PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine

Background Anti-programmed cell death protein 1 and programmed cell death ligand 1 (PD-L1) agents are broadly used in first-line and second-line treatment across different tumor types. While immunohistochemistry-based assays are routinely used to assess PD-L1 expression, their clinical utility remai...

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Main Authors: Mariaelena Pierobon, Johann De Bono, Ruth Riisnaes, Giuseppe Giaccone, Matthew Blackburn, Lance Liotta, Vienna Ludovini, Neil J Shah, Giuseppina Improta, Antonella Ravaggi, Niven Mehra, Angelo Sidoni, Elisa Baldelli, K Alex Hodge, Guido Bellezza, Guido Gambara, Martina Mandarano, Chamodya Ruhunusiri, Bryant Dunetz, Maysa Abu-Khalaf, Julia Wulfkuhle, Rosa I Gallagher, Franco Odicino, Maria Isabella Sereni, Angela Zupa, Perry Demsko, Lucio Crino', Emanuel F Petricoin
Format: Article
Language:English
Published: BMJ Publishing Group 2021-10-01
Series:Journal for ImmunoTherapy of Cancer
Online Access:https://jitc.bmj.com/content/9/10/e002179.full
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author Mariaelena Pierobon
Johann De Bono
Ruth Riisnaes
Giuseppe Giaccone
Matthew Blackburn
Lance Liotta
Vienna Ludovini
Neil J Shah
Giuseppina Improta
Antonella Ravaggi
Niven Mehra
Angelo Sidoni
Elisa Baldelli
K Alex Hodge
Guido Bellezza
Guido Gambara
Martina Mandarano
Chamodya Ruhunusiri
Bryant Dunetz
Maysa Abu-Khalaf
Julia Wulfkuhle
Rosa I Gallagher
Franco Odicino
Maria Isabella Sereni
Angela Zupa
Perry Demsko
Lucio Crino'
Emanuel F Petricoin
author_facet Mariaelena Pierobon
Johann De Bono
Ruth Riisnaes
Giuseppe Giaccone
Matthew Blackburn
Lance Liotta
Vienna Ludovini
Neil J Shah
Giuseppina Improta
Antonella Ravaggi
Niven Mehra
Angelo Sidoni
Elisa Baldelli
K Alex Hodge
Guido Bellezza
Guido Gambara
Martina Mandarano
Chamodya Ruhunusiri
Bryant Dunetz
Maysa Abu-Khalaf
Julia Wulfkuhle
Rosa I Gallagher
Franco Odicino
Maria Isabella Sereni
Angela Zupa
Perry Demsko
Lucio Crino'
Emanuel F Petricoin
author_sort Mariaelena Pierobon
collection DOAJ
description Background Anti-programmed cell death protein 1 and programmed cell death ligand 1 (PD-L1) agents are broadly used in first-line and second-line treatment across different tumor types. While immunohistochemistry-based assays are routinely used to assess PD-L1 expression, their clinical utility remains controversial due to the partial predictive value and lack of standardized cut-offs across antibody clones. Using a high throughput immunoassay, the reverse phase protein microarray (RPPA), coupled with a fluorescence-based detection system, this study compared the performance of six anti-PD-L1 antibody clones on 666 tumor samples.Methods PD-L1 expression was measured using five antibody clones (22C3, 28–8, CAL10, E1L3N and SP142) and the therapeutic antibody atezolizumab on 222 lung, 71 ovarian, 52 prostate and 267 breast cancers, and 54 metastatic lesions. To capture clinically relevant variables, our cohort included frozen and formalin-fixed paraffin-embedded samples, surgical specimens and core needle biopsies. Pure tumor epithelia were isolated using laser capture microdissection from 602 samples. Correlation coefficients were calculated to assess concordance between antibody clones. For two independent cohorts of patients with lung cancer treated with nivolumab, RPPA-based PD-L1 measurements were examined along with response to treatment.Results Median-center PD-L1 dynamic ranged from 0.01 to 39.37 across antibody clones. Correlation coefficients between the six antibody clones were heterogeneous (range: −0.48 to 0.95) and below 0.50 in 61% of the comparisons. In nivolumab-treated patients, RPPA-based measurement identified a subgroup of tumors, where low PD-L1 expression equated to lack of response.Conclusions Continuous RPPA-based measurements capture a broad dynamic range of PD-L1 expression in human specimens and heterogeneous concordance levels between antibody clones. This high throughput immunoassay can potentially identify subgroups of tumors in which low expression of PD-L1 equates to lack of response to treatment.
