- Email: [email protected]
F-40 A novel metabolic action for the multikinase inhibitor sorafenib: targeting tumor cells through mitochondrial damage
S44
Abstracts of the A.I.S.F. Annual Meeting 2012 / Digestive and Liver Disease 44S (2012), S1–S48
marker of tumour hypoxia, neo-angiogenesis and worse prognosis. However data about HCC are lacking particularly in the clinical setting of patients undergoing transarterial-chemoembolization (TACE) in whom hypoxia and neo-angiogenesis may represent a molecular key to treatment failure. Aim of our analysis was to evaluate the role of lactate dehydrogenase (LDH) pre-treatment levels in determining clinical outcome for patients with HCC receiving TACE. One-hundred and fourteen patients were available for our analysis. For all patients LDH values were collected within one month before the procedure. For study purposes we divided our patients into two groups, according to LDH serum concentration registered before TACE (first group: LDH ≤450U/l 84 patients; second group: LDH >450U/l 30 patients). Patients were, also, classified according to the variation in LDH serum levels preand post-treatment (increased: 62 patients vs decreased 52 patients). Patients proved homogeneous for all clinical characteristics analyzed. In patients with LDH values below 450 U/l median time to progression (TTP) was 16.3 months, whereas it was of 10.1 months in patients above the cut-off (p=0.0085). Accordingly median overall survival (OS) was 22.4 months and of 11.7 months (p=0.0049). In patients with decreased LDH values after treatment median TTP was 12.4 months, and median OS was 22.1 months, whereas TTP was 9.1 months and OS was 9.5 in patients with increased LDH levels (TTP: p=0.0087, Figure 3; OS: p<0.0001). In our experience, LDH seemed able to predict clinical outcome in terms of TTP and OS for HCC patients undergoing TACE. Given the correlation between LDH levels and tumour angiogenesis we can speculate that patients with high LDH pretreatment levels may be optimal candidates for clinical trial exploring a multimodality treatment approach with TACE and anti-VEGF inhibitors in order to improve TTP and OS.
F-39 Management of symptomatic osteoporosis after liver transplantation in a real-world setting G. Valente 1 , G. Italiano 2 , L. Rinaldi 1 , G. Forte 3 , A. Gargiulo 4 , G. Piai 1 1 UOSD
SATTE; 2 UOS Mal. Met. Osseo e Reumatologia; 3 UOC Gastroenterologia; 4 UOC Medicina Interna, AO S. Anna San Sebastiano, Caserta Osteoporosis and vertebral fractures, painful and with functional limitations, are common complications after orthotopic liver transplantation (OLT). Most bone loss occurs in the first 6-12 months after OLT and later follow up should comprise monitoring and treatment of osteoporosis, but this is not always feasible in a practical setting out of a research context. Our purpose was to evaluate how the management of osteoporosis in transplanted patients is approached in the real setting of a gastro-hepatology team peripheral to Transplant Centers. We admitted 113 patients for follow-up after OLT in the period June 2005June 2010. An anamnesis of known bone loss or common risk factors for osteoporosis or back pain and functional limitations prompted to perform, in 69 of them, DXA (N=8) or calcaneal QUS (N=61), to measure bone mass, and spine x-rays to diagnose prevalent fractures. All patients were immunosuppressed (Cyc. 36, FK 25, other 8). They all had been treated with corticosteroids after OLT, but no one was still in therapy. Osteopenia was treated with calcium/vitamin D and osteoporosis or fragility fractures with bisphosphonates. The median follow up of patients after OLT was 38 months (range 14-232). Osteoporosis (T score <-2,5) was found in 37 (54%), osteopenia (T score >-2.5 <-1) in 19 (27%), normal (T score >-1) in 13 patients (19%). Twentynine patients (42%) had vertebral fractures: 2 of them (7%) had normal densitometry and 6 (21%) had values of osteopenia. After 12 months of treatment osteopenia values were found in 20/37 (54%) patients with previous osteoporosis values, while normal values were measured in 2/37 (5%) patients with previous osteoporosis values and in 5/19 (26%) with previous osteopenia. No one had worsened. Our on-field experience showed that: 1) evaluation of osteoporosis was prompted mainly by symptoms, even if impaired bone density and vertebral fractures are still frequent years after transplant, in some cases with no close relationship between altered bone density and severity of osteoporosis. 2) Common treatment with antiresorptive bisphosphonate drugs is effective in a
large part of patients. 3) Any protocol of follow-up of transplanted patients must include the systematic evaluation of osteoporosis, either symptomatic or not.
