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Ribavirin
904 TELAPREVIR-BASED REGIMENS SUBSTANTIALLY IMPROVED SVR RATES ACROSS ALL IL28B GENOTYPES IN THE ADVANCE TRIAL Ira M. Jacobson, Ian M. Catlett, Patrick Marcellin, Natalie H. Bzowej, Andrew J. Muir, Nathalie Adda, Shelia Seepersaud, Ravi Ramachandran, Katherine Sussky, Leif Bengtsson, Shelley George, Robert S. Kauffman, Martyn C. Botfield
873 Paired Box Gene 5 is a Novel Tumor Suppressor Involved in the Pathogenesis of Hepatocellular Carcinoma Through Interaction With p53 Signaling Pathway Weili Liu, Xiaoxing Li, Eagle SH Chu, Minnie Y. Go, Joseph J. Sung, Jun Yu
Background & Aim: Single nucleotide polymorphisms (SNPs) near the IL28B gene have been strongly associated with the likelihood of SVR in genotype 1 HCV patients treated with peginterferon/ribavirin (PR). During the evaluation of an exploratory diagnostic test that characterizes genetic polymorphisms near the IL28B gene, the impact of rs12979860 on SVR in telaprevir (T)-based regimens was evaluated. Methods: IL28B genotype was determined in de-identified left-over specimens available from treatment-naïve genotype 1 HCV patients from ADVANCE U.S. sites. Because of the limited number of patients of nonCaucasian race and the requirements of the de-identification procedure, only samples from Caucasian patients were tested. Results: The diagnostic assay developed provided consistent, unambiguous genotype calls and was considered suitable for research. 454/1088 (42%) patients had IL28B test results available. 150/454 (33%) were CC, 224/454 (49%) CT, and 80/454 (18%) TT. SVR rates for each subgroup by arm are shown in the Table. 72%, 54% and 48% of CC, CT and TT telaprevir patients, respectively had undetectable HCV RNA at weeks 4 and 12 (eRVR) compared with 16%, 3% and 0% of PR patients. Among eRVR telaprevir patients, 91% achieved SVR (97% of CC, 88% of CT/TT) with 24 weeks of therapy whereas 45% of non-eRVR telaprevir patients had SVR (67% of CC, 38% CT/TT) with 48 weeks of therapy. Conclusions: Telaprevir-based therapy improved eRVR and SVR rates across all IL28B genotypes. Specifically, telaprevir-based therapy more than doubled the rates of SVR in CT/TT patients, and substantially increased SVR rates in those with CC genotype, as compared with PR therapy alone. Non-attainment of eRVR was associated with lower SVR rates across all IL28B genotypes, with the largest decrement in CT/TT patients. SVR rates
Background: The paired box 5 (PAX5) is a member of PAX transcription factors family associated with the regulation of embryonic development, however the role of PAX5 in carcinogenesis is largely unclear. We identified PAX5 is involved in human cancer by methylation-sensitive representational difference analysis. We analyzed the epigenetic inactivation, biological functions and related molecular mechanisms of PAX5 in hepatocellular carcinoma (HCC). Methods: Promoter methylation of PAX5 was evaluated by methylationspecific PCR and bisulfite genomic sequencing (BGS). The functions of ectopic PAX5 expressions were determined by viability assays, colony formation and cell cycle analyses, along with In Vivo tumorigenicity assay. PAX5 target signal pathway was identified by promoter luciferase assay, chromosome immunoprecipitation (ChIP) and pathway PCR array analyses. Results: PAX5 is expressed in normal human liver tissue, but silenced or down-regulated in 83% (10/12) of HCC cell lines. Real-time PCR analysis showed that PAX5 expression is significantly down-regulated in primary HCCs as compared with their corresponding adjacent normal tissues (P=0.0008). The promoter methylation status contributes to the inactivity of PAX5 in HCC cell lines, as evidenced by demethylating treatment and BGS. Restoring PAX5 expression in Hep3B or HepG2 HCC cell line suppressed cell viability (P<0.0001), colony formation (down to 44-54% of vector control, P<0.01), induced apoptosis (24.75% ± 2.09% vs., 33.11% ± 2.06%, P<0.05) In Vitro, and inhibited tumor growth in nude mice (P<0.0001). Pathway luciferase reporter assays indicated that PAX5 activated p53 and p21 signalings. ChIP assay demonstrated that PAX5 directly bound to the p53 promoter. The anti-tumorigenic function of PAX5 were at least by up-regulating p53 and its downstream targets including tumor necrosis factor, Fas ligand, leucine-rich repeats and death domain containing, poly(rC) binding protein4, p21 and growth arrest and DNA-damage-inducible, alpha. Conclusions: PAX5 is frequently inactivated by promoter methylation in HCCs. PAX5 appears to be a functional tumor suppressor involved in liver carcinogenesis through directly regulating p53 signaling pathway. Acknowledgment The project was supported by Research Grants Council Competitive Earmarked Research Grant CUHK (GRF 473008, GRF 478108, CUHK/CRF/ 09 and scheme B MK/KT/fis0910/0543/10yc).
