Differential gene expression profile of first-generation and second-generation rapamycin-resistant allogeneic T cells

Background aimsWe completed a phase II clinical trial evaluating rapamycin-resistant allogeneic T cells (T-rapa) and now have evaluated a T-rapa product manufactured in 6 days (T-rapa6) rather than 12 days (T-Rapa12).MethodsUsing gene expression microarrays, we addressed our hypothesis that the two products would express a similar phenotype. The products had similar phenotypes using conventional comparison methods of cytokine secretion and surface markers.ResultsUnsupervised analysis of 34,340 genes revealed that T-rapa6 and T-rapa12 products clustered together, distinct from culture input CD4+ T cells. Statistical analysis of T-rapa6 products revealed differential expression of 19.3% of genes (n = 6641) compared with input CD4+ cells; similarly, 17.8% of genes (n = 6147) were differentially expressed between T-rapa12 products and input CD4+ cells. Compared with input CD4+ cells, T-rapa6 and T-rapa12 products were similar in terms of up-regulation of major gene families (cell cycle, stress response, glucose catabolism, DNA metabolism) and down-regulation (inflammatory response, immune response, apoptosis, transcriptional regulation). However, when directly compared, T-rapa6 and T-rapa12 products showed differential expression of 5.8% of genes (n = 1994; T-rapa6 vs. T-rapa12).ConclusionsSecond-generation T-rapa6 cells possess a similar yet distinct gene expression profile relative to first-generation T-rapa12 cells and may mediate differential effects after adoptive transfer.