High efficacy of Sofosbuvir plus Simeprevir in a large cohort of Spanish cirrhotic patients infected with genotypes 1 and 4

[Abstract] Background and Aims. Hepatitis C (HCV) therapy with Sofosbuvir (SOF)/Simeprevir (SMV) in clinical trials and real‐world clinical practice, showed high rates of sustained virological response (SVR) in non‐cirrhotic genotype (GT)‐1 and GT‐4 patients. These results were slightly lower in cirrhotic patients. We investigated real‐life effectiveness and safety of SOF/SMV with or without ribavirin (RBV) in a large cohort of cirrhotic patients. Methods. This collaborative multicentre study included data from 968 patients with cirrhosis infected with HCV‐GT1 or 4, treated with SOF/SMV±RBV in 30 centres across Spain between January‐2014 and December‐2015. Demographic, clinical, virological and safety data were analysed. Results. Overall SVR was 92.3%; the majority of patients were treated with RBV (62%) for 12 weeks (92.4%). No significant differences in SVR were observed between genotypes (GT1a:94.3%; GT1b:91.7%; GT4:91.1%). Those patients with more advanced liver disease (Child B/C, MELD≥10) or portal hypertension (platelet count≤100×109/L, transient elastography≥21 Kpa) showed significantly lower SVR rates (84.4%‐91.9%) than patients with less advanced liver disease (93.8%‐95.9%, P<.01 in all cases). In the multivariate analysis, the use of RBV, female gender, baseline albumin≥35 g/L, MELD<10 and lack of exposure to a triple therapy regimen were independent predictors of SVR (P<.05). Serious adverse events (SAEs) and SAE‐associated discontinuation events occurred in 5.9% and 2.6%. Conclusions. In this large cohort of cirrhotic patients managed in the real‐world setting in Spain, SOF/SMV±RBV yielded to excellent SVR rates, especially in patients with compensated liver cirrhosis. In addition, this combination showed to be safe, with low rates of SAEs and early discontinuations.Instituto de Salud Carlos III; PI15/0015

Safety and Efficacy of Sofosbuvir plus Simeprevir in a Spanish Cohort of 622 Cirrhotic Patients Infected with Genotypes 1 or 4

Presentation Poste

Efficacy of Sofosbuvir and Velpatasvir, With and Without Ribavirin, in Patients With Hepatitis C Virus Genotype 3 Infection and Cirrhosis.

In phase 3 trials and real-world settings, smaller proportions of patients with genotype 3 hepatitis C virus (HCV) infection and cirrhosis have a sustained virologic response 12 weeks after treatment (SVR12) with the combination of sofosbuvir and velpatasvir than in patients without cirrhosis. It is unclear whether adding ribavirin to this treatment regimen increases SVRs in patients with genotype 3 HCV infection and cirrhosis. We performed a phase 2 trial of 204 patients with genotype 3 HCV infection and compensated cirrhosis (mean age 51 ± 7.4 years) at 29 sites in Spain from August 19, 2016 through April 18, 2017. Patients were assigned to groups given sofosbuvir and velpatasvir for 12 weeks (n = 101) or sofosbuvir and velpatasvir plus ribavirin for 12 weeks (n = 103). The primary efficacy end point was SVR12. The overall rates of SVR12 were 91% (92 of 101; 95% CI 84-96) for the sofosbuvir-velpatasvir group and 96% (99 of 103; 95% CI 90-99) for the sofosbuvir-velpatasvir plus ribavirin group. In the sofosbuvir-velpatasvir group, a smaller proportion of patients with baseline resistance-associated substitutions (RASs) in nonstructural protein 5A (NS5A) achieved an SVR12 (84%) than did patients without (96%). In the sofosbuvir-velpatasvir plus ribavirin group, baseline RASs had less effect on the proportion of patients with an SVR12 (96% for patients with baseline RASs; 99% for patients without). The most common adverse events (which occurred in ≥10% of patients) were asthenia (12%) in the sofosbuvir-velpatasvir group and asthenia (27%), headache (24%), and insomnia (12%) in the sofosbuvir-velpatasvir plus ribavirin group. Consistent with findings from previous studies, a high rate of patients (91% and 96%) with genotype 3 HCV infection and compensated cirrhosis achieved an SVR12 with sofosbuvir and velpatasvir, with or without ribavirin. Of patients treated with sofosbuvir and velpatasvir without ribavirin, fewer patients with baseline NS5A RASs achieved an SVR12 compared with patients without baseline NS5A. ClinicalTrials.govNCT02781558

Role of p63 and p73 isoforms on the cell death in patients with hepatocellular carcinoma submitted to orthotopic liver transplantation

