ESP/ACSP session Precision medicine, cancer diagnosis and treatment selection/monitoring
Circulating Free DNA or Circulating Tumour Cells in Lung Cancer What to choose?
Paul Hofman Laboratory of Clinical and Experimental Pathology Liquid Biopsy Laboratory & FHU OncoAge University of Nice Sophia Antipolis, Nice, Fr (www.oncoage.org)
• Short background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
• Short background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
The right drug
The right patient
The right time
Stratified Medicine
The right approach
The right cost
The right turnaround time! The right drug
The right patient
The right time
Stratified Medicine The right sample! solid and/or liquid biopsies? The right approach
The right cost
Liquid biopsy: Low volume of whole blood sample (10/20 ml) for assessment of prognosis and predictive biomarkers
Liquid biopsy is more than a substantial overturning in lung cancer patient taking care
The different compartments of a liquid biopsy
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Hofman et al, Curr Mol Med , 2014
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Exosomes
Free DNA Free long RNA Free microRNA
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Hofman et al, Curr Mol Med , 2014
Free tumor DNA
Exosomes
Free DNA Free « germinal » DNA
Free RNA Free microRNA
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Hofman et al, Curr Mol Med , 2014
Liquid biopsy CTCs vs. cf DNA ? What is the good choice for lung cancer patient management ?
Current challenges for detection of circulating tumor cells and cell-free circulating nucleic acids, and their characterization in NSCLC. What is the best blood substrate for personalized medicine? Ilie M and Hofman P, Ann Transl Med. 2014;2(11):107.
• Short background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
CTC detection (enrichment)
1 First challenge for CTC detection
Sensitivity: how to detect a rare event into the blood stream ?
Epithelio-mesenchymal phenomenon
2 Second challenge for CTC detection
Specificity: how to avoid pitfalls (false negative results) due to cell population heterogeneity ?
CellSearch An indirect method
ISET A direct method
Hofman et al, Clin Cancer Res 2011 Hofman et al, Int J Cancer, 2011 Hofman et al, Am J Clin Pathol, 2011 Hofman et al, Cytopathology, 2011 Hofman et al, Cancer Cytopathol, 2016
• Background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
Technology to assess cfDNA Luis Diaz, AACR 2016 •
Digital PCR – Best for individual point mutations but can be used for crude copy number analysis – Sensitivity is dependent on specific mutation and assay optimization – Multiplexing assay is possible – Fast and highly reproducible-results in hours – Minimal bioinformatics needs – Inexpensive
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FDA-test approved (companion diagnostic testing) •
• •
Therascreen EGFR RGQ PCR Kit for EGFR deletions 19 and L858R (Qiagen) – Rotor-Gene Q MDx • Roche Cobas EGFR Mutation test V2 Kit for deletions 19, L858 R and T790M – Cobas Z480 Others Next-generation sequencing
– Has been used for point mutations, rearrangements, genomic amplifications, aneuploidy, whole exome and whole exome sequencing – High false discovery rate that requires pre-sequencing barcoding and post-sequencing bioinformatics for error suppression – Quite Expensive – Turnaround time ?
• Background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
Advantages and limitations of the detection of circulating tumor cells Advantages
Limitations
Advantages and limitations of the detection of circulating free DNA
Advantages
Limitations
Alternative or complementary ?... Focus on one example T790M (acquired resistance EGFR mutation) detection in lung cancer patients 40 EGFR positive patients and progression under TKI treatment ctDNA
T790M+
CTCs T790M-
ctDNA & CTCs
Biopsy
N/A
Re biopsy
Tissue and liquid biopsies are not concordant at 100% (spatio-temporal tumour heterogeneity) No gold standard at the end? Combination of ctDNA and CTCs and tissue biopsy increases the efficiency!
Detection of T790M, the acquired resistance EGFR mutation, by tumor biopsy versus noninvasive blood-based analyses. Sundaresan TK, et al. Clin Cancer Res. 2015
We need to be aware! Can tissue biopsy be replaced by liquid biopsy? Ilié M and Hofman P, Transl Lung Cancer Res, 2016 Aug;5(4):420-3
Liquid biopsy versus tissue biopsy…What to choose in lung cancer patients..?! Liquid Biopsy
Tissue Biopsy
P Hofman & HH Popper, Virchow Archivs, 2016 P Hofman, Curr Opin Oncol, 2016 in press
• Short background • Detection and characterisation of CTCs in lung cancer patients • Detection and characterisation of cfDNA in lung cancer patients • What to choose ? • Conclusion
• cfDNA – In clinical routine practice for identifying actionable genetic alterations – For patients lack tissue for molecular analysis – For patients whose tumors have evolved over time and treatment (too risky to perform or after relapse when biopsies are not routine) • Discordance between mutations in primary/metastases lesions • Acquired resistance (e.g. patients who develop resistance to EGFR blockade)
• CTC – – – –
Exploring the pathophysiology of lung cancer diseases Translational research project Xenograft establishment Patients follow up under treatment (prognosis)
• Complementary approaches of both cfDNA and CTC detection – Detection of occult disease (early detection, minimal residual and screening test) – Potentiality for better detection of some genetic alterations ?
A breath of relaxation Take off, Nice 25 September 2016, 11.45 AM
ESP/ACSP session Precision medicine, cancer diagnosis and treatment selection/monitoring
Circulating Free DNA or Circulating Tumour Cells in Lung Cancer What to choose?
Paul Hofman Laboratory of Clinical and Experimental Pathology Liquid Biopsy Laboratory & FHU OncoAge University of Nice Sophia Antipolis, Nice, Fr (www.oncoage.org)
