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P2.02-047 Comparison of PD-L1 Immunohistochemistry Assays and Response to PD-L1 Inhibitors
S2116
Journal of Thoracic Oncology
Vol. 12 No. 11S2
into lung cancer pathogenesis, and immunotherapeutic strategies targeting this important immune checkpoint protein. The aim was to investigate the correlation between multiplex immunofluorescence (mIF) expression of PD-L1, density and nature of tumor infiltrating immune cells in non-small cell lung carcinomas (NSCLC), and correlate those profiles with clinical and pathological variables including patient outcome. Method: We studied 194 stage II/III patients that underwent pulmonary resection, including 98 adenocarcinoma (ADC), 59 squamous cell carcinoma (SqCC), 15 large cells carcinomas (LCC) and 22 neuroendocrine carcinomas (NEC), primary tumors. Formalin-fixed and paraffin embedded (FFPE) tissue microarrays were constructed with five 1.5 mm cores representative of histologic patterns found in each tumor. mIF was performed using the Opal 7-color fIHC KitTM, scanning in the VectraTM multispectral microscope and analyzed using the inFormTM software (Perkin Elmer, Waltham, MA). The markers studied were grouped in two 6-antibody panels: Panel 1, AE1/AE3 pancytokeratins, PD-L1 (clone E1L3N), PD1, CD3, CD8 and CD68; and Panel 2, AE1/AE3, Granzyme B, CD45RO and CD57, FOXP3, and CD20. General linear model was used to evaluate the interaction among primary vs metastatic tumors, histologic type and TAICs and Cox’s proportional hazard model for overall survival (OS). Result: Fifty-eight % out of 164 tumors were positive for PDL-1+ expression (5% cut-off) in malignant cells (EA1/EA3+). Significant higher levels of PD-L1+ expression were detected in NEC compared with other histologies (ADC, SqCC and LCC) (P¼0.006). In the same way, we observed higher densities of cytotoxic T lymphocytes (CD3+CD8+) in NEC when compared with the lowest expression in SqCC (P¼0.02). Large cell carcinomas presented high levels of memory/regulatory T cells (CD3+FOXP3+CD45RO+) compared with other histologic types but the difference didn’t achieve statistical significance. No difference was found for CD3+PD-L1+, CD68+PD-L1+, natural killer T lymphocytes (CD3+CD57+) and B lymphocytes (CD20+) among the histologic types. Difference between primary and metastatic tumors was found only for naive/memory T lymphocytes (CD3+ CD45RO+) (P¼0.04). High CD3+FOXP3+CD45RO+ and CD3+PDL1+ expression were independent favorable prognostic factor for DFS and OS adjusted by smoking, primary vs metastatic, and histologic type [HR 2.68, 95% (CI 1.37e5.24), P¼0.004; HR 2.11 (CI 1.07-4.18, P¼0.03]. Conclusion: High abundance of CD3+PD-L1+ cells and memory/regulatory T cells CD3+FOXP3+CD54RO are favorable prognostic factors for resected NSCLC, highlighting the importance of comprehensive assessment of both tumor and immune cells. Supported by CNPQ P246042/2012-5 e CNPQ 301411/2016-6; FAPESP 2013/10113-7. Keywords: immune cells, lung cancer, PD-L1
and to investigate the association between PD-L1 assays and response to PD-L1 inhibitors. Method: 189 sequential lung tumor samples from patients with advanced NSCLC were prospectively stained with Ventana SP263 and Dako 22C3 clones. Both antibodies were optimized for use on the automated Ventana BenchMark ULTRA platform, and validated against corresponding FDA approved assays. Scoring algorithms for staining of the tumor cells approved by matched FDA assays were applied to all samples. Best overall response (BOR) for 43 patients treated with either nivolumab or pembrolizumab were assessed using RECIST v.1.1 and correlated with PD-L1 expression with SP263 and 22C3 using cut off points of 1%, 5% and 10%, 1-49% and 50%. Result: Ventana SP263 and Dako 22C3 LDTs showed good agreement with a concordance correlation coefficient of 0.86 (95% CI 0.82-0.90). Comparing the assays using the cutoffs of 1%, 5% or 10% for SP263 and the two cutoffs of 1% and 50% for 22C3 showed an association between the two assays as well (p<0.0001). There were no differences in BOR between pembrolizumab and nivolumab (p¼0.12). For SP263, BOR was associated with a cutoff point of 10% (Fisher’s p¼0.030); while the 1% (Fisher’s p¼0.09)and 5 % (Fisher’s p¼0.054) cut off points were not associated with response. In contrast, for 22C3, BOR was associated with a cutoff point of 1% (Fisher’s exact test p-value ¼ 0.006), 5% (p 0.006) and 10% (0.006) Conclusion: Similar to FDA approved assays, Ventana SP263 and Dako 22C3 LDT, if properly validated, demonstrate good concordance, but are not interchangeable. Dako 22C3 is more sensitive assay and its expression shows better association with therapeutic response. Larger studies evaluating associations of alternative staining assays and a response to specific therapy are needed. Keywords: PD-L1 assay response
P2.02-047 Comparison of PD-L1 Immunohistochemistry Assays and Response to PD-L1 Inhibitors
Background: Tumor suppressor gene TP53 mutation is common in human cancers, especially playing an important role in lung cancer tumor genesis. Some clinical studies have shown that TP53 alterations in non-small cell lung carcinoma (NSCLC) carry a worse prognosis and may relatively more resistant to chemotherapy and radiation. We conducted this study to evaluate the impact of TP53 assessed by limited targeted profiling, correlating with PD-L1 tumor expression and clinicopathological variables in resected NSCLC. Method: NSCLC patients who underwent curative resection between 1998 and 2006 at our institution were included. PD-L1 status was assessed using Ventana SP142 antibody on archival FFPE surgical tumor specimens, arrayed on tissue microarrays (TMAs) with triplicate 0.6 mm cores. PD-L1 was scored as positive if membranous staining was present in >1% of tumor cells aggregated across the replicate cores to address heterogeneity. In collaboration with the Lung Cancer Genomics Ireland Study, a targeted panel of 49 genes was assessed by Sequenom MassArray including TP53 and genes in MAPK and PI3K pathways. Clinicopathological data was obtained from
S. Dacic,1 K. Ancevski,2 S. Aberrbock,2 C. Herbst,1 L. Villaruz2 1 Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA/US, 2 Hematology Oncology, University of Pittsburgh Hillman Cancer Center, Pittsburgh, PA/US Background: The FDA has approved different companion and complementary PD-L1 assays for selection of patients for different PD-L1 inhibitors. As a result, laboratories intending to implement FDA approved assays face significant operating and capital expenses. Several studies have demonstrated technical concordance between different PD-L1 assays, but their clinical validity in terms of the response to treatment has not entirely been explored. The aim of our study is to prospectively assess the staining performance of laboratory developed tests (LDT) for Ventana SP263 (nivolumab) and Dako 22C3 (pembrolizumab) clones on formalin fixed paraffin embedded samples,
P2.02-048 Survival Correlation Between TP53 Gene and PD-L1 Tumor Expression in Resected Non-Small Cell Lung Carcinoma J. Sui,1 M.Y. Teo,2 S. Twomey,3 S. Rafee,1 J. Mcfadden,4 K. Gately,5 M. Barr,5 S. Gray,5 B. Hennessy,6 K. O’Byrne,7 S. Cuffe,1 S. Finn8 1 Medical Oncology, St. James’s Hospital, Dublin/IE, 2Memorial Sloan Kettering, New York, NY/US, 3Department of Molecular Medicine, Rcsi, Beaumont Hospital, Dublin/IE, 4Department of Histopathology, St. James’s Hospital, Dublin/IE, 5Trinity Translational Medicine Institute, Trinity Centre of Health Science, St. James’s Hospital, Dublin/IE, 6 Department of Medical Oncology, Beaumont Hospital, Dublin/IE, 7 Princess Alexandra Hospital and Queensland University of Technology, Brisbane/AU, 8Cancer Molecular Diagnostics, St James’s Hospital, Dublin/IE
