emerging tissue and serum markers for immune checkpoint …gbcc2016.gbcc.kr/upload/kyong hwa...
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Emerging Tissue and Serum Markers for Immune Checkpoint Inhibitors
Kyong Hwa Park MD, PhD
Medical Oncology
Korea University College of Medicine
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Contents
• Immune checkpoint inhibitors in clinical practice
• Biomarkers for immune checkpoint inhibitors
- Tissue or blood based multi-level assays
• Immunotherapy in breast cancer: recent signatures
• Emerging biomarkers
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Atezolizumab(Genentech/Roche)
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Current Approval Status of ITA by USFDA
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Immunologic Armaments in Oncology Clinic
Anti-CTLA4Anti-CD137 agonistAnti-OX40 agonistAnti-CD27 agonist
IL-2IL-12
CARs
Anti-VEGF
VaccineIFN-alphaGM-CSF
Anti-CD40 agonistTLR agonist
ChemotherapyRadiation therapyTargeted therapy
Anti-PD1Anti-PD-L1
IDO inhibitors
Nat Rev Genetics Aug 2016
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Combination immunotherapy: More trials, Better targeted
2017 Nature
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PD-1/PD-L1 Blockades in MBC
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Pembrolizumab in TNBC:Phase Ib KEYNOTE-012 Study
• Response rate: 18.5%
• No response in LDH > 2X UNL
J Clin Oncol. 2016 May 2
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Avelumab in Metastatic Breast Cancer
All MBC Patients (N=111)ORR: 3%
TNBC Patients (N=46)ORR: 5%
PD-L1+ PD-L1-
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Hyperprogressors!
Clin Cancer Res; 2017
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Metastatic Cancer Remission
• Patient selection ?
• Anti-tumor immune response?• How long treat?• Combination approach?• Toxicities?
• Still immune response?• Retreat?
Clinical Questions that you might have….
Immune Targeted Agents
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• PD-L1 expression status by IHC: anti–PD-L1 antibody clone 22C3 (Merck)
• Cells counted: neoplastic and intercalated mononuclear inflammatory cells
KEYNOTE-001: Phase Ib study ofPembrolizumab for the Treatment of NSCLC
N Engl J Med 2015;372:2018-28.
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Mechanisms of PD-1 Expression
Nature Immunology 14, 1212–1218 (2013)
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PD-1/PD-L1: Exhaustion of T cells
1) Antagonizing TCR signaling
2) Decreased survival, proliferation, altered metabolism
3) Reduced proliferation via RAS path
4) Altered transcription
5) Altered T cell motility
Trends in Immunology April 2015, Vol. 36, No. 4
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• There are no harmonized criteria to define PD-L1 positivity by IHC1
• Various PD-L1 IHC assays differ in terms of the cells assessed, scoring methods, and thresholds used:
Technical Complexity With PD-L1:Different Thresholds for Expression
NivolumabDako Assay2
PembrolizumabDako Assay3,4
AtezolizumabVentana Assay5,6
DurvalumabVentana Assay7
Cells assessed
Tumour cellsTumour cells and
immune cellsTumour cells and immune cells Tumour cells
Ab clone 28-8 22C3 SP142 SP263
Scoring method
Tumour score: Percentage of cells with membrane staining at any intensity
Proportion score: Percentage of cells with membrane staining at any intensity
Tumour cell (TC) score: staining percentage of tumour cells
Immune cell (IC) score: staining percentage of tumour area
Percentage tumour cells with membrane staining
PD-L1 Expression Levels
<1% or ≥1%, <5% or ≥5%,
<10% or ≥10%≥50%
TC3 or IC3: TC ≥50% or IC ≥10%TC2/3 or IC2/3: TC or IC ≥5%TC1/2/3 or IC1/2/3: TC or IC ≥1%TC0 and IC0: TC and IC <1%
≥25%
Ab, antibody; IHC, immunohistochemistry; PD-L1, programmed death ligand-1. 1. Heskamp S et al. Cancer Res. 2015. [Epub ahead of print]. 2. Phillips T et al. Appl Immunohistochem Mol Morphol. 2015. doi:10.1097/PAI.0000000000000256. 3. Dolled-Filhart M et al. Poster presentation at ASCO 2015. 11065. 4. Rizvi N et al. Poster presentation at ASCO 2015. 8026. 5. Spira AI et al. Oral presentation at ASCO 2015. 8010. 7. Spigel DR et al. Poster presentation at ASCO 2015. 8028. 7. Rebelatto MC et al. Poster presentation at ASCO 2015. 8033.
