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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：2 型糖尿病影响着全球 3.66 亿人，仅美国就有 2580 万人，尽管我们对这个问题的严重程度有基本了解，但它仍然是 21 世纪的主要公共卫生和经济负担之一。对于 2 型糖尿病患者来说，疾病本身是不完全的，运动可以降低胰岛素抵抗。 R21 应用的目的是探索骨骼肌收缩与胰腺胰岛素分泌的下游调节之间的细胞和分子联系。为了检验这一假设，我们将从新诊断的 2 型糖尿病患者、年龄和性别匹配的非糖尿病患者的股外侧肌中产生原代细胞培养物。控制，并使用电脉冲刺激 (EPS) 测量肌因子分泌组对收缩的反应，我们将利用最先进的蛋白质组学分析，包括 2D 电泳和质谱法来识别非靶向肌因子，并利用免疫分析来识别靶向肌因子。我们将使用计算过滤来识别分泌的肌因子，我们的研究结果的整体生理验证将根据这些肌因子对葡萄糖刺激的集体效应进行评估。分离的人胰岛的胰岛素分泌根据我们对 2 型糖尿病的初步人体研究，运动训练后胰岛素分泌增加，我们预计在 EPS 培养基中培养的胰岛中，葡萄糖刺激的胰岛素分泌和刺激分泌耦合将会增加。预计 EPS 期间分泌的肌因子会增加胰腺胰岛素分泌。我们将使用化学抑制、siRNA 和免疫细胞化学来探索内质网-高尔基体。这些研究将为对临床相关的 2 型糖尿病患者群体中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补充分析提供初步的指导。我们期望这些原理验证研究将导致确定。这些发现也将进一步加深我们对骨骼肌作为内分泌器官及其在调节 2 型糖尿病葡萄糖代谢和胰岛素分泌中的作用的了解。