The Role of Mitochondrial TNAP in Adaptive Thermogenesis

线粒体 TNAP 在适应性产热中的作用

• 批准号: 10591696
• 负责人: Yizhi Sun
• 金额: $ 12.9万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-01 至 2025-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10591696
• 关键词: Ablation; Acceleration; Address; Adipocytes; Adipose tissue; Advisory Committees; Alkaline Phosphatase; Amino Acid Sequence; Animals; Area; Automobile Driving; Binding Proteins; Biochemical; Biological; Biology; Biophysics; Cardiovascular Diseases; Cells; Chemicals; Consumption; Creatine; Cysteine; Dana-Farber Cancer Institute; Data; Diabetes Mellitus; Disease; Doctor of Philosophy; Elements; Energy Metabolism; Ensure; Environment; Event; Fatty Liver; Fatty acid glycerol esters; Futile Cycling; Generations; Genes; Genetic; Goals; Hydrolysis; Immunoprecipitation; Kidney Diseases; Laboratories; Laboratory Research; Link; Malignant Neoplasms; Mediator; Medical; Membrane Microdomains; Metabolic; Metabolic Diseases; Metabolism; Methods; Mitochondria; Mitochondrial Proteins; Molecular; Mouse Strains; Mus; Muscle Fibers; Mutation Analysis; Obesity; Oxidative Phosphorylation; Pathway interactions; Phosphocreatine; Phosphorylation; Physiological; Play; Post-Translational Protein Processing; Prevention; Process; Protein Dephosphorylation; Proteins; Proteome; Proteomics; Public Health; Reactive Oxygen Species; Regulation; Reporter; Research; Research Personnel; Respiration; Role; Signal Transduction; Specificity; Testing; Therapeutic; Thermogenesis; Tissue Engineering; Tissues; Transgenic Mice; Transportation; Travel; Variant; Visualization; Work; adipocyte biology; career; cellular imaging; diet-induced obesity; gain of function; genetic approach; imaging approach; imaging study; innovation; medical schools; metabolic abnormality assessment; novel; obesity prevention; obesity treatment; operation; pharmacologic; polypeptide; protein transport; red fluorescent protein; therapeutic target; trafficking

Abstract Thermogenic adipocytes and adaptive thermogenesis are promising therapeutic targets for treating and preventing obesity and obesity-linked diabetes. We recently demonstrated that the mitochondrial tissue- nonspecific alkaline phosphatase, TNAP, plays a crucial role in the futile creatine cycle (FCC) and adaptive thermogenesis by hydrolyzing phosphocreatine. Pharmacological inhibition or genetic ablation of TNAP in mice reduces systemic energy expenditure. Genetic ablation of TNAP in mice also causes rapid-onset obesity. TNAP assumes a mitochondrial localization specifically in thermogenic adipocytes, which might ensure the cell- selectivity of the phosphocreatine hydrolysis events and the FCC operation. Here, we propose to study the metabolic effects of gain-of- function of TNAP in the adipose tissues in mice, as well as the molecular mechanism and regulation of its mitochondrial localization. Using a transgenic mouse strain that artificially expresses TNAP in the adipose tissues, we will investigate its effects on obesity, diabetes, fatty liver, and energy expenditure. To study the TNAP localization, we will test whether its cell specificity lies in the TNAP polypeptide or its trafficking pathway, or both. I will first determine whether there are any molecular elements (e.g., PTMs or amino acid sequences) on the TNAP polypeptide crucial for its mitochondrial localization. This will be done using a combination of biochemical and cell imaging approaches. In addition, we will investigate the cellular pathway of TNAP trafficking to mitochondria. We will interrogate the role of lipid rafts in TNAP localization. This might allow us to identify key regulators and mediators of the mitochondrial localization of TNAP. Finally, we will identify other mitochondrial proteins that share the localization pathway of TNAP and study their functions. The candidate, Dr. Yizhi Sun, has a strong track record of innovative research with a focus on the molecular mechanisms of diseases. The candidate’s career goal is to become an independent academic investigator with a research laboratory oriented towards understanding and reversing obesity and obesity-linked metabolic disorders. The proposed research will be conducted in the laboratory of Bruce Spiegelman, PhD at Dana-Farber Cancer Institute and Harvard Medical School, who is a leader in the fields of molecular metabolism and adipocyte biology. The proposed studies will also bring together leading laboratories of the advisory committee that have expertise in cell imaging, protein trafficking, and mitochondrial biology. All of these, together with the ideal research environment in the Longwood Medical Area, will maximize applicant’s potential to successfully transition to an independent investigator.

抽象的 产热脂肪细胞和适应性产热是有希望的治疗靶点 预防肥胖和与肥胖相关的糖尿病。我们最近证明线粒体组织- 非特异性碱性磷酸酶 (TNAP) 在无效肌酸循环 (FCC) 和适应性中起着至关重要的作用 通过水解磷酸肌酸产生热量。小鼠体内 TNAP 的药理抑制或基因消除 减少全身能量消耗。小鼠体内 TNAP 的基因消除也会导致快速肥胖。 TNAP 假设线粒体定位于产热脂肪细胞，这可能确保细胞 磷酸肌酸水解事件和 FCC 操作的选择性。在此，我们建议研究 TNAP功能获得对小鼠脂肪组织代谢的影响及分子机制 及其线粒体定位的调节。使用人工表达 TNAP 的转基因小鼠品系 在脂肪组织中，我们将研究其对肥胖、糖尿病、脂肪肝和能量消耗的影响。到 研究TNAP定位，我们将测试其细胞特异性是否在于TNAP多肽或其运输 途径，或两者兼而有之。我将首先确定是否存在任何分子元素（例如 PTM 或氨基酸） TNAP 多肽上的序列）对其线粒体定位至关重要。这将使用 生物化学和细胞成像方法的结合。此外，我们还将研究细胞途径 TNAP 转运至线粒体。我们将探讨脂筏在 TNAP 定位中的作用。这可能允许 我们确定了 TNAP 线粒体定位的关键调节因子和介导因子。最后，我们将识别 其他共享 TNAP 定位途径的线粒体蛋白并研究其功能。 候选人孙益智博士在专注于分子生物学的创新研究方面有着良好的记录。 疾病的机制。候选人的职业目标是成为一名独立的学术研究者 一个致力于了解和逆转肥胖以及与肥胖相关的代谢的研究实验室 失调。拟议的研究将在丹娜—法伯癌症研究所 Bruce Spiegelman 博士的实验室进行 癌症研究所和哈佛医学院，分子代谢和脂肪细胞领域的领导者 生物学。拟议的研究还将汇集咨询委员会的主要实验室，这些实验室已经 细胞成像、蛋白质运输和线粒体生物学方面的专业知识。所有这些，连同理想 朗伍德医学区的研究环境将最大限度地发挥申请人成功的潜力 过渡为独立调查员。