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PD-L1 as a Biomarker?: Issues
• Tumor heterogeneity
• Interval between biopsy and treatment
• Primary vs metastatic disease
• Ab and staining conditions
• Defining a positive result (cut-offs)
- Immune cells vs tumor cells
- Location of expression: intracellular vs surface vs stroma
- Intensity, percent of positive cells
- Distribution
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HER-2 type Breast Cancer: PANACEA
2017 SABCS Loi et al
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Genomic Signatures of TIL is Prognostic in HER2 Type
2018 BCRT, doi: 10.1007/s10549-017-4502-3
Luminal low Luminal high
HER-2 TNBC
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Clinical Development in Cancers with High mutational burden
Science 3 April 2015
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Mutational Landscape of Melanoma according to the CTLA-4 blockade Response
Snyder A et al. N Engl J Med 2014;371:2189-2199.
Mutational Load
Survival in Discovery set
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Mutation burden, clinical response, and factors contributing to mutation burden: NSCLC
Science 3 April 2015
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Neoantigen burden & Intratumoral Heterogeneity (ITH)
Science MAR 2016
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Clonal Heterogeneity and Immune Signature in TNBC
N=193 TNBC with follow up data @ TCGAJAMA Oncol. 2017;3(12):1707-1711. doi:10.1001/jamaoncol.2017.2140
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TIL Infiltration and Response to anti–PD-1: Melanoma
J Clin Invest. 2016;126(9):3447-3452. doi:10.1172/JCI87324.
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Tumoral CD8+ T cell Infiltration after Pembrolizumab
Nature 515, 568–571 (27 November 2014)
Response (n=22)
Progression (n=24)
Proliferating T cells
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Change in TCR Repertoire
Genes and Immunity (2016)Nature (2014)
• Theoretical repertoire of 1015–1020 different TCRs that could be generated in humans• Effective anti-tumor immunity increased TCR clonality
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T cell Diversity and Mutational Burden
Science 3 April 2015Nat Genetics 2016
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What we should know for Rational application of Immune Targeted Agents (ITA)
• Immune responses with standard anti-cancer treatment
• Indicators to judge Immune-related Response
- Pseudo-progression
- Delayed but prolonged response, delayed K-M curve separation
- Mixed responses
• Predictors for Immune-related Adverse Events
• Timing for Re-treatment: monitoring immunosuppressive TME
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Best Information is in Tissue
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Peter Lee, 2016 SABCS
Multiplex IHC NanoString Analysis: KEYNOTE-001
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Assessment of Tumor Immune Microenvironment
Static markers Dynamic markers
• Genomic analysis (WES, targeted seq)• IHC for molecular & immune markers• Flow/CyTOF for phenotyping• RNA seq for profiling the transcriptome• Single cell (TCR seq, RNAseq)
• Analysis of germline SNPs• Flow/CyTOF for phenotyping (best if paired with tumor)• Cytokine profiling in serum• Exosome analysis (WES, RNAseq, paired with tumor)• Single cell (TCRseq, RNAs eq)
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Blood mirrors anti-tumor immune responses!
- Immune cell profiles- Cytokines
- PBMC Immunomics
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F/81, Metastatic Breast Cancer (TNBC)
Baseline Treatment week #18 Treatment week #16
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Cell-based Analysis Flow-cytometry (FACS)
Mass Cytometry (Cy-TOF)
• Easy accessibility• High-throughput• High signal overlap• ~16 parameters
• No signal overlap• >40 parameters• High cost• Low-throughput• Limited availability
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MDSC in Ipilimumab Treated Melanoma Patients
Cancer Immunol Immunother (2014) 63:247–257
MDSC with Treatment
MDSC vs LDH MDSC vs M stage
MDSC vs Response
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Change in circulating Lymphocytes after Ipilimumab
Early change in ALC Delayed change in CD8
Delayed change in EØ Delayed change in CD4
2016 CCR*Early: 2-8 weeks after Tx, Delayed: 8-18 weeks after Tx.
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Anti-PD1 Response: T cell invigoration & Tumor burden
2017 Nature doi:10.1038/nature22079
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Anti-PD1 Response: T cell invigoration/Tumor burden ratio
2017 Nature doi:10.1038/nature22079
PennTumor burden (pre) PD-1+CD8 (pre)
MSKCC
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Gut microbiome influences efficacy of PD-1–based immunotherapy
Science 05 Jan 2018: DOI: 10.1126/science.aan3706
All cancer
NSCLC
Frequency of
Akkermansia
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Dynamic immune monitoring is necessary in
• Development of new IO-based cancer treatment.
: Best combination between ITA-ITA, SOC-ITA.
Endpoint measures
• Decision for treatment, retreatment – Public issue
• Neoantigen discovery & precision medicine
Need collaborative effort in academic society!
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Bio-specimens reflecting Anti-tumor Immune Responses
Annu. Rev. Med. 2014.65:185-202
• Translational research for optimal immune monitoring
• Validation of immunoassays using clinical samples
• Establish the most feasible, standardized assays for immune monitoring
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AACR Tweeter
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Smart Research Cures Cancer!