*T12PR = T+PR 12 weeks, then PR 12 or 36 weeks depending on eRVR status **T8PR = T+PR 8 weeks, then PR 16 or 40 weeks depending on eRVR status §In patients tested for IL28B, SVR was defined as undetectable HCV RNA at EOT and at 24 weeks after last actual dose of study drug †In the ADVANCE study, SVR was defined as undetectable HCV RNA at EOT and at 24 weeks after last planned dose of study drug
903 High Sustained Virologic Response (SVR) Among Genotype 1 Previous NonResponders and Relapsers to Peginterferon/Ribavirin When Re-Treated with Boceprevir (BOC) Plus Peginterferon Alfa-2A/Ribavirin Steven L. Flamm, Eric J. Lawitz, Ira M. Jacobson, Raymond A. Rubin, Marc Bourliere, C. Hezode, J. Vierling, Claus Niederau, Morris Sherman, Venkata S. Goteti, Regis A. Vilchez, Clifford A. Brass, Janice K. Albrecht, Fred Poordad
905 Obtaining SVR To Antiviral Therapy Highly Protects Patients With HCV Recurrence On The Graft After Liver Transplantation From Liver Related Death: Insights from The AISF-RECOLT-C GROUP. Maria Rendina, Nicola M. Castellaneta, Stefano Fagiuoli, Francesca R. Ponziani, Chiara Vigano', Rosa Maria Iemmolo, Maria F. Donato, Pierluigi Toniutto, Luisa Pasulo, Maria Cristina Morelli, Patrizia Burra, Lucia Miglioresi, Valerio Giannelli, Daniele Di Paolo, Alfredo Di Leo
Background: The RESPOND-2 trial demonstrated significantly increased SVR for prior nonresponders and relapsers when BOC was added to peginterferon alfa-2b/ribavirin (66% vs. 21% control). We assessed SVR with BOC combined with peginterferon alfa-2a (PEG2a) and ribavirin (R) in patients who met identical entry criteria. Methods: This double-blind, placebo-controlled trial randomized 201 genotype-1 relapsers and non-responders to two arms (1:2 ratio, Table). Arm 1 (control) received a 4-week lead-in of PEG2a/R followed by placebo + PEG2a/R for 44 weeks. Arm 2 received a 4-week lead-in of PEG2a/R followed by BOC + PEG2a/R for 44 weeks. Therapy was discontinued if HCV-RNA was detectable (undetectable HCV RNA <9.3 IU/mL [Roche TaqMan, LLD]) at week 12. Primary endpoint: SVR 24-weeks post-therapy. Results: Baseline demographics: 70% male; 10% black; 16% cirrhotic. The addition of BOC after a 4-week lead-in with PEG2a/R significantly increased SVR: 21% in Arm 1 vs. 64% in Arm 2 (p<0.0001). SVR for patients with poor interferon responsiveness (<1-log10 decrease in HCV-RNA after 4-week lead-in) was 0% in Arm 1 and 39% in Arm 2. For patients responsive to interferon (≥1-log10 decrease in HCV-RNA after 4-week lead-in), SVR was 25% in Arm 1 and 71% in Arm 2. Discontinuation due to adverse events (AEs) occurred in 4% and 17% of patients in Arms 1 and 2. Rates of serious AEs were 10% in Arm 1 and 13% in Arm 2. The frequencies of anemia (<10.0 g/dL) in Arms 1 and 2 were 27% vs. 49%; neutropenia (WHO grade 3-4 [<750/mm3]) 21% vs. 43%; erythropoietin use 30% vs. 47%. There were no serious AEs due to anemia and one discontinuation due to anemia (Arm 2). Conclusions: Lead-in with PEG2a and ribavirin followed by addition of boceprevir resulted in high SVR rates similar to that observed using an identical treatment regimen with peginterferon alfa-2b. Therapy was generally welltolerated. These are the first large trials to demonstrate a direct acting antiviral agent may be combined with either PEG2a or PEG2b to significantly increase SVR in patients who failed prior therapy.
