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: 10634739
• 负责人: Patrick Robert Griffin
• 金额: $ 66.56万
• 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: 2021-09-30 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634739
• 关键词: Adipocytes; Affect; Age-Related Bone Loss; Aging; Agonist; American; Applications Grants; Bioenergetics; Bone Formation Inhibition; 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; Mitochondria; Molecular; Mus; Nuclear Receptors; Organ; Organ Culture Techniques; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Osteoporosis; PPAR gamma; Pathway interactions; Physiologic calcification; 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; insight; mouse model; novel; osteoblast differentiation; paracrine; pharmacologic; 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.

衰老的能量代谢中的骨质流失和损害会导致骨折和糖尿病。作为 器官，骨骼是骨骼重塑和维护所需的最大能量消费者之一 骨体内平衡。骨重塑的骨细胞控制，占骨细胞的90-95％，并且 哪个生物能计划在很大程度上未知。新证据表明核接收器PPARG， 是能源代谢和药物治疗高血糖的全球调节剂，调节 骨细胞能量代谢及其功能，包括生产Sclerostin，这是Wnt途径的抑制剂 活性和关键蛋白质重塑骨重塑。本应用程序中提出的研究将提供 深入了解骨细胞生物能学之间的联系，其在骨骼代谢和全身性中的功能 能量代谢以及骨骼是否通过骨细胞内分泌活性充当身体“能量”。 主要假设是随着衰老的变化，PPARG活性改变了骨细胞功能和生物能力 导致骨形成的简单减少并减少能量代谢。这个假设将会 以三个特定目标进行测试。 AIM 1将决定燃料选择和衰老如何影响骨细胞功能和 这些过程是否取决于PPARG。使用老化的C57BL/6和γotko小鼠，骨骼器官 培养物和骨细胞样的MLO-Y4细胞，具有下调的PPARG，结合转录组学， 定量蛋白质组学和共培养实验将定义PPARG对骨细胞衰老的贡献。 AIM 2将对比骨细胞的PPARG蛋白相互作用与脂肪细胞的相互作用组对比 为了定义最佳的响应，对全部激动剂的罗斯列酮或反激动剂SR10171的响应 PPARG调节剂对骨骼和代谢有益作用。该分析将在骨细胞上进行 像MLO-Y4细胞一样使用定量串联质量标签蛋白质组学。 AIM 3将定义PPARG的动力学 在SOST启动子中存在的PPRE序列上的互动组作为衰老的函数，并响应于 PPARG活性的药理调节。此应用程序的前提是为精确设置阶段 在骨细胞中使用PPARG活性的药理操作，以同时改善骨骼和 代谢功能障碍较长。