Mitochondrial Membrane Potential Identifies Cells with Enhanced Stemness for Cellular Therapy

Long-term survival and antitumor immunity of adoptively transferred CD8+ T cells is dependent on their metabolic fitness, but approaches to isolate therapeutic T cells based on metabolic features are not well established. Here we utilized a lipophilic cationic dye tetramethylrhodamine methyl ester (TMRM) to identify and isolate metabolically robust T cells based on their mitochondrial membrane potential (ΔΨm). Comprehensive metabolomic and gene expression profiling demonstrated global features of improved metabolic fitness in low-ΔΨm-sorted CD8+ T cells. Transfer of these low-ΔΨm T cells was associated with superior long-term in vivo persistence and an enhanced capacity to eradicate established tumors compared with high-ΔΨm cells. Use of ΔΨm-based sorting to enrich for cells with superior metabolic features was observed in CD8+, CD4+ T cell subsets, and long-term hematopoietic stem cells. This metabolism-based approach to cell selection may be broadly applicable to therapies involving the transfer of HSC or lymphocytes for the treatment of viral-associated illnesses and cancer. [Display omitted] •ΔΨm-based sorting segregates short-lived effector from memory T cell precursors•Low-ΔΨm CD8+ T cells demonstrate decreased oxidative stress•Low-ΔΨm T cells demonstrate superior antitumor activity•Low-ΔΨm marks self-renewing hematopoietic stem cells Metabolic fitness is required for long-term function of T cells and HSC. Sukumar et al. describe a simple and clinically feasible method to isolate such metabolically robust cells, using a single parameter—mitochondrial membrane potential (ΔΨm)—for long-term survival, antitumor immunity, and hematopoietic reconstitution.