PD-L1 Testing German Experience. P. Schirmacher for QuIP

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1 PD-L1 Testing German Experience P. Schirmacher for QuIP

2 Quality Management Molecular Diagnostics Method/Inter-Center-Optimisation/Validation Preclinical Validation/internal QM Accreditation Institutes (DAkkS; ISO 9001 DIN 17020) National Round Robins (QuIP)

3 QuiP (DGP) Independent Self administered Non commercial Expert guided (Panels) Extensive RR-Expertise (> 300 RRs) High acceptance (diagnostic community, clinicians, industry, politics) International (in selected indications) ESP-LOA and cooperation

4 Principle: Free Choice of Methods Free Choice of Method, e.g. Sanger Pyro-Sequencing Mutation-spezifische PCR NGS (Deep/Panel Sequencing) Reflection of technology Evaluation by diagnostic result

5 Constant Round Robins RfB BRAF 600 BRAF 600 BRAF 600 BRAF 600 RfB CD117 CD117 CD117 CD117 CD117 CD117 CD117 CD117 RfB EGFR EGFR EGFR RfB Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRez RfB GIST GIST GIST GIST GIST RfB HER2n IHC HER2n IHC HER2n IHC HER2n IHC HER2n IHC RfB HER2n ISH HER2n ISH HER2n ISH HER2n ISH HER2n ISH RfB Keratine Keratine Keratine Keratine Keratine Keratine Keratine Keratine RfB KlonML KlonML KlonML KlonML KlonML KlonML KlonML RfB KRAS RfB Lymphome Lymphome Lymphome Lymphome Lymphome Lymphome Lymphome Lymphome RfB MMRD MMRD MMRD MMRD MMRD MMRD MMRD MMRD RfB MSI MSI MSI MSI MSI MSI MSI RfB NEM NEM NEM NEM NEM NEM NEM NEM RfB RAS RAS RAS RAS multiblock ER ER ER ER ER ER ER ER multiblock PR PR PR PR PR PR PR PR multiblock Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM multiblock Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH multiblock Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 provitro HPV HPV HPV HPV HPV HPV HPV HPV provitro TBc TBc TBc TBc TBc TBc TBc TBc RfB T790M T790M RfB PD-L1MM RfB PD-L1 NSCLC PD-L1 NSCLC EGFR IHC RfB sqnsclc RfB BRCA1/2 BRCA1/2 RfB BRAF 600 Lunge

6 Prototypic Round Robins Round Robin Status Round Robin Status T790M 2016 PD-L1 NSCLC EGFR IHC sqnsclc PD-L1 Melanoma (2) finished final report final report final report MLH1 ROS1 IDH1(R132H) Consideration QuIP-Board Consideration QuIP-Board Consideration QuIP-Board T790M 2017 (2) ongoing Q1-Q2 PD-L1 NSCLC (2) ongoing Q1-Q2 BRAF V600 (2) BRCA 1/2 (2) planned Q3-Q4 planned Q3-Q4

7 Implementation of new Diagnostic Assays Information about approval of new drug with predictive molecular testing (1-3 months ahead) Implementation Validation and optimisation of testing modalities Solutions for open scientific issues Diagnostic recommendations, information, discussion with clinical societies Diagnostics for early accession/ compassionate use programs Broad/nationwide availability (methodology, expertise, training) Agreement on reimbursement issues Local issues Others (e.g. legal issues) Quality Management Panel-establishment and -validation (> 3 m) Establishment of expert panel (n~5-8; Lead- Institutes; other Panel centers) Consent about approach (methodology, time schedule, testing conditions etc.) Identification and selection of suitable test cases Cross validation by lead institutes Testing/validation of panel centers; evaluation; set-up of RR conditions National/international round robin (> 2 m) announcement/feed-back; application preparation by Lead-institutes, RfB Probe mailing, testing, test results mailed Evaluation and publication Implementation without regulatory, financial and intellectual framework but decisive for drug/therapy success (system relevant). Requires Competence Center System (Peers) and more attention by industry and regulatory authorities

8 PD-L1 IHC German Strategy for Roll-out and QM National Harmonisation Study (DGP) Validation Harmonisation Recommendation National QA Measures (QuIP) Quality Assurance Teaching and Training National Improvement Activity (nngm/dkh)

9 PD-L1 IHC in NSCLC National Harmonisation-Study Pathology: Reinhard Büttner Manfred Dietel Lukas Heukamp Korinna Jöhrens Thomas Kirchner Simone Reu Josef Rüschoff Andreas Scheel Hans-Ulrich Schildhaus Peter Schirmacher Markus Tiemann Arne Warth Wilko Weichert Industrial Partners BMS MSD Roche AstraZeneca Ventana Dako Targos

