Clinical implementation of circulating tumour DNA analysis for ......Circulating Tumor DNA From...
Transcript of Clinical implementation of circulating tumour DNA analysis for ......Circulating Tumor DNA From...
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Radical treatments for gynaecological cancers: HOPE or HYPE?
Dr Stephen Wong
Peter MacCallum Cancer Centre
Clinical implementation of circulating tumour
DNA analysis for personalised oncology:
From blocks to blood
@stephenqwong
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ctDNAfragment size ~160bp
Characteristics of ctDNA
Adapted from D. Leja, NHGRI
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ctDNA as a “Liquid Biopsy”Tumour (blocks) sampling
Plasma (blood) sampling
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Tumour (blocks) sampling
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Plasma (blood) sampling
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Methods of ctDNA detection
Increasing sensitivity
Increasing genomic coverage
Single Locus Assays
(eg Digital PCR)
Targeted sequencing approaches
(eg hybrid capture,
amplicon)
Whole genome
sequencing
Quantitative Analysis
• Tumour burden• Disease
monitoring
Genomic Analysis
• Molecular profiling
• Clonal evolution
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ctDNA testing workflowBlood taken Blood Spin
Issue report
EDTA tube (within 1 hour)Streck tube (within 14 days)
NGS technology
Plasma storage
Plasma DNA extraction
10-50 mls of blood
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Adapted from Wan et al, Nature Reviews Clinical Oncology, 2017
Cancer detection:Screening or early diagnosis
Risk stratification and prognostication
Molecular profiling and treatment selection
Surveillance: Detection of minimal residual disease
Monitoring response to therapy and clonal evolution
Clinical Applications
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ctDNA and minimal residual disease
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Minimal Residual Disease in Stage III melanoma
Tan et. al., Annals of Oncology, 2019
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Distant metastatic free survival: Baseline
Distant metastatic free survival: Postoperative
Minimal Residual Disease detection and Monitoring
during Adjuvant Therapy in Stage III Melanoma
HR 4.9, 95% CI 2.4-10,
p=0.22x10-4
HR 14, 95% CI 5.7-
36, p=0.16x10-7
Postoperative ctDNA detected (n=15)
Postoperative ctDNA not detected (n=48)
Tan et. al., Annals of Oncology, 2019
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Adapted from Wan et al, Nature Reviews Clinical Oncology, 2017
Cancer detection:Screening or early diagnosis
Risk stratification and prognostication
Molecular profiling and treatment selection
Surveillance: Detection of minimal residual disease
Monitoring response to therapy and clonal evolution
Clinical Applications
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Biopsy available
Molecular profiling
(long waiting time)
Molecular profiling
of advanced cancer
Tumour tissue Plasma
Biopsy not available
Standard treatment
Blood always available
1-2 day results
Targeted Therapy ctDNA
SEGMENT and M-BCB (Metastatic Breast Circulating Biomarker Study)
Molecular Profiling in Breast Cancer
Digital PCR NGS
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Molecular profiling using ctDNA analysis in Breast Cancer
• Cost• Turn around time• Performance• Reproducibility
Research Diagnostics
Screening for PIK3CA mutations enables enrolment on PI3K inhibitor clinical trial
Andjelija ZivanovicChen-Fang WengMiriam YeungUnpublished
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Number of patients enrolled
Identification of targetable mutations
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Cancer detection:Screening or early diagnosis
Risk stratification and prognostication
Molecular profiling and treatment selection
Surveillance: Detection of minimal residual disease
Monitoring response to therapy and clonal evolution
Clinical Applications
Adapted from Wan et al, Nature Reviews Clinical Oncology, 2017
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Disease monitoring using ctDNAMelanoma molecular subtypes
BRAF mut
NRAS mut
BRAF/NRAS WT TERT promoter mutations
Mar V, Wong SQ et al, Clin Can Res 2013
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BRAF mutant pt. NRAS mutant pt.
BRAF/NRAS WT pt. TERT mutant pt.
Disease monitoring using ctDNA
Wong et. al., Journal of Clinical Oncology - Precision Oncology, 2017
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Identifying treatment resistance using ctDNA
Reversion of BRCA1/2 Germline Mutations Detected in Circulating Tumor DNA From Patients With High-Grade Serous Ovarian Cancer
Christie et al, JCO 2017
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Is ctDNA ready for prime timePre analytical variability
Standardisation of sample processing
Analytical validity
Different assay performance characteristics
Assay validation: quality control, robustness
Lower limit of detection ranging from 1%
What is the minimal analytical sensitivity and specificity to maximise clinical utility
Cross platform comparisons
Clinical Validity
Concordance b/w tissue and plasma; ? discordance due to biological or analytical factors
How consistently and accurately the test detects or predicts the intermediate or final outcome of interest; PPV and NPV
Clinical utility
How likely is the test to significantly improve patient outcomes?
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Endometrial cancer ctDNA study
CODEC Study – Circulating tumOur Dna in endometrial cancer
A prospective, pilot, “proof-of-concept” ctDNA study
45 patients from all stages
Aim: Determine if ctDNA analysis:
-can prognosticate for RFS/OS
-can be used as a MRD tool
-can reflect tumour responses in comparison to other biomarkers, e.g. CA125, HE4, PET scans
Rachel Delahunty, Petermac, Epworth, RWH
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Acknowledgements
Sarah-Jane Dawson
(and lab)
Mark Dawson
(and lab)
Grant McArthur
Con Tam
Andrew Roberts
John Seymour
Michael Dickinson
Shahneen Sandhu
Jeanette Raleigh
Athena Hatzimihalis
Rod Hicks
Jason Callahan
Tony Papenfuss
Jason Li
Ismael Vergara
Petermac Molecular
Genomics core
Sherene Loi
Geoff Lindeman
Stephen Fox
Molecular Pathology
Ben Solomon
Richard Marias
CASCADE investigators
MRV investigators
Collaborators
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Faculty Disclosure
Honoraria/
Expenses
Consulting/
Advisory
Board
Funded
Research
Royalties/
Patent
Stock
Options
Ownership
/ Equity
Position
EmployeeOther
(please specify)
March 30 - April 2, 2014Sheraton Sonoma CountyPetaluma, California
Off-Label Product Use
Will you be presenting or referencing off-label or investigational use of a therapeutic product?
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X No, nothing to disclose
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