Abstract
Circulating tumor cells (CTCs) hold promise as biomarkers to aid in patient treatment stratification and disease monitoring. Because the number of cells is a critical parameter for exploiting CTCs for predictive biomarker’s detection, we developed a FISH (fluorescent in situ hybridization) method for CTCs enriched on filters (filter-adapted FISH [FA-FISH]) that was optimized for high cell recovery. To increase the feasibility and reliability of the analyses, we combined fluorescent staining and FA-FISH and developed a semi-automated microscopy method for optimal FISH signal identification in filtration-enriched CTCs . Here we present these methods and their use for the detection and characterization of ALK-, ROS1-, RET-rearrangement in CTCs from non-small-cell lung cancer and ERG-rearrangements in CTCs from prostate cancer patients.
Cyril Catelain and Emma Pailler are contributed equally to the manuscript.
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Competing Interests
The authors declare that they have no competing interests.
Funding
CC is supported by the Agence Nationale pour la Recherche (ANR-CE17-0006-01). EP is supported by the LabEx LERMIT (grant no ANR-10-LABX-0033-LERMIT) and the Fondation pour la Recherche Médicale (grant no FDT20150532072). VF is supported by the Fondation pour la Recherche Médicale (grants no FDT20160435543). The authors are grateful for the research support of the Fondation de France (grant no 201300038317), the Fondation ARC pour la Recherche sur le Cancer (grant no 20131200417), Innovative Medicines Initiative 11th Call CANCER ID (IMI-JU-11-2013, 115749), Institut National du Cancer (PRT-K14-032) and Agence Nationale de la Recherche (ANR-CE17-0006-01).
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Manuscript drafted by FF and CC. All authors have read and approved the final version of the manuscript.
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Catelain, C., Pailler, E., Oulhen, M., Faugeroux, V., Pommier, AL., Farace, F. (2017). Detection of Gene Rearrangements in Circulating Tumor Cells: Examples of ALK-, ROS1-, RET-Rearrangements in Non-Small-Cell Lung Cancer and ERG-Rearrangements in Prostate Cancer. In: Magbanua, M., Park, J. (eds) Isolation and Molecular Characterization of Circulating Tumor Cells. Advances in Experimental Medicine and Biology, vol 994. Springer, Cham. https://doi.org/10.1007/978-3-319-55947-6_9
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DOI: https://doi.org/10.1007/978-3-319-55947-6_9
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