F-40 A novel metabolic action for the multikinase inhibitor sorafenib: targeting tumor cells through mitochondrial damage V. Tesori 1 , A.C. Piscaglia 1 , M. Barba 1 , M.A. Puglisi 1 , G. Gasbarrini 2 , G. Pani 3 , A. Gasbarrini 1 1 Institute
of Internal Medicine and Gastroenterology, Gemelli Hospytal, Rome, Italy; 2 Medical Research Foundation ONLUS, Bologna, Italy; 3 Institute of General Pathology, Gemelli Hospytal, Rome Italy Background and Aim: Although the multikinase inhibitor Sorafenib specifically targets tumor cells with V590E mutation of B-RAF, additional mechanisms likely account for the elevated anticancer activity of the drug in tumors lacking this mutation, and for the emergence of chemoresistance. In the present study a metabolic effect of the multikinase inhibitor Sorafenib (SFB) was investigated, alone and in combination with the glycolytic inhibitor 2-deoxyglucose (2dg), in a liver tumor model, the LCSC-2 (Liver Cancer Stem Cells) cell line. Materials and Methods: Drug toxicity was assessed by Propidium Iodide (PI) assay. Gene expression was assessed by microarray analysis. Protein expression was evaluated by western blotting. Mitochondrial activity was assessed by Respirometric Analysis. Mitochondrial potential was measured with JC-1 probe. Results: In LCSC-2 cells exposure to Sfb increases intracellular reactive oxygen species. At a molecular level SFB induces the phosphorylation, and presumably the activation, of the AMP-activated protein kinase (AMPK); taken together, ROS increase and AMPK induction, are consistent with mitochondrial damage. Accordingly, SFB led to a substantial reduction of oxygen consumption, cellular ATP level and mitochondrial transmembrane potential, and isolated mitochondria from rat liver were depolarized by the drug in vitro, indicating a direct effect of Sfb on the organelle. In keeping with its inhibitory action on mitochondrial respiration, Sfb killed much more efficiently LCSC-2 cells, as well as mouse B16F10 melanoma cells when associated with the glycolysis inhibitor 2DG. Conclusion: Taken together our results identify in mitochondrial damage and generation of ROS a novel modality of cytotoxicity by Sfb; moreover, the synergistic action of SFB plus 2DG outline a novel combined metabolic therapy to eradicate liver tumors.
F-41 Mixed cryoglobulinemia syndrome is a negative prognostic factor in the standard of care (Peg-IFN+ribavirin) anti-HCV therapy A. Piluso 1 , C. Giannini 1 , E. Fognani 1 , A. Petrarca 1 , C. Iannacone 2 , L. Gragnani 1 , J. Ranieri 1 , E. Triboli 1 , E. Pellegrini 1 , T. Urraro 1 , U. Arena 1 , M. Monti 1 , G. Laffi 1 , A.L. Zignego 1 1 MASVE 2 ITT,
Center and Dept of Internal Medicine, University of Florence, Italy; Florence, Italy; 1 SPARC Consulting, Milan, Italy
HCV leads to chronic hepatitis (CHC) and mixed cryoglobulinemia (MC). MC is observed in the majority of HCV pts, but is clinically evident (MC syndrome, MCS) in a minority of cases. Antiviral therapy is the first therapeutic option for both CHC and MCS. In previous reports, generally based on retrospective analyses and/or limited populations, no difference was observed in MC/MCS virological response. This is an elusive condition (the diagnosis is not well standardized and the MC impossible to determine in stored samples), and the possibility that some MC/MCS pts were included in the negative control group cannot be ruled out. In this study, we prospectively analyzed MC/MCS data and virological response of a large population of HCV pts treated with anti-HCV SoC (Peg-IFN+RBV). Pts were consecutively recruited from July 2003 to July 2010. Results: 424 pts were enrolled. These included: 121 pts with MCS (HCV+MCS), 132 with MC without MCS (HCV+MC) and 158 without
Abstracts of the A.I.S.F. Annual Meeting 2012 / Digestive and Liver Disease 44S (2012), S1–S48
