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P3.03-034 Comprehensive Immunophenotyping of the Blood and Pleural Fluid from Patients with Malignant Pleural Mesothelioma by Flow Cytometry
January 2017
Anthony Linton,1 Matthew Soeberg,1 Steven Kao,2 Richard Broome,3 Nico Van Zandwijk1 1Asbestos Diseases Research Institute, Sydney/NSW/Australia, 2 Medical Oncology, Chris O’Brien Lifehouse, Sydney/ Australia, 3Public Health Observatory, Sydney/NSW/ Australia Background: Whilst the impact of clinico-pathological factors on the prognosis of malignant pleural mesothelioma (MPM) is well understood, socio-economic and geographic factors have received less attention. Although the majority of Australians reside in major cities, a dispersed population lives in regional and remote areas, where access to clinical services may be limited. We investigated the association between geographic and socio-economic factors and treatment provision and survival in a large series of patients from New South Wales. Methods: All patients awarded compensation by the NSW Dust Diseases Board (2002e2009) following diagnosis with MPM were assessed. Geographic remoteness, distance from oncological multidisciplinary teams (MDT) and socioeconomic status according to the index of relative socio-economic advantage and disadvantage (IRSAD), were assessed with known prognostic factors using Kaplan Meir and Cox-regression analysis. Chi-square testing compared categorical variables to analyse the impact of these factors upon clinical features and treatment received. Cancer Registry incidence data was assessed to allow comparison of the compensated DDB cohort to all NSW MPM cases. Results: We assessed 910 patients: Geographic remoteness (major city 67%; regional/remote 33%), distance to MDT (<10km 65%, <50km 92%). Geographic distribution was comparable to cancer registry data. Median overall survival was 10.0 months. On multivariate analysis, non-epithelioid histological subtype (HR.2.19); male gender (HR¼1.37); age >70 (HR¼1.39) and IRSAD status by decreasing quintile (HR¼1.07) were independent prognostic factors, with a pronounced survival difference between highest and lowest IRSAD quintiles (8.4 vs 12.8 months). A trend to improved survival when residing in major cities (10.6 vs 8.8 months; p¼0.162) and within 50km of MDT (10.3 vs 7.8 months;p¼0.539) was noted. Patients geographic location and distance to MDT affected the use of palliative radiotherapy (p<0.05) however did not impact chemotherapy, adjuvant radiotherapy or extrapleural pneumonectomy provision. Socioeconomically disadvantaged patients were less likely to receive chemotherapy (40.3% vs 47.7%; p¼0.032), with pronounced disparity between the most socioeconomically advantaged and disadvantaged quintiles (54.2% vs 37.6%;p¼0.001).
Abstracts
S1365
Conclusion: Despite ‘universal’ health care and the support of a compensation scheme, socioeconomic disadvantage was an independent prognostic factor for MPM in NSW Australia. A significant reduction in chemotherapy utilization was noted, particularly in highly socioeconomically deprived areas. Furthermore, a trend to improved survival was noted in patients residing in major cities within closer proximity to oncology units, though treatment provision did not differ. Prospective research analyzing specific factors including comorbidity, income, and individual preference will be required to better understand these findings in both compensated and non-compensated individuals. Keywords: Mesothelioma, Prognosis
P3.03-034 Comprehensive Immunophenotyping of the Blood and Pleural Fluid from Patients with Malignant Pleural Mesothelioma by Flow Cytometry Topic: Mesothelioma Clinical Tobias Peikert, Virginia Van Keulen, Svetlana Bornschlegl, Allen Dietz, Mike Gustafson Mayo Clinic, Rochester/MN/United States of America Background: Malignant pleural mesothelioma (MM) remains an almost universally fatal disease with limited treatment options. Immunotherapy represents a rapidly emerging therapeutic strategy for multiple malignancies including MM. However durable therapeutic responses occur in a minority (w30%) of patients. Better understanding of the quantities and immunophenotype of circulating and intrapleural immune cells is essential to design and apply personalized immunotherapeutic strategies. Methods: We used a comprehensive flow cytometry panel (PLoS One. 2015 Mar 23;10 (3):e0121546) to prospectively characterize circulating and pleural fluid immune cells in patients with MPM (n¼12) and normal volunteers (circulating cells only, n¼50). Matched blood and pleural samples were available from 11 patients, including samples from 9 patients enrolled into a Phase I study investigating the intrapleural administration of the modified vaccine strain measles virus (MV-NIS), MC1023. Pre- and post-treatments samples were available from 7 patients. The immune cell counts and cell fractions were compared using a false discovery rate (FDR) of 10% and the non-parametric Mann-Whitney test and the Wilcoxon matched-pairs signed rank test (blood versus pleural fluid and different time points) (p 0.05).