BACKGROUND & AIMS: Patients with hepatocellular carcinoma (HCC) submitted to orthotopic liver transplantation (OLT) have a variable 5-year survival rate limited mostly by tumor recurrence. The etiology, age, sex, alcohol, Child-Pugh, and the immunesuppressor have been associated with tumour recurrence. The expression of ΔNp73 is related to the reduced survival of patients with HCC. The study evaluated the expression of p63 and p73 isoforms and cell death receptors, and their relation to tumour recurrence and survival. The results were in vitro validated in HCC cell lines. METHODS: HCC sections from patients submitted to OLT were used. The in vitro study was done in differentiated hepatitis B virus (HBV)-expressing Hep3B and control HepG2 cells. The expression of cell death receptors and cFLIPS/L, caspase-8 and -3 activities, and cell proliferation were determined in control and p63 and p73 overexpressing HCC cells. RESULTS: The reduced tumor expression of cell death receptors and TAp63 and TAp73, and increased ΔNp63 and ΔNp73 expression were associated with tumor recurrence and reduced survival. The in vitro study demonstrated that HBV-expressing Hep3B vs HepG2 cells showed reduced expression of p63 and p73, cell death receptors and caspase activation, and increased cFLIPL/cFLIPS ratio. The overexpression of TAp63 and TAp73 exerted a more potent pro-apoptotic and anti-proliferative effects in Hep3B than HepG2-transfected cells which was related to cFLIPL upregulation. CONCLUSIONS: The reduction of TAp63 and TAp73 isoforms, rather than alteration of ΔN isoform expression, exerted a significant functional repercussion on cell death and proliferation in HBV-expressing HepB cells

TAp63 and ΔNp63 (B), TAp73 and ΔNp73 (C) protein expression in cytoplasm and nuclear fractions, and their corresponding TAp63/ΔNp63 (D) and TAp73/ΔNp73 (E) protein ratio obtained from HepG2 and Hep3B cells.

<p>The specificity of antibodies against the different TA and ΔN p63 and p73 isoforms was assessed by western-blot analysis in transfected HepG2 using TA and ΔN p63 and p73 overexpressing plasmids (A). The protein expression was assessed by western-blot analysis following the procedure described in Material and Methods. Data are expressed as mean ± SEM of the densitometry analysis referred to the corresponding loading control (β-actin and histone H3 in cytoplasm and nuclear fractions, respectively). The ponceau staining is also included (B and C). The asterisks indicate statistical significance compared with their corresponding control group (HepG2) (*p≤0.05). The groups with different letter (a, b, c or d) were significantly different among them (p≤0.05).</p

TNF-R1 (A), CD95 (B) and TRAIL-R1 (C) expression obtained from HepG2 and Hep3B.

<p>The protein expression of cell death receptors was assessed by western-blot analysis following the procedure described in Material and Methods. Data are expressed as mean ± SEM of the densitometry analysis referred to the corresponding loading control (β-actin). The asterisks indicate statistical significance compared with their corresponding group (HepG2) (***p≤0.001).</p

TAp63, ΔNp63, TAp73 and ΔNp73 expression in well-differentiated (A) and moderately-poorly differentiated HCC (B) in patients subject to OLT, and their correlation to survival or death as a consequence of tumor recurrence or other causes such as pneumonia, fungal sepsis, heart complications and septic shock (C).

<p>The protein expression of p53 gene family members was assessed by immunohistochemistry following the procedure described in Material and Methods. Data are expressed as mean ± SEM of the densitometry analysis. The asterisks indicate statistical significance (*p≤0.05, **p≤0.01 or ***p≤0.001) compared with their corresponding control. The images are representative of the corresponding group. Image magnification x10.</p

TNF-R1 (A), CD95 (B) and TRAIL-R1 (C) expression, caspase-8 (D) and caspase-3 (E) activities and cell proliferation (F) in transfected HepG2 and Hep3B cells with TAp63α, ΔNp63α, TAp73α and ΔNp73α overexpressing plasmids.

<p>The parameters were assessed 48 hours after cell transfections. The protein expression of cell death receptors was assessed by western-blot analysis following the procedure described in Material and Methods. Caspase-8 and -3 activities were determined using commercial assays described in Material and Methods. Cell proliferation was determined by BrdU incorporation administered 2 hours before cell collection following the procedure described in Material and Methods. Data are expressed as mean ± SEM either from the densitometry analysis of blots referred to the corresponding loading control (stain-free procedure) or to caspase activity referred to a calibration curve developed using recombinant purified enzyme. The asterisks indicate statistical significance (*p≤0.05, **p≤0.01 or ***p≤0.001) compared with their corresponding empty vector transfected cell line. The groups denoted by different letters (a, b, c or d) are significantly different among them (p≤0.05).</p

Distribution of all pairwise variables and the corresponding linear equations in the three-studied populations (alive, dead by HCC recurrence and dead by other causes).

<p>The values of R² lineal of the linear equation were superior between ΔN isoforms and cell death receptors (Fig 3D-3F and 3K-3M) that those between ΔN isoforms and cell death receptors (Fig 3A-3C and 3H-3J).</p

Values of Pearson correlation coefficient (r) and the significance of the correlation (<i>p</i>) between the expression of p63 and p73 isoforms, and cell death receptors in studied population.

<p>Values of Pearson correlation coefficient (r) and the significance of the correlation (<i>p</i>) between the expression of p63 and p73 isoforms, and cell death receptors in studied population.</p