Markers for HCV-related HCC: plasma profiling, targeted strategies and validation

描述（由申请方提供）：全球范围内报告了肝细胞癌（HCC）发生率增加的趋势，这与人群中丙型肝炎病毒（HCV）感染的高患病率有关。迫切需要更好的HCC诊断方法。我们先前的研究提供了证据，证明肝细胞癌中潜在肝脏疾病的病因与基因/蛋白表达模式的差异之间存在关联。他们还提供了新的见解蛋白质亚型，在HCC的发展中可能是重要的。我们最近开发了一种方法来全面和定量分析复杂样品的蛋白质组，我们已经将这种方法应用于肝脏和血浆。该方法基于完整蛋白质的广泛分级分离，导致在肝脏中鉴定出约9，000种蛋白质，以高置信度鉴定，包括低丰度蛋白质，如细胞因子，趋化因子和受体。基于光谱计数的丰度得分归因于每种鉴定的蛋白质。计算的丰度分数似乎是蛋白质丰度的良好估计。在血浆中，鉴定了生物标志物浓度范围（pg/ml）内的许多低丰度蛋白，并且在肝组织以及血浆中鉴定了对疾病阶段（纤维化和早期HCC）特异的蛋白。有趣的是，对于随疾病阶段变化的选择性蛋白质，在组织中观察到的丰度变化与在血浆中观察到的丰度变化之间没有相关性。总之，该方法在蛋白质组学分析方面达到了以前没有报道过的深度，用于复杂的生物混合物，如哺乳动物组织和血浆，允许鉴定与疾病相关的蛋白质变化，并表明基于组织的发现和基于血浆的发现研究可能会导致不同的结果。本提案的目的是利用这种方法鉴定HCV相关HCC的蛋白质生物标志物，可用于早期检测和诊断。我们将应用这种方法来鉴定蛋白质及其亚型，这些蛋白质及其亚型在最近进展为HCC的HCV相关肝硬化患者和无HCC的HCV相关肝硬化患者的血浆中的表达水平不同。在本研究的发现部分中确定的最有希望的候选者将被作为验证的目标。我们将建立这些蛋白质生物标志物的敏感性和特异性，单独和组合，用于早期检测HCC。公共卫生相关性：肝细胞癌（HCC）的发病率在全球范围内呈上升趋势，这与人群中丙型肝炎病毒（HCV）感染的高患病率有关。迫切需要更好的HCC诊断方法。该提案的目的是开发一套强大的HCC生物标志物，可用于早期检测和诊断。

DESCRIPTION (provided by applicant): A trend of increasing rates of hepatocellular carcinoma (HCC) has been reported worldwide, that is related to the high prevalence of hepatitis C virus (HCV) infection in the population. Better means for HCC diagnosis are urgently needed. Our prior studies provided evidence for an association between the etiology of the underlying liver disease and differences in patterns of gene/protein expression in HCC. They also offered new insights into protein isoforms that are potentially important in the development of HCC. We recently developed a method to comprehensively and quantitatively profile the proteome of a complex sample and we have applied this method to the liver and to plasma. This method, based on extensive fractionation of intact proteins, resulted in the identification of approximately 9,000 proteins in the liver, identified with high confidence and including low-abundance proteins such as cytokines, chemokines and receptors. An abundance score based on spectral counts was attributed to each identified protein. The calculated abundance scores appeared to be a good estimate of protein abundance. In the plasma, numerous low-abundance proteins in the biomarker concentration range (pg/ml) were identified and proteins specific to disease stage (fibrosis and early HCC) were identified in liver tissue as well as in plasma. Interestingly, there was no correlation between the abundance changes observed in the tissues and the abundance changes observed in the plasma for selective proteins changing with disease stage. In conclusion, this method reached a depth in proteomic profiling not previously reported for complex biological mixtures such as mammalian tissue and plasma, allowed for the identification of protein changes associated with disease and suggested that tissue-based discovery and plasma-based discovery studies may lead to different results. The purpose of this proposal is to utilize this method for the identification of protein biomarkers for HCV-related HCC that could be used for early detection and diagnosis. We will apply this approach to identify proteins and their isoforms that differ in expression levels between plasma obtained from patients with HCV-related cirrhosis that have recently progressed to HCC and patients with HCV-related cirrhosis with no HCC. The most promising candidates identified in the discovery component of this research will be targeted for validation. We will establish the sensitivity and specificity of these protein biomarkers individually and in combination, for detecting HCC early. PUBLIC HEALTH RELEVANCE: A trend of increasing rates of hepatocellular carcinoma (HCC) has been reported worldwide, that is related to the high prevalence of hepatitis C virus (HCV) infection in the population. Better means for HCC diagnosis are urgently needed. The purpose of this proposal is the development of a robust set of biomarkers for HCC that could be used for early detection and diagnosis.