A novel approach integrating proteomics and metabolomics to understand diabetic cardiomyopathy

整合蛋白质组学和代谢组学来了解糖尿病心肌病的新方法

基本信息

批准号：
10617277
负责人：
Benjamin Wancewicz
金额：
$ 3.7万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10617277
关键词：
Acetylation Adrenergic Agents Animals Attention Back Branched-Chain Amino Acids Cardiac Cardiac Myocytes Ceramides Complex Contractile Proteins Cyclic AMP-Dependent Protein Kinases Data Detection Development Diabetic mouse Echocardiography Energy-Generating Resources Enzymes Epidemic Functional disorder Future Head Heart Hexoses High Fat Diet Hyperinsulinism Hypertrophy Impairment Insulin Knowledge Length Link Lipids Mass Spectrum Analysis Measurement Measures Metabolic Metabolic Control Methods Molecular Morbidity - disease rate Mus Muscle Cells Myocardial Non-Insulin-Dependent Diabetes Mellitus Obesity Pathway interactions Patients Phosphorylation Phosphorylcholine Polyunsaturated Fatty Acids Post-Translational Protein Processing Prevalence Proteins Proteomics Protocols documentation Regulation Relaxation Research Risk Risk Factors Role Sampling Sarcomeres Serial Extraction Signal Transduction Testing Therapeutic Tissue Sample Tissues Triglycerides Troponin Troponin I United States acylcarnitine adverse outcome comparison control diabetic diabetic cardiomyopathy fatty acid-binding proteins feeding genetic regulatory protein glycemic control insight lipidomics liquid chromatography mass spectrometry metabolomics mortality mouse model new therapeutic target novel novel strategies response success surfactant two-dimensional ultra high resolution

项目摘要

Project Summary/Abstract Diabetic cardiomyopathy (DCM) is a major cause of morbidity and mortality in the United States and is a rising global epidemic in parallel with the worldwide prevalence of obesity and type-2 diabetes mellitus (T2DM). However, the underlying molecular mechanisms in DCM remain poorly understood. It is believed that hyperinsulinemia might promote adverse consequences in the DCM hearts with T2DM and obesity. Recently, our collaborator, Dr. Kevin Xiang’s lab has discovered a novel mechanism that hyperinsulinemia might impair myocardial contractility by inhibiting adrenergic signaling and identified a direct interaction between insulin and adrenergic pathways of the heart in a T2DM C57J/B6 mouse model generated by feeding a high fat diet (HFD). Importantly, they have shown that the PKA phosphorylation of cardiac troponin I (cTnI, a critical sarcomeric regulatory protein) in response to adrenergic stimulation was impaired in HFD animals. Although post- translational modifications (PTM) of the sarcomeric proteins are known to be an important mechanism in the regulation of cardiac contraction and relaxation, its specific role in DCM hearts remains largely uncharacterized. Given that diastolic dysfunction and cardiomyocyte hypertrophy were ameliorated by a greater glycemic control in diabetic mice, I hypothesize that metabolic proteins will undergo various PTMs, like acetylation and phosphorylation, leading to a less efficient metabolic state and a reduced energy state, which will ultimately culminate in multiple altered sarcomeric PTMs and reduced contractility. Herein, I propose to utilize a novel ultra- high resolution mass spectrometry (MS)-based top-down proteomics platform to comprehensively characterize the sarcomeric proteins PTMs in DCM hearts. Moreover, I propose to investigate the interplay between sarcomeric PTMs and the metabolic state in DCM. The specific aims of this proposal include: 1) Identify PTM changes of key regulatory sarcomeric proteins in response to T2DM-associated hyperinsulinemia in heart tissue using a top-down proteomics strategy and link the sarcomeric changes with alterations in cardiac contractility found by echocardiography measurements of HFD and normal chow (control) mice. 2) Identify changes in metabolic energy stores in parallel with lipid energy stores with attention to specific acyl chain information like acyl length, degrees of unsaturation, and lipid head group by utilizing a comprehensive metabolomics and lipidomics extraction protocol in DCM samples. 3) Identify PTM changes of metabolic proteins using a two- dimensional liquid chromatography mass spectrometry-based top-down proteomics platform and link the measured changes to the contractile dysfunction. The success of the proposed research will offer new insights into the molecular mechanism underlying DCM and may identify new therapeutic targets.

项目概要/摘要糖尿病心肌病 (DCM) 是美国发病率和死亡率的主要原因，并且呈上升趋势全球流行病与肥胖和 2 型糖尿病 (T2DM) 的全球流行同时发生。然而，人们相信 DCM 的潜在分子机制仍然知之甚少。最近，高胰岛素血症可能会导致 DCM 心脏出现 T2DM 和肥胖的不良后果。我们的合作者向凯文博士的实验室发现了一种新的机制，高胰岛素血症可能会损害通过抑制肾上腺素信号传导来调节心肌收缩力，并确定胰岛素和胰岛素之间的直接相互作用通过喂养高脂肪饮食 (HFD) 产生的 T2DM C57J/B6 小鼠模型中心脏的肾上腺素能通路。重要的是，他们已经证明心肌肌钙蛋白 I (cTnI，一种关键的肌细胞) 的 PKA 磷酸化 HFD 动物对肾上腺素能刺激的反应受到损害。众所周知，肌节蛋白的翻译修饰（PTM）是一个重要的机制。尽管其对心脏收缩和舒张的调节，但其在 DCM 心脏中的具体作用在很大程度上仍然未知。鉴于舒张功能障碍和心肌细胞肥大可以通过更好的血糖控制得到改善在糖尿病小鼠中，我挣扎着代谢蛋白质会经历各种 PTM，如乙酰化和磷酸化，导致代谢状态效率降低和能量状态降低，最终将最终导致多个肌节 PTM 改变和收缩性降低。在此，我建议利用一种新型的超- 基于高分辨率质谱 (MS) 的自上而下蛋白质组学平台，可全面表征此外，我建议研究 DCM 心脏中的肌节蛋白 PTM 之间的相互作用。肌节 PTM 和 DCM 中的代谢状态该提案的具体目标包括： 1) 识别 PTM。心脏组织中关键调节肌节蛋白响应 T2DM 相关高胰岛素血症的变化使用自上而下的蛋白质组学策略并将肌节变化与心肌收缩力的变化联系起来通过对 HFD 和正常饲料（对照）小鼠进行超声心动图测量发现 2) 识别变化。代谢能量存储与脂质能量存储并行，并注意特定的酰基链信息，例如酰基长度、不饱和度和脂质头基，通过利用综合代谢组学和 DCM 样品中的脂质组学提取方案 3) 使用两种方法识别代谢蛋白的 PTM 变化。基于多维液相色谱质谱的自上而下的蛋白质组学平台，并将所提出的研究的成功将提供新的见解。深入了解 DCM 的分子机制，并可能确定新的治疗靶点。