Biomarkers of Opisthorchis viverrini-induced cholangiocarcinoma

描述（由申请人提供）：胆管癌(CCA) -胆管癌-与晚期相关，对诊断构成挑战，死亡率高，这些特征突出了对生物标志物的需求，而这些生物标志物不能在早期和可获得的样本（如血浆）中测量。然而，尽管进行了广泛的研究，努力未能产生具有足够诊断准确性和CCA实用性的生物标志物。我们将通过在全球CCA中心泰国Khon Kaen省开展生物标志物项目来解决以前发现CCA生物标志物的局限性，泰国Khon Kaen省是世界上肝内CCA发病率最高的地区。这一建议成功的一个关键因素是，虽然在西方CCA的致病因子仍然不清楚，但泰国肝内CCA的一个最重要的危险因素早已确定-感染肝吸虫（OV）。根据世界卫生组织国际癌症研究机构的确定，人类恶性肿瘤与真核病原体之间没有比CCA与OV感染之间更强的联系。我们将利用这种寄生虫感染与癌症之间的良好联系来发现和验证血浆中CCA的生物标志物。使用定量蛋白质组学方法，我们建议扫描30个冷冻切除的肝肿瘤组织，这些组织来自确诊的ov诱导的CCA病例，以在疾病部位近端组装一组蛋白质（候选生物标志物）。我们将用CCA细胞系的扫描来补充这一分析，与冷冻肝脏切片不同，CCA细胞系可以测量分泌/膜蛋白（分泌组）的表达。从这两种来源中鉴定出的潜在生物标志物将在与30个冷冻切除的肝组织配对的血浆样本中进行验证，这些组织来自确诊的ov诱导的CCA患者。候选生物标志物将在niaid赞助的纵向研究中在有CCA风险的OV感染个体的血浆中进行验证，该研究追踪从慢性O. viverrini感染到CCA的胆管癌发生。使用这组特殊的样本将使我们能够解决以前CCA生物标志物发现的限制，如下所示。首先，我们可以通过确定OV感染来可靠地测量暴露风险。其次，孔敬研究地点是世界上CCA发病率最高的地方，这使我们能够获得大量的CCA样本。第三，使用niaid赞助的纵向研究，我们可以沿着整个连续体追踪发病机制：从感染OV到诊断为CCA。第四，我们计划在肿瘤（切除的肝脏）附近的组织中发现生物标志物，然后在这些相同的CCA患者的血浆中进行验证，然后在我们的队列研究中对“有风险”的患者进行验证。因此，该提案的主要创新之处在于，我们将利用一个人类致癌的模型，在这个模型中，主要的风险因素，以及通往CCA的途径上的许多中间阶段，已经被明确定义。

DESCRIPTION (provided by applicant): Cholangiocarcinoma (CCA) - bile duct cancer - is associated with late presentation, poses challenges for diagnosis and has high mortality, features that highlight the need for biomarkers than can be measured early and in accessible samples such as plasma. However, despite extensive investigations, efforts have failed to yield biomarkers with adequate diagnostic accuracy and utility for CCA. We will address previous limitations in the discovery of CCA biomarker(s) by undertaking a biomarker program in the global epicenter of CCA, Khon Kaen province, Thailand, which has highest incidence of intrahepatic CCA in the world. A key factor in the success of this proposal is that, while the causative agent for CCA in the West remains obscure, the single most important risk factor for intrahepatic CCA in Thailand has long been established - infection with the liver fluke Opisthorchis viverrini (OV). As determined by the WHO's IARC, no stronger link between a human malignancy and a eukaryotic pathogen exists than between CCA and infection with OV. We will utilize this well-established link between a parasite infection and cancer for the discovery and verification of biomarkers for CCA in plasma. Using a quantitative proteomic approach, we propose to scan 30 frozen, resected liver tumor tissues from confirmed OV-induced CCA cases to assemble a suite of proteins (candidate biomarkers) proximal to the disease site. We will complement this analysis with a scan of CCA cell lines which, unlike the frozen liver sections, measure the expression of secreted/membrane proteins (the secretome). Potential biomarkers identified from these two sources will be verified in plasma samples paired with the 30 frozen, resected liver tissues from confirmed OV-induced CCA patients. The candidate biomarkers will then be verified in the plasma of OV- infected individuals at risk for CCA in a case control study from our NIAID-sponsored longitudinal study that traces cholangiocarcinogenesis from chronic O. viverrini infection to CCA. The use of this exceptional set of samples will enable us to address previous limitations to CCA biomarker discovery as follows. First, we can reliably measure risk for exposure by determining infection with OV. Second, the Khon Kaen study site has the highest incidence of CCA in the world, giving us access to large numbers of CCA samples. Third, using our NIAID-sponsored longitudinal study, we can trace pathogenesis along the entire continuum: from infection with OV to diagnosis with CCA. Fourth, we plan biomarker discovery in banked tissue proximal to the tumor (resected liver) followed by verification in plasma from these same CCA patients and then in "at risk" patients in our cohort study. Hence, the overarching innovation of this proposal is that we will utilize a model of human carcinogenesis in which the major risk factor, as well as many of the intermediate stages on the pathway, to CCA have been well-defined. PUBLIC HEALTH RELEVANCE: Cholangiocarcinoma (CCA) is a form of liver cancer with a devastatingly poor prognosis. In East Asia, unlike in the West, long term infection with a parasitic worm leads to CCA. Because of access to numerous cases of CCA in Thailand and nearby countries, and because it is feasible to monitor the development of CCA from the time of infection with the parasite, we propose a biomarker discovery program using CCA samples from liver fluke infected persons in Thailand. This could eventuate in tools for early diagnosis of CCA, which are not available at present, and thereby allow for treatment, not only in Asian populations but for people at risk of CCA in the US and other western countries.