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PD-L1 TestingGerman Experience
P. Schirmacherfor
QuIP
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Quality Management Molecular Diagnostics
• Method/Inter-Center-Optimisation/Validation• Preclinical Validation/internal QM • Accreditation Institutes (DAkkS; ISO 9001 DIN
17020)• National Round Robins (QuIP)
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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
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Free Choice of Method, e.g.• Sanger• Pyro-Sequencing• Mutation-spezifische PCR• NGS (Deep/Panel
Sequencing)Reflection of technologyEvaluation by diagnostic result
Principle: Free Choice of Methods
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Constant Round Robins2010 2011 2012 2013 2014 2015 2016 2017 2018
RfB BRAF 600 BRAF 600 BRAF 600 BRAF 600RfB CD117 CD117 CD117 CD117 CD117 CD117 CD117 CD117RfB EGFR EGFR EGFRRfB Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRez Erez/PRezRfB GIST GIST GIST GIST GISTRfB HER2n IHC HER2n IHC HER2n IHC HER2n IHC HER2n IHCRfB HER2n ISH HER2n ISH HER2n ISH HER2n ISH HER2n ISHRfB Keratine Keratine Keratine Keratine Keratine Keratine Keratine KeratineRfB KlonML KlonML KlonML KlonML KlonML KlonML KlonMLRfB KRASRfB Lymphome Lymphome Lymphome Lymphome Lymphome Lymphome Lymphome LymphomeRfB MMRD MMRD MMRD MMRD MMRD MMRD MMRD MMRDRfB MSI MSI MSI MSI MSI MSI MSIRfB NEM NEM NEM NEM NEM NEM NEM NEMRfB RAS RAS RAS RASmultiblock ER ER ER ER ER ER ER ERmultiblock PR PR PR PR PR PR PR PRmultiblock Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IM Her2-IMmultiblock Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISH Her2-ISHmultiblock Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67 Ki-67provitro HPV HPV HPV HPV HPV HPV HPV HPVprovitro TBc TBc TBc TBc TBc TBc TBc TBcRfB T790M T790MRfB PD-L1MMRfB PD-L1 NSCLC PD-L1 NSCLC
RfBEGFR IHC sqNSCLC
RfB BRCA1/2 BRCA1/2RfB BRAF 600 Lunge
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Round Robin Status
T790M 2016 finished
PD-L1 NSCLC final report
EGFR IHC sqNSCLC
final report
PD-L1 Melanoma(2)
final report
T790M 2017 (2) ongoing Q1-Q2
PD-L1 NSCLC (2) ongoing Q1-Q2
BRAF V600 (2) planned Q3-Q4
BRCA 1/2 (2) planned Q3-Q4
Prototypic Round Robins
Round Robin Status
MLH1ConsiderationQuIP-Board
ROS1ConsiderationQuIP-Board
IDH1(R132H) ConsiderationQuIP-Board
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Implementation of new Diagnostic Assays
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
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)
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
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PD-L1 IHCGerman 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)
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Pathology: Industrial PartnersReinhard Büttner BMSManfred Dietel MSDLukas Heukamp RocheKorinna Jöhrens AstraZenecaThomas Kirchner VentanaSimone Reu DakoJosef Rüschoff TargosAndreas ScheelHans-Ulrich SchildhausPeter SchirmacherMarkus TiemannArne WarthWilko Weichert
PD-L1 IHC in NSCLCNational Harmonisation-Study
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Response to Anti PD-1 (Pembrolizumab) and Expression of PD-L1 in NSCLC
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Different Antibodies and Antibody Reactivities
Scheel et al., 2016
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Tumorimmunotherapy: PDL-1-Testing
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Besse et al, Oral Presentation at ECC 2015
BIRCH: PD-L1 TC and IC Selection Criteria
• 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
Intrinsic PD-L1 expression in tumor cells (TC)
Adaptive PD-L1 expression in tumor-infiltrating immune
cells (IC)
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 2015.
IC2/3TC2/3
34% of screened patients between
Jan. 2014 and Dec. 2014
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A Global NSCLC-PDL1 Score?
