Mapping kinase-dependent tumor immune adaptation with multiplexed single-cell CRISPR screens

Area of Science:

• Cancer immunology
• Genomics
• Single-cell analysis

Background:

• Tumor cells employ intrinsic programs to evade immune attack.
• Direct tumor-immune interactions can model these evasion mechanisms in vitro.

Purpose of the Study:

• To map tumor-intrinsic regulation of T cell-driven immune pressure in glioblastoma (GBM).
• To identify genetic perturbations that alter tumor state and adaptive responses to T cell engagement.

Main Methods:

• High-throughput single-cell CRISPR screening (CRISPR interference and activation) focused on the protein kinome.
• Allogeneic GBM-T cell co-culture with immune-matched antigen specificity.
• Massively multiplexed single-cell transcriptomics and deep generative models for data analysis.

Main Results:

• Resolved distinct modules of immune evasion, including antigen presentation, interferon/NF-κB signaling, oxidative stress resilience, and cytokine programs.
• Identified genetic perturbations that alter tumor transcriptional trajectories under T cell pressure.
• Discovered kinase targets (EPHA2, PDGFRA) involved in GBM immune evasion phenotypes.

Conclusions:

• The developed CRISPR screening framework provides a scalable blueprint for dissecting tumor-immune interactions.
• Inhibition of identified kinase targets enhances T cell-mediated killing of glioblastoma.
• This research elucidates genetic control mechanisms underlying tumor immune evasion.