Development of a programmed cell death ligand-1 immunohistochemical assay validated for analysis of non-small cell lung cancer and head and neck squamous cell carcinoma

BackgroundA high-quality programmed cell-death ligand 1 (PD-L1) diagnostic assay may help predict which patients are more likely to respond to anti-programmed cell death-1 (PD-1)/PD-L1 antibody-based cancer therapy. Here we describe a PD-L1 immunohistochemical (IHC) staining protocol developed by Ventana Medical Systems Inc. and key analytical parameters of its use in formalin-fixed, paraffin-embedded (FFPE) samples of non-small cell lung cancer (NSCLC) and head and neck squamous cell carcinoma (HNSCC).MethodsAn anti-human PD-L1 rabbit monoclonal antibody (SP263) was optimized for use with the VENTANA OptiView DAB IHC Detection Kit on the automated VENTANA BenchMark ULTRA platform. The VENTANA PD-L1 (SP263) Assay was validated for use with FFPE NSCLC and HNSCC tissue samples in a series of studies addressing sensitivity, specificity, robustness, and precision. Samples from a subset of 181 patients from a Phase 1/2 study of durvalumab (NCT01693562) were analyzed to determine the optimal PD-L1 staining cut-off for enriching the probability of responses to treatment. The scoring algorithm was defined using statistical analysis of clinical response data from this clinical trial and PD-L1 staining parameters in HNSCC and NSCLC tissue. Inter-reader agreement was established by three pathologists who evaluated 81 NSCLC and 100 HNSCC samples across the range of PD-L1 expression levels.ResultsThe VENTANA PD-L1 (SP263) Assay met all pre-defined acceptance criteria. For both cancer types, a cut-off of 25 % of tumor cells with PD-L1 membrane staining of any intensity best discriminated responders from nonresponders. Samples with staining above this value were deemed to have high PD-L1 expression, and those with staining below it were deemed to have low or no PD-L1 expression. Inter-reader agreement on PD-L1 status was 97 and 92 % for NSCLC and HNSCC, respectively.ConclusionsThese results highlight the robustness and reproducibility of the VENTANA PD-L1 (SP263) Assay and support its suitability for use in the evaluation of NSCLC and HNSCC FFPE tumor samples using the devised ≥25 % tumor cell staining cut-off in a clinical setting. The clinical utility of the PD-L1 diagnostic assay as a predictive biomarker will be further validated in ongoing durvalumab studies.Trial registrationClinicalTrials.gov: NCT01693562

[1]  S. Gettinger,et al.  First-Line Monotherapy With Nivolumab (Anti-PD-1; BMS-936558, ONO-4538) in Advanced Non-Small Cell Lung Cancer (NSCLC): Safety, Efficacy, and Correlation of Outcomes With PD-L1 Status , 2014 .

[2]  Sofamor Danek,et al.  SUMMARY OF SAFETY AND EFFECTIVENESS DATA (SSED) , 2004 .

[3]  Yih-Leong Chang,et al.  Programmed cell death-ligand 1 expression in surgically resected stage I pulmonary adenocarcinoma and its correlation with driver mutations and clinical outcomes. , 2014, European journal of cancer.

[4]  S Senan,et al.  Metastatic non-small cell lung cancer: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. , 2018, Annals of oncology : official journal of the European Society for Medical Oncology.

[5]  M. Ahn,et al.  Association Between PD-L1 and HPV Status and the Prognostic Value of PD-L1 in Oropharyngeal Squamous Cell Carcinoma , 2015, Cancer research and treatment : official journal of Korean Cancer Association.

[6]  F. Hirsch,et al.  Programmed Death Ligand-1 Immunohistochemistry: Friend or Foe? , 2016, Archives of pathology & laboratory medicine.

[7]  G. Freeman,et al.  PD-L1 and PD-L2 modulate airway inflammation and iNKT-cell-dependent airway hyperreactivity in opposing directions , 2010, Mucosal Immunology.