S1366
Results: Cell counts and immune phenotype of circulating immune cells differ between of patients with MPM differ from normal volunteers (31 of 86 cell types). Patients with MPM had fewer B-lymphocytes, more proinflammatory monocytes (CD14+CD16+) and exhausted CD4 and CD8 T-lymphocytes (CD4 and CD8 PD1+TIM3+ double positive cells). As expected we observed characteristic differences between blood and pleural fluid in MPM patients. For example there are a higher numbers of antigen experienced, PD-1 expressing CD4 and CD8memory cells within the pleural fluid compared to the peripheral blood. The intrapleural administration of a modified vaccine strain measles (MV-NIS) triggered changes in pleural and circulating immune cells. While these changes varied among patients, we observed increased CD80 and CD86 expression on CD14+ monocytes in the pleural fluid down. Conclusion: Comprehensive flow-cytometric immunophenotying of blood and pleural fluid is feasible in patients with MPM. This approach may help to identify patients who may respond favorably to specific immunotherapeutic interventions such as immune checkpoint inhibitors or facilitate longitudinal immune monitoring during clinical trials. Additional data is needed and we are continuing to prospectively analyze samples from patients with MPM. Keywords: flow cytometry, pleural fluid, Mesothelioma, immunophenotyping
P3.03-035 Prognostic Role of hENT1 and RRM1 in Patients with Advanced Pleural Mesothelioma Treated with Second Line Gemcitabine Based Regimens Topic: Mesothelioma Clinical Andrés Cardona,1 Oscar Arrieta,2 Leonardo Rojas,3 Luis Corrales,4 Beatriz Wills,5 George Oblitas,6 Ludwing Bacon,7 Claudio Martin,8 Mauricio Cuello,9 Luis Mas,10 Carlos Vargas,1 Hernan Carranza,1 Jorge Otero,1 Maria Perez,11 Lisde González,6 Luis Chirinos,12 Rafael Rosell13 1Medical Oncology, Clinical and Traslational Oncology Group, Institute of Oncology, Clínica Del Country, Bogota/Colombia, 2 Thoracic Oncology Unit and Laboratory of Personalized Medicine, Instituto Nacional de Cancerologia, Mexico City/Mexico, 3Centro Javeriano de Oncología, Hospital Universitario San Ignacio, Bogotá/Colombia, 4Clinical Oncology Department, Hospital San Juan de Dios (San José, Costa Rica), San Jose/Costa Rica, 5Internal Medicine Department, Johns Hopkins Hospital, Baltimore,
Journal of Thoracic Oncology
Vol. 12 No. 1S
Maryland/United States of America, 6Hospital Oncológico Luis Razetti, Caracas, Venezuela, Caracas/Venezuela, 7 Oncology Department, Hospital Roberto Calderón, Managua, Nicaragua, Managua/Nicaragua, 8Department of Clinical Oncology, Instituto Alexander Fleming, Buenos Aires/Argentina, 9Medical Oncology Department, Udelar (Montevideo, Uruguay), Montevideo/Uruguay, 10Medical Oncology, Instituto Nacional de Enfermedades Neoplasicas, Lima/Peru, 11Medical Oncology, Arsuve, Caracas, Venezuela, Caracas/Venezuela, 12Clínica de Prevención Del Cáncer, Caracas, Venezuela, Caracas/ Venezuela, 13Hospital Germans Trias I Pujol, Catalan Institute of Oncology, Barcelona/Spain Background: Nucleoside transporter proteins mediates the transport of nucleosides and nucleoside analog drugs across the plasma membrane. The human equilibrative nucleoside transporter 1 (hENT1) is a nucleoside transporter protein that mediates cellular entry of gemcitabine, cytarabine, and fludarabine. The hENT1 expression has been demonstrated to be related with prognosis and activity of gemcitabine-based therapy in breast, ampullary, lung, and pancreatic cancer. In the same way, RRM1 encodes the regulatory subunit of ribonucleotide reductase and is a molecular target of gemcitabine. Previous studies showed increased RRM1 expression on continuous exposure of cell lines to gemcitabine and suggested improved survival for patients with low RRM1 expression. Methods: We measured hENT1 