PPARG regulates osteocyte bioenergetics and function during aging

PPARG 在衰老过程中调节骨细胞生物能和功能

• 批准号: 10426408
• 负责人: Patrick Robert Griffin
• 金额: $ 2.64万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10426408
• 关键词: Adipocytes; Affect; Age-Related Bone Loss; Aging; Agonist; American; Applications Grants; Bioenergetics; Bone Resorption; Bone remodeling; C57BL/6 Mouse; CRISPR/Cas technology; Cell physiology; Cells; Chronic Disease; Coculture Techniques; Diabetes Mellitus; Diagnosis; Disease; Elderly; Endocrine; Energy Metabolism; Enhancers; Exhibits; Fracture; Genetic; Genetic Transcription; Homeostasis; Hormonal; Hyperglycemia; Impairment; In Vitro; Knowledge; Lead; Maintenance; Metabolic Diseases; Metabolic dysfunction; Metabolism; Minerals; Mitochondria; Molecular; Mus; Nuclear Receptors; Organ; Organ Culture Techniques; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; PPAR gamma; Pathway interactions; Pharmacology; Population; Process; Production; Proteins; Proteomics; Quality of life; Regulation; Research; Role; Signal Transduction; Skeleton; TNFSF11 gene; Testing; aged; aging population; bone; bone cell; bone loss; bone mass; bone metabolism; bone quality; effective therapy; experimental study; improved; in vivo Model; inhibitor/antagonist; insight; mouse model; novel; osteoblast differentiation; paracrine; preference; programs; promoter; protein complex; response; rosiglitazone; skeletal; transcriptomics

Bone loss and impairment in energy metabolism with aging prevalently lead to fractures and diabetes. As an organ, the skeleton is one of the largest consumers of energy needed for bone remodeling and maintenance of bone homeostasis. Bone remodeling is under control of osteocytes, which constitute 90-95% of bone cells, and which bioenergetic program is largely unknown. New evidence indicates that nuclear receptor PPARG, which is a global regulator of energy metabolism and pharmacological target to treat hyperglycemia, regulates osteocyte energy metabolism and their function including production of sclerostin, an inhibitor of WNT pathway activity and a key protein regulating bone remodeling. Research proposed in this application will provide an insight into the connection between osteocyte bioenergetics, its function in bone metabolism and systemic energy metabolism, and whether bone acts as the body “energostat” via osteocyte endocrine activities. The leading hypothesis is that changes in PPARG activity with aging alter osteocyte function and bioenergetics resulting in simultaneous decrease in bone formation and decrease in energy metabolism. This hypothesis will be tested in three specific aims. Aim 1 will determine how fuel choice and aging affect osteocyte function and whether these processes are PPARG dependent. With the use of aged C57BL/6 and γOTKO mice, bone organ cultures, and osteocyte-like MLO-Y4 cells with down-regulated PPARG, in combination with transcriptomics, quantitative proteomics, and coculture experiments, PPARG contribution to osteocyte aging will be defined. Aim 2 will contrast the PPARG protein interactome of osteocytes with the interactome of adipocytes, in response to either full agonist rosiglitazone or the inverse agonist SR10171, in order to define an optimal PPARG modulator for its beneficial effects on bone and metabolism. The analysis will be done on osteocyte- like MLO-Y4 cells using quantitative Tandem Mass Tag Proteomics. Aim 3 will define dynamics of PPARG interactome on PPRE sequences present in Sost promoter as a function of aging and in response to pharmacological modulation of PPARG activity. The premise of this application is to set a stage for precise pharmacologic manipulation with PPARG activities in osteocytes to simultaneously improve skeletal and metabolic dysfunction in elderly.

随着年龄的增长，骨丢失和能量代谢受损会导致骨折和糖尿病。作为 骨骼是骨骼重建和维持骨骼健康所需能量的最大消耗者之一。 骨稳态骨重建受骨细胞控制，骨细胞占骨细胞的90-95%， 哪一种生物能量计划在很大程度上是未知的新的证据表明，核受体PPARG， 是能量代谢和治疗高血糖药理学靶点的全局调节剂，调节 骨细胞能量代谢及其功能，包括WNT途径抑制剂sclerostin的产生 活性和调节骨重建的关键蛋白质。本申请中提出的研究将提供 深入了解骨细胞生物能量学，其在骨代谢中的功能和全身 能量代谢，以及骨是否通过骨细胞内分泌活动作为机体的“能量调节器”。的 主要假设是PPARG活性随年龄的变化改变了骨细胞的功能和生物能量学 导致骨形成的同时减少和能量代谢的减少。这一假设将 在三个具体目标中进行测试。目标1将确定燃料选择和老化如何影响骨细胞功能， 这些过程是否依赖于PPARG。使用老年C57 BL/6和γOTKO小鼠，骨器官 培养物和具有下调的PPARG的骨细胞样MLO-Y 4细胞，与转录组学组合， 定量蛋白质组学和共培养实验，PPARG对骨细胞老化的贡献将被定义。 目的2将比较骨细胞和脂肪细胞的PPARG蛋白相互作用组， 对完全激动剂罗格列酮或反向激动剂SR 10171的反应，以确定最佳的 PPARG调节剂对骨和代谢的有益作用。分析将在骨细胞上进行- 如MLO-Y 4细胞，使用定量串联质量标签蛋白质组学。目标3将定义PPARG的动态 Sost启动子中存在PPRE序列上的相互作用基因组作为衰老的函数和响应于 PPARG活性的药理学调节。这一应用的前提是为精确的 在骨细胞中具有PPARG活性的药理学操作， 老年人代谢功能障碍。