Figure 5

PF-04449913 induced cell cycle activation enhances TKI sensitivity. a. Schematic of in vivo experiments. RAG2^−/−γ_c ^−/− pups were transplanted intrahepatically with 50,000 CD34⁺ BC CML cells within 48 hours of birth. Engrafted mice were treated daily for 14 days by oral gavage with vehicle, PF-04449913 (100 mg/kg), Dasatinib (50 mg/kg) or the combination. b. Graph of myeloid sarcoma count in blast crisis CML engrafted mice in vehicle (n =13, blue), PF-04449913 (n=7, purple), dasatinib (n =6, red) and combination (n =3, black) treated mice +/− SEM; *p < 0.05 and *p < 0.01 by ANOVA and Tukey post-hoc analysis c. FACS analysis showing percentage of marrow engrafted blast crisis progenitor LSC (n = 3 patients) after 14-day treatment with vehicle (n =31, blue), PF-04449913 (n =25, purple), dasatinib (n =27, maroon) and combination (n=27, grey). *p < 0.05 by ANOVA and Tukey post-hoc analysis d. BCR-ABL transcripts in the blast crisis CML engrafted marrow mice after 14 days of treatment. Graph shows normalized BCR-ABL expression (HPRT) +/− SEM; *p < 0.05 by ANOVA and Tukey post-hoc analysis e. Hedgehog pathway gene expression in FACS purified human progenitor cells from blast crisis LSC engrafted mouse marrow treated with vehicle (n = 3, blue), PF-04449913 (n = 4, purple) dasatinib (n = 4, maroon), combination (n=3, dark grey). Expression levels of seven genes were significantly altered by synergistic effect of PF-04449913 and Dasatinib (NUMB, PRKACB, CTNNB1, FKBP8, CSNK1A1, CSNK1D and STK36), where five represent SHH regulatory genes (graphed). f. Mice serially transplanted with FACS purified human progenitors from LSC engrafted mice treated with vehicle (n=12, green), PF-04449913 (n=12, purple), dasatinib (n = 8, maroon) or combination (n=7, grey) were examined for myeloid sarcomas; *p < 0.05 and *p < 0.01 by ANOVA and Tukey post-hoc analysis.