EQA for PD-L1 IHC staining: is it a conundrum?
Keith Miller
Director
UK National External Quality Assessment Scheme for
Immunohistochemistry & In-situ Hybridisation
The dangers of using laboratory developed tests as a CDx and especially in an NSCLC setting
Keith Miller
Director
UK National External Quality Assessment Scheme for
Immunohistochemistry & In-situ Hybridisation
New Host for the UKNEQAS-ICC&ISH
Bart’s HEALTH NHS Trust
External Quality Assessment Services Cancer Diagnosticsfor
- Community Interest Company
“not for profit”
Professor Peter Isaacson
• Fellow of the Royal Society of Medicine 2009
• Others who are Fellows of the Royal Society in London include Charles Darwin, Louis Pasteur
The UK National External Quality Assessment Scheme for
Immunocytochemistry and ISH
UKNEQAS.org.uk [email protected]
Fully Accredited under ISO 17043
Strict Code of Practice
“Not for Profit”
Commercially neutral
Monitor poor performance and assisting laboratories
UK NEQAS central services are a registered charity
Breast HER2 IHC
Range of HER2 immunostaining
Negative Equivocal Positive
→ FISH analysis
HER2 FISH
• Ratio HER2:Chr17 ≥ 2.00 is considered positive
(20 – 60 non-overlapping nuclei assessed)
– 1.8 < 2.0 borderline negative – 2.0 < 2.2 borderline positive
Chr 17 centromere, , HER2.
Breast Cancer Module: HER-2
Two diagnostic techniques AND two UK NEQAS EQA modules:
Subjective scoring criteria
0 No staining / very slight partial membrane staining, < 10% of tumour cells 1+ Faint barely perceptible partial membrane staining, >10% of tumour cells 2+ Weak - moderate complete membrane staining, > 10% of tumour cells 3+ Strong complete membrane staining, > 10% of tumour cells
• IHC: Cell surface HER-2 protein expression • FISH: Gene expression
3+ 2+ 1+ 0
Each of the cell lines for HER-2 have different levels of expression
They are grown and processed to paraffin- wax under very strict conditions
The thickness of each section is measured with an interferometer
UKNEQAS.org.uk [email protected]
External EQA helps improve procedures over time HER2 IHC example UK laboratory Pass Rates:
2003-2012
The IHC Instruments available today
All can also support either FISH or Chromogenic ISH protocols too
Breast HER2 IHC: Kit v Laboratory Developed Test (HB) OS & UK (2007-2013 combined data)
53% 47%
92%
8%
70%
38%
88%
73%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
0%
10%
20%
30%
40%
50%
60%
70%
80%
90%
100%
Kit-OS (n=2,483) HB-OS (n=1,574) Kit-UK (n=1,297) HB-UK (n=130)
% Users % Pass rate UK
P<0.0001
P<0.0001
Users of
Approved CDx
Users of
Approved CDx Users of
LDTs
Users of LDTs
OS
ALK in NSCLC
Detected in tumour specimens using:
• Immunohistochemistry (IHC)
• Fluorescence in situ hybridization (FISH)
• Reverse transcription polymerase chain reaction of cDNA (RT-PCR) or NGS
With positive cases confirmed by
UK NEQAS EQA for ALK includes cell lines and patient
cancers
ALK Immunohistochemistry in Non-Small Cell Lung Carcinoma (NSCLC):
A. Lessons Learned From External Quality Control OR
B. Discordant Staining and Potential Impact On Patient Treatment Regimen C. Discordant Staining Can Impact Patient Treatment Regimen
Merdol Ibrahim
1*, Suzanne Parry
1, Dawn Wilkinson
1, Neil Bilbe
1, David Allen
2, Steven Forrest
3, Perry Maxwell
5, Anthony O’Grady
6, Jane Starczynski
7, Phillipe Taniere
1, John Gosney
3, Keith Kerr
1, Keith Miller
1& Erik Thunnissen
91 UK NEQAS ICC & SH, Research Department of Pathology, University College London, London UK 2 UCLAD, Research Department of Pathology, University College London, London UK
3 Royal Liverpool University Hospital, Liverpool, UK
4 Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands 5 Royal Victoria Hospital, Northern Ireland, UK
6 Royal College of Surgeons in Ireland, Beaumont Hospital, Dublin, Ireland 7 Birmingham Heartlands Hospital, Birmingham, UK
8 Queen Elizabeth Hospital, Birmingham, UK 9 Aberdeen Royal Infirmary, Aberdeen, Scotland
Journal of Thoracic Oncology: July 2016
Average Pass Rates for ALK IHC
Dochotomous ALK-IHC is a better Predictor for
ALK inhibition outcome than traditional ALK-FISH in advanced non-small cell lung cancer
van der Wekken A, Pelgrim R, 't Hart N, et al.
Clinical Cancer Research 2017 Feb 9.
Main points:
Results: Twenty-nine consecutive patients with ALK-positive advanced NSCLC diagnosed by FISH and/or IHC on small biopsies or fine needle aspirations (FNA) were treated
with ALK inhibitors (Crizotinib).
All ALK-IHC-positive patients responded to crizotinib except three with primary resistance.