Scheel et al., 2016
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Interobserver-Validation (Phase I)
Scheel et al., 2016
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Harmonisation Study: Immune Cells
Scheel AH et al., Modern Pathology 2016
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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
Phase 2
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- Trial analog protocolsDako 22C3, 28-8Ventana SP142, SP263
- Open protocol: 22C3, 28-8(local protocols)
- ≥3 institutes per protocol
TargosMolecular Pathology;
8 PathologyInstitutes
Phase 2
- Project management Phase 2
- NSCLC-TMAs (project specific):- 'TMA-Master' (3DHistotech)- 1.5 mm cores
- Central provision of slides
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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
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Prototypic Round RobinPD-L1-NSCLC (1)
28.4.16 Decision for RR, Lead- andPanel Institutes6.16 Coordination of Lead Institutes, selection of cases, trilateral validation15.7-26.7. Testing of the Panel Institutes (internal RR)27.7. -29.7. Evaluation of internal RR-results, Lead/Panel-Institute; meeting, decision for open RR15.9 Public PD-L1 training meeting(Charite)End of 9.16 Open RR
Lead Institutes: Berlin, Cologne, GöttingenPanel Institutes: Hannover, Heidelberg, Jena, Munich TU, Munich LMUComposition: 10 cases; 2 pointseach, 18 points necessary forcertificateNo methodical restriction, sectionsprovidedOptional reporting of methodologyOption to challenge and foreducational
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Prototypic Round RobinPD-L1-NSCLC (1)
• Participants: 83 (76 D, 4 A, 3 CH)
• Successful: 60
• Success rate: 72%
Composition
< 1%: 3 cases
1%-49%: 2 cases
50-100 %: 5 cases
Diagramm1
836023
gesamt
mit Erfolg teilgenommen
teilgenommen
Sheet1
gesamt83
mit Erfolg teilgenommen60
teilgenommen23
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PD-L1-NSCLC Round RobinAntibodies
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PD-L1-NSCLC Challenge
Challenged result: 18 Institutes
• 1 Institute post hoc success
• 4 Institutes: Interpretation problems
• 13 Institutes: Staining problems
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Prototypic Round RobinMalignant Melanoma (1)
3.8 Panel-Institutes.16 Decision forRR22.8.16 Decision on Lead- and Panel Institutes9/10.16 Coordination of Lead Institutes, Case selection andreciprocal testing14-28.10 Testing of Panel Institutes (internal RR)9/10 Announcement of open RR9.-25.11.16 open RR28.11-2.12. Evaluation of open RR; announcement of results
Lead Institutes: Berlin, Heidelberg, TU MunichPanel Institutes: Hannover, MunichLMU, Cologne, Hamburg, Göttingen, WürzburgComposition: 10 cases; 2 pointseach, 18 points necessary forcertificateNo methodical restriction, sectionsprovidedOptional reporting of methodologyOption to challenge and foreducational
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Prototypic Round RobinPD-L1-Melanoma
• Participants: 44
• Successful: 37
• Success rate: 84%
Composition
5%: 5 cases
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PD-L1-Melanoma Round RobinAntibodies
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PD-L1-NSCLC and MM Round RobinOther Conclusions
• Diverse use of primary antibodies
• 28-8 and SP263 stain stronger
• 22C3 stains rather weaker
• Discrepancies are rather towards lower stainingintensities
• Establishment using tonsil tissue suggested
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Future and Ongoing Activities
• PD-L1-NSCLC (2) ongoing (until 15.5.)• PD-L1 Bladder Cancer – in preparation• PD-L1 MM (2) – in discussion
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Other/Better Markers!?
• PD-L1 Gene CNV/expression• MSI• Mutational load• Immunophenotyping• Combinations• RESISTANCE MARKERS
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PD-L1 Amplifications/Deletions and/or Mutational Load
Increasing number of mutations
n=78n=40
Budczies et al. 2016
WGS/WES?
Panels?
Others?
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Immune Cell Infiltration and Response to Conventional and Personalized Treatment
Lasitschka, F, Schirmacher P, Jäger D, Halama N, et al.
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Constant Round Robins
RR
Participants 2016
RR
Participants 2016
BRAF 58 ER 186
CD117 66 PR 177
GIST 39 Her2-IM 198
Her2nlHC 88 Her2-ISH 103
Her2nlSH 52 Ki-67 177
RAS 71
Lymphone 71 HPV 7
MMRD 84 TBC 14
MSI 45
Keratine 88
KlonML 29
NEM 69
EGFR 48
PD-L1 Testing�German ExperienceQuality Management Molecular DiagnosticsQuiP (DGP)Diapositiva numero 4Constant Round Robins�Diapositiva numero 6Diapositiva numero 7PD-L1 IHC�German Strategy for Roll-out and QMPD-L1 IHC in NSCLC�National Harmonisation-StudyDiapositiva numero 10Different Antibodies and Antibody ReactivitiesTumorimmunotherapy: PDL-1-TestingDiapositiva numero 13A Global NSCLC-PDL1 Score?Interobserver-Validation (Phase I)Diapositiva numero 16Diapositiva numero 17Diapositiva numero 18PD-L1 TestingPrototypic Round Robin�PD-L1-NSCLC (1)�Prototypic Round Robin�PD-L1-NSCLC (1)�PD-L1-NSCLC Round Robin�AntibodiesPD-L1-NSCLC �ChallengePrototypic Round Robin�Malignant Melanoma (1)�Prototypic Round Robin�PD-L1-MelanomaPD-L1-Melanoma Round Robin�AntibodiesPD-L1-NSCLC and MM Round Robin�Other ConclusionsFuture and Ongoing ActivitiesOther/Better Markers!?PD-L1 Amplifications/Deletions and/or Mutational LoadImmune Cell Infiltration and Response to Conventional and Personalized TreatmentDiapositiva numero 32Diapositiva numero 33Constant Round Robins