TY - JOUR
T1 - Assessment of the mutational status of NSCLC using hypermetabolic circulating tumor cells
AU - Turetta, Matteo
AU - Bulfoni, Michela
AU - Brisotto, Giulia
AU - Fasola, Gianpiero
AU - Zanello, Andrea
AU - Biscontin, Eva
AU - Mariuzzi, Laura
AU - Steffan, Agostino
AU - Di Loreto, Carla
AU - Cesselli, Daniela
AU - Del Ben, Fabio
PY - 2018/8/14
Y1 - 2018/8/14
N2 - Molecular characterization is currently a key step in NSCLC therapy selection. Circulating tumor cells (CTC) are excellent candidates for downstream analysis, but technology is still lagging behind. In this work, we show that the mutational status of NSCLC can be assessed on hypermetabolic CTC, detected by their increased glucose uptake. We validated the method in 30 Stage IV NSCLC patients: peripheral blood samples were incubated with a fluorescent glucose analog (2-NBDG) and analyzed by flow cytometry. Cells with the highest glucose uptake were sorted out. EGFR and KRAS mutations were detected by ddPCR. In sorted cells, mutated DNA was found in 85% of patients, finding an exact match with primary tumor in 70% of cases. Interestingly, in two patients multiple KRAS mutations were detected. Two patients displayed different mutations with respect to the primary tumor, and in two out of the four patients with a wild type primary tumor, new mutations were highlighted: EGFR p.746_750del and KRAS p.G12V. Hypermetabolic CTC can be enriched without the need of dedicated equipment and their mutational status can successfully be assessed by ddPCR. Finally, the finding of new mutations supports the possibility of probing tumor heterogeneity.
AB - Molecular characterization is currently a key step in NSCLC therapy selection. Circulating tumor cells (CTC) are excellent candidates for downstream analysis, but technology is still lagging behind. In this work, we show that the mutational status of NSCLC can be assessed on hypermetabolic CTC, detected by their increased glucose uptake. We validated the method in 30 Stage IV NSCLC patients: peripheral blood samples were incubated with a fluorescent glucose analog (2-NBDG) and analyzed by flow cytometry. Cells with the highest glucose uptake were sorted out. EGFR and KRAS mutations were detected by ddPCR. In sorted cells, mutated DNA was found in 85% of patients, finding an exact match with primary tumor in 70% of cases. Interestingly, in two patients multiple KRAS mutations were detected. Two patients displayed different mutations with respect to the primary tumor, and in two out of the four patients with a wild type primary tumor, new mutations were highlighted: EGFR p.746_750del and KRAS p.G12V. Hypermetabolic CTC can be enriched without the need of dedicated equipment and their mutational status can successfully be assessed by ddPCR. Finally, the finding of new mutations supports the possibility of probing tumor heterogeneity.
KW - CTC
KW - Glucose uptake
KW - Liquid biopsy
KW - Metabolism
KW - Non-small cell lung cancer
UR - http://www.scopus.com/inward/record.url?scp=85052512903&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85052512903&partnerID=8YFLogxK
U2 - 10.3390/cancers10080270
DO - 10.3390/cancers10080270
M3 - Article
VL - 10
JO - Cancers
JF - Cancers
SN - 2072-6694
IS - 8
M1 - 270
ER -