Related Experiment Video
Updated: Nov 27, 2025
Identification of Mediators of T-cell Receptor Signaling via the Screening of Chemical Inhibitor Libraries
Published on: January 22, 2019
Pharmacologic Screening Identifies Metabolic Vulnerabilities of CD8+ T Cells
Jefte M Drijvers1,2,3, Jacob E Gillis1,3, Tara Muijlwijk1,3
1Department of Immunology, Blavatnik Institute and Ludwig Center at Harvard, Harvard Medical School, Boston, Massachusetts.
Abstract:
Metabolic constraints in the tumor microenvironment constitute a barrier to effective antitumor immunity and similarities in the metabolic properties of T cells and cancer cells impede the specific therapeutic targeting of metabolism in either population. To identify distinct metabolic vulnerabilities of CD8+ T cells and cancer cells, we developed a high-throughput in vitro pharmacologic screening platform and used it to measure the cell type-specific sensitivities of activated CD8+ T cells and B16 melanoma cells to a wide array of metabolic perturbations during antigen-specific killing of cancer cells by CD8+ T cells. We illustrated the applicability of this screening platform by showing that CD8+ T cells were more sensitive to ferroptosis induction by inhibitors of glutathione peroxidase 4 (GPX4) than B16 and MC38 cancer cells. Overexpression of ferroptosis suppressor protein 1 (FSP1) or cytosolic GPX4 yielded ferroptosis-resistant CD8+ T cells without compromising their function, while genetic deletion of the ferroptosis sensitivity-promoting enzyme acyl-CoA synthetase long-chain family member 4 (ACSL4) protected CD8+ T cells from ferroptosis but impaired antitumor CD8+ T-cell responses. Our screen also revealed high T cell-specific vulnerabilities for compounds targeting NAD+ metabolism or autophagy and endoplasmic reticulum (ER) stress pathways. We focused the current screening effort on metabolic agents. However, this in vitro screening platform may also be valuable for rapid testing of other types of compounds to identify regulators of antitumor CD8+ T-cell function and potential therapeutic targets.
Insights
Metabolic vulnerabilities in tumors hinder immune responses. Researchers developed a screening platform to find unique metabolic weaknesses in CD8+ T cells and cancer cells, aiding targeted therapies.
Area of Science:
- Immunology
- Metabolic pathways
- Cancer biology
Background:
- Metabolic constraints within the tumor microenvironment limit effective antitumor immunity.
- Similar metabolic profiles of T cells and cancer cells complicate targeted metabolic therapies.
- Identifying distinct metabolic vulnerabilities is crucial for enhancing cancer immunotherapy.
Purpose of the Study:
- To develop and apply a high-throughput screening platform for identifying cell type-specific metabolic vulnerabilities.
- To compare the metabolic sensitivities of CD8+ T cells and cancer cells (B16 melanoma).
- To investigate the role of specific metabolic pathways and regulators in CD8+ T cell function and ferroptosis.
Main Methods:
- Developed a high-throughput in vitro pharmacologic screening platform.
- Assessed cell type-specific sensitivities to metabolic perturbations during antigen-specific killing.
- Utilized genetic manipulation (overexpression, deletion) to study ferroptosis regulators (GPX4, FSP1, ACSL4).
Main Results:
- CD8+ T cells exhibited higher sensitivity to ferroptosis induction by GPX4 inhibitors compared to cancer cells.
- Overexpression of FSP1 or cytosolic GPX4 conferred ferroptosis resistance to CD8+ T cells without impairing function.
- ACSL4 deletion protected CD8+ T cells from ferroptosis but diminished their antitumor responses.
- The screen identified T cell-specific vulnerabilities in NAD+ metabolism, autophagy, and ER stress pathways.
Conclusions:
- The developed screening platform effectively identifies distinct metabolic vulnerabilities in CD8+ T cells and cancer cells.
- Targeting specific metabolic pathways, like ferroptosis, offers potential for modulating CD8+ T cell function in cancer.
- The platform can be extended to test various compounds for identifying novel regulators of antitumor immunity and therapeutic targets.
Related Concept Videos
02:46Genetic Screens
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which...
01:29Therapeutic Drug Monitoring: Affecting Factors