[en] In the past 20 years, the interest for the tumor microenvironment (TME) has exponentially increased. Indeed, it is now commonly admitted that the TME plays a crucial role in cancer development, maintenance, immune escape and resistance to therapy. This stands true for hematological malignancies as well. A considerable amount of newly developed therapies are directed against the cancer-supporting TME instead of targeting tumor cells themselves. However, the TME is often not clearly defined. In addition, the unique phenotype of each tumor and the variability among patients limit the success of such therapies. Recently, our group took advantage of the mass cytometry technology to unveil the specific TME in the context of chronic lymphocytic leukemia (CLL) in mice. We found the enrichment of LAG3 and PD1, two immune checkpoints. We tested an antibody-based immunotherapy, targeting these two molecules. This combination of antibodies was successful in the treatment of murine CLL. In this methods article, we provide a detailed protocol for the staining of CLL TME cells aiming at their characterization using mass cytometry. We include panel design and validation, sample preparation and acquisition, machine set-up, quality control, and analysis. Additionally, we discuss different advantages and pitfalls of this technique.
Disciplines :
Hematology
Author, co-author :
GONDER, Susanne ✱; University of Luxembourg ; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
Fernandez Botana, Iria ✱; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Wierz, Marina ✱; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
Pagano, Giulia; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg ; Faculty of Science, Technology and Medicine, University of Luxembourg, Esch-sur-Alzette, Luxembourg
Gargiulo, Ernesto; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
Cosma, Antonio; National Cytometry Platform, Quantitative Biology Unit, Transversal Activities, Luxembourg Institute of Health, Luxembourg, Luxembourg
MOUSSAY, Etienne ; University of Luxembourg ; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
PAGGETTI, Jerome ; University of Luxembourg ; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
Largeot, Anne; Tumor Stroma Interactions, Department of Oncology, Luxembourg Institute of Health, Luxembourg, Luxembourg
✱ These authors have contributed equally to this work.
External co-authors :
no
Language :
English
Title :
Method for the Analysis of the Tumor Microenvironment by Mass Cytometry: Application to Chronic Lymphocytic Leukemia.
Fonds De La Recherche Scientifique - FNRS Fonds De La Recherche Scientifique - FNRS Fonds De La Recherche Scientifique - FNRS Fonds De La Recherche Scientifique - FNRS Fonds De La Recherche Scientifique - FNRS Fonds De La Recherche Scientifique - FNRS Fonds National de la Recherche Luxembourg Fonds National de la Recherche Luxembourg
Funding text :
This work was supported by grants from FNRS “Télévie” to SG, IF, MW, GP, and AL (7.4502.19, 7.4529.19, 7.6504.18, 7.4501.18, 7.4502.17, and 7.4503.19), from FNR Luxembourg to EG and JP (PRIDE15/10675146/CANBIO and INTER/DFG/16/11509946) and from Plooschter Projet.
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