Objectives of the Workpackage

This work package is to investigate CTC and plasma nucleic acids in Non Small Cell Lung Cancer (NSCLC) and has four main objectives:

•    CTC enumeration with different approaches
•    CTC characterization
•    Analysis of plasma nucleic acids
•    Clinical validation in NSCLC

Description of the tasks

CTC enumeration with different approaches

CTC will be enumerated in NSCLC patients using 1. the “gold standard” CellSearch (Lead Leon Terstappen); 2. CellSearch blood depleted from EpCAM+ CTC will be investigated for the presence of EpCAM- CTC (Lead Leon Terstappen). 3. The CellCollector needle (Lead Klaus Pantel), 4. Large blood volumes obtained by leukapheresis and processed with different technologies (Lead Nickolas Stoecklein), 5. Technologies identified as being promising. CTC from all approaches will be scored using the Open Source CTC scoring algorithm.

CTC characterization

This objective is divided up in three tasks. 1.Comparison of isolation methods to obtain single CTCs. Each participating site will process blood spiked with cell lines and report results (Lead Nickolas Stoecklein). 2. Standardization of quantitative protein expression analysis. The expression of EGFR in NSCLC and Her2 in Her2RMBC on CTC will be used to compare results between sites and technologies using the Open Source computer program (Lead Leon Terstappen). 3. Functional analysis of viable CTCs using the in vitro dual fluoro-EPISPOT assay and an in vivo xeno-transplant lung cancer CTC model (Lead Catherine Alix-Panabières).

Analysis of plasma nucleic acids

Standard Operating Procedures for nucleic acid analysis will be generated. This will include protocols for: 1. gene mutation, rearrangement and amplification analyses in CTCs and ctDNA (Lead Michael Speicher). 2 copy number analyses and gene expression in CTCs and ctDNA ctDNA (Lead Michael Speicher). 3. novel biomarker identification using exome and whole genome sequencing(Lead Hans Lehrach). 4. cfmiRNA analyses in plasma(Lead Klaus Pantel).

Clinical validation in NSCLC

The selected methods will be validated in clinical studies and compared to the results from tissue biopsies. The results will be correlated with therapy response in cohorts of 150 NSCLC patients in total coming from the following studies:

1. NVALT-17 (Lead: Harry Groen) a randomized phase III study in the Netherlands of erlotinib compared to intercalated erlotinib with cisplatinum pemetrexed as first-line therapy for advanced EGFR mutated NCSLC.

Find more information here: http://www.trialregister.nl/trialreg/admin/rctview.asp?TC=4410

2. TRACERx (Lead: Caroline Dive) a study in the UK enrolling patients with resectable NSCLC cancer aiming to define the evolutionary trajectories of lung cancer in both space and time in relation to selection pressures imposed by cancer therapies when disease recurs.

Find more information here: http://www.cruklungcentre.org/Research/TRACERx

3. SPECTAlung (Lead Stephane Lejeune) study aiming to identify predictive and resistance biomarkers to targeted therapies where molecular profiling of tumor biopsies or resected NSCLC tumors will be performed. 

Find more information here: http://www.eortc.org/news/spectalung-open-to-patient-entry/ 

4. NSCLC patients receiving standard of care in the participating institutions

>> WP 2: Small indication: Her2-therapy resistant metastatic breast cancer

Participants