Background: Results of antiviral therapy for HCV recurrence on the graft are controversial. Sustained virological response (SVR) to antiviral is strongly associated with reduced liverrelated mortality in chronic hepatitis and cirrhosis. In liver transplant (LT) setting, however, a huge number of factors and co-morbidities could frustrate the benefit provided by the elimination of the virus. Aim of the Study: To determine the impact of achieving SVR on liver related and non-liver related mortality in HCV recurrent pts, analysis was made of a large retrospective multicentre database (from 12 LT Centres in Italy. Patients and methods: Data on 464 pts (transplant between 1989 and 2008) were retrospectively analyzed. All pts, after virological and histological diagnosis of HCV recurrence, underwent antiviral therapy with standard or peginterferon (24% and 76%) plus ribavirin . Mean age at LT 53.7±7.9 yrs; male/female: 348/116; genotype 1-4: 75% pts; diabetes: 50%. Mean interval from LT to therapy: 24±28 months; median duration of therapy: 44.8±35 weeks. Results: Overall, the SVR rate was 35% (164/464). SVR pts had: younger donor age (p< 0.009), longer treatment duration and higher cumulative IFN dose (p<0.002), lower drop-out rate and less diabetes (p<0.001). No differences in immunosuppression. During post-treatment F-up of 4.4 years 120 patients died or were re-transplanted. Mortality rate in SVR and non-SVR pts was 10% and 34% respectively (p< 0.001). The incidence rate of liver-related mortality/100 persons/year of follow up was significantly different between SVR and no-SVR (0.1 vs 6.3, p< 0.001); no differences in the non-liver related mortality (1.8 vs 1.4; P 0.4). At univariate Cox regression, risk factors for death/re-transplantation were older donor age (P<0.001; HR 1.03), diabetes (P< 0.001; HR 2.3), genotype 1-4 (P 0.03; HR 1.7) and lack of SVR (P< 0.001; HR 3.9). At multivariate analysis, failure in obtaining SVR remained significantly associated with mortality (HR 3.4 p=0.001) Conclusion: Despite significant drawbacks in conducting antiviral therapy in HCV recurrent liver transplant patients, strategies of HCV
S-145
AGA Abstracts
AGA Abstracts
RASSF10 in silenced gastric cancer cell lines could be restored after treating with demethylation agent, 5-aza-deoxycytidine. We further tested the biological function of RASSF10 in human gastric cancer cells. Stable transfection of RASSF10 in AGS and MKN45 cancer cells reduced colony formation from 100% to 51.6% in AGS (P<0.01) and to 56.0% in MKN45 (P<0.01). Ectopic expression of RASSF10 in AGS and MKN45 cells significantly suppressed cell viability (P<0.0001 for both cell lines), induced cell apoptosis (P<0.05 for both cell lines), and repressed cell migration and invasion (P<0.0001 for both cell lines). We further found that RASSF10 up-regulated the expression of pro-apoptotic gene TNF and metastasis suppressor MTSS1, and down-regulated the expression of multiple oncogenes including AKT1, ERBB2, FOS and Jun. RASSF10 hypermethylation was detected in 54% (53/99) of primary gastric cancers, but only in 6% (1/18) of non-cancer tissues (P<0.0001). Conclusions: Our data show that RASSF10 is a functional tumor suppressor frequently methylated in gastric cancers. Acknowledgment The project was supported by Research Grant ERG CUHK (GRF 473008).