Journal of Thoracic Oncology
Vol. 12 No. 11S2
into lung cancer pathogenesis, and immunotherapeutic strategies targeting this important immune checkpoint protein. The aim was to investigate the correlation between multiplex immunofluorescence (mIF) expression of PD-L1, density and nature of tumor infiltrating immune cells in non-small cell lung carcinomas (NSCLC), and correlate those profiles with clinical and pathological variables including patient outcome. Method: We studied 194 stage II/III patients that underwent pulmonary resection, including 98 adenocarcinoma (ADC), 59 squamous cell carcinoma (SqCC), 15 large cells carcinomas (LCC) and 22 neuroendocrine carcinomas (NEC), primary tumors. Formalin-fixed and paraffin embedded (FFPE) tissue microarrays were constructed with five 1.5 mm cores representative of histologic patterns found in each tumor. mIF was performed using the Opal 7-color fIHC KitTM, scanning in the VectraTM multispectral microscope and analyzed using the inFormTM software (Perkin Elmer, Waltham, MA). The markers studied were grouped in two 6-antibody panels: Panel 1, AE1/AE3 pancytokeratins, PD-L1 (clone E1L3N), PD1, CD3, CD8 and CD68; and Panel 2, AE1/AE3, Granzyme B, CD45RO and CD57, FOXP3, and CD20. General linear model was used to evaluate the interaction among primary vs metastatic tumors, histologic type and TAICs and Cox’s proportional hazard model for overall survival (OS). Result: Fifty-eight % out of 164 tumors were positive for PDL-1+ expression (5% cut-off) in malignant cells (EA1/EA3+). Significant higher levels of PD-L1+ expression were detected in NEC compared with other histologies (ADC, SqCC and LCC) (P¼0.006). In the same way, we observed higher densities of cytotoxic T lymphocytes (CD3+CD8+) in NEC when compared with the lowest expression in SqCC (P¼0.02). Large cell carcinomas presented high levels of memory/regulatory T cells (CD3+FOXP3+CD45RO+) compared with other histologic types but the difference didn’t achieve statistical significance. No difference was found for CD3+PD-L1+, CD68+PD-L1+, natural killer T lymphocytes (CD3+CD57+) and B lymphocytes (CD20+) among the histologic types. Difference between primary and metastatic tumors was found only for naive/memory T lymphocytes (CD3+ CD45RO+) (P¼0.04). High CD3+FOXP3+CD45RO+ and CD3+PDL1+ expression were independent favorable prognostic factor for DFS and OS adjusted by smoking, primary vs metastatic, and histologic type [HR 2.68, 95% (CI 1.37e5.24), P¼0.004; HR 2.11 (CI 1.07-4.18, P¼0.03]. Conclusion: High abundance of CD3+PD-L1+ cells and memory/regulatory T cells CD3+FOXP3+CD54RO are favorable prognostic factors for resected NSCLC, highlighting the importance of comprehensive assessment of both tumor and immune cells. Supported by CNPQ P246042/2012-5 e CNPQ 301411/2016-6; FAPESP 2013/10113-7. Keywords: immune cells, lung cancer, PD-L1
and to investigate the association between PD-L1 assays and response to PD-L1 inhibitors. Method: 189 sequential lung tumor samples from patients with advanced NSCLC were prospectively stained with Ventana SP263 and Dako 22C3 clones. Both antibodies were optimized for use on the automated Ventana BenchMark ULTRA platform, and validated against corresponding FDA approved assays. Scoring algorithms for staining of the tumor cells approved by matched FDA assays were applied to all samples. Best overall response (BOR) for 43 patients treated with either nivolumab or pembrolizumab were assessed using RECIST v.1.1 and correlated with PD-L1 expression with SP263 and 22C3 using cut off points of 1%, 5% and 10%, 1-49% and 50%. Result: Ventana SP263 and Dako 22C3 LDTs showed good agreement with a concordance correlation coefficient of 0.86 (95% CI 0.82-0.90). Comparing the assays using the cutoffs of 1%, 5% or 10% for