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spelling doaj-art-b20bc5349766429ca47d7ba6932e22802025-08-20T03:04:47ZengBMJ Publishing GroupJournal for ImmunoTherapy of Cancer2051-14262021-10-0191010.1136/jitc-2020-002179PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicineMariaelena Pierobon0Johann De Bono1Ruth Riisnaes2Giuseppe Giaccone3Matthew Blackburn4Lance Liotta5Vienna Ludovini6Neil J Shah7Giuseppina Improta8Antonella Ravaggi9Niven Mehra10Angelo Sidoni11Elisa Baldelli12K Alex Hodge13Guido Bellezza14Guido Gambara15Martina Mandarano16Chamodya Ruhunusiri17Bryant Dunetz18Maysa Abu-Khalaf19Julia Wulfkuhle20Rosa I Gallagher21Franco Odicino22Maria Isabella Sereni23Angela Zupa24Perry Demsko25Lucio Crino'26Emanuel F Petricoin27School of Systems Biology, George Mason University, Manassas, Virginia, USAregius professor of cancer research and professor in experimental cancer medicineThe Institute of Cancer Research, London, UKDepartment of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, USADepartment of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USADivision of Medical Oncology, S. Maria della Misericordia Hospital, Perugia, ItalyDepartment of Oncology, Lombardi Comprehensive Cancer Center, Georgetown University, Washington, District of Columbia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USAAngelo Nocivelli Institute of Molecular Medicine, Division of Gynecologic Oncology, University of Brescia and ASST Spedali Civili di Brescia, Brescia, ItalyThe Institute of Cancer Research, London, UKDepartment of Experimental Medicine, Section of Anatomic Pathology and Histology, University of Perugia, Perugia, ItalyCenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USADepartment of Experimental Medicine, Section of Anatomic Pathology and Histology, University of Perugia, Perugia, ItalyCenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USADepartment of Experimental Medicine, Section of Anatomic Pathology and Histology, University of Perugia, Perugia, ItalyCenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USASide Out Foundation, Fairfax, Virginia, USADepartment of Medical Oncology, Sidney Kimmel Cancer Center at Jefferson Health, Thomas Jefferson University, Philadelphia, Pennsylvania, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USADepartment of Clinical and Experimental Sciences, University of Brescia, Brescia, ItalyCenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USACenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USADepartment of Medical Oncology, Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) IRCCS, Meldola, ItalyCenter for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia, USABackground Anti-programmed cell death protein 1 and programmed cell death ligand 1 (PD-L1) agents are broadly used in first-line and second-line treatment across different tumor types. While immunohistochemistry-based assays are routinely used to assess PD-L1 expression, their clinical utility remains controversial due to the partial predictive value and lack of standardized cut-offs across antibody clones. Using a high throughput immunoassay, the reverse phase protein microarray (RPPA), coupled with a fluorescence-based detection system, this study compared the performance of six anti-PD-L1 antibody clones on 666 tumor samples.Methods PD-L1 expression was measured using five antibody clones (22C3, 28–8, CAL10, E1L3N and SP142) and the therapeutic antibody atezolizumab on 222 lung, 71 ovarian, 52 prostate and 267 breast cancers, and 54 metastatic lesions. To capture clinically relevant variables, our cohort included frozen and formalin-fixed paraffin-embedded samples, surgical specimens and core needle biopsies. Pure tumor epithelia were isolated using laser capture microdissection from 602 samples. Correlation coefficients were calculated to assess concordance between antibody clones. For two independent cohorts of patients with lung cancer treated with nivolumab, RPPA-based PD-L1 measurements were examined along with response to treatment.Results Median-center PD-L1 dynamic ranged from 0.01 to 39.37 across antibody clones. Correlation coefficients between the six antibody clones were heterogeneous (range: −0.48 to 0.95) and below 0.50 in 61% of the comparisons. In nivolumab-treated patients, RPPA-based measurement identified a subgroup of tumors, where low PD-L1 expression equated to lack of response.Conclusions Continuous RPPA-based measurements capture a broad dynamic range of PD-L1 expression in human specimens and heterogeneous concordance levels between antibody clones. This high throughput immunoassay can potentially identify subgroups of tumors in which low expression of PD-L1 equates to lack of response to treatment.https://jitc.bmj.com/content/9/10/e002179.full
spellingShingle Mariaelena Pierobon
Johann De Bono
Ruth Riisnaes
Giuseppe Giaccone
Matthew Blackburn
Lance Liotta
Vienna Ludovini
Neil J Shah
Giuseppina Improta
Antonella Ravaggi
Niven Mehra
Angelo Sidoni
Elisa Baldelli
K Alex Hodge
Guido Bellezza
Guido Gambara
Martina Mandarano
Chamodya Ruhunusiri
Bryant Dunetz
Maysa Abu-Khalaf
Julia Wulfkuhle
Rosa I Gallagher
Franco Odicino
Maria Isabella Sereni
Angela Zupa
Perry Demsko
Lucio Crino'
Emanuel F Petricoin
PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
Journal for ImmunoTherapy of Cancer
title PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
title_full PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
title_fullStr PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
title_full_unstemmed PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
title_short PD-L1 quantification across tumor types using the reverse phase protein microarray: implications for precision medicine
title_sort pd l1 quantification across tumor types using the reverse phase protein microarray implications for precision medicine
url https://jitc.bmj.com/content/9/10/e002179.full
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