Abstract 2500: Urinary metabolites are diagnostic biomarkers of liver cancer
Authors: Haznadar Majda , Krausz Kristopher W. , Diehl Christopher M. , Bowman Elise D. , Budhu Anuradha S. , Chaisaingmongkol Jittiporn , Rabibhadana Siritida , Forgues Marshonna , Oike Takahiro , Ruchirawat Mathuros , Gonzalez Frank J. , Wang Xin W. , Harris Curtis C. 
Published: July 2017

Abstract
Liver cancer is third leading cause of cancer-related deaths worldwide. A tremendous burden of this disease is evident in that there is an estimated rise in liver cancer incidence from 39,239 in 2016 to 95,374 by 2020. This bleak statistic serves as a motivation to search for early diagnosis biomarkers, especially considering that the 5-year relative survival rate for people with localized disease is ~30.5%, and for regional stage is only 10.7%. Hepatocellular Carcinoma (HCC) is the most common primary malignant tumor of the liver, and is considered to be the third leading cause of all cancer-related deaths and fifth common cancer worldwide. Hepatitis virus B and C infections, fatty and alcoholic liver disease are established risk factors for developing HCC. Cholangiocarcinoma (CCA) is the second most common primary hepatic malignancy. Liver fluke, a chronic parasitic disease of the bile ducts, is a risk factor for developing CCA; otherwise not enough is known about the etiology of this form of liver cancer. We initially conducted a pilot study in the NCI-MD cohort comprising 98 HCC cases, 101 high risk subjects and 95 controls, to evaluate whether previously identified urinary metabolite biomarkers of lung cancer were also predictive of liver cancer, given preliminary evidence that they were universally elevated in several cancer types. Thus, we leveraged ultraperformance liquid chromatography coupled to tandem mass-spectrometry (UPLC-MS/MS) for quantitation of creatine riboside, N-acetylneuraminic acid, cortisol sulfate, and a lipid molecule designated as 561+. This study resulted in findings that all four aforementioned metabolites are significantly increased in HCC cases compared to both, population controls and, more importantly, high risk subjects. We next evaluated whether our previously described findings from the NCI-MD liver case-control study validate in the TIGER-LC cohort (n=370 cases, 471 high risk subjects, 251 controls), where CCA is highly prevalent. We confirmed that indeed the four metabolites were significantly increased in HCC cases compared to both, controls and high risk subjects in TIGER-LC. Most importantly, we also showed that these metabolites are also deregulated in CCA, as their levels are significantly elevated in CCA cases when compared to both, controls and high risk subjects. Furthermore, we showed in the Receiver Operating Characteristic (ROC) analysis a robust ability of the four-metabolite profile to classify both, HCC (AUC=0.75) and CCA (AUC=0.81) compared to high risk subjects. Additionally, the four-metabolite profile performed better in classifying CCA than a clinically utilized CCA tumor marker, CA19-9, and their combination led to a significantly improved classifier (AUC=0.88, P=9.4E-7). Currently, we are investigating a number of pesticides and herbicides used in the areas of Thailand from where TIGER-LC subjects were recruited, to investigate whether these putative carcinogens participate in the etiology of CCA.
Citation Format: Majda Haznadar, Kristopher W. Krausz, Christopher M. Diehl, Elise D. Bowman, Anuradha S. Budhu, Jittiporn Chaisaingmongkol, Siritida Rabibhadana, Marshonna Forgues, Takahiro Oike, Mathuros Ruchirawat, Frank J. Gonzalez, Xin W. Wang, Curtis C. Harris. Urinary metabolites are diagnostic biomarkers of liver cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2500. doi:10.1158/1538-7445.AM2017-2500