No tumor response was observed in
13 ALK-FISH-positive but ALK-IHC-negative
patients
This was confirmed in an external cohort of 16 patients. ROC curves for ALK- IHC and ALK-FISH compared to treatment outcome, showed that dichotomous ALK- IHC outperforms ALK-FISH22C3
SP263 SP142
Pharm-Dx
28.8
LDTs
(laboratory developed tests)
SP142 22C3 28.8
SP263
off-label use of Pharm-Dx!
Blueprint studies
German harmonisation studies
CD, Poundbury Cancer Inst.
PD-L1 CDx Landscape
Ventana
(SP142) Ventana
(SP263) Dako
(28-8) Dako
(22C3)
Type Rabbit
Monoclonal Rabbit
Monoclonal Rabbit
Monoclonal Rabbit Monoclonal Commercial
Availability CE IVD analytical claim CE-IVD predictive claim CE IVD predictive
claim CE IVD predictive claim, CDx Pharma Partner Roche / Genentech AstraZeneca /
MedImmune BMS Merck
Pharma
Drug name Atezolizumab Durvalumab
Nivolumab Nivolumab Pembrolizumab
CDx Configuration
• BenchMark Series
• RTU Antibody
• OptiView Detection
• OptiView Amp
• Complementary Dx
• BenchMark Series
• RTU Antibody
• OptiView Detection
• Complementary Dx
• Dako Autostainer
• Dako Link
• RTU antibody
• Complementary Dx
• Dako Autostainer
• Dako Link
• RTU antibody
• Companion Dx
PD-1/PD-L1: the target of many drug
EQA for PD-L1
Technical
Interpretive
PD-L1
A mixture of appropriate tissues and cell lines that cover the main decision points within each of the respective assays.
PD-L1: Some of our assessors
PD-L1 Protocols/Antibodies employed by our participants
At least 50% of
participants are
using laboratory
developed tests
PD-L1 EQA on lung cancer & cell lines
PD-L1 IHC -Dako Approved 22C3/28.8 CDx PD-L1 IHC - Ventana Approved SP263 CDx
Differences between the different assays for PD-L1
Roche-Ventana SP142
Roche-Ventana SP263 Dako 28-8
Dako 22C3
Scores for Clones SP263/22C3/28.8
Scores for Clones SP263/22C3/28.8
PD-L1 Laboratory Developed Test where there was a significant reduction of positive cells in all samples
Seen with both SP263 and 22C3 antibodies
PD-L1: Approved v LDT
Approved PD-L1 IHC CDx using SP263 on strong expressing cell line.
80%-100% of cells are positive
Laboratory Developed Test using SP263 on strong
expressing cell line.
<20% of cells are positive
PD-L1: Approved v LDT
Approved CDx using SP263 on Tonsil
Laboratory developed test for SP263 on Tonsil
RTD Multiplexing Capabilities in Lung
Esteban Roberts, Adrian Murillo, Teresa Ruffalo and Larry Morrison
All images shown in this presentation are meant to
serve as examples of the capability of the technology
and are not to be interpreted as a medical claim or
source of medical information
EQA for PD-L1
PD1/PD-L1
An online service is being planned
Interpretive
PD-L1
Creating a digital slide resource for all
EQA’s across the world to access for
their local Educational EQA activities
IQN-Path Digital Resource +Ad Board
Journal Publication
PD-L1 Digital Education Resource Pilot
Academia
Industry providerEQA
providerEQA
Industry Academia
providerEQA
IQN-Path Digital Education Resource Working Group
Working Group
Keith Miller UK-NEQAS Chair Colin Tristram - Histiocyte John Garratt – cIQc
Jacqueline Hall (IQN Path) Committee Members
Hans Juergen Grote (Merck), Els Dequeker (ESP EQA), Erik Thunissen (ESP EQA), Julia Pagliuso (RCPAQAP), Rudy Hovelinck (Astra Zeneca), Mogens Vyberg (NordiQC), Soren Nielsen (NordiQC), Robert Lott (Roche), Beth
Sheppard (Roche), Zenobia Haffajee (RCPAQAP), Jessica Whiteley (RCPAQAP), Helle Fisker (Visiopharm), Daniel Martinez (SEAP).
Manfred Dietel (QUIP), Colin Tristram (Histocyte), Xueying Shi’s (CCP)
Digital Education Resource from IQN-Path
STAGE 1
DEVELOPMENT STAGE 2
PD-L1 PILOT STAGE 3
FULL
IMPLEMENTATION
Is this slide positive for Red Marbles?
What is the Percentage of Red
Marbles?
Based at:
The Poundbury Cancer Institute, Newborough House,
Queen Mother Square, Poundbury, Dorset, UK
A unique combination in a single building:
• A dedicated independent slide-based Cancer Testing Laboratory
• An Education Centre for the pathology community
Teaching and Training
Advanced Course for Pathologists and Scientists at Poundbury
Professor Keith Kerr giving a seminar on Lung Cancer
Is this
something for
IQNPath to get
Involved in?
And finally to our PD-L1/PD1 IHC Trainer!
Another successful PD-L1 trainee!
ACKNOWLEDGEMENTS
•Dr Merdol Ibrahim
•Suzanne Parry
•Neil Bilbe
•Ailin Rhodes
•Clara Lynch
•Dawn Wilkinson
•Seema Dhanjal
•Professor Bharat Jasani
•Dr Corrado D’Arrigo
•Dr Sarah Wedden
•David Allen
•Josep Linares
•Numerous assessors
•Roche-Ventana
•Menarini
•Visiopharm and many more
[email protected]
& John Garratt from CIQC