and RRM1 messenger RNA (mRNA) levels by quantitative real-time reverse transcription-polymerase chain reaction in tumor samples from 29 patients with advanced pleural mesothelioma (APM) treated in second line with gemcitabine based chemotherapy correlating these data with clinical parameters and disease outcomes (overall response rate-ORR, progression free survival-PFS and overall survival-OS). Results: The median age was 60-yo (r, 33-70 years), 15 patients were males (51%), 34% of cases undergone debulking surgery and the median follow-up was 13.4 months (95% CI 6.4-34). All patients were treated with Pem based chemotherapy in first line achieving a PFS of 8.1 months (95% CI 3.7-12.6), while PFS with secondline Gem based chemotherapy was 6.3 months (95% CI 3.6-8.8). 62% and 34.5% of patients had low levels of mRNA for hENT1 and RRM1, respectively. On univariate survival analysis, the hENT1 expression was associated with OS (p¼0.001) and PFS (p¼0.022). Specifically, those patients with overexpression of hENT1 showed a shorter OS p¼0.021) and a shorter PFS (p¼0.033). In contrast, mRNA expression of RRM1 did not influence the OS (p ¼ 0.44) but it modifies positively the PFS (p¼0.034). Multivariate analysis found that combined
Anthony Linton,1 Matthew Soeberg,1 Steven Kao,2 Richard Broome,3 Nico Van Zandwijk1 1Asbestos Diseases Research Institute, Sydney/NSW/Australia, 2 Medical Oncology, Chris O’Brien Lifehouse, Sydney/ Australia, 3Public Health Observatory, Sydney/NSW/ Australia Background: Whilst the impact of clinico-pathological factors on the prognosis of malignant pleural mesothelioma (MPM) is well understood, socio-economic and geographic factors have received less attention. Although the majority of Australians reside in major cities, a dispersed population lives in regional and remote areas, where access to clinical services may be limited. We investigated the association between geographic and socio-economic factors and treatment provision and survival in a large series of patients from New South Wales. Methods: All patients awarded compensation by the NSW Dust Diseases Board (2002e2009) following diagnosis with MPM were assessed. Geographic remoteness, distance from oncological multidisciplinary teams (MDT) and socioeconomic status according to the index of relative socio-economic advantage and disadvantage (IRSAD), were assessed with known prognostic factors using Kaplan Meir and Cox-regression analysis. Chi-square testing compared categorical variables to analyse the impact of these factors upon clinical features and treatment received. Cancer Registry incidence data was assessed to allow comparison of the compensated DDB cohort to all NSW MPM cases. Results: We assessed 910 patients: Geographic remoteness (major city 67%; regional/remote 33%), distance to MDT (<10km 65%, <50km 92%). Geographic distribution was comparable to cancer registry data. Median overall survival was 10.0 months. On multivariate analysis, non-epithelioid histological subtype (HR.2.19); male gender (HR¼1.37); age >70 (HR¼1.39) and IRSAD status by decreasing quintile (HR¼1.07) were independent prognostic factors, with a pronounced survival difference between highest and lowest IRSAD quintiles (8.4 vs 12.8 months). A trend to improved survival when residing in major cities (10.6 vs 8.8 months; p¼0.162) and within 50km of MDT (10.3 vs 7.8 months;p¼0.539) was noted. Patients geographic location and distance to MDT affected the use of palliative radiotherapy (p<0.05) however did not impact chemotherapy, adjuvant radiotherapy or extrapleural pneumonectomy provision. Socioeconomically disadvantaged patients were less likely to receive chemotherapy (40.3% vs 47.7%; p¼0.032), with pronounced disparity between the most socioeconomically advantaged and disadvantaged quintiles (54.2% vs 37.6%;p¼0.001).
