AUTHOR=Martinez Ramiro , Finocchiaro Chiara , Delhaye Louis , Gysens Fien , Anckaert Jasper , Trypsteen Wim , Versteven Maarten , Lion Eva , Van Lint Sandra , Vermaelen Karim , de Bony Eric James , Mestdagh Pieter TITLE=A co-culture model to study modulators of tumor immune evasion through scalable arrayed CRISPR-interference screens JOURNAL=Frontiers in Immunology VOLUME=15 YEAR=2024 URL=https://www.frontiersin.org/journals/immunology/articles/10.3389/fimmu.2024.1444886 DOI=10.3389/fimmu.2024.1444886 ISSN=1664-3224 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.