Extended Data Fig. 9: Suppression of metastasis by DON is enhanced by co-treatment with ASNase in a primary PDAC orthotopic mouse model.
(a) Tumor weights of 779E orthotopic tumors from animals treated with vehicle (Ctrl), ASNase (3U/g), DON (5 mg/kg), or combination of DON + ASNase for 2 weeks. Data are presented as mean ± SEM of n = 9, n = 7, n = 8, or n = 9 mice per group. Ctrl vs ASNase, P = 0.5836. Ctrl vs DON, P = 0.042. Ctrl vs DON+ASNase, P = 0.0491. (b) Characterization of macrometastases in animals with 779E orthotopic tumors treated as indicated. Representative images of macrometastases in the different tissues are shown. The white arrows point to macrometastases. The number of mice with macrometastases in each organ site was quantified as indicated in the table. n = 9, n = 7, n = 8, or n = 9 mice per group. (c) Quantification of the percentage of mice with 779E-derived primary orthotopic tumors that presented with macrometastases to any organ (all sites), the liver, intestines, or diaphragm. n = 9, n = 7, n = 8, or n = 9 mice per group. All Sites: Ctrl vs DON, P = 0.029; Ctrl vs DON+ASNase, P = 0.009. Liver: Ctrl vs DON, P = 0.0498; Ctrl vs DON+ASNase, P = 0.0034. Diaphragm: Ctrl vs DON, P = 0.0498; Ctrl vs DON+ASNase, P = 0.347. (d) Quantification of the metastatic tumors in the liver using image-based computation. The total area of tumor tissue per area of liver tissue was determined. Data are presented as Min to Max box-and-whisker plot with median indicated at center line for n = 16, n = 14, n = 12, or n = 18 images per condition from a total of 9, 7, 6 or 9 mice per group. Ctrl vs ASNase, P = 0.347. Ctrl vs DON, P = 0.016. Ctrl vs DON+ASNase, P < 0.0001. DON vs DON+ASNase, P = 0.013. Statistical significance was calculated using one-way ANOVA followed by Tukey’s multiple comparisons (a), two-tailed chi-square test (c), or unpaired two-tailed Student’s t test (d). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns = not significant.
