Metabolomic Analysis by 1H-NMR Spectroscopy of Human Tissues reveals different metabolic biomarkers of Hepato-Cellular Carcinoma according to the underlying liver disease

Hepato-Cellular Carcinoma (HCC) accounts for the third cause of cancer mortality worldwide. HCC developed in Non Alco-holic Fatty Liver Disease (NAFLD) occurs in 40% of cases in the absence of cirrhosis and therefore may escape detection enabled by systematic screening of cirrhotic patients. Thus, there is a special need to identify new biomarkers for early diagnosis of HCC arising in patients with non-cirrhotic NAFLD. The aim of this metabolomic study is to discover new biomark-ers by identifying either an abnormal metabolite or a metabolic signature. A non-targeted metabolomics strategy was applied. The study was approved by the ethics committee. The analysis included 24 pairs of Human liver Tumor Tissue (TT) and Distant Uninvolved Tissue (DUT) collected from patients undergoing hepatectomy. Aqueous and lipid tissue extracts were analyzed by 1H-Nuclear Magnetic Resonance (NMR) spectroscopy at 400 MHz. Multivariate Statistical Analysis of spectral data and metabolites quantification were performed. In the non-targeted multivariate analysis of the 24 pairs of tissues, spectral regions having the largest discriminant value for the TT and DUT groups corresponded to lactate and glucose respectively. The quantitative data revealed in the TT an increased level of lactate, amino acids (valine, leucine, glutamine, tyrosine and phenylal-anine), ascorbate and a decreased level of glucose, glycogen and Krebs cycle metabolites such as succinate and fumarate, compared with DUT ilcoxon test, p<0.05). HCC of the 24 patients were associated with either underlying cirrhosis (n=7)(cirrhosis due to viral hepatitis for n=2, alcohol for n=4, NAFLD for n=1) or non-cirrhotic NAFLD (n=17). HCC developed in NAFLD showed a significant decrease of total cholesterol and esterified cholesterol, and a significant increase of glutamine compared to HCC developed in cirrhosis (Mann-Whitney test, p