Protein Expression in Failing Human Hearts

描述（由申请人提供）：本R21申请中的实验旨在使用强大的多维质谱（MS）方法分析从衰竭的人类心脏收集的心脏样本中的蛋白质表达。本研究的总体目的是鉴定具有大规模表达变化的蛋白质，然后关注经典蛋白激酶C（PKC）亚型的蛋白质表达以及与这些亚型相关的蛋白质在衰竭的人类心脏中的表达。将分析现有的一组样本，以比较衰竭与非衰竭心脏中的蛋白质表达。还将在衰竭的人类心脏中评价响应于心室辅助装置治疗的机械卸载的蛋白质表达和蛋白质的区域表达的改变。定量蛋白质组学策略将用于使用液相色谱聚焦、反相HPLC和电喷雾飞行时间串联质谱法来鉴定具有大规模表达变化的蛋白质。经典PKC亚型和相关蛋白的蛋白表达将通过这些组分的蛋白质微阵列分析进行分析。这些研究的结果有望为人类心力衰竭期间全球和PKC蛋白表达的变化提供深入了解，并确定在心力衰竭动物模型中探索的关键蛋白和未来治疗靶点。公共卫生相关性分析蛋白质表达和PKC信号级联中蛋白质表达的大规模变化，预计将为人类心力衰竭发生的蛋白质表达的一些关键变化提供新的见解。这些实验将提供在心力衰竭动物模型中测试新假设所需的基本信息，并设计未来治疗心力衰竭的治疗靶点。

DESCRIPTION (provided by applicant): Experiments in this R21 application are designed to analyze protein expression in cardiac samples collected from failing human hearts using a powerful multi-dimensional mass spectrometry (MS) approach. The overall aim of this study is to identify proteins with large scale changes in expression, and then focus on protein expression of classical protein kinase C (PKC) isoforms and proteins associated with these isoforms in failing human hearts. An existing set of samples will be analyzed to compare protein expression in failing versus non-failing hearts. Protein expression in response to mechanical unloading by ventricular assist device therapy and alterations in regional expression of proteins also will be evaluated in failing human hearts. A quantitative proteomic strategy will be used to identify proteins with large-scale changes in expression using liquid phase chromatofocusing, reverse phase HPLC and electrospray time-of-flight tandem mass spectrometry. Protein expression of classical PKC isoforms and associated proteins will be analyzed by protein microarray analysis of these fractions. Results from these studies are expected to provide insight into the changes in global and PKC protein expression during human heart failure, and identify key proteins and future therapeutic targets to explore in animal models of heart failure. PUBLIC HEALTH RELEVANCE Analysis of the large scale changes in protein expression and in the expression of protein within the PKC signaling cascade are expected to provide new insights into some critical changes in protein expression that occur with human heart failure. These experiments will provide fundamental information needed to test new hypotheses in animal models of heart failure, and design future therapeutic targets for treating heart failure.