Mellitus; Obesity; PTGS2 gene; Pathway interactions; Play; Population; Process; Protein Binding; Proteins; Reagent; Receptor Signaling; Recruitment Activity; Regulation; Regulatory Element; Reporting; Research Personnel; Risk Factors; Role; Serum; Signal Pathway; Signal Transduction; Skeletal Muscle; Small Interfering RNA; Subcutaneous Tissue; Symptoms; Testing; Testosterone; Time; Tissues; Transforming Growth Factors; Transgenes; Transgenic Mice; Transgenic Mouse Facility; Triglycerides; United States National Institutes of Health; Western Blotting; Wild Type Mouse; Work; X-Ray Computed Tomography; adipocyte biology; adiponectin; base; burden of illness; disability; energy balance; experience; extracellular; fatty acid oxidation; glucose disposal; glucose metabolism; glucose tolerance; improved; in vitro Model; in vivo; in vivo Model; insulin sensitivity; knock-down; lipid metabolism; male; muscle form; myostatin; novel; novel therapeutics; obesity treatment; prevent; programs; promoter; protein expression; response; treatment program

DESCRIPTION (provided by applicant): Obesity is a major health problem spreading at an epidemic pace throughout the world without any sign of abatement. Development of obesity, which is often associated with insulin resistance and diabetes, results from an excess of energy intake over expenditure. Brown adipose tissues (BAT) have the unique ability to burn excess calories, and facilitate triglyceride clearance and glucose disposal. Previously, we identified Follistatin (Fst) as a direct target of testosterone that regulates skeletal muscle mass and inhibits transforming growth factor-β (TGF-β) signaling. Based on our preliminary findings, we hypothesize that Fst promotes overall thermogenic program and improves symptoms of obesity and metabolic disorder by regulating overall Mst/TGF-β /BMP/Myf5/PRDM16- signaling pathways. We will test our hypothesis with the following Specific Aims- Aim 1: We will demonstrate the essential role of Fst during brown fat differentiation and the regulation of thermogenic program in vitro and in vivo. Aim 2: We will determine the molecular mechanisms by which Fst regulates the overall thermogenic program and Aim 3: We will generate mice expressing Fst under the control of adiponectin or UCP1 regulatory elements, and compare their metabolic parameters and response to high fat chow diet with control littermates. We will determine the effect of endogenous and exogenous Fst on protein and gene expression profiles of key thermogenic markers, and overall cellular bioenergetics using both in vitro and in vivo models under basal and β-adrenoceptor agonist (CL316,248) stimulated conditions. Involvement of Myf5/PRDM16, Mst/pSmad2/3/BMP/COX 2, insulin receptor and AMPK/PGC-1α signaling pathway in in vitro models as well as in adiponectin-Fst and UCP1-Fst transgenic mice will be analyzed by Affymetrix gene expression and quantitative western blot analysis. Expression levels of Fst, Myf5 and Smad3 in mouse preadipocytes and MEF cultures will be inhibited by siRNAs and their thermogenic capabilities will be determined. We will generate Fst-transgenic mice using adiponectin (Adipoq) and UCP1-specific promoters, and test the effect of high fat diet on their body composition (Micro CT), energy expenditure (indirect calorimetric analysis), insulin sensitivity and glucose tolerance under both basal and β-adrenoceptor agonist (CL 316,248) stimulated conditions. Serum levels of Fst, adiponectin and lipid profiles will be analyzed by ELISA. Evaluating the critical role of Fst during its regulation of overall thermogenic process will provide rationale for novel therapeutic drug design for the treatment of obesity and related metabolic diseases.

描述（由申请人提供）：肥胖是一个主要的健康问题，在世界范围内以流行病的速度蔓延，没有任何减少的迹象。肥胖通常与胰岛素抵抗和糖尿病有关，是能量摄入超过消耗的结果。棕色脂肪组织（BAT）具有燃烧多余卡路里的独特能力，并促进甘油三酯的清除和葡萄糖的处理。在此之前，我们发现Follistatin （Fst）是睾酮调节骨骼肌质量和抑制转化生长因子-β (TGF-β)信号传导的直接靶点。基于我们的初步研究结果，我们假设Fst通过调节Mst/TGF-β /BMP/Myf5/PRDM16-信号通路，促进整体产热程序，改善肥胖和代谢紊乱症状。我们将通过以下具体目标来验证我们的假设-目标1：我们将在体外和体内证明Fst在棕色脂肪分化和产热程序调节中的重要作用。目的2：我们将确定Fst调控整个产热程序的分子机制。目的3：我们将产生在脂联素或UCP1调控元件控制下表达Fst的小鼠，并将其代谢参数和对高脂饲料的反应与对照组进行比较。我们将确定内源性和外源性Fst对关键产热标志物的蛋白质和基因表达谱的影响，以及在基础和β-肾上腺素能受体激动剂（CL316,248）刺激条件下的体外和体内模型的整体细胞生物能量学。通过Affymetrix基因表达和定量western blot分析Myf5/PRDM16、Mst/pSmad2/3/BMP/COX 2、胰岛素受体和AMPK/PGC-1α信号通路在体外模型以及脂联素- fst和UCP1-Fst转基因小鼠中的参与情况。Fst、Myf5和Smad3在小鼠前脂肪细胞和MEF培养物中的表达水平将被sirna抑制，其产热能力将被确定。我们将使用脂联素（Adipoq）和ucp1特异性启动子培育转基因小鼠，并在基础和β-肾上腺素受体激动剂（CL 316,248）刺激的条件下，测试高脂肪饮食对小鼠体成分（Micro CT）、能量消耗（间接量热分析）、胰岛素敏感性和葡萄糖耐量的影响。血清Fst水平、脂联素和脂质谱将通过ELISA分析。评估Fst在整个产热过程中的关键作用，将为设计治疗肥胖和相关代谢疾病的新型治疗药物提供依据。