Adipose Tissue Angiogenesis and Metabolic Disease

脂肪组织血管生成和代谢疾病

基本信息

批准号：
10523517
负责人：
Silvia Corvera
金额：
$ 50.78万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-01 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10523517
关键词：
Adipocytes Adipose tissue Affect Affinity Alleles Biochemical Calories Cell Nucleus Cell Respiration Cells Code Complex Cytoplasm Development Diabetes Mellitus Diagnostic Fatty Acids Fatty acid glycerol esters Feedback Funding Genes Genetic Transcription Goals Growth Health Heart Diseases Homeostasis Hormone secretion Human Image Impairment Individual Interphase Kinetics Label Lipids Lipolysis Mass Spectrum Analysis Mediating Mesenchymal Stem Cells Metabolic Metabolic Diseases Metabolism Mitochondria Mitochondrial Proteins Modeling Molecular Mus Nature Non-Insulin-Dependent Diabetes Mellitus Obesity Onset of illness Pathway interactions Physiological Primates Proteins RNA RNA-Protein Interaction Repression Research Respiration Role Stimulus Synapses Techniques Testing Transplantation Untranslated RNA Vascularization Work adipocyte differentiation angiogenesis blood glucose regulation crosslink experimental study fat burning gene function genetic regulatory protein glucose metabolism high resolution imaging improved in vivo knock-down mutant nerve supply novel strategies overexpression oxidation physiologic model protein complex protein protein interaction therapeutic target transcriptomics

项目摘要

The long-term objective of this project is to understand the mechanisms that distinguish human thermogenic adipocyte metabolism and contribute to metabolic health. During the past funding periods, we have been successful in three significant lines of research, as follows: a, We have developed a model to interrogate human adipose tissue physiological function in vivo in mice. b, We have discovered at least 4 distinct adipocyte subtypes differentiating from human mesenchymal progenitors, associated with specific adipose tissue functions and differentially enriched in human adipose tissue depots, and c, We have discovered LINC00473, a primate-specific long non-coding RNA specifically expressed in thermogenic adipocytes and involved in lipolysis and mitochondrial oxidative metabolism. LINC00473 levels are decreased in obesity and Type 2 Diabetes, consistent with an important physiological role. We will now further elucidate molecular mechanisms by which LINC00473 regulates these pathways and its contribution to systemic energy homeostasis. Aim 1. To elucidate the mechanism by which LINC00473 interacts with lipid droplet and mitochondrial proteins and the effect of these interactions on fatty acid and mitochondrial oxidative metabolism. Using affinity isolation and mass spectrometry, we have identified lipid droplet and mitochondrial proteins that interact with LINC00473. We will now: 1A. Define direct and indirect interactions between LINC00473 and candidate proteins, using molecular approaches including ChIRP and APEX2 proximity labeling. 1B. Define the kinetics of assembly of LINC00473-protein complexes and their relationship with stimulated lipolysis and respiration, and 1C. Define the role of specific interactions though expression of LINC00473 mutant constructs. Aim 2. To test the hypothesis that LINC00473 mediates a feedback mechanism to control lipolysis. We shall: 2A. Determine whether inhibition of LINC00473 expression upon PLIN1 depletion is due to enhanced basal lipolysis. 2B. Determine whether impaired stimulation of lipolysis in PLIN1 depleted cells is due LINC00473 supprssion. 2C. Determine whether PLIN1 depletion suppresses LINC00473 through decreased transcription, enhanced degradation or both. Aim 3. We will test the role of LINC00473 on thermogenic adipose tissue development and systemic glucose metabolism. Using techniques to generate human adipose tissue in mice we shall: 3A. Assess the effects of overexpression or knockdown of LINC00473 on dynamics of adipose tissue growth, vascularization, and innervation. 3B. Determine how adipose tissue developed from adipocytes expressing high or low levels of LINC00473 responds to physiological stimuli such as cold. 3C. Determine whether adipose tissue developed from adipocytes expressing high or low levels of LINC00473 differentially affects systemic glucose metabolism. This work will reveal molecular mechanisms that define human thermogenic adipocyte function, and how these mechanisms operate systemically and could impact metabolic disease.

该项目的长期目标是了解区分人类生热脂肪细胞代谢和促进代谢健康的机制。在过去的资助期间，我们在三个重要的研究领域取得了成功，如下：a，我们开发了一种模型来探究小鼠体内人类脂肪组织的生理功能。 b，我们发现了至少 4 种与人类间充质祖细胞不同的不同脂肪细胞亚型，与特定的脂肪组织功能相关，并且在人类脂肪组织库中差异富集，并且 c，我们发现了 LINC00473，一种灵长类特异性长非编码 RNA在产热脂肪细胞中表达并参与脂肪分解和线粒体氧化代谢。 LINC00473 水平在肥胖和 2 型糖尿病中降低，一致具有重要的生理作用。我们现在将进一步阐明 LINC00473 调节这些途径的分子机制及其对系统能量稳态的贡献。目的 1. 阐明 LINC00473 与脂滴和线粒体蛋白相互作用的机制以及这些相互作用对脂肪酸和线粒体氧化代谢的影响。使用亲和分离和质谱分析，我们鉴定了与 LINC00473 相互作用的脂滴和线粒体蛋白。我们现在将：1A。使用 ChIRP 和 APEX2 邻近标记等分子方法定义 LINC00473 与候选蛋白质之间的直接和间接相互作用。 1B.定义 LINC00473-蛋白质复合物的组装动力学及其与刺激脂肪分解和呼吸的关系，以及 1C。通过 LINC00473 突变体构建体的表达定义特定相互作用的作用。目标 2. 检验 LINC00473 介导控制脂肪分解的反馈机制的假设。我们应：2A。确定 PLIN1 耗尽后 LINC00473 表达的抑制是否是由于基础脂肪分解增强所致。 2B。确定 PLIN1 耗尽细胞中脂肪分解刺激受损是否是由于 LINC00473 抑制所致。 2C。确定 PLIN1 缺失是否通过减少转录、增强降解或两者同时抑制 LINC00473。目标 3. 我们将测试 LINC00473 对生热脂肪组织发育和全身葡萄糖代谢的作用。使用在小鼠体内生成人类脂肪组织的技术，我们将： 3A。评估 LINC00473 的过表达或敲低对脂肪组织生长、血管化和神经支配动力学的影响。 3B.确定由表达高水平或低水平 LINC00473 的脂肪细胞发育而来的脂肪组织如何响应生理刺激（例如寒冷）。 3C。确定由表达高水平或低水平 LINC00473 的脂肪细胞发育而来的脂肪组织是否对全身葡萄糖代谢产生不同的影响。这项工作将揭示定义人类产热脂肪细胞功能的分子机制，以及这些机制如何系统地运作并可能影响代谢疾病。