Abstracts
S1365
Conclusion: Despite ‘universal’ health care and the support of a compensation scheme, socioeconomic disadvantage was an independent prognostic factor for MPM in NSW Australia. A significant reduction in chemotherapy utilization was noted, particularly in highly socioeconomically deprived areas. Furthermore, a trend to improved survival was noted in patients residing in major cities within closer proximity to oncology units, though treatment provision did not differ. Prospective research analyzing specific factors including comorbidity, income, and individual preference will be required to better understand these findings in both compensated and non-compensated individuals. Keywords: Mesothelioma, Prognosis
P3.03-034 Comprehensive Immunophenotyping of the Blood and Pleural Fluid from Patients with Malignant Pleural Mesothelioma by Flow Cytometry Topic: Mesothelioma Clinical Tobias Peikert, Virginia Van Keulen, Svetlana Bornschlegl, Allen Dietz, Mike Gustafson Mayo Clinic, Rochester/MN/United States of America Background: Malignant pleural mesothelioma (MM) remains an almost universally fatal disease with limited treatment options. Immunotherapy represents a rapidly emerging therapeutic strategy for multiple malignancies including MM. However durable therapeutic responses occur in a minority (w30%) of patients. Better understanding of the quantities and immunophenotype of circulating and intrapleural immune cells is essential to design and apply personalized immunotherapeutic strategies. Methods: We used a comprehensive flow cytometry panel (PLoS One. 2015 Mar 23;10 (3):e0121546) to prospectively characterize circulating and pleural fluid immune cells in patients with MPM (n¼12) and normal volunteers (circulating cells only, n¼50). Matched blood and pleural samples were available from 11 patients, including samples from 9 patients enrolled into a Phase I study investigating the intrapleural administration of the modified vaccine strain measles virus (MV-NIS), MC1023. Pre- and post-treatments samples were available from 7 patients. The immune cell counts and cell fractions were compared using a false discovery rate (FDR) of 10% and the non-parametric Mann-Whitney test and the Wilcoxon matched-pairs signed rank test (blood versus pleural fluid and different time points) (p 0.05).
S1366
Results: Cell counts and immune phenotype of circulating immune cells differ between of patients with MPM differ from normal volunteers (31 of 86 cell types). Patients with MPM had fewer B-lymphocytes, more proinflammatory monocytes (CD14+CD16+) and exhausted CD4 and CD8 T-lymphocytes (CD4 and CD8 PD1+TIM3+ double positive cells). As expected we observed characteristic differences between blood and pleural fluid in MPM patients. For example there are a higher numbers of antigen experienced, PD-1 expressing CD4 and CD8memory cells within the pleural fluid compared to the peripheral blood. The intrapleural administration of a modified vaccine strain measles (MV-NIS) triggered changes in pleural and circulating immune cells. While these changes varied among patients, we observed increased CD80 and CD86 expression on CD14+ monocytes in the pleural fluid down. Conclusion: Comprehensive flow-cytometric immunophenotying of blood and pleural fluid is feasible in patients with MPM. This approach may help to identify patients who may respond favorably to specific immunotherapeutic interventions such as immune checkpoint inhibitors or facilitate longitudinal immune monitoring during clinical trials. Additional data is needed and we are continuing to prospectively analyze samples from patients with MPM. Keywords: flow cytometry, pleural fluid, Mesothelioma, immunophenotyping
