Early Diagnostics of Hepatocellular Carcinoma in Patients With Liver Cirrhosis by Novel Molecular Spectrometric Biomarkers in Blood Plasma

Subjects: Patient recruitment will be performed at the Department of Gastroenterology, Hepatology and Metabolism at the Internal Clinic of the First Faculty of Medicine of the Charles University and the Military University Hospital in Prague (MUH Prague). The selection of sample set sizes is clarified below (subsection Data collection and statistical analysis).

Group A: ~100 patients with liver cirrhosis with the HCC diagnosis confirmed according to standard diagnostic criteria published recently by European Association for the study of the liver (EASL). Patients of all HCC stages according to Barcelona Clinic Liver Cancer (BCLC), in our department ~40-50 patients per year, will be included.

Group B: ~100 patients with liver cirrhosis without HCC or dysplastic nodules at the time of examination and within the next 12 months of follow-up. The patients will be monitored by regular clinical visits with general laboratory exams and liver ultrasound in 6-month intervals (recommended surveillance for patients with liver cirrhosis in risk of HCC development).

Group C: ~50 healthy volunteers without liver cirrhosis (Czech Army recruits, Czech Army active military personnel).

Routinely examined biochemistry (e.g. total protein, albumin, alpha feto protein, etc.) of all studied groups will be correlated with the newly identified spectroscopic/metabolomic biomarkers. A signed informed consent will be secured from each subject included in the study (Inclusion/exclusion criteria listed below). Plasma preparation: Following the already established procedure for sample collection, preparation and analysis, the whole anticoagulant-treated (K3EDTA) blood of all subjects will be collected by venipuncture and centrifuged at 1500 × g for 10 minutes. The plasma fractions will be frozen immediately (stored at -80 °C) and transported to the UCT Prague for spectroscopic and metabolomics analyses. Prior to each analysis, the samples will be thawed at room temperature (~25 °C) and centrifuged through a 0.45 μm membrane filter at 13,000 × g for 10 minutes. Subsequently, the spectral measurements will be carried out (in the case of ROA and Raman, a kinetic fluorescence quencher will be added to suppress the spectrally broad background of large amplitude, which overlaps the signals of interest and is caused by a high content of fluorescing molecules in blood plasma).