Fig. 4
Dual targeting of CP-CML CD34+ cells reduces cell proliferation, CD34+ cell and colony numbers. Ai-iv CTV proliferation analysis of CP-CML cells treated with IM,SC or DOR and the combination (IM = 1 µM, SC = 1 µM, 2 µM, 5 µM; DOR = 2.5µM; Combo = 1 µM TKI (IM or SC) + 2.5 µM DOR) in absence or presence of BMP4 (20 ng/ml) or in co-culture with HS-5 at 72 h. Aii-iv Proliferation analysis of CP-CML samples indicates that DOR in combination with TKIs synergistically inhibits proliferation compared to single treatment. SC + DOR alone display the biggest fold change compared to NDC in all culture conditions. Bi-iii TKIs and DOR reduce CD34+ cell numbers in CP-CML samples and to a lower extent in normal samples. Bi Biggest reduction was observed with SC 5 µM, DOR and combinatorial treatments. Bii Similar results were obtained in presence of BMP4. Biii Co-cultures displayed a decrease for all treatments but to a lower extent compared to monocultures. Microscopic analysis revealed a reduction in Ci + iii total colony counts and (Cii + iv) types but no visible change in (Cv) colony morphology. TKI and DOR alone and in combination reduced (Ci + ii) total colony counts and affected different (Cii + iv) colony types independent of BMP4 presence in CP-CML samples. Data are expressed as mean ± SD (n = 3). CTVmax and division 1 were compared using Two-Way-ANOVA, normalized CD34.+ values and colony counts were compared using Two-Way-ANOVA (**** p < 0.0001, *** p 0.001 to 0.0001, ** p 0.01 to 0.001, * p 0.05 to 0.01). See also Figure S3 and S4