873 Paired Box Gene 5 is a Novel Tumor Suppressor Involved in the Pathogenesis of Hepatocellular Carcinoma Through Interaction With p53 Signaling Pathway Weili Liu, Xiaoxing Li, Eagle SH Chu, Minnie Y. Go, Joseph J. Sung, Jun Yu
Background & Aim: Single nucleotide polymorphisms (SNPs) near the IL28B gene have been strongly associated with the likelihood of SVR in genotype 1 HCV patients treated with peginterferon/ribavirin (PR). During the evaluation of an exploratory diagnostic test that characterizes genetic polymorphisms near the IL28B gene, the impact of rs12979860 on SVR in telaprevir (T)-based regimens was evaluated. Methods: IL28B genotype was determined in de-identified left-over specimens available from treatment-naïve genotype 1 HCV patients from ADVANCE U.S. sites. Because of the limited number of patients of nonCaucasian race and the requirements of the de-identification procedure, only samples from Caucasian patients were tested. Results: The diagnostic assay developed provided consistent, unambiguous genotype calls and was considered suitable for research. 454/1088 (42%) patients had IL28B test results available. 150/454 (33%) were CC, 224/454 (49%) CT, and 80/454 (18%) TT. SVR rates for each subgroup by arm are shown in the Table. 72%, 54% and 48% of CC, CT and TT telaprevir patients, respectively had undetectable HCV RNA at weeks 4 and 12 (eRVR) compared with 16%, 3% and 0% of PR patients. Among eRVR telaprevir patients, 91% achieved SVR (97% of CC, 88% of CT/TT) with 24 weeks of therapy whereas 45% of non-eRVR telaprevir patients had SVR (67% of CC, 38% CT/TT) with 48 weeks of therapy. Conclusions: Telaprevir-based therapy improved eRVR and SVR rates across all IL28B genotypes. Specifically, telaprevir-based therapy more than doubled the rates of SVR in CT/TT patients, and substantially increased SVR rates in those with CC genotype, as compared with PR therapy alone. Non-attainment of eRVR was associated with lower SVR rates across all IL28B genotypes, with the largest decrement in CT/TT patients. SVR rates
Background: The paired box 5 (PAX5) is a member of PAX transcription factors family associated with the regulation of embryonic development, however the role of PAX5 in carcinogenesis is largely unclear. We identified PAX5 is involved in human cancer by methylation-sensitive representational difference analysis. We analyzed the epigenetic inactivation, biological functions and related molecular mechanisms of PAX5 in hepatocellular carcinoma (HCC). Methods: Promoter methylation of PAX5 was evaluated by methylationspecific PCR and bisulfite genomic sequencing (BGS). The functions of ectopic PAX5 expressions were determined by viability assays, colony formation and cell cycle analyses, along with In Vivo tumorigenicity assay. PAX5 target signal pathway was identified by promoter luciferase assay, chromosome immunoprecipitation (ChIP) and pathway PCR array analyses. Results: PAX5 is expressed in normal human liver tissue, but silenced or down-regulated in 83% (10/12) of HCC cell lines. Real-time PCR analysis showed that PAX5 expression is significantly down-regulated in primary HCCs as compared with their corresponding adjacent normal tissues (P=0.0008). The promoter methylation status contributes to the inactivity of PAX5 in HCC cell lines, as evidenced by demethylating treatment and BGS. Restoring PAX5 expression in Hep3B or HepG2 HCC cell line suppressed cell viability (P<0.0001), colony formation (down to 44-54% of vector control, P<0.01), induced apoptosis (24.75% ± 2.09% vs., 33.11% ± 2.06%, P<0.05) In Vitro, and inhibited tumor growth in nude mice (P<0.0001). Pathway luciferase reporter assays indicated that PAX5 activated p53 and p21 signalings. ChIP assay demonstrated that PAX5 directly bound to the p53 promoter. The anti-tumorigenic function of PAX5 were at least by up-regulating p53 and its downstream targets including tumor necrosis factor, Fas ligand, leucine-rich repeats and death domain containing, poly(rC) binding protein4, p21 and growth arrest and DNA-damage-inducible, alpha. Conclusions: PAX5 is frequently inactivated by promoter methylation in HCCs. PAX5 appears to be a functional tumor suppressor involved in liver carcinogenesis through directly regulating p53 signaling pathway. Acknowledgment The project was supported by Research Grants Council Competitive Earmarked Research Grant CUHK (GRF 473008, GRF 478108, CUHK/CRF/ 09 and scheme B MK/KT/fis0910/0543/10yc).