[8]  Identification and Characterization of MEDI4736, an Antagonistic Anti–PD-L1 Monoclonal Antibody , 2015, Cancer Immunology Research.

[9]  Ming-Sound Tsao,et al.  Programmed Death-Ligand 1 Immunohistochemistry in Lung Cancer: In what state is this art? , 2015, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[10]  S. Ou,et al.  Safety and efficacy of MEDI4736, an anti-PD-L1 antibody, in patients from a squamous cell carcinoma of the head and neck (SCCHN) expansion cohort. , 2015 .

[11]  J. Cheville,et al.  B7-H1 blockade augments adoptive T-cell immunotherapy for squamous cell carcinoma. , 2003, Cancer research.

[12]  R. Herbst,et al.  Overview of the efficacy of cetuximab in recurrent and/or metastatic squamous cell carcinoma of the head and neck in patients who previously failed platinum‐based therapies , 2008, Cancer.

[13]  A. Tarhini.  Tremelimumab: a review of development to date in solid tumors. , 2013, Immunotherapy.

[14]  David C. Smith,et al.  Overall Survival and Long-Term Safety of Nivolumab (Anti-Programmed Death 1 Antibody, BMS-936558, ONO-4538) in Patients With Previously Treated Advanced Non-Small-Cell Lung Cancer. , 2015, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[15]  J. Lunceford,et al.  Pembrolizumab for the treatment of non-small-cell lung cancer. , 2015, The New England journal of medicine.

[16]  H. Kohrt,et al.  Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients , 2014, Nature.

[17]  Patrick Schöffski,et al.  Safety and clinical activity of MEDI4736, an anti-programmed cell death-ligand 1 (PD-L1) antibody, in patients with non-small cell lung cancer (NSCLC). , 2015 .

[18]  J. Taube,et al.  Evidence for a role of the PD-1:PD-L1 pathway in immune resistance of HPV-associated head and neck squamous cell carcinoma. , 2013, Cancer research.

[19]  G. Freeman,et al.  Blockade of Programmed Death-1 Ligands on Dendritic Cells Enhances T Cell Activation and Cytokine Production 1 , 2003, The Journal of Immunology.

[20]  Jill Walker,et al.  Abstract LB-094: A comparative study of PD-L1 diagnostic assays and the classification of patients as PD-L1 positive and PD-L1 negative , 2016 .

[21]  Ping Yang,et al.  Tumor B7-H1 and B7-H3 expression in squamous cell carcinoma of the lung. , 2013, Clinical lung cancer.

[22]  S. Husby,et al.  Alternative splice variants of the human PD-1 gene. , 2005, Cellular immunology.

[23]  A. Kulkarni,et al.  PD-1 blockade attenuates immunosuppressive myeloid cells due to inhibition of CD47/SIRPα axis in HPV negative head and neck squamous cell carcinoma , 2015, Oncotarget.

[24]  G. Freeman,et al.  Association of PD-L1 expression on tumor-infiltrating mononuclear cells and overall survival in patients with urothelial carcinoma. , 2015, Annals of oncology : official journal of the European Society for Medical Oncology.

[25]  J. Madore,et al.  PD-L1 expression is a favorable prognostic factor in early stage non-small cell carcinoma. , 2015, Lung cancer.

[26]  D. Siegmund,et al.  Maximally Selected Chi Square Statistics , 1982 .

[27]  D. Pardoll,et al.  B7-DC induced by IL-13 works as a feedback regulator in the effector phase of allergic asthma. , 2008, Biochemical and biophysical research communications.

[28]  S. Gettinger,et al.  Safety and Efficacy of First-Line Nivolumab (Anti-PD-1; BMS-936558, ONO-4538) and Ipilimumab in Non-Small Cell Lung Cancer (NSCLC) , 2014 .

[29]  R. Herbst,et al.  Programmed death ligand-1 expression in non-small cell lung cancer , 2014, Laboratory Investigation.

