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）方法分析从衰竭人类心脏收集的心脏样品中的蛋白质表达。本研究的总体目的是鉴定具有大规模表达变化的蛋白，然后重点研究经典蛋白激酶C （PKC）亚型及其相关蛋白在衰竭人类心脏中的表达。现有的一组样本将被分析，以比较衰竭和非衰竭心脏中的蛋白质表达。在衰竭的人类心脏中，通过心室辅助装置治疗的机械卸载反应的蛋白质表达和蛋白质区域表达的改变也将被评估。定量蛋白质组学策略将使用液相色谱聚焦、反相高效液相色谱和电喷雾飞行时间串联质谱技术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部分的蛋白微阵列分析进行分析。这些研究的结果有望深入了解人类心力衰竭过程中全局和PKC蛋白表达的变化，并确定心力衰竭动物模型中的关键蛋白和未来的治疗靶点。对PKC信号级联蛋白表达的大规模变化和蛋白表达的分析有望为人类心力衰竭时发生的蛋白表达的一些关键变化提供新的见解。这些实验将为检验心力衰竭动物模型的新假设提供必要的基础信息，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.