*T12PR = T+PR 12 weeks, then PR 12 or 36 weeks depending on eRVR status **T8PR = T+PR 8 weeks, then PR 16 or 40 weeks depending on eRVR status §In patients tested for IL28B, SVR was defined as undetectable HCV RNA at EOT and at 24 weeks after last actual dose of study drug †In the ADVANCE study, SVR was defined as undetectable HCV RNA at EOT and at 24 weeks after last planned dose of study drug
903 High Sustained Virologic Response (SVR) Among Genotype 1 Previous NonResponders and Relapsers to Peginterferon/Ribavirin When Re-Treated with Boceprevir (BOC) Plus Peginterferon Alfa-2A/Ribavirin Steven L. Flamm, Eric J. Lawitz, Ira M. Jacobson, Raymond A. Rubin, Marc Bourliere, C. Hezode, J. Vierling, Claus Niederau, Morris Sherman, Venkata S. Goteti, Regis A. Vilchez, Clifford A. Brass, Janice K. Albrecht, Fred Poordad
905 Obtaining SVR To Antiviral Therapy Highly Protects Patients With HCV Recurrence On The Graft After Liver Transplantation From Liver Related Death: Insights from The AISF-RECOLT-C GROUP. Maria Rendina, Nicola M. Castellaneta, Stefano Fagiuoli, Francesca R. Ponziani, Chiara Vigano', Rosa Maria Iemmolo, Maria F. Donato, Pierluigi Toniutto, Luisa Pasulo, Maria Cristina Morelli, Patrizia Burra, Lucia Miglioresi, Valerio Giannelli, Daniele Di Paolo, Alfredo Di Leo
Background: The RESPOND-2 trial demonstrated significantly increased SVR for prior nonresponders and relapsers when BOC was added to peginterferon alfa-2b/ribavirin (66% vs. 21% control). We assessed SVR with BOC combined with peginterferon alfa-2a (PEG2a) and ribavirin (R) in patients who met identical entry criteria. Methods: This double-blind, placebo-controlled trial randomized 201 genotype-1 relapsers and non-responders to two arms (1:2 ratio, Table). Arm 1 (control) received a 4-week lead-in of PEG2a/R followed by placebo + PEG2a/R for 44 weeks. Arm 2 received a 4-week lead-in of PEG2a/R followed by BOC + PEG2a/R for 44 weeks. Therapy was discontinued if HCV-RNA was detectable (undetectable HCV RNA <9.3 IU/mL [Roche TaqMan, LLD]) at week 12. Primary endpoint: SVR 24-weeks post-therapy. Results: Baseline demographics: 70% male; 10% black; 16% cirrhotic. The addition of BOC after a 4-week lead-in with PEG2a/R significantly increased SVR: 21% in Arm 1 vs. 64% in Arm 2 (p<0.0001). SVR for patients with poor interferon responsiveness (<1-log10 decrease in HCV-RNA after 4-week lead-in) was 0% in Arm 1 and 39% in Arm 2. For patients responsive to interferon (≥1-log10 decrease in HCV-RNA after 4-week lead-in), SVR was 25% in Arm 1 and 71% in Arm 2. Discontinuation due to adverse events (AEs) occurred in 4% and 17% of patients in Arms 1 and 2. Rates of serious AEs were 10% in Arm 1 and 13% in Arm 2. The frequencies of anemia (<10.0 g/dL) in Arms 1 and 2 were 27% vs. 49%; neutropenia (WHO grade 3-4 [<750/mm3]) 21% vs. 43%; erythropoietin use 30% vs. 47%. There were no serious AEs due to anemia and one discontinuation due to anemia (Arm 2). Conclusions: Lead-in with PEG2a and ribavirin followed by addition of boceprevir resulted in high SVR rates similar to that observed using an identical treatment regimen with peginterferon alfa-2b. Therapy was generally welltolerated. These are the first large trials to demonstrate a direct acting antiviral agent may be combined with either PEG2a or PEG2b to significantly increase SVR in patients who failed prior therapy.
