Germany

(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)

Free Choice of Method, e.g.

• Sanger

• Pyro-Sequencing

• Mutation-spezifische PCR

• NGS (Deep/Panel Sequencing)

Reflection of technology Evaluation by diagnostic result

Principle: Free Choice of

Methods

(5)

Constant Round Robins

2010 2011 2012 2013 2014 2015 2016 2017 2018

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

RfB EGFR IHC

sqNSCLC

RfB BRCA1/2 BRCA1/2

RfB BRAF 600 Lunge

(6)

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

MLH1 Consideration

QuIP-Board

ROS1 Consideration

QuIP-Board IDH1(R132H) Consideration

QuIP-Board

(7)

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

(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)

Pathology: Industrial Partners

Reinhard Büttner BMS

Manfred Dietel MSD

Lukas Heukamp Roche

Korinna Jöhrens AstraZeneca Thomas Kirchner Ventana

Simone Reu Dako

Josef Rüschoff Targos

Andreas Scheel

Hans-Ulrich Schildhaus Peter Schirmacher Markus Tiemann Arne Warth

Wilko Weichert

PD-L1 IHC in NSCLC

National Harmonisation-Study

(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)

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.

IC2/3 TC2/3

34% of screened patients between Jan. 2014 and Dec. 2014

(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)

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

(18)

- Trial analog protocols Dako 22C3, 28-8

Ventana SP142, SP263 - Open protocol: 22C3, 28-8

(local protocols)

- ≥3 institutes per protocol

Targos

Molecular Pathology;

8 Pathology Institutes

Phase 2

- Project management Phase 2 - NSCLC-TMAs (project specific):

- 'TMA-Master' (3DHistotech) - 1.5 mm cores

- Central provision of slides

(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)

28.4.16 Decision for RR, Lead- and Panel Institutes

6.16 Coordination of Lead Institutes, selection of cases, trilateral

validation

15.7-26.7. Testing of the Panel Institutes (internal RR)

27.7. -29.7. Evaluation of internal RR- results, Lead/Panel-Institute;

meeting, decision for open 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 PD-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

(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

22.8.16 Decision on Lead- and Panel Institutes

9/10.16 Coordination of Lead Institutes, Case selection and reciprocal testing

14-28.10 Testing of Panel Institutes (internal RR)

9/10 Announcement of open RR 9.-25.11.16 open RR

28.11-2.12. 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=78n=40

Budczies et al. 2016 WGS/WES?

Panels?

Others?

(31)

Immune Cell Infiltration and Response to Conventional and Personalized Treatment

Lasitschka, F, Schirmacher P, Jäger D, Halama N, et al.

(32)

(33)

(34)

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