Frontiers in Immunology (Oct 2024)
A co-culture model to study modulators of tumor immune evasion through scalable arrayed CRISPR-interference screens
- Ramiro Martinez,
- Ramiro Martinez,
- Ramiro Martinez,
- Chiara Finocchiaro,
- Louis Delhaye,
- Louis Delhaye,
- Louis Delhaye,
- Louis Delhaye,
- Fien Gysens,
- Fien Gysens,
- Fien Gysens,
- Jasper Anckaert,
- Jasper Anckaert,
- Jasper Anckaert,
- Wim Trypsteen,
- Wim Trypsteen,
- Wim Trypsteen,
- Maarten Versteven,
- Eva Lion,
- Sandra Van Lint,
- Sandra Van Lint,
- Sandra Van Lint,
- Karim Vermaelen,
- Karim Vermaelen,
- Karim Vermaelen,
- Eric James de Bony,
- Eric James de Bony,
- Eric James de Bony,
- Pieter Mestdagh,
- Pieter Mestdagh,
- Pieter Mestdagh
Affiliations
- Ramiro Martinez
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Ramiro Martinez
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Ramiro Martinez
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Chiara Finocchiaro
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Louis Delhaye
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Louis Delhaye
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Louis Delhaye
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Louis Delhaye
- Center for Medical Biotechnology, Flanders Institute for Biotechnology – UGENT (VIB-UGENT), Ghent, Belgium
- Fien Gysens
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Fien Gysens
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Fien Gysens
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Jasper Anckaert
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Jasper Anckaert
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Jasper Anckaert
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Wim Trypsteen
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Wim Trypsteen
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Wim Trypsteen
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Maarten Versteven
- Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, Antwerp, Belgium
- Eva Lion
- Laboratory of Experimental Hematology, Vaccine and Infectious Disease Institute (VAXINFECTIO), Faculty of Medicine and Health Sciences, University of Antwerp, Antwerp, Belgium
- Sandra Van Lint
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Sandra Van Lint
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Sandra Van Lint
- Tumor Immunology Laboratory, Department of Pulmonary Medicine, Ghent University, Ghent, Belgium
- Karim Vermaelen
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Karim Vermaelen
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Karim Vermaelen
- Tumor Immunology Laboratory, Department of Pulmonary Medicine, Ghent University, Ghent, Belgium
- Eric James de Bony
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Eric James de Bony
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Eric James de Bony
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- Pieter Mestdagh
- OncoRNALab, Center for Medical Genetics (CMGG), Ghent University, Ghent, Belgium
- Pieter Mestdagh
- Cancer Research Institute Ghent (CRIG), Ghent University, Ghent, Belgium
- Pieter Mestdagh
- Department of Biomolecular Medicine, Ghent University, Ghent, Belgium
- DOI
- https://doi.org/10.3389/fimmu.2024.1444886
- Journal volume & issue
-
Vol. 15
Abstract
Cancer cells effectively evade immune surveillance, not only through the well-known PD-1/PD-L1 pathway but also via alternative mechanisms that impair patient response to immune checkpoint inhibitors. We present a novel co-culture model that pairs a reporter T-cell line with different melanoma cell lines that have varying immune evasion characteristics. We developed a scalable high-throughput lentiviral arrayed CRISPR interference (CRISPRi) screening protocol to conduct gene perturbations in both T-cells and melanoma cells, enabling the identification of genes that modulate tumor immune evasion. Our study functionally validates the co-culture model system and demonstrates the performance of the CRISPRi-screening protocol by modulating the expression of known regulators of tumor immunity. Together, our work provides a robust framework for future research aimed at systematically exploring mechanisms of tumor immune evasion.
Keywords
