Detection of Circulating Tumor Cells Harboring a Unique ALK Rearrangement in ALK-Positive Non–Small-Cell Lung Cancer

• Published in: Journal of clinical oncology Vol. 31; no. 18; pp. 2273 - 2281
• Main Authors: PAILLER, Emma, ADAM, Julien, SORIA, Jean Charles, VIELH, Philippe, BESSE, Benjamin, FARACE, Françoise, BARTHELEMY, Amélie, OULHEN, Marianne, AUGER, Nathalie, VALENT, Alexander, BORGET, Isabelle, PLANCHARD, David, TAYLOR, Melissa, ANDRE, Fabrice
• Format: Journal Article
• Language: English
• Published: Alexandria, VA American Society of Clinical Oncology 20.06.2013
• Subjects: Adult; Aged; Biological and medical sciences; Cadherins - genetics; Cadherins - metabolism; Carcinoma, Non-Small-Cell Lung - drug therapy; Carcinoma, Non-Small-Cell Lung - genetics; Carcinoma, Non-Small-Cell Lung - pathology; Cell Line, Tumor; Female; Gene Rearrangement; HeLa Cells; Humans; Immunohistochemistry; In Situ Hybridization, Fluorescence; Keratins - genetics; Keratins - metabolism; Lung Neoplasms - drug therapy; Lung Neoplasms - genetics; Lung Neoplasms - pathology; Male; MCF-7 Cells; Medical sciences; Middle Aged; Multiple tumors. Solid tumors. Tumors in childhood (general aspects); Neoplastic Cells, Circulating - metabolism; Pneumology; Protein Kinase Inhibitors - therapeutic use; Pyrazoles - therapeutic use; Pyridines - therapeutic use; Receptor Protein-Tyrosine Kinases - genetics; Receptor Protein-Tyrosine Kinases - metabolism; Tumors; Tumors of the respiratory system and mediastinum; Vimentin - genetics; Vimentin - metabolism; Lung disease; Cancerology; Respiratory disease; Lung cancer; Bronchus disease; Malignant tumor; Diagnosis; non-small cell lung carcinoma; Detection; Anaplastic lymphoma kinase; Tumor cell; Cancer
• ISSN: 0732-183X; 1527-7755
• DOI: 10.1200/JCO.2012.44.5932

The diagnostic test for ALK rearrangement in non-small-cell lung cancer (NSCLC) for crizotinib treatment is currently done on tumor biopsies or fine-needle aspirations. We evaluated whether ALK rearrangement diagnosis could be performed by using circulating tumor cells (CTCs). The presence of an ALK rearrangement was examined in CTCs of 18 ALK-positive and 14 ALK-negative patients by using a filtration enrichment technique and filter-adapted fluorescent in situ hybridization (FA-FISH), a FISH method optimized for filters. ALK-rearrangement patterns were determined in CTCs and compared with those present in tumor biopsies. ALK-rearranged CTCs and tumor specimens were characterized for epithelial (cytokeratins, E-cadherin) and mesenchymal (vimentin, N-cadherin) marker expression. ALK-rearranged CTCs were monitored in five patients treated with crizotinib. All ALK-positive patients had four or more ALK-rearranged CTCs per 1 mL of blood (median, nine CTCs per 1 mL; range, four to 34 CTCs per 1 mL). No or only one ALK-rearranged CTC (median, one per 1 mL; range, zero to one per 1 mL) was detected in ALK-negative patients. ALK-rearranged CTCs harbored a unique (3'5') split pattern, and heterogeneous patterns (3'5', only 3') of splits were present in tumors. ALK-rearranged CTCs expressed a mesenchymal phenotype contrasting with heterogeneous epithelial and mesenchymal marker expressions in tumors. Variations in ALK-rearranged CTC levels were detected in patients being treated with crizotinib. ALK rearrangement can be detected in CTCs of patients with ALK-positive NSCLC by using a filtration technique and FA-FISH, enabling both diagnostic testing and monitoring of crizotinib treatment. Our results suggest that CTCs harboring a unique ALK rearrangement and mesenchymal phenotype may arise from clonal selection of tumor cells that have acquired the potential to drive metastatic progression of ALK-positive NSCLC.