Biomarkers of Opisthorchis viverrini-induced cholangiocarcinoma

描述（由申请人提供）：胆管癌（CCA）-胆管癌-与晚期表现相关，对诊断提出了挑战，死亡率高，这些特征突出了对生物标志物的需求，而这些生物标志物可以在早期和可获得的样本（如血浆）中进行测量。然而，尽管进行了广泛的研究，但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。我们将通过在全球肝内CCA发病率最高的泰国孔敬省CCA的全球中心开展生物标志物项目，解决以前发现CCA生物标志物的局限性。这一提议成功的一个关键因素是，虽然西方CCA的病原体仍然不清楚，但泰国肝内CCA的单一最重要风险因素早已确定-肝吸虫后睾吸虫（OV）感染。正如WHO的IARC所确定的，人类恶性肿瘤和真核病原体之间的联系没有比CCA和OV感染之间的联系更强。我们将利用寄生虫感染和癌症之间的这种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个冷冻的，切除的肝肿瘤组织，从确认的OV诱导的CCA情况下，组装一套蛋白质（候选生物标志物）接近疾病部位。我们将通过扫描CCA细胞系来补充这一分析，与冷冻肝脏切片不同，CCA细胞系测量分泌/膜蛋白（分泌蛋白）的表达。从这两个来源中鉴定的潜在生物标志物将在与来自确认的OV诱导的CCA患者的30个冷冻、切除的肝组织配对的血浆样本中进行验证。然后，在来自我们NIAID赞助的追踪慢性O型肝炎的胆管癌发生的纵向研究的病例对照研究中，在有CCA风险的OV感染个体的血浆中验证候选生物标志物。灵猫感染至CCA。使用这组特殊的样品将使我们能够解决以前对CCA生物标志物发现的限制，如下所述。首先，我们可以通过确定OV感染来可靠地测量暴露风险。第二，孔敬研究地点的CCA发病率是世界上最高的，这使我们能够获得大量的CCA样本。第三，使用我们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OV感染到CCA诊断。第四，我们计划在肿瘤近端的库存组织（切除的肝脏）中发现生物标志物，然后在来自这些相同CCA患者的血浆中进行验证，然后在我们的队列研究中的“风险”患者中进行验证。因此，该提案的总体创新在于，我们将利用人类致癌作用模型，其中已明确定义了CCA的主要风险因素以及途径上的许多中间阶段。

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.