[30]  Xiaoling Zhang,et al.  Development of an Automated PD-L1 Immunohistochemistry (IHC) Assay for Non–Small Cell Lung Cancer , 2015, Applied immunohistochemistry & molecular morphology : AIMM.

[31]  Keunchil Park,et al.  Atezolizumab versus docetaxel for patients with previously treated non-small-cell lung cancer (POPLAR): a multicentre, open-label, phase 2 randomised controlled trial , 2016, The Lancet.

[32]  K. Kelly,et al.  3090 Avelumab (MSB0010718C), an anti-PD-L1 antibody, in patients with metastatic or recurrent non-small-cell lung cancer progressing after platinum-based chemotherapy: A phase Ib trial , 2015 .

[33]  Y. Shentu,et al.  Pembrolizumab versus docetaxel for previously treated, PD-L1-positive, advanced non-small-cell lung cancer (KEYNOTE-010): a randomised controlled trial , 2016, The Lancet.

[34]  Drew M. Pardoll,et al.  The blockade of immune checkpoints in cancer immunotherapy , 2012, Nature Reviews Cancer.

[35]  J. Wolchok,et al.  Immune Checkpoint Blockade in Cancer Therapy. , 2015, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[36]  F. Khuri,et al.  Advances in the management of recurrent or metastatic squamous cell carcinoma of the head and neck , 2013, Head & neck.

[37]  G. Freeman,et al.  PD-1 and its ligands in tolerance and immunity. , 2008, Annual review of immunology.

[38]  Xian-hui He,et al.  Identification of a novel splice variant of human PD-L1 mRNA encoding an isoform-lacking Igv-like domain , 2005, Acta Pharmacologica Sinica.

[39]  J. T. Jørgensen.  Companion diagnostic assays for PD-1/PD-L1 checkpoint inhibitors in NSCLC , 2016, Expert review of molecular diagnostics.

[40]  W. Youden,et al.  Index for rating diagnostic tests , 1950, Cancer.

[41]  Marius Ilie,et al.  Assessment of the PD-L1 status by immunohistochemistry: challenges and perspectives for therapeutic strategies in lung cancer patients , 2016, Virchows Archiv.

[42]  E. Wardelmann,et al.  PD-1 and PD-L1 Expression in NSCLC Indicate a Favorable Prognosis in Defined Subgroups , 2015, PloS one.

[43]  K. Steele,et al.  Development of a PD-L1 companion diagnostic assay for treatment with MEDI4736 in NSCLC and SCCHN patients. , 2015 .

[44]  Lieping Chen,et al.  Inhibitory B7-family molecules in the tumour microenvironment , 2008, Nature Reviews Immunology.

[45]  D. Pardoll,et al.  B7-DC regulates asthmatic response by an IFN-gamma-dependent mechanism. , 2004, Journal of immunology.

[46]  Matteo Brunelli,et al.  Differential Activity of Nivolumab, Pembrolizumab and MPDL3280A according to the Tumor Expression of Programmed Death-Ligand-1 (PD-L1): Sensitivity Analysis of Trials in Melanoma, Lung and Genitourinary Cancers , 2015, PloS one.

[47]  M. Dolled-Filhart,et al.  Safety and Clinical Activity of MK-3475 as Initial Therapy in Patients With Advanced Non-Small Cell Lung Cancer (NSCLC) , 2014 .

[48]  G. Freeman,et al.  PD-L1 Antibodies to Its Cytoplasmic Domain Most Clearly Delineate Cell Membranes in Immunohistochemical Staining of Tumor Cells , 2015, Cancer Immunology Research.

[49]  F. Cappuzzo,et al.  Activity and safety of nivolumab, an anti-PD-1 immune checkpoint inhibitor, for patients with advanced, refractory squamous non-small-cell lung cancer (CheckMate 063): a phase 2, single-arm trial. , 2015, The Lancet. Oncology.