10 Response to Anti PD-1 (Pembrolizumab) and Expression of PD-L1 in NSCLC

11 Different Antibodies and Antibody Reactivities Scheel et al., 2016

12 Tumorimmunotherapy: PDL-1-Testing

13 BIRCH: PD-L1 TC and IC Selection Criteria TC2/3 IC2/3 Intrinsic PD-L1 expression in tumor cells (TC) 34% of screened patients between Jan and Dec Adaptive PD-L1 expression in tumor-infiltrating immune cells (IC) BIRCH enrolled patients with tumors that were PD-L1 TC2/3 or IC2/3 VENTANA SP142 IHC assay was used to determine PD-L1 expression on both TC and IC Archival or freshly collected tumor specimens were required for PD-L1 testing at a central laboratory PD-L1 as a predictive biomarker: PD-L1 expression on TC and IC was independently predictive of response in patients with previously treated NSCLC (i.e., POPLAR) 1,2 TC3 or IC3 = TC 50% or IC 10% PD-L1+ cells; TC2/3 or IC2/3 = TC or IC 5% PD-L1+ cells, respectively. 1. Horn L, et al. ASCO 2015; 2. Vansteenkiste J, et al. ECC Besse et al, Oral Presentation at ECC 2015

14 A Global NSCLC-PDL1 Score? Scheel et al., 2016

15 Interobserver-Validation (Phase I) Scheel et al., 2016

16 Harmonisation Study: Immune Cells Scheel AH et al., Modern Pathology 2016

17 Phase 2 Assays: - Performance in multiple Institutes / Reliability - Staining pattern - validation of Phase 1; fixed assay vs. open protocol - Inter-lab concordance Evaluation: - Validation proportional score (6-step) - Local vs. central evaluation

18 Phase 2 Targos Molecular Pathology; - Project management Phase 2 - NSCLC-TMAs (project specific): - 'TMA-Master' (3DHistotech) mm cores - Central provision of slides 8 Pathology Institutes - Trial analog protocols Dako 22C3, 28-8 Ventana SP142, SP263 - Open protocol: 22C3, 28-8 (local protocols) - 3 institutes per protocol

19 PD-L1 Testing + Robust, broadly available technology (IHC) Good antibodies; alternatives exist Robust, relatively stable assay Consented algorithm - Stand-alone assay/ no methodical convergence Singular analyses Algorithm/reporting complexity; dynamic Heterogenous expression; 1% threshold problematic Insufficient refunding

20 Prototypic Round Robin PD-L1-NSCLC (1) Decision for RR, Lead- and Panel Institutes 6.16 Coordination of Lead Institutes, selection of cases, trilateral validation Testing of the Panel Institutes (internal RR) Evaluation of internal RRresults, Lead/Panel-Institute; meeting, decision for open RR 15.9 Public PD-L1 training meeting (Charite) End of 9.16 Open RR Lead Institutes: Berlin, Cologne, Göttingen Panel Institutes: Hannover, Heidelberg, Jena, Munich TU, Munich LMU Composition: 10 cases; 2 points each, 18 points necessary for certificate No methodical restriction, sections provided Optional reporting of methodology Option to challenge and for educational

21 Prototypic Round Robin Participants: 83 (76 D, 4 A, 3 CH) Successful: 60 Success rate: 72% PD-L1-NSCLC (1) Composition < 1%: 3 cases 1%-49%: 2 cases %: 5 cases

22 PD-L1-NSCLC Round Robin Antibodies

23 PD-L1-NSCLC Challenge Challenged result: 18 Institutes 1 Institute post hoc success 4 Institutes: Interpretation problems 13 Institutes: Staining problems

24 Prototypic Round Robin Malignant Melanoma (1) 3.8 Panel-Institutes.16 Decision for RR Decision on Lead- and Panel Institutes 9/10.16 Coordination of Lead Institutes, Case selection and reciprocal testing Testing of Panel Institutes (internal RR) 9/10 Announcement of open RR open RR Evaluation of open RR; announcement of results Lead Institutes: Berlin, Heidelberg, TU Munich Panel Institutes: Hannover, Munich LMU, Cologne, Hamburg, Göttingen, Würzburg Composition: 10 cases; 2 points each, 18 points necessary for certificate No methodical restriction, sections provided Optional reporting of methodology Option to challenge and for educational

25 Prototypic Round Robin PD-L1-Melanoma Participants: 44 Successful: 37 Success rate: 84% Composition <5%: 5 cases >5%: 5 cases

26 PD-L1-Melanoma Round Robin Antibodies

27 PD-L1-NSCLC and MM Round Robin Other Conclusions Diverse use of primary antibodies 28-8 and SP263 stain stronger 22C3 stains rather weaker Discrepancies are rather towards lower staining intensities Establishment using tonsil tissue suggested

28 Future and Ongoing Activities PD-L1-NSCLC (2) ongoing (until 15.5.) PD-L1 Bladder Cancer in preparation PD-L1 MM (2) in discussion

29 Other/Better Markers!? PD-L1 Gene CNV/expression MSI Mutational load Immunophenotyping Combinations RESISTANCE MARKERS

30 PD-L1 Amplifications/Deletions and/or Mutational Load Increasing number of mutations n=78 n=40 WGS/WES? Panels? Others? Budczies et al. 2016

31 Immune Cell Infiltration and Response to Conventional and Personalized Treatment Lasitschka, F, Schirmacher P, Jäger D, Halama N, et al.

32

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34 Constant Round Robins Participants 2016 Participants 2016 RR RR BRAF 58 ER 186 CD PR 177 GIST 39 Her2-IM 198 Her2nlHC 88 Her2-ISH 103 Her2nlSH 52 Ki RAS 71 Lymphone 71 HPV 7 MMRD 84 TBC 14 MSI 45 Keratine 88 KlonML 29 NEM 69 EGFR 48

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