Romanian Society of Pharmaceutical Sciences

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OPTIMIZATION OF A FLOW CYTOMETRY PROTOCOL FOR PD-1/CTLA-4 IMMUNE CHECKPOINTS RECEPTORS DETECTION IN COLORECTAL CANCER TUMOUR MICROENVIRONMENT

MARIUS ZAMFIR 1,2, ARIANA HUDIȚĂ 3, MARA MARDARE 1,2, IRINA BONDOC 2, ANDREI VĂCĂRAȘU 2, BOGDAN COSMIN TĂNASE 2, GEORGE TRAIAN ALEXANDRU BURCEA-DRAGOMIROIU 4, MIRCEA LIȚESCU 5, THEODOR VOIOSU 6, BIANCA GĂLĂȚEANU 3*, CLAUDE LAMBERT 7, OCTAV GINGHINĂ 2,8

1Doctoral School of Medicine, “Carol Davila” University of Medicine and Pharmacy Bucharest, Romania
2Department of Surgery 3, “Prof. Dr. Al. Trestioreanu” Institute of Oncology Bucharest, Romania
3Department of Biochemistry and Molecular Biology, University of Bucharest, Bucharest, Romania
4Department of Drug Control, Faculty of Pharmacy, “Carol Davila” University of Medicine and Pharmacy Bucharest, Romania
5Department of Surgery, “Sf. Ioan” Clinical Emergency Hospital, Bucharest, Romania
6Department of Gastroenterology, Colentina Clinical Hospital, Bucharest, Romania
7Immunology lab, Central University Hospital, Saint-Etienne, France
8Department II, Faculty of Dental Medicine, “Carol Davila” University of Medicine and Pharmacy Bucharest, Romania

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Considering the individual heterogeneity of the tumours, a personalized approach in oncology is crucial to design a particular dynamic treatment for the individual, from the diagnostic, to monitoring and therapy. In particular, the cellular makeup of the tumour microenvironment consists of diverse immune cells, which include tumour-infiltrating lymphocytes (TILs), as well as tumour-associated macrophages (TAMs) and tumour-associated neutrophils (TANs). These immune cells express various immune checkpoint molecules such as PD-1 and CTLA-4 that play a crucial role in modulating the colorectal cancer microenvironment, controlling the disease's development and response to therapy. Considering the huge potential of PD-1/PD-L1 and CTLA-4 as actionable targets for immunotherapy in colorectal cancer as well as the advantages of the flow cytometry technique, we propose here a flow cytometry method to characterize the colorectal cancer microenvironment in terms of cells populations and their associate immune checkpoint molecular profile.