Background: Results of antiviral therapy for HCV recurrence on the graft are controversial. Sustained virological response (SVR) to antiviral is strongly associated with reduced liverrelated mortality in chronic hepatitis and cirrhosis. In liver transplant (LT) setting, however, a huge number of factors and co-morbidities could frustrate the benefit provided by the elimination of the virus. Aim of the Study: To determine the impact of achieving SVR on liver related and non-liver related mortality in HCV recurrent pts, analysis was made of a large retrospective multicentre database (from 12 LT Centres in Italy. Patients and methods: Data on 464 pts (transplant between 1989 and 2008) were retrospectively analyzed. All pts, after virological and histological diagnosis of HCV recurrence, underwent antiviral therapy with standard or peginterferon (24% and 76%) plus ribavirin . Mean age at LT 53.7±7.9 yrs; male/female: 348/116; genotype 1-4: 75% pts; diabetes: 50%. Mean interval from LT to therapy: 24±28 months; median duration of therapy: 44.8±35 weeks. Results: Overall, the SVR rate was 35% (164/464). SVR pts had: younger donor age (p< 0.009), longer treatment duration and higher cumulative IFN dose (p<0.002), lower drop-out rate and less diabetes (p<0.001). No differences in immunosuppression. During post-treatment F-up of 4.4 years 120 patients died or were re-transplanted. Mortality rate in SVR and non-SVR pts was 10% and 34% respectively (p< 0.001). The incidence rate of liver-related mortality/100 persons/year of follow up was significantly different between SVR and no-SVR (0.1 vs 6.3, p< 0.001); no differences in the non-liver related mortality (1.8 vs 1.4; P 0.4). At univariate Cox regression, risk factors for death/re-transplantation were older donor age (P<0.001; HR 1.03), diabetes (P< 0.001; HR 2.3), genotype 1-4 (P 0.03; HR 1.7) and lack of SVR (P< 0.001; HR 3.9). At multivariate analysis, failure in obtaining SVR remained significantly associated with mortality (HR 3.4 p=0.001) Conclusion: Despite significant drawbacks in conducting antiviral therapy in HCV recurrent liver transplant patients, strategies of HCV
S-145
AGA Abstracts
AGA Abstracts
RASSF10 in silenced gastric cancer cell lines could be restored after treating with demethylation agent, 5-aza-deoxycytidine. We further tested the biological function of RASSF10 in human gastric cancer cells. Stable transfection of RASSF10 in AGS and MKN45 cancer cells reduced colony formation from 100% to 51.6% in AGS (P<0.01) and to 56.0% in MKN45 (P<0.01). Ectopic expression of RASSF10 in AGS and MKN45 cells significantly suppressed cell viability (P<0.0001 for both cell lines), induced cell apoptosis (P<0.05 for both cell lines), and repressed cell migration and invasion (P<0.0001 for both cell lines). We further found that RASSF10 up-regulated the expression of pro-apoptotic gene TNF and metastasis suppressor MTSS1, and down-regulated the expression of multiple oncogenes including AKT1, ERBB2, FOS and Jun. RASSF10 hypermethylation was detected in 54% (53/99) of primary gastric cancers, but only in 6% (1/18) of non-cancer tissues (P<0.0001). Conclusions: Our data show that RASSF10 is a functional tumor suppressor frequently methylated in gastric cancers. Acknowledgment The project was supported by Research Grant ERG CUHK (GRF 473008).