SP263 and the two cutoffs of 1% and 50% for 22C3 showed an association between the two assays as well (p<0.0001). There were no differences in BOR between pembrolizumab and nivolumab (p¼0.12). For SP263, BOR was associated with a cutoff point of 10% (Fisher’s p¼0.030); while the 1% (Fisher’s p¼0.09)and 5 % (Fisher’s p¼0.054) cut off points were not associated with response. In contrast, for 22C3, BOR was associated with a cutoff point of 1% (Fisher’s exact test p-value ¼ 0.006), 5% (p 0.006) and 10% (0.006) Conclusion: Similar to FDA approved assays, Ventana SP263 and Dako 22C3 LDT, if properly validated, demonstrate good concordance, but are not interchangeable. Dako 22C3 is more sensitive assay and its expression shows better association with therapeutic response. Larger studies evaluating associations of alternative staining assays and a response to specific therapy are needed. Keywords: PD-L1 assay response
P2.02-047 Comparison of PD-L1 Immunohistochemistry Assays and Response to PD-L1 Inhibitors
Background: Tumor suppressor gene TP53 mutation is common in human cancers, especially playing an important role in lung cancer tumor genesis. Some clinical studies have shown that TP53 alterations in non-small cell lung carcinoma (NSCLC) carry a worse prognosis and may relatively more resistant to chemotherapy and radiation. We conducted this study to evaluate the impact of TP53 assessed by limited targeted profiling, correlating with PD-L1 tumor expression and clinicopathological variables in resected NSCLC. Method: NSCLC patients who underwent curative resection between 1998 and 2006 at our institution were included. PD-L1 status was assessed using Ventana SP142 antibody on archival FFPE surgical tumor specimens, arrayed on tissue microarrays (TMAs) with triplicate 0.6 mm cores. PD-L1 was scored as positive if membranous staining was present in >1% of tumor cells aggregated across the replicate cores to address heterogeneity. In collaboration with the Lung Cancer Genomics Ireland Study, a targeted panel of 49 genes was assessed by Sequenom MassArray including TP53 and genes in MAPK and PI3K pathways. Clinicopathological data was obtained from
S. Dacic,1 K. Ancevski,2 S. Aberrbock,2 C. Herbst,1 L. Villaruz2 1 Pathology, University of Pittsburgh Medical Center, Pittsburgh, PA/US, 2 Hematology Oncology, University of Pittsburgh Hillman Cancer Center, Pittsburgh, PA/US Background: The FDA has approved different companion and complementary PD-L1 assays for selection of patients for different PD-L1 inhibitors. As a result, laboratories intending to implement FDA approved assays face significant operating and capital expenses. Several studies have demonstrated technical concordance between different PD-L1 assays, but their clinical validity in terms of the response to treatment has not entirely been explored. The aim of our study is to prospectively assess the staining performance of laboratory developed tests (LDT) for Ventana SP263 (nivolumab) and Dako 22C3 (pembrolizumab) clones on formalin fixed paraffin embedded samples,
P2.02-048 Survival Correlation Between TP53 Gene and PD-L1 Tumor Expression in Resected Non-Small Cell Lung Carcinoma J. Sui,1 M.Y. Teo,2 S. Twomey,3 S. Rafee,1 J. Mcfadden,4 K. Gately,5 M. Barr,5 S. Gray,5 B. Hennessy,6 K. O’Byrne,7 S. Cuffe,1 S. Finn8 1 Medical Oncology, St. James’s Hospital, Dublin/IE, 2Memorial Sloan Kettering, New York, NY/US, 3Department of Molecular Medicine, Rcsi, Beaumont Hospital, Dublin/IE, 4Department of Histopathology, St. James’s Hospital, Dublin/IE, 5Trinity Translational Medicine Institute, Trinity Centre of Health Science, St. James’s Hospital, Dublin/IE, 6 Department of Medical Oncology, Beaumont Hospital, Dublin/IE, 7 Princess Alexandra Hospital and Queensland University of Technology, Brisbane/AU, 8Cancer Molecular Diagnostics, St James’s Hospital, Dublin/IE