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.

National Institute of Health (NIH)
National Cancer Institute (NCI)
Research Project (R01)
Project #
Application #
Study Section
Epidemiology of Cancer Study Section (EPIC)
Program Officer
Ogunbiyi, Peter
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
George Washington University
Schools of Medicine
United States
Zip Code
Plieskatt, Jordan L; Rinaldi, Gabriel; Feng, Yanjun et al. (2014) Distinct miRNA signatures associate with subtypes of cholangiocarcinoma from infection with the tumourigenic liver fluke Opisthorchis viverrini. J Hepatol 61:850-8
Plieskatt, Jordan; Rinaldi, Gabriel; Brindley, Paul J et al. (2014) Bioclojure: a functional library for the manipulation of biological sequences. Bioinformatics 30:2537-9
Vale, Nuno; Gouveia, Maria João; Botelho, Mónica et al. (2013) Carcinogenic liver fluke Opisthorchis viverrini oxysterols detected by LC-MS/MS survey of soluble fraction parasite extract. Parasitol Int 62:535-42
Plieskatt, Jordan L; Deenonpoe, Raksawan; Mulvenna, Jason P et al. (2013) Infection with the carcinogenic liver fluke Opisthorchis viverrini modifies intestinal and biliary microbiome. FASEB J 27:4572-84
Gouveia, Maria Joao; Brindley, Paul J; Santos, Lucio Lara et al. (2013) Mass spectrometry techniques in the survey of steroid metabolites as potential disease biomarkers: a review. Metabolism 62:1206-17
Saichua, Prasert; Sithithaworn, Paiboon; Jariwala, Amar R et al. (2013) Microproteinuria during Opisthorchis viverrini infection: a biomarker for advanced renal and hepatobiliary pathologies from chronic opisthorchiasis. PLoS Negl Trop Dis 7:e2228
Mulvenna, Jason; Yonglitthipagon, Ponlapat; Sripa, Banchob et al. (2012) Banking on the future: biobanking for "omics" approaches to biomarker discovery for Opisthorchis-induced cholangiocarcinoma in Thailand. Parasitol Int 61:173-7
Sithithaworn, Paiboon; Andrews, Ross H; Nguyen, Van De et al. (2012) The current status of opisthorchiasis and clonorchiasis in the Mekong Basin. Parasitol Int 61:10-6
Yonglitthipagon, Ponlapat; Pairojkul, Chawalit; Bhudhisawasdi, Vajarabhongsa et al. (2012) Proteomics-based identification of ýý-enolase as a potential prognostic marker in cholangiocarcinoma. Clin Biochem 45:827-34
Nair, Sujit S; Bommana, Anitha; Pakala, Suresh B et al. (2011) Inflammatory response to liver fluke Opisthorchis viverrini in mice depends on host master coregulator MTA1, a marker for parasite-induced cholangiocarcinoma in humans. Hepatology 54:1388-97