Fig. 6: Therapeutic vaccination specifically eliminated MC38 WT tumor cells bearing the targeted neo-mutations.
Mice were engrafted s.c. with 2.5 × 105 MC38 WT cells at day 0 and were then vaccinated on day 6 with 1 × 109 TU of various LV (n = 7). DNA and RNA were extracted from the tumors. Neo-mutation loci were amplified by PCR on genomic DNA, and VAF was analyzed by NGS (d–f). Transcript levels of mutated genes were quantified by RT-qPCR on RNA-derived cDNA (g). a Experiment timeline. b Tumor growth curves. c Kaplan–Meier survival analysis. d VAF comparison for neo-mutations in healthy colon tissue from female C57BL6/j mice (n = 2), cultured MC38 (n = 1), and MC38 tumors from the mice treated with Ctrl LV (n = 7). e VAF of neo-mutations in tumors treated with each of the LV vaccines. f Mean percentage loss of VAF (red) for each neo-mutation in tumors from vaccinal LV-treated mice relative to tumors from Ctrl LV-treated mice. g Heatmap of transcript expression changes for neo-mutated genes, normalized on β2m transcripts and compared to the Ctrl LV group. Statistical significance was determined by log-rank test for survival (c), and by one-way ANOVA for VAF comparisons (e). (* = p < 0.05, ** = p < 0.01, *** = p < 0.001, **** = p < 0.0001).
