HCV INFECTION AND HCV/HIV CO-INFECTION IN LIVER TISSUE

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。所列机构为 中心，不一定是研究者的机构。 该项目将利用最先进的蛋白质组学技术来推进对HCV感染过程和肝病进展的研究，包括肝硬化和肝细胞癌的发展。 我们先前已经完成了几个人肝细胞群体的全球蛋白质组分析，确定了干扰素（IFN）处理的肝细胞系中差异表达的蛋白质，并表征了含有全长HCV复制子的高度允许的Huh-7.5细胞系中蛋白质丰度的变化。 最值得注意的是，我们已经成功地证明了R中心独有的超灵敏纳米蛋白质组学功能的应用，以分析可以从人类肝脏活检标本中获得的有限数量的蛋白质。 这代表了临床蛋白质组学的重大进步，现在可以直接从感染组织中研究HCV相关蛋白质水平变化的临床相关性。 本文概述的具体目标描述了一种策略，该策略涉及利用蛋白质组学仪器和方法的持续进步，对大量临床样本进行超灵敏、高通量分析，以获得具有高统计置信度的生物学意义的结论。 这些研究产生的数据将提供从HCV感染和HCV/HIV合并感染到终末期肝病进展期间发生的蛋白质丰度变化的详细分子画像，并可能产生改进的诊断方法，疾病进展的标志物和治疗干预的新方法。 具体目标是： 1)获得人类肝活检标本的超灵敏、高通量定量蛋白质谱研究 2)利用技术和方法的进步来表征NIDA中心可用的新模型系统和临床样本类型

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This project will utilize cutting-edge proteomic technologies to advance studies on the process of HCV infection and progression of liver disease, including the development of cirrhosis and hepatocellular carcinoma. We have previously completed global proteome analyses of several human liver cell populations, identified differentially expressed proteins in interferon (IFN)-treated hepatocyte cell lines, and characterized protein abundance changes in the highly permissive Huh-7.5 cell line containing a full length HCV replicon. Most notably, we have successfully demonstrated the application of an ultra-sensitive nanoproteomics capabilities uniquely available at the R Center to analyze the limited amounts of protein that can be obtained from human liver biopsy specimens. This represents a significant advancement in clinical proteomics now making it possible to investigate the clinical relevance of HCV-associated changes in protein level directly from infected tissue. The specific aims outlined here describe a strategy that involves utilizing continued advances in proteomics instrumentation and methodologies to perform ultra-sensitive, high-throughput analysis of the large numbers of clinical samples necessary to make biologically meaningful conclusions with high statistical confidence. The data generated from these studies will provide a detailed molecular portrait of the protein abundance changes that occur during the progression from HCV infection and HCV/HIV co-infection to end-stage liver disease and are likely to yield improved diagnostic methods, markers of disease progression, and novel approaches to therapeutic intervention. The Specific Aims are to: 1) Obtaining ultra-sensitive, high-throughput quantitative protein profiling studies of human liver biopsy specimens 2) Utilize advances in technologies and methodologies to characterize new model systems and types of clinical samples available to the NIDA Center