Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Vernieuwing en diagnostiek bij NSCLC:

Immunotherapy: PD-L1 analyse: waar staan we

Wim Timens Professor and Chair Dept. Pathology and Medical Biology, University Medical Center Groningen

“9e avondsymposium: "Nieuwe ontwikkelingen in de behandeling van NSCLC" 9 november 2016, UMCG

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Immunopathology in lung cancer immune checkpoint diagnostics

• PD-L1 testing

• Other immune checkpoints

• New/other checkpoint (companion) diagnostics?

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Pardoll, Nature Rev Cancer 2012

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Immune checkpoint blockade

Drake, C. G. et al. (2013) Nat. Rev. Clin. Oncol.

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

PD-L1 testing: present state and problems

From the FDA-AACR-ASCO Public Workshop, March 24, 2015, (Reena Philip, Ph.D.):

• Why Companion Diagnostics: Companion Diagnostics are those tests that provides information that is essential for the safe and effective use of a corresponding drug or biological product.

• Issues - The Case of PD-L1

• Complications

• First results (2016)

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Agent Nivolumab Pembrolizumab Durvalumab Atezolizumab

Antibody Dako 28-8 Ventana SP263

Dako 22C3 Ventana SP263

Ventana SP142

Cut-off(s) Tested (NSCLC)

1%, 5% or 10% (TC)

≥ 25% TC TC ≥ 50% (and 1% any stroma)

≥ 25% TC

TC 1%, 5%, 50% IC 1%, 5%, 10%

Analytically validated assays used in clinical studies

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Issues - The Case of PD-L1

• Performance of each IHC antibody optimized for a particular protocol and platform

• Is the sensitivity and specificity between clones the same?

• Is the reactivity in tumor cells and TILs the same?

• Can laboratories apply one protocol to the same clone for all uses?

• Can laboratories adequately assess concordance with an adequate number of specimens?

Rx/Dx Industry PD-L1 Blueprint Proposal

FDA-AACR-ASCO Public Workshop 24 March 2015

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Complications

• Running a different test for every drug is impractical

– Limited tumor tissue

– Turnaround time

• Using one test for every drug is equally impractical

– All tests will not run on all platforms

– Each test has different performance characteristics

– Scoring and interpretation guidelines are not harmonized

– Each drug may have different clinical response based on biologic, chemistry and MOA differences

• There is potential for harm to patients if:

– FDA-approved IVD’s and drugs are cross-matched by end users in the absence of FDA reviewed and approved claims of clinical and analytical concordance.

Rx/Dx Industry PD-L1 Blueprint Proposal

FDA-AACR-ASCO Public Workshop 24 March 2015

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

PD-L1 immunohistochemistry comparison and harmonisation

• Blueprint project (22C3, 28-8, SP142, SP263, cf validation protocol: on DAKO Link Autostainer AS-48 and Ventana Benchmark Ultra) n=39, path=3

• Ratcliffe study (SP263, 28-8, 22C3 cf validation protocol) n=500? 81? Path=?

• Scheel et al., Mod. Pathol 2016 (22C3, 28-8, SP142, SP263, cf validation protocol; SP142 and E1L3N lab devel. assay on Leica) n=15, path=9

Hirsch, Blueprint, AACR 2016

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

PD-L1 IHC variation

Different PD-L1 clones on consecutive sections of one tumor

Scheel et al. Modern Pathol 2016

Hirsch, Blueprint,

AACR 2016

Hirsch, Blueprint, AACR 2016

Hirsch, Blueprint, AACR 2016

Hirsch, Blueprint, AACR 2016

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Conclusions Astra-Zeneca study (Ratcliffe)

• overall percent agreement (OPA) of 96% between Dako 28-8 at the 10% cut-off and the Ventana SP263 assay (positive percent agreement [PPA]; 91%, negative percent agreement [NPA]; 98%).

• similar results between SP263 and 22C3 tests at the 50% cut-off mark.

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Results/Conclusions Scheel-study

• The assays 28-8 and 22C3 stained similar proportions of carcinoma cells in 12 of 15 cases.

• SP142 stained fewer carcinoma cells compared to 28-8, 22C3, and SP263 in four cases, whereas SP263 stained more carcinoma cells in nine cases.

• SP142 and SP263 stained immune cells more intensely.

• The data indicate that carcinoma cells can be reproducibly scored in PD-L1 immunohistochemistry for pulmonary adenocarcinoma and squamous-cell carcinoma.

• No differences in interobserver concordance were noticed among the tested assays.

• The scoring of immune cells yielded low concordance rates and might require specific standardization.

Scheel et al. Modern Pathol 2016

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Key Points

• Question: Do the 4 drug-matched companion diagnostic antibodies for PD-1 axis therapies all produce the same results?

• Findings: In this study, 3 key components of diagnostic tests were assessed: the primary antibody, assay-specific variables related to the staining platform, and immunohistochemical assessment of tissue. All antibodies tested were concordant.

• Meaning: Discordance of the companion diagnostic test is not attributable to the antibody but rather to inherent tumor heterogeneity or assay- or platform-specific variables

JAMA Oncol. doi:10.1001/jamaoncol.2016.3015

Published online August 18, 2016

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Variable expression of PD-L1 within one tumor / within one patient

Cree et al, Histopathology 2016

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Pitfalls of using PDL1 immunohistochemistry as a biomarker test for anti-PD1–PDL1 therapy

• Focal programmed cell death 1 ligand 1 (PDL1) expression in some tumours may be missed in small biopsy specimens, such as needle biopsies

• PDL1 expression among multiple tumour lesions from individual patients can vary over time and by anatomical site

• PDL1 expression in tumour biopsies collected months or years earlier might not accurately reflect PDL1 status at the time of treatment initiation; therapies given after biopsy but before administration of programmed cell death protein 1 (PD1) pathway blockade (radiation therapy, chemotherapy or kinase inhibitors) may alter PDL1 expression

• PDL1 epitopes detected by some antibodies are potentially unstable with prolonged specimen fixation or inadequate tissue handling before fixation (see NCI guidelines for tissue handling)

• Antibodies used for PDL1 detection have different affinities and specificities

• PDL1 protein expression can be membranous and/or cytoplasmic; however, only membranous PDL1 is functionally relevant, by contacting PD1+ T cells

• PDL1 can be expressed by multiple cell types within the tumour microenvironment, which poses challenges for scoring and interpretation

Topalian et al. Nature Rev Cancer 2016

Pardoll, Nature Rev Cancer 2012

Immune checkpoints

Kerr, USCAP 2015

Eur Respir J 2015; 46: 1762–

1772

O’Callaghan et al.

Eur Respir J 2015;

CD3

CD8 FoxP3

Pathology UNIVERSITY MEDICAL CENTER GRONINGEN

Conclusions:

• PD-L1 testing evaluation is encouraging with respect to better and simpler, reliable routine application in pathology

• Since PD-L1 alone is not the optimal biomarker for PD-1 therapies, considerable effort is and should be spent in examining (combination with) others such as tumor-infiltrating immune cells, mutational load, gene signatures

• In the future, it is likely that it will be an optimal combination of biomarkers to be used to determine, with a sufficient degree of certainty, whether a particular patient will benefit from anti-PD-1/PD-L1 therapy and future immune checkpoint blocking drugs to come

Thank you for your

attention