Translational regulation of PGC1alpha and oxidative metabolism

PGC1α 和氧化代谢的翻译调控

• 批准号: 10214944
• 负责人: Phillip Anthony Dumesic
• 金额: $ 9万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10214944
• 关键词: 3' Untranslated Regions; Adipocytes; Adrenergic Agents; Affect; Affinity Chromatography; American; Area; Attention; Award; Basic Science; Biogenesis; Bioinformatics; Biotinylation; Case Study; Cell Respiration; Cells; Chemicals; Complement; Cytoplasmic Granules; Dana-Farber Cancer Institute; Data; Data Set; Diabetes Mellitus; Disease; Economic Burden; Elements; Energy Metabolism; Environment; Fatty acid glycerol esters; Fibrinogen; Follow-Up Studies; Foundations; Functional disorder; Gene Expression; Genetic Transcription; Genetic Translation; Goals; Health; Homeostasis; Hormones; Human; In Vitro; Instruction; Lead; Leadership; Mass Spectrum Analysis; Mentors; Mentorship; Messenger RNA; Metabolic; Metabolic Diseases; Metabolism; Methods; Mitochondria; Molecular; Mus; Mutation; Norepinephrine; Obesity; Output; Oxidative Phosphorylation; Pathway interactions; Pharmacology; Phase; Phenotype; Physiological; Physiology; Play; Post-Transcriptional Regulation; Prevalence; Process; Property; Proteins; Proteomics; RNA; RNA-Binding Proteins; Regulation; Research; Ribosomes; Rodent Model; Role; Signal Transduction; System; Technology; Testing; Therapeutic; Thermogenesis; Tissues; Training; Translational Regulation; Translations; United States National Institutes of Health; Work; Writing; adipocyte biology; adipocyte differentiation; cell type; cis acting element; design; experimental study; genetic manipulation; helicase; hormone regulation; in vivo; insight; knock-down; medical schools; metabolic phenotype; mouse model; novel; obesity treatment; preference; prevent; programs; ribosome profiling; skills; stem; stress granule; transcriptomics; translation factor; translational impact; uncoupling protein 1

PROJECT SUMMARY As the prevalence of obesity and associated disorders rises, manipulation of adipocyte energy expenditure has gained attention as a potential means to treat metabolic disease. This strategy leverages the physiology of thermogenic adipocytes, a cell type that relies on oxidative phosphorylation metabolism to drive futile chemical cycles that release energy as heat. Recent work has suggested the potential for ribosomal control of thermogenic gene expression. Elucidating the responsible mechanisms may reveal novel means to manipulate adipocyte metabolism for the treatment of obesity and diabetes. This proposal tests the hypothesis that the distinct metabolism of thermogenic fat is enabled by specialized mRNA translation preferences inherent in this cell type. A first set of studies focuses on a single mRNA, PPARGC1A, and aims to purify the proteins that confer its cell-type-specific translational output. This mRNA encodes PGC1α, a dominant regulator of mitochondrial biogenesis. A second set of studies develops an in vivo mouse model to test the hypothesis that the helicase DDX3X, a regulator of mRNA translation and cytoplasmic stress granules, is a thermogenic-fat- selective translational regulator that supports expression of genes critical for oxidative metabolism. A third set of studies uses ribosome profiling technology to define the global mRNA translation dynamics associated with thermogenic activation. Together, these studies expand the scope of my work while generating foundational datasets and mouse models for my scientific independence. They will be carried out in the lab of a recognized leader in the field of molecular metabolism, Dr. Bruce Spiegelman, at the Dana-Farber Cancer Institute and Harvard Medical School. In this rich environment, I will benefit from a mentorship team committed to instruct me in: adipocyte biology, including in vivo genetic manipulation; mouse metabolic phenotyping; mass spectrometry; and bioinformatics. The technical training is complemented by formal instruction in human metabolic pathophysiology, as well as activities for honing my leadership, speaking, and writing skills. In this way the NIH Pathway to Independence Award supports my transition from mentored work to an independent stage in which I intend to use cutting-edge methods to broadly define the translational dynamics of thermogenic fat, identify the responsible regulators, and test how post-transcriptional dynamics such as stress granule assembly may contribute. My ultimate goal is to lead an academic research group that investigates how the gene expression programs that define cellular identity and metabolism are established, how they are perturbed in metabolic disease, and how they may be therapeutically manipulated to prevent or treat metabolic disease.

项目概要 随着肥胖和相关疾病患病率的上升，对脂肪细胞能量消耗的控制已经 作为治疗代谢疾病的潜在手段而受到关注。该策略利用了生理学 产热脂肪细胞，一种依赖氧化磷酸化代谢来驱动无用化学物质的细胞类型 以热能形式释放能量的循环。最近的工作表明核糖体控制的潜力 产热基因表达。阐明负责任的机制可能会揭示新的操纵手段 脂肪细胞代谢用于治疗肥胖和糖尿病。该提议检验了以下假设： 生热脂肪的独特代谢是通过其固有的专门 mRNA 翻译偏好实现的 细胞类型。第一组研究重点关注单个 mRNA PPARGC1A，旨在纯化能够 赋予其细胞类型特异性的翻译输出。该 mRNA 编码 PGC1α，PGC1α 是 线粒体生物发生。第二组研究开发了体内小鼠模型来检验以下假设： 解旋酶 DDX3X 是 mRNA 翻译和细胞质应激颗粒的调节剂，是一种产热脂肪 支持氧化代谢关键基因表达的选择性翻译调节因子。第三组 的研究使用核糖体分析技术来定义与相关的全局 mRNA 翻译动态 生热激活。总之，这些研究扩大了我的工作范围，同时产生了基础 数据集和小鼠模型以实现我的科学独立性。它们将在公认的实验室中进行 分子代谢领域的领导者，布鲁斯·斯皮格曼 (Bruce Spiegelman) 博士，达纳-法伯癌症研究所 哈佛医学院。在这个丰富的环境中，我将受益于致力于指导的导师团队 我在：脂肪细胞生物学，包括体内基因操作；小鼠代谢表型分析；大量的 光谱测定；和生物信息学。技术培训辅以正式的人力指导 代谢病理生理学，以及磨练我的领导力、口语和写作技能的活动。在这个 NIH 独立之路奖支持我从指导工作过渡到独立工作 我打算使用尖端方法来广泛定义转化动力学的阶段 生热脂肪，确定负责的调节因子，并测试转录后动态（例如压力） 颗粒组装可能有所贡献。我的最终目标是领导一个学术研究小组来调查 定义细胞身份和代谢的基因表达程序是如何建立的，它们是如何建立的 代谢疾病的扰动，以及如何通过治疗操纵它们来预防或治疗代谢疾病 疾病。