The role of p300/CBP and acetylation towards skeletal muscle insulin action

p300/CBP 和乙酰化对骨骼肌胰岛素作用的作用

• 批准号: 9975145
• 负责人: Vitor Fernandes Martins
• 金额: $ 5.05万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9975145
• 关键词: Acetylation; Acetyltransferase; Actins; Address; Adipocytes; Alleles; Binding Proteins; Cell membrane; Cell model; Complication; Consensus; Cyclic AMP-Responsive DNA-Binding Protein; Development; EP300 gene; F-Actin; Flow Cytometry; Fluorescence Microscopy; Functional disorder; Genetic Transcription; Glucose Transporter; Health; Human; Impairment; Insulin; Insulin Receptor; Insulin Resistance; Intuition; Knock-out; Knowledge; Lead; Left; Link; LoxP-flanked allele; Lysine; Measures; Mechanics; Mediating; Metabolic; Modeling; Molecular; Morbidity - disease rate; Movement; Mus; Muscle; Myoblasts; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Outcome; Pharmacology; Phosphatidylinositols; Phosphorylation; Phosphotransferases; Protein Acetylation; Proteins; Proteomics; Quality of life; Regulation; Research; Role; Signal Transduction; Signaling Protein; Skeletal Muscle; Validation; Work; aged; basal insulin; base; clinical care; experimental study; glucose transport; glucose uptake; imaging capabilities; improved; in vivo; inhibitor/antagonist; innovation; insulin regulation; insulin sensitivity; insulin signaling; mortality risk; mouse model; novel; novel strategies; novel therapeutics; polymerization; predictive modeling; protein transport; response; therapy development; trafficking

PROJECT SUMMARY Insulin resistance is a common metabolic complication in aged and obese skeletal muscle that is central to the pathophysiology of type 2 diabetes (T2D). In order to develop strategies to treat insulin resistance it is necessary to define the signaling transduction steps that lead to insulin-stimulated glucose uptake, particularly in skeletal muscle and adipocytes; remarkably, these steps have yet to be fully elucidated. Thus, the long-term objective of this research is to expand the knowledge of the molecular regulation of insulin-stimulated glucose transport. Currently, phosphorylation-based signaling from the insulin receptor through phosphoinositide 3- kinase (PI3K)-Akt, to Akt substrate of 160 kDa (AS160) and actin remodeling-related proteins, is considered the major mechanism regulating insulin-stimulated GLUT4 translocation to the plasma membrane and glucose uptake. In contrast to this `classical' model, herein this project proposes a new model in which lysine acetylation (in combination with phosphorylation-based signaling) of key insulin signaling and GLUT4 trafficking proteins is necessary for insulin-stimulated glucose uptake. Accordingly, the primary objective of this proposal is to elucidate the importance of the acetyltransferases (KATs), p300 (E1A binding protein p300) and CBP (cAMP response element-binding protein [CREB] binding protein), to insulin-stimulated glucose transport. The central hypothesis of this proposal is that acetylation, by p300 and/or CBP, of actin remodeling-related proteins are required for insulin-stimulated GLUT4 translocation and, therefore, the insulin-mediated increase in glucose uptake. To address this hypothesis, insulin-induced skeletal muscle glucose uptake, actin remodeling and GLUT4 translocation will be measured in murine and cell models in which p300 and CBP activity are manipulated. This proposal predicts that loss of both p300 and CBP activity will abolish insulin- stimulated actin reorganization, GLUT4 translocation and glucose transport. Specifically, in Aim #1 the necessity and redundancy of p300 and CBP for insulin-stimulated glucose uptake in skeletal muscle will be elucidated, whilst in Aim #2, the importance of p300/CBP to actin remodeling and polymerization, and GLUT4 translocation will be assessed. Overall, these studies will broaden the understanding of the contribution of p300, CBP and acetylation to skeletal muscle glucose uptake in response to insulin. Ultimately, knowledge gathered from this work is expected to manifest novel targets for the development of therapies to improve insulin sensitivity and treat type 2 diabetes, with enhancement of human health and quality of life being anticipated outcomes.

项目总结 胰岛素抵抗是老年和肥胖骨骼肌中常见的代谢并发症，它是 2型糖尿病的病理生理学(T2D)为了开发治疗胰岛素抵抗的策略， 有必要确定导致胰岛素刺激的葡萄糖摄取的信号转导步骤，特别是 在骨骼肌和脂肪细胞中；值得注意的是，这些步骤尚未完全阐明。因此，从长远来看， 本研究的目的是扩大对胰岛素刺激的葡萄糖的分子调控的认识。 运输。目前，来自胰岛素受体的基于磷酸化的信号通过磷脂酰肌醇3- 激酶(PI3K)-Akt，到Akt底物160 kDa(AS160)和肌动蛋白重塑相关的蛋白质 调节胰岛素刺激的GLUT4转位到质膜和葡萄糖的主要机制 领悟。与这个经典的模型不同，这个项目在这里提出了一个新的模型，在这个模型中赖氨酸 关键胰岛素信号和GLUT4的乙酰化(与基于磷酸化的信号相结合) 转运蛋白是胰岛素刺激的葡萄糖摄取所必需的。因此，这一行动的主要目标是 建议阐明乙酰转移酶(KATS)、p300(E1a结合蛋白p300)和 CBP(cAMP反应元件结合蛋白[CREB]结合蛋白)，以促进胰岛素刺激的葡萄糖转运。 这一提议的中心假设是p300和/或CBP对肌动蛋白重塑相关的乙酰化。 蛋白质是胰岛素刺激的GLUT4移位所必需的，因此，胰岛素介导的增加 在葡萄糖摄取方面。为了解决这一假设，胰岛素诱导的骨骼肌葡萄糖摄取，肌动蛋白 将在p300和CBP的小鼠和细胞模型中测量重塑和GLUT4移位 活动是被操纵的。这项提议预测，p300和CBP活性的丧失将使胰岛素- 刺激肌动蛋白重组、GLUT4易位和葡萄糖转运。具体地说，在目标1中 P300和CBP对骨骼肌胰岛素刺激的葡萄糖摄取的必要性和冗余性将是 在目标2中，阐明了p300/CBP对肌动蛋白重塑和聚合的重要性，以及GLUT4 移位将被评估。总体而言，这些研究将扩大对 P300、CBP和乙酰化对骨骼肌葡萄糖摄取的响应。归根结底，知识 从这项工作中收集的信息有望显示出新的靶点，用于开发改进的治疗方法 胰岛素敏感性和治疗2型糖尿病，提高人类健康和生活质量 预期的结果。