Immunogenetic Metabolomics Reveals Key Enzymes That Modulate CAR T-cell Metabolism and Function

Area of Science:

• Immunology
• Cancer Biology
• Metabolic Engineering

Background:

• Immune evasion by tumors is a major challenge for T cell immunotherapies.
• Tumor microenvironments (TME) often suppress T cell function metabolically.
• Genetic reprogramming of T cells offers a potential strategy to overcome immune evasion.

Purpose of the Study:

• To investigate if T cells can be genetically reprogrammed to overcome tumor-induced metabolic suppression.
• To identify metabolic regulators that enhance T cell function against cancer.
• To evaluate the therapeutic potential of engineered T cells in preclinical cancer models.

Main Methods:

• In silico screening to identify metabolic regulators (ADA, PDK1).
• Genetic engineering of chimeric antigen receptor (CAR) T cells for overexpression (OE) or deficiency of ADA and PDK1.
• In vitro assessment of T cell cytolysis, proliferation, and exhaustion.
• High-throughput transcriptomics and metabolomics analysis.
• In vivo studies using a colorectal cancer model.

Main Results:

• Overexpression of ADA and PDK1 enhanced CAR T cell-mediated cancer cell killing.
• ADA-overexpressing CAR T cells showed improved function in the presence of adenosine, an immunosuppressive metabolite.
• Genetic modifications altered global gene expression and metabolic profiles of CAR T cells.
• ADA-OE improved CAR T cell proliferation, reduced exhaustion, enhanced tumor infiltration, and promoted tumor clearance in vivo.

Conclusions:

• Metabolic reprogramming of CAR T cells by targeting ADA and PDK1 is a viable strategy to enhance anti-cancer immunity.
• Engineered CAR T cells demonstrate improved efficacy in preclinical models.
• These findings provide novel targets for improving the effectiveness of CAR T cell therapy.