P3.03-035 Prognostic Role of hENT1 and RRM1 in Patients with Advanced Pleural Mesothelioma Treated with Second Line Gemcitabine Based Regimens Topic: Mesothelioma Clinical Andrés Cardona,1 Oscar Arrieta,2 Leonardo Rojas,3 Luis Corrales,4 Beatriz Wills,5 George Oblitas,6 Ludwing Bacon,7 Claudio Martin,8 Mauricio Cuello,9 Luis Mas,10 Carlos Vargas,1 Hernan Carranza,1 Jorge Otero,1 Maria Perez,11 Lisde González,6 Luis Chirinos,12 Rafael Rosell13 1Medical Oncology, Clinical and Traslational Oncology Group, Institute of Oncology, Clínica Del Country, Bogota/Colombia, 2 Thoracic Oncology Unit and Laboratory of Personalized Medicine, Instituto Nacional de Cancerologia, Mexico City/Mexico, 3Centro Javeriano de Oncología, Hospital Universitario San Ignacio, Bogotá/Colombia, 4Clinical Oncology Department, Hospital San Juan de Dios (San José, Costa Rica), San Jose/Costa Rica, 5Internal Medicine Department, Johns Hopkins Hospital, Baltimore,
Journal of Thoracic Oncology
Vol. 12 No. 1S
Maryland/United States of America, 6Hospital Oncológico Luis Razetti, Caracas, Venezuela, Caracas/Venezuela, 7 Oncology Department, Hospital Roberto Calderón, Managua, Nicaragua, Managua/Nicaragua, 8Department of Clinical Oncology, Instituto Alexander Fleming, Buenos Aires/Argentina, 9Medical Oncology Department, Udelar (Montevideo, Uruguay), Montevideo/Uruguay, 10Medical Oncology, Instituto Nacional de Enfermedades Neoplasicas, Lima/Peru, 11Medical Oncology, Arsuve, Caracas, Venezuela, Caracas/Venezuela, 12Clínica de Prevención Del Cáncer, Caracas, Venezuela, Caracas/ Venezuela, 13Hospital Germans Trias I Pujol, Catalan Institute of Oncology, Barcelona/Spain Background: Nucleoside transporter proteins mediates the transport of nucleosides and nucleoside analog drugs across the plasma membrane. The human equilibrative nucleoside transporter 1 (hENT1) is a nucleoside transporter protein that mediates cellular entry of gemcitabine, cytarabine, and fludarabine. The hENT1 expression has been demonstrated to be related with prognosis and activity of gemcitabine-based therapy in breast, ampullary, lung, and pancreatic cancer. In the same way, RRM1 encodes the regulatory subunit of ribonucleotide reductase and is a molecular target of gemcitabine. Previous studies showed increased RRM1 expression on continuous exposure of cell lines to gemcitabine and suggested improved survival for patients with low RRM1 expression. Methods: We measured hENT1 and RRM1 messenger RNA (mRNA) levels by quantitative real-time reverse transcription-polymerase chain reaction in tumor samples from 29 patients with advanced pleural mesothelioma (APM) treated in second line with gemcitabine based chemotherapy correlating these data with clinical parameters and disease outcomes (overall response rate-ORR, progression free survival-PFS and overall survival-OS). Results: The median age was 60-yo (r, 33-70 years), 15 patients were males (51%), 34% of cases undergone debulking surgery and the median follow-up was 13.4 months (95% CI 6.4-34). All patients were treated with Pem based chemotherapy in first line achieving a PFS of 8.1 months (95% CI 3.7-12.6), while PFS with secondline Gem based chemotherapy was 6.3 months (95% CI 3.6-8.8). 62% and 34.5% of patients had low levels of mRNA for hENT1 and RRM1, respectively. On univariate survival analysis, the hENT1 expression was associated with OS (p¼0.001) and PFS (p¼0.022). Specifically, those patients with overexpression of hENT1 showed a shorter OS p¼0.021) and a shorter PFS (p¼0.033). In contrast, mRNA expression of RRM1 did not influence the OS (p ¼ 0.44) but it modifies positively the PFS (p¼0.034). Multivariate analysis found that combined