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Multi-phenotype CRISPR-Cas9 Screen Identifies p38 Kinase as a Target for Adoptive Immunotherapies
Devikala Gurusamy1, Amanda N Henning1, Tori N Yamamoto2
1Surgery Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD 20892, USA; Center for Cell-based Therapy, Center for Cancer Research, National Institutes of Health (NIH), Bethesda, MD 20892, USA.
Abstract:
T cells are central to all currently effective cancer immunotherapies, but the characteristics defining therapeutically effective anti-tumor T cells have not been comprehensively elucidated. Here, we delineate four phenotypic qualities of effective anti-tumor T cells: cell expansion, differentiation, oxidative stress, and genomic stress. Using a CRISPR-Cas9-based genetic screen of primary T cells we measured the multi-phenotypic impact of disrupting 25 T cell receptor-driven kinases. We identified p38 kinase as a central regulator of all four phenotypes and uncovered transcriptional and antioxidant pathways regulated by p38 in T cells. Pharmacological inhibition of p38 improved the efficacy of mouse anti-tumor T cells and enhanced the functionalities of human tumor-reactive and gene-engineered T cells, paving the way for clinically relevant interventions.
Insights
Effective anti-tumor T cells possess four key traits. Targeting p38 kinase enhances T cell function, improving cancer immunotherapy efficacy in preclinical models and human cells.
Area of Science:
- Immunology
- Cancer Biology
- Cellular Biology
Background:
- T cells are crucial for cancer immunotherapy effectiveness.
- Key characteristics of therapeutically effective anti-tumor T cells remain incompletely understood.
- Identifying these characteristics is vital for advancing cancer treatment.
Purpose of the Study:
- To identify phenotypic qualities of effective anti-tumor T cells.
- To investigate the role of T cell receptor-driven kinases in regulating these phenotypes.
- To explore p38 kinase as a potential therapeutic target in cancer immunotherapy.
Main Methods:
- Utilized a CRISPR-Cas9 genetic screen on primary T cells.
- Assessed the impact of kinase disruption on T cell phenotypes: expansion, differentiation, oxidative stress, and genomic stress.
- Employed pharmacological inhibition of p38 kinase.
Main Results:
- Delineated four critical phenotypes of effective anti-tumor T cells.
- Identified p38 kinase as a central regulator of these four phenotypes.
- Uncovered p38-regulated transcriptional and antioxidant pathways within T cells.
- Demonstrated that p38 inhibition enhances anti-tumor T cell efficacy in mouse models.
- Showed improved functionality of human tumor-reactive and gene-engineered T cells upon p38 inhibition.
Conclusions:
- p38 kinase is a key regulator of T cell phenotypes critical for anti-tumor immunity.
- Pharmacological targeting of p38 kinase represents a promising strategy to enhance cancer immunotherapies.
- This research provides a foundation for developing novel, clinically relevant interventions to boost T cell-mediated cancer treatment.
