Urinary Metabolites Diagnostic and Prognostic of Intrahepatic Cholangiocarcinoma

Abstract

Background: Liver cancer is the second leading cause of cancer-related deaths worldwide. With a predicted 2.4-fold rise in liver cancer incidence by 2020, there is an urgent need for early, inexpensive diagnostic biomarkers to deploy in the clinic. Methods: We employed ultraperformance liquid chromatography tandem mass-spectrometry (UPLC/MS-MS) for the quantitation of four metabolites, creatine riboside (CR), N-acetylneuraminic acid (NANA), cortisol sulfate, and a lipid molecule designated as 561þ, in urine samples from the NCI-MD cohort comprising 98 hepatocellular carcinoma (HCC) cases, 101 high-risk subjects, and 95 controls. Validation was carried out in the TIGER-LC cohort [n ¼ 370 HCC and intrahepatic cholangiocarcinoma (ICC) cases, 471 high-risk subjects, 251 controls], where ICC, the second most common primary hepatic malignancy, is highly prevalent. Metabolite quantitation was also conducted in TIGER-LC tissue samples (n ¼ 48 ICC; n ¼ 51 HCC). Results: All profiled metabolites were significantly increased in liver cancer when compared with high-risk subjects and controls in the NCI-MD study. In the TIGER-LC cohort, the four-metabolite profile was superior at classifying ICC than a clinically utilized marker, CA19-9, and their combination led to a significantly improved model (AUC ¼ 0.88, P ¼ 4E-8). Metabolites CR and NANA were significantly elevated in ICC when compared with HCC cases in both urine and tissue samples. High levels of CR were associated with poorer prognosis in ICC. Conclusions: Four metabolites are significantly increased in HCC and ICC and are robust at classifying ICC in combination with the clinically utilized marker CA19-9. Impact: Noninvasive urinary metabolite biomarkers hold promise for diagnostic and prognostic evaluation of ICC.

Creatine riboside is a cancer cell–derived metabolite associated with arginine auxotrophy

“…Our analysis identifies CR as a tumor-derived metabolite detectable in serum and plasma biospecimens, building on previous studies of CR as a urinary biomarker of risk and prognosis in lung, liver, and adrenocortical carcinoma ( 21 – 24 ). The strong positive correlation of urinary CR levels with tumoral CR levels enabled us to dissect the metabolic profile of CR hi tumors directly at the tissue level.…”

“…While the ability of cells to produce CR is not confined to cancer cells, the reprogramming of these metabolic pathways in cancer cells enables them to produce higher levels of this metabolite than nontransformed cells. By integrating the pentose phosphate and urea cycle pathways, CR levels amplify the disparity between tumoral and healthy metabolism, thereby underpinning its potential utility as a sensitive diagnostic and prognostic biomarker ( 21 , 22 , 24 ).…”

“…Lung tumor and nontumor biospecimens from the NCI-MD cohort were collected according to procedures approved by the NCI, NIH, as described previously ( 21 ) (IRB OH98-C-N027). Liver cancer biospecimens and data were obtained from the TIGER-LC cohort as described previously ( 24 , 67 ). Written informed consent from patients was received prior to participation in these studies.…”

“…Recently, urinary CR has also been identified as a biomarker of risk and prognosis in adrenocortical ( 23 ) and liver ( 24 ) cancer. The metabolic reprogramming of these different tumor types remains diverse ( 4 , 17 ), yet current data suggest that CR may act as a surrogate marker for a common metabolic program associated with tumorigenesis and tumor progression.…”

Pre- and Post-Resection Urine Metabolic Profiles of Bladder Cancer Patients: Results of Preliminary Studies on Time Series Metabolomics Analysis

“…It is worth noting the fact that few of the determined metabolites (N-acetylneuraminic acid, creatine riboside or indoxyl sulfate) were previously reported as possible markers of intrahepatic cholangiocarcinoma, lung cancer or adrenal neoplasms [ 42 , 43 , 44 ]. In addition, the elevated level of glutarylcarnitine in cancer is consistent with the observations regarding the increased level of pentanedioic acid [ 13 ].…”