Clinically relevant T cell expansion media activate distinct metabolic programs uncoupled from cellular function.

Area of Science:

• Immunology and Cell Biology
• Metabolic Engineering
• Cancer Immunotherapy

Background:

• Current ex vivo T-cell expansion methods for immunotherapy may induce metabolic phenotypes hindering in vivo therapeutic efficacy.
• Understanding T-cell metabolism during expansion is crucial for optimizing cell-based therapies.

Purpose of the Study:

• To compare the impact of five different culture media on T-cell expansion outcomes.
• To investigate metabolic adaptations of expanded T-cells and their relationship with function.
• To assess the influence of ovarian cancer ascites on T-cell function post-expansion.

Main Methods:

• Comparative analysis of five distinct clinical T-cell expansion media.
• Assessment of T-cell proliferation, differentiation, function, activation, and mitochondrial phenotypes.
• Metabolic profiling using glucose and glutamine uptake assays and glucose isotope labeling.
• Functional assessment of T-cells cultured in ovarian cancer patient ascites.

Main Results:

• Culture media, not stimulation conditions, determined T-cell proliferation and function.
• Expanded T-cells dynamically adapted their metabolism (glucose/glutamine uptake) to different media formulations.
• Metabolic phenotype adoption was uncoupled from T-cell function.
• T-cells expanded in ovarian cancer ascites exhibited suppressed mitochondrial activity and function, independent of prior expansion media.

Conclusions:

• Ex vivo expansion media profoundly influence T-cell metabolism and function.
• Metabolic adaptations during expansion do not guarantee improved in vivo therapeutic efficacy.
• Tumor microenvironment factors, like ascites, can override ex vivo culture effects on T-cell function.