marker of tumour hypoxia, neo-angiogenesis and worse prognosis. However data about HCC are lacking particularly in the clinical setting of patients undergoing transarterial-chemoembolization (TACE) in whom hypoxia and neo-angiogenesis may represent a molecular key to treatment failure. Aim of our analysis was to evaluate the role of lactate dehydrogenase (LDH) pre-treatment levels in determining clinical outcome for patients with HCC receiving TACE. One-hundred and fourteen patients were available for our analysis. For all patients LDH values were collected within one month before the procedure. For study purposes we divided our patients into two groups, according to LDH serum concentration registered before TACE (first group: LDH ≤450U/l 84 patients; second group: LDH >450U/l 30 patients). Patients were, also, classified according to the variation in LDH serum levels preand post-treatment (increased: 62 patients vs decreased 52 patients). Patients proved homogeneous for all clinical characteristics analyzed. In patients with LDH values below 450 U/l median time to progression (TTP) was 16.3 months, whereas it was of 10.1 months in patients above the cut-off (p=0.0085). Accordingly median overall survival (OS) was 22.4 months and of 11.7 months (p=0.0049). In patients with decreased LDH values after treatment median TTP was 12.4 months, and median OS was 22.1 months, whereas TTP was 9.1 months and OS was 9.5 in patients with increased LDH levels (TTP: p=0.0087, Figure 3; OS: p<0.0001). In our experience, LDH seemed able to predict clinical outcome in terms of TTP and OS for HCC patients undergoing TACE. Given the correlation between LDH levels and tumour angiogenesis we can speculate that patients with high LDH pretreatment levels may be optimal candidates for clinical trial exploring a multimodality treatment approach with TACE and anti-VEGF inhibitors in order to improve TTP and OS.
F-39 Management of symptomatic osteoporosis after liver transplantation in a real-world setting G. Valente 1 , G. Italiano 2 , L. Rinaldi 1 , G. Forte 3 , A. Gargiulo 4 , G. Piai 1 1 UOSD
SATTE; 2 UOS Mal. Met. Osseo e Reumatologia; 3 UOC Gastroenterologia; 4 UOC Medicina Interna, AO S. Anna San Sebastiano, Caserta Osteoporosis and vertebral fractures, painful and with functional limitations, are common complications after orthotopic liver transplantation (OLT). Most bone loss occurs in the first 6-12 months after OLT and later follow up should comprise monitoring and treatment of osteoporosis, but this is not always feasible in a practical setting out of a research context. Our purpose was to evaluate how the management of osteoporosis in transplanted patients is approached in the real setting of a gastro-hepatology team peripheral to Transplant Centers. We admitted 113 patients for follow-up after OLT in the period June 2005June 2010. An anamnesis of known bone loss or common risk factors for osteoporosis or back pain and functional limitations prompted to perform, in 69 of them, DXA (N=8) or calcaneal QUS (N=61), to measure bone mass, and spine x-rays to diagnose prevalent fractures. All patients were immunosuppressed (Cyc. 36, FK 25, other 8). They all had been treated with corticosteroids after OLT, but no one was still in therapy. Osteopenia was treated with calcium/vitamin D and osteoporosis or fragility fractures with bisphosphonates. The median follow up of patients after OLT was 38 months (range 14-232). Osteoporosis (T score <-2,5) was found in 37 (54%), osteopenia (T score >-2.5 <-1) in 19 (27%), normal (T score >-1) in 13 patients (19%). Twentynine patients (42%) had vertebral fractures: 2 of them (7%) had normal densitometry and 6 (21%) had values of osteopenia. After 12 months of treatment osteopenia values were found in 20/37 (54%) patients with previous osteoporosis values, while normal values were measured in 2/37 (5%) patients with previous osteoporosis values and in 5/19 (26%) with previous osteopenia. No one had worsened. Our on-field experience showed that: 1) evaluation of osteoporosis was prompted mainly by symptoms, even if impaired bone density and vertebral fractures are still frequent years after transplant, in some cases with no close relationship between altered bone density and severity of osteoporosis. 2) Common treatment with antiresorptive bisphosphonate drugs is effective in a
large part of patients. 3) Any protocol of follow-up of transplanted patients must include the systematic evaluation of osteoporosis, either symptomatic or not.
