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The mutational profile of hepatocellular carcinoma (HCC) is still poorly understood, hampering the development of target therapies for these heterogeneous cancers. Few studies found CTNNB1 and TP53 mutations to correlate with HCC prognosis, while even less is known about the role of the promoter TERT gene, despite the high frequency of its mutations. To identify the prognostic role of the mutations in TERT, with or without concomitant CTNNB1 and TP53, in HCC after surgical resection. This is a prospective monocentric study on ex vivo tissue; 65 patients were enrolled so far, undergone to liver resection for histologically-proven HCC (not previously treated). After routine histological analysis, next-generation sequencing (NGS) was carried out with a laboratory-developed multi-gene panel using Gene-Studio S5 sequencer. The incidence of TERT, TP53 and CTNNB1 mutations was 63%, 35% and 26%, respectively; as expected, TP53 and CTNNB1 mutations were mutually exclusive. TP53-mutated HCCs had a significantly higher incidence of poorly differentiated grade (Edmondson's G4, p =0.039 chi-square, OR 3.5) and of macrotrabecular or solid architecture (p =0.032 chi-square, OR 3.4). TERT mutation showed to correlate with post-resection HCC recurrence on both multivariate (Exp(B): 39, Cox regression analysis) and univariate (p =0.041, log rank) analysis: in particular, 27.5% TERT-mutated HCCs experienced tumor recurrence, while no cases of tumor recurrence were recorded among the cases without mutations. The concomitant mutations of both TERT and TP53 showed an even more dramatic recurrence rate than TERT-mutated alone cases (p =0.039, OR 6.4). Mutation of the TERT promoter correlates with a higher risk of recurrence after surgical resection. TP53 mutation characterizes the histologically advanced HCCs, with high grade and macrotrabecular or solid architecture; combined TERT and TP53 mutations worsen the recurrence rate. The possibility of identifying these mutations before surgery opens new scenarios for a personalized therapy for advanced HCC. [ABSTRACT FROM AUTHOR]

The epidemiology of hepatocellular carcinoma (HCC) in industrialized countries is increasingly related to metabolic syndrome and non-alcoholic steatohepatitis (NASH). Despite the approval of molecular targeted drugs, including immune checkpoint inhibitors, the lack of circulating biomarkers still impacts patient responsiveness to treatments, as evidenced by reduced sensitivity to immunotherapy in NASH-associated HCCs. In this context, there is an urgent need to identify new biomarkers to help stratify patients according to personalized therapeutic regimens. The aims of this work are to study metabolic mechanisms underlying miR-494 overexpression in HCC preclinical models and to assess miR-494 as a circulating biomarker associated with metabolic features and sorafenib resistance in HCC patients. We identified miR-494 metabolic targets through bioinformatics analysis. QPCR analysis of glucose 6-phosphatase catalytic subunit (G6pc) was performed in preclinical models and HCC patients. Functional and metabolic analyses assessed G6pc targeting and miR-494 involvement in metabolic change, mitochondrial dysfunction, and ROS production in HCC cells. Live-imaging analysis elucidated the involvement of miR-494/G6pc axis in cell proliferation of HCC cells under stressful conditions. Circulating miR-494 levels were assayed in sorafenib-treated HCC patients and DEN-HCC rats. We demonstrated that miR-494 promote a metabolic shift of HCC cells towards a glycolytic phenotype trough G6pc targeting. The miR-494/G6pc axis exhibits an active role in the metabolic plasticity of tumor cells promoted by glycogen and lipid droplets accumulation to be used under detrimental conditions. High miR-494 serum levels are predictive of sorafenib resistance in a small cohort of HCCs and identify patients with metabolic derangements. AntagomiR-494-based strategies showed a synergic effect with metabolic inhibitors and sorafenib in HCC cells. Our preliminary data suggest miR-494 as a possible target for combined treatments and a promising biomarker for the identification of patients with dysmetabolic HCCs who may be refractory to currently approved treatments. [ABSTRACT FROM AUTHOR]

Objectives Hepatocyte growth factor (HGF), the c-Met receptor, and hypoxia-inducible factor (HIF) are crucial for regenerative processes including ischemic wound healing. The aims of the present study are (a) to analyze the tissue c-Met and HIF-1α expression in skin from patients with critical limb ischemia (CLI); (b) to compare the serum HGF levels of CLI and control subjects. Methods This is a prospective, controlled, single-center study. Thirty-seven patients were enrolled. A skin sample adjacent to the ischemic lesion was taken from 20 patients with CLI; skin samples were taken from the surgical wounds of 17 patients surgically treated for abdominal aortic aneurysm as healthy controls. Serum samples were taken in all cases. Samples were formalin fixed, paraffin embedded, and routinely processed. Tissue inflammation was histologically assessed. Immunohistochemistry was performed with antibodies against total c-Met receptor, activated Met (p-Met), and HIF-1α. RT-polymerase chain reaction was used to quantify HIF-1α mRNA. The enzyme-linked immunosorbent assay was performed to evaluate serum HGF levels. Results With immunohistochemistry, while total c-Met was unchanged, different patterns of p-Met positivity were observed between CLI and control cases ( p < .001). In particular, CLI skin showed a total negativity or membrane positivity for p-Met (19/20 cases), while control skin mainly showed cytoplasmic positivity in the epidermal basal layer (16/17 cases). HIF-1α was diffusely lost in CLI, but HIF-1α mRNA was threefold higher than in controls. Finally, mean serum HGF levels were 590.5 pg/mL and 2380.0 pg/mL in CLI and control groups respectively ( p < .001). Conclusions In CLI patients a significant decrease in serum HGF levels, concomitant with a loss of skin HIF-1α stabilization and a lack of c-Met phosphorylation were seen, probably driving a decrease in wound-healing functions. The next hypothesis is that HGF application might reactivate the c-Met receptor, stabilizing the normal wound healing process. [ABSTRACT FROM AUTHOR]

Abstract: Background: Progression of recurrent hepatitis C is accelerated in liver transplant recipients, leading to special need of non-invasive validated methods to estimate liver fibrosis. Aim: To assess the efficacy of liver stiffness measurement by transient elastography (Fibroscan) and serum parameters in predicting fibrosis stage in HCV-infected transplant recipients. Methods: The correlation between liver fibrosis, assessed at liver histology on bioptic specimens obtained for clinical indications, and stiffness or clinico-serological indexes (Benlloch, APRI, Forns, Fibrotest and Doppler resistance index), was investigated in transplant recipients with recurrence of HCV chronic hepatitis. A total of 56 patients (of which 36 with all clinico-serological indexes), presenting with the following METAVIR fibrosis stage F1=38, F2=9, F3=8, F4=1, were enrolled in the study population. Differences between fibrosis stages were calculated by non-parametric analysis. The best cut-off for identifying significant fibrosis (F2–F4) was assessed by ROC curve analysis. Results: Stiffness (median and range) was 7.7KPa (range 4.2–13.9) in F1 and 17.0KPa (range 6.8–36.3) in ≥F2 (p <0.001). A stiffness cut-off of 10.1KPa revealed 94% Sensitivity, 89% Specificity, 81% PPV and 94% NPV in differentiating F1 from F2–F4. The area under the receiver operator curve in the assessment of fibrosis was significantly higher for Liver stiffness (AUROC 0.943) than for any of the other non-invasive indexes (AUROCs ranging 0.591–0.815). Conclusions: Transient elastography of the liver provides good accuracy in identifying patients with significant fibrosis and performs better than non-invasive indexes based on clinico-serological parameters in transplant recipients. [Copyright &y& Elsevier]