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OPTIMIZATION OF A FLOW CYTOMETRY METHOD FOR THE APPROACH OF LIQUID BIOPSY AS A THERAPY MODULATION TOOL IN PATIENTS WITH COLORECTAL CANCER

ARIANA HUDIȚĂ ^1#, VLAD IOANA-LAVRIC ^1#, ANCA ZAMFIR ^2#, LAURA BUBURUZAN ^2#, OCTAV GINGHINĂ ^3#*, CAROLINA NEGREI ^4#, GEORGE TRAIAN ALEXANDRU BURCEA DRAGOMIROIU ^5#, MARIETA COSTACHE ^1#, CARMEN ARDELEANU ^2#, EUGEN RADU ^6#, DANIELA ELENA POPA ^5#, MARIA BÂRCĂ ^5#, NICULAE IORDACHE ^3#, IULIANA CEAUȘU ^7#, BIANCA GĂLĂȚEANU ^1#

1.Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Bucharest, 91-95 Splaiul Independenței, 050095, Bucharest, Romania
2.OncoTeam Diagnostic, 313A Splaiul Unirii Street, 030138, Bucharest, Romania
3.Department of Surgery, “Sf. Ioan” Clinical Emergency Hospital, Faculty of Dental Medicine, “Carol Davila” University of Medicine and Pharmacy, 13 Vitan Bârzești Road, 042122, Bucharest, Romania
4.Departament of Toxicology, Faculty of Pharmacy, “Carol Davila” University of Medicine and Pharmacy, 6 Traian Vuia Street, 020956, Bucharest, Romania
5.Departament of Drug Control, Faculty of Pharmacy, “Carol Davila” University of Medicine and Pharmacy, 6 Traian Vuia Street, 020956, Bucharest, Romania
6.“Molimagex” Molecular Biology and Pathology Research Lab, University Hospital Bucharest, 169 Splaiul Independenței Street, 050098 Bucharest, Romania
7.Faculty of Medicine, University of Medicine and Pharmacy “Carol Davila”, Department of Obstetrics and Gynaecology, “Dr. I. Cantacuzino” Hospital, 5-7 Ion Movilă Street, Bucharest, Romania

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Personalized medicine in oncology aims to tailor a particular dynamic treatment for the individual, starting from the diagnostic throughout therapy. To guide the appropriate treatment decisions, liquid biopsy is being used as a real time monitoring analysis targeting the detection and analysis of: (i) circulating tumour cells that shed from the tumours and circulate through the blood stream, (ii) circulating tumour DNA (cell free DNA originated from apoptotic and necrotic tumour cells) and (iii) exosomes of tumour origin. Many techniques were developed to isolate cells of epithelial origin in whole blood based on the expression of cell-surface markers like EpCAM and panCK. However, due to their low number (1 - 10 cells/mL of whole blood) as compared to normal blood cells, enrichment strategies are to be developed for optimum results. In this context, the aim of this study was to develop a flow cytometry protocol to detect the circulating tumour cells in patients with colon cancer, with high impact on therapy modulation.