CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D. Localized lung cancer COPD Screening Metastatic lung cancer ALK liquid biopsy Technical Insights into Highly Sensitive Isolation and Molecular Characterization of circulating tumor cells for early detection of tumor invasion. Patrizia Paterlini Bréchot, MD, Ph.D. Professor of Cellular and Molecular Biology University Paris Descartes– Paris Lucile Broncy, Msc, Ph.D. student (bourse CIFRE) University Paris Descartes– Paris
11
Embed
Technical Insights into Highly Sensitive Isolation and ... allows single tumor cells genetic characterization for theranostic purposes ! Ongoing developments for single tumor cell
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
Localized lung cancer
COPD Screening
Metastatic lung cancer ALK liquid biopsy
Technical Insights into Highly Sensitive Isolation and Molecular Characterization of circulating tumor cells for early
detection of tumor invasion.
Patrizia Paterlini Bréchot, MD, Ph.D. Professor of Cellular and Molecular Biology University Paris Descartes– Paris
Lucile Broncy, Msc, Ph.D. student (bourse CIFRE)
University Paris Descartes– Paris
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
Circulating Tumor Cells, more than a biomarker. Actor of invasion
ü Epithelial Tumor cells ü ≠ Normal Epithelial cells ü Mesenchymal Tumor cells ü Hybrid epithelial- mesenchymal Tumor cells ü Tumor stem cells ≠ Normal stem cells ü Circulating Tumor Microemboli (CTM)
Heterogeneous
ü Few per ml, mixed with ü 10 millions WBC, 5 billion RBC
• 17 control with benign disease: – 0/17 with CCC/CCM
Mazzini et al, Cancers, 2014
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
An independently funded, 6-year-long monocentric study, published in 2014, demonstrated that ISET® was able to make a very early diagnosis of lung cancer.
Project Air: Based on these results, a multicenter study with ISET® of 600 patients funded by independent Public National and Regional sources has started in November 2015 to confirm the results.
Project Detector: Another multicenter study using ISET® will start in December 2016 focusing on screening of tobacco-induced cancers. This independent study is part of the “Priorité Cancers Tabac” national research plan and has received significant financing from INCa (Institut national du cancer)
Major Clinical Trials M
ultic
ente
r Stu
dies
• 254 patients including 168 patients with COPD followed by ISET and low dose CT-scan screening
– 5 of 168 presented CCC detected by ISET – Per 10 mL of blood, 19 to 67 isolated CCC and 1 to
3 CCM, Vimentin expression • In these 5 patients,
– CT-scan detected a lung nodule 1 to 4 years after CCC detection by ISET leading to surgical resection and early diagnosis of Lung Cancer (stage 1A)
– Follow up of the 5 patients at 18 months after surgery: no tumor recurrence, no CCC.
Mon
ocen
tric
Stu
dy
Ilie et al. Plos One 2014
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
Lower Limit Of Detection (LLOD): 1 Tumor cell /10 ml of blood; sensitivity at LLOD: 100%
n=2 n=13
n=9
n=30
n=6 n=6
y = 0.9991x R2 = 0.96263
1
10
100
1000
1 10 100 1000 Num
ber o
f A54
9 ce
lls d
etec
ted
by IS
ET®
(log
sca
le)
Number of spiked A549 cells in 10 mL of blood or equivalent (log scale)
y = 1.0128x R! = 0.99514
0
20
40
60
80
100
120
0 20 40 60 80 100 120
Num
ber o
f HeL
a ce
lls d
etec
ted
by IS
ET®
Number of spiked HeLa cells in 1 mL of blood
Laget et al, Plos One, 2017
Capture of individual cells by micropipetting
Incubation with Rarecells® Buffer
Fixed cancer cells on the filter
Sens
itivi
ty te
sts
ISET®
Filtration
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
Culture of tumor cells and cytoskeleton analysis after live isolation from blood
Laget et al, Plos One, 2017
Blood Collection
Incubation with Rarecells® Live Cells Buffer (Ref 540208) Li
ve C
ells
CTC culture and functional assays
CTC-derived xenografts
RNA / Transcriptomic DNA / NGS analyses
ISET®
Filtration
CONFIDENTIAL – NOT FOR REDISTRIBUTION Patrizia Paterlini Bréchot, MD, Ph.D.
Targeted and global Next Generation Sequencing results obtained on single cancer cells isolated by ISET
Ion AmpliSeq Hotspot Cancer Panel V2: 2800 theranostic mutations in 50 genes K-RAS, PIK3CA and SMO: non-silent mutations found in single HCT116 cells
No mutations detected in ABL1, APC, FGFR3 and STK11 genes : these specific variants are not in the list of mutations of the panel
Rarecells Diagnostics • Ms. Katia Hormigos • Dr Sophie Laget • Lucile Broncy INSERM / Paris Descartes University • Lucile Broncy • Dr Basma Ben Jima • Dr Thierry Capiod • Dr Fatima Ben Mohamed • Pr Patrizia Paterlini Bréchot
Scientific collaborators • Dr Stephen Jackson and Dahlia Dhingra from Thermofisher Scientific • Dr Filip Van Nieuwerburgh and Lieselot Deleye from Ghent University