F-40 A novel metabolic action for the multikinase inhibitor sorafenib: targeting tumor cells through mitochondrial damage V. Tesori 1 , A.C. Piscaglia 1 , M. Barba 1 , M.A. Puglisi 1 , G. Gasbarrini 2 , G. Pani 3 , A. Gasbarrini 1 1 Institute
of Internal Medicine and Gastroenterology, Gemelli Hospytal, Rome, Italy; 2 Medical Research Foundation ONLUS, Bologna, Italy; 3 Institute of General Pathology, Gemelli Hospytal, Rome Italy Background and Aim: Although the multikinase inhibitor Sorafenib specifically targets tumor cells with V590E mutation of B-RAF, additional mechanisms likely account for the elevated anticancer activity of the drug in tumors lacking this mutation, and for the emergence of chemoresistance. In the present study a metabolic effect of the multikinase inhibitor Sorafenib (SFB) was investigated, alone and in combination with the glycolytic inhibitor 2-deoxyglucose (2dg), in a liver tumor model, the LCSC-2 (Liver Cancer Stem Cells) cell line. Materials and Methods: Drug toxicity was assessed by Propidium Iodide (PI) assay. Gene expression was assessed by microarray analysis. Protein expression was evaluated by western blotting. Mitochondrial activity was assessed by Respirometric Analysis. Mitochondrial potential was measured with JC-1 probe. Results: In LCSC-2 cells exposure to Sfb increases intracellular reactive oxygen species. At a molecular level SFB induces the phosphorylation, and presumably the activation, of the AMP-activated protein kinase (AMPK); taken together, ROS increase and AMPK induction, are consistent with mitochondrial damage. Accordingly, SFB led to a substantial reduction of oxygen consumption, cellular ATP level and mitochondrial transmembrane potential, and isolated mitochondria from rat liver were depolarized by the drug in vitro, indicating a direct effect of Sfb on the organelle. In keeping with its inhibitory action on mitochondrial respiration, Sfb killed much more efficiently LCSC-2 cells, as well as mouse B16F10 melanoma cells when associated with the glycolysis inhibitor 2DG. Conclusion: Taken together our results identify in mitochondrial damage and generation of ROS a novel modality of cytotoxicity by Sfb; moreover, the synergistic action of SFB plus 2DG outline a novel combined metabolic therapy to eradicate liver tumors.
F-41 Mixed cryoglobulinemia syndrome is a negative prognostic factor in the standard of care (Peg-IFN+ribavirin) anti-HCV therapy A. Piluso 1 , C. Giannini 1 , E. Fognani 1 , A. Petrarca 1 , C. Iannacone 2 , L. Gragnani 1 , J. Ranieri 1 , E. Triboli 1 , E. Pellegrini 1 , T. Urraro 1 , U. Arena 1 , M. Monti 1 , G. Laffi 1 , A.L. Zignego 1 1 MASVE 2 ITT,
Center and Dept of Internal Medicine, University of Florence, Italy; Florence, Italy; 1 SPARC Consulting, Milan, Italy
HCV leads to chronic hepatitis (CHC) and mixed cryoglobulinemia (MC). MC is observed in the majority of HCV pts, but is clinically evident (MC syndrome, MCS) in a minority of cases. Antiviral therapy is the first therapeutic option for both CHC and MCS. In previous reports, generally based on retrospective analyses and/or limited populations, no difference was observed in MC/MCS virological response. This is an elusive condition (the diagnosis is not well standardized and the MC impossible to determine in stored samples), and the possibility that some MC/MCS pts were included in the negative control group cannot be ruled out. In this study, we prospectively analyzed MC/MCS data and virological response of a large population of HCV pts treated with anti-HCV SoC (Peg-IFN+RBV). Pts were consecutively recruited from July 2003 to July 2010. Results: 424 pts were enrolled. These included: 121 pts with MCS (HCV+MCS), 132 with MC without MCS (HCV+MC) and 158 without