Fig. 5: LNP^IT carrying PNP mRNA, combined with fludarabine phosphate, induces tumor suppression of HNSCC patient-derived xenografts.

a E. coli PNP transfection by LNP^IT was quantified in PDX tumors via an HPLC-based PNP enzymatic assay. b PNP activity was measured in PDX tumors harvested at different timepoints to determine the pharmacokinetics of PNP mRNA expression (n = 5–8 experimental replicates, mean +/− SD). Data analyzed by two-way ANOVA with Tukey post-hoc test for multiple comparisons. c Dose response of the LNP^IT-PNP when concentrated and administered at 40 µg per PDX tumor (n = 5–7 experimental replicates, mean +/− SD). Data analyzed by ordinary one-way ANOVA. d On day 14 after tumor inoculation, LNP^IT-PNP (or PBS for control groups) was injected at 20 µg/tumor in the morning and in the afternoon (40 µg total). The next day, fludarabine phosphate (or DMSO vehicle control) was injected in the morning and afternoon, and tumor volume measured twice a week. e Tumor volume was diminished for the group treated with LNP^IT-PNP and fludarabine, compared to the control groups (n = 6 experimental replicates, mean +/− SD). Data analyzed by two-way ANOVA with Tukey post-hoc test for multiple comparisons (p₁₂ for group 1 vs 2, p₁₃ for group 1 vs 3, p₁₄ for group 1 vs 4). f Higher probability of survival was observed in mice (N = 6) treated with the LNP^IT-PNP and fludarabine combination therapy (p_log-rank calculated with the log-rank/Mantel-Cox test, p_GBW calculated with the Gehan-Breslow-Wilcoxon test). Source data are provided as a Source Data file. Figure 5a was created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license (https://creativecommons.org/licenses/by-nc-nd/4.0/deed.en).