Role of the skeletal muscle/pancreatic axis in type 2 diabetes

骨骼肌/胰轴在 2 型糖尿病中的作用

• 批准号: 9014518
• 负责人: JOHN P. KIRWAN
• 金额: $ 17.32万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-02-13 至 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9014518
• 关键词: Affect; Age; Bathing; Beta Cell; Biochemical; Biopsy Specimen; Brefeldin A; Cell Culture Techniques; Cell physiology; Cells; Chemicals; Conditioned Culture Media; Contracts; Coupling; Data; Disease; Economic Burden; Electrophoresis; Endocrine Glands; Endoplasmic Reticulum; Exercise; Gender; Glucose; Goals; Golgi Apparatus; Health; Human; Immunoassay; Incubated; Individual; Insulin Resistance; Interleukin-6; Islets of Langerhans; Lead; Link; Liquid substance; Mass Spectrum Analysis; Measures; Mediating; Metabolic; Metabolic Pathway; Metabolism; Molecular; Muscle; Muscle Cells; Muscle Contraction; Muscle Fibers; Newly Diagnosed; Non-Insulin-Dependent Diabetes Mellitus; Outcome; Pancreas; Pathway interactions; Patients; Peptides; Phenotype; Physiologic pulse; Physiological; Play; Population; Primary Cell Cultures; Protein Isoforms; Proteins; Proteomics; Public Health; Regulation; Reporting; Rest; Role; Sampling; Skeletal Muscle; Small Interfering RNA; Stimulus; System; Testing; Validation; Vesicle; Work; base; blood glucose regulation; clinically relevant; diabetes management; diabetic; drug development; drug discovery; exercise training; genetic regulatory protein; glucose metabolism; health economics; immunocytochemistry; improved; in vitro Model; inhibitor/antagonist; insulin secretion; islet; non-diabetic; novel; pancreatic islet function; response; skeletal; vastus lateralis

DESCRIPTION (provided by applicant): Type 2 diabetes affects 366 million people globally, and 25.8 million in the US alone. It is one of the major public health and economic burdens of the 21st century. Despite the scale of the problem our fundamental understanding of the disease itself is incomplete. In patients with type 2 diabetes, exercise reduces insulin resistance in muscle and increases glucose sensitivity in the pancreas. The objective of this R21 application is to explore the cellular and molecular links between skeletal muscle contraction and downstream regulation of pancreatic insulin secretion. Our central hypothesis is that contracting skeletal muscle secretes myokines that target β-cells and participate in the regulation of glucose stimulated insulin secretion. To test this hypothesis we will generate primary cell cultures from vastus lateralis muscle of newly diagnosed patients with type 2 diabetes and non-diabetic age and gender matched controls, and measure the myokine secretome response to contraction using electrical pulse stimulation (EPS). We will utilize state-of-the-art proteomic analyses, including 2D electrophoresis and mass spectrometry to identify non-targeted myokines, and immunoassays to identify a targeted myokine signature in the media from muscle cells that undergo contraction. We will use computational filtering to identify the secreted myokines. Global physiological validation of our findings will be assessed from the collective effect of these myokines on glucose stimulated insulin secretion from isolated human pancreatic islets. Based on our preliminary human studies in type 2 diabetes, insulin secretion was increased after exercise training. We expect that glucose stimulated insulin secretion and stimulus-secretory- coupling will be increased in islets incubated in media from EPS-conditioned cells. It is expected that myokines secreted during EPS mediate increased pancreatic insulin secretion. We will use chemical inhibition, siRNA and immunocytochemistry to explore the endoplasmic reticulum-Golgi network as a likely secretory pathway for contraction-induced myokine secretion. These studies will provide the initial steps towards a comprehensive and complementary analysis of a biochemical link between skeletal muscle contraction and downstream pancreatic insulin secretion in a clinically relevant population of individuals with type 2 diabetes. We expect that these proof-of-principle studies will lead to the identification of specific myokines that can be used for drug development and diabetes management. The discoveries will also further our understanding of skeletal muscle as an endocrine organ and its role in the regulation of glucose metabolism and insulin secretion in type 2 diabetes.

描述(申请人提供)：2型糖尿病影响全球3.66亿人，仅美国就有2580万人。它是21世纪主要的公共卫生和经济负担之一。尽管问题的规模很大，但我们对疾病本身的基本了解是不完整的。在2型糖尿病患者中，运动可降低胰岛素抵抗 并增加胰腺对葡萄糖的敏感度。此R21应用的目的是探索骨骼肌收缩和胰腺胰岛素分泌下游调节之间的细胞和分子联系。我们的中心假设是，骨骼肌收缩会分泌以β细胞为靶点的肌细胞因子，并参与葡萄糖刺激的胰岛素分泌的调节。为了验证这一假设，我们将从新诊断的2型糖尿病患者和非糖尿病年龄和性别匹配的对照组的股外侧肌进行原代细胞培养，并使用电脉冲刺激(EPS)测量肌肉因子分泌体对收缩的反应。我们将利用最先进的蛋白质组学分析，包括2D电泳法和质谱仪来识别非靶向肌动蛋白，并利用免疫分析来识别介质中发生收缩的肌肉细胞中的靶向肌动蛋白信号。我们将使用计算过滤来识别分泌的肌动蛋白。我们发现的全球生理学验证将从这些肌动蛋白对分离的人胰岛葡萄糖刺激的胰岛素分泌的集体效应来评估。根据我们对2型糖尿病的初步人类研究，运动训练后胰岛素分泌增加。我们预计，在EPS条件细胞的培养液中孵育的胰岛将增加葡萄糖刺激的胰岛素分泌和刺激-分泌-偶联。预计在EPS过程中分泌的肌动因子介导了胰腺胰岛素分泌的增加。我们将使用化学抑制、siRNA和免疫细胞化学来探索内质网-高尔基网络作为收缩诱导的肌细胞因子分泌的可能的分泌途径。这些研究将为在临床相关的2型糖尿病患者群体中对骨骼肌收缩和下游胰腺胰岛素分泌之间的生化联系进行全面和互补的分析提供初步步骤。我们预计，这些原则证明研究将导致确定 可用于药物开发和糖尿病治疗的特定肌动蛋白。这些发现还将进一步加深我们对骨骼肌作为内分泌器官的理解，以及它在2型糖尿病患者葡萄糖代谢和胰岛素分泌调节中的作用。