A co-culture model to study modulators of tumor immune evasion through scalable arrayed CRISPR-interference screens

Autor: Ramiro Martinez, Chiara Finocchiaro, Louis Delhaye, Fien Gysens, Jasper Anckaert, Wim Trypsteen, Maarten Versteven, Eva Lion, Sandra Van Lint, Karim Vermaelen, Eric James de Bony, Pieter Mestdagh
Jazyk: angličtina
Rok vydání: 2024
Předmět:
Zdroj: Frontiers in Immunology, Vol 15 (2024)
Druh dokumentu: article
ISSN: 1664-3224
DOI: 10.3389/fimmu.2024.1444886
Popis: 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.
Databáze: Directory of Open Access Journals