Fig. 6: CPZ has antileukemic activities against AML cells with MT-RTKs in vivo.

a NOG mice were transplanted with BMMNCs from case #2 with FLT3 ITD⁺ AML. These mice were treated with 10 mg/kg of CPZ or normal saline (as a control) from 10 days after transplantation. Nine weeks after transplantation, femurs were collected from CPZ-treated (n = 3, right panel) or control mice (n = 3, left panel). Representative photographs of the collected specimens are shown. b BM cells were subjected to flow cytometric analyses. Engrafted cells were detected as a hCD45⁺ fraction. In this fraction, engrafted AML cells were further identified as hCD33⁺hCD56⁺ cells. c NOG mice were transplanted with primary AML cells from patients with FLT3 ITD (cases #1- #8), KIT D816V (case #10), or FLT3 WT/KIT WT (cases #11- #20). Each AML sample was injected into 6–12 mice and these animals were treated with 10 mg/kg of CPZ or normal saline (as a control). Eight to ten weeks after transplantation, mice were euthanized and specimens were collected. The proportion of hCD45⁺ cells in the BM or PB was assessed by flow cytometry. Figures depict the mean ± SEM of the %hCD45⁺ cells in each group (n = 3 mice per group for case1, 2, 3, 4, 5, 6, 7, 11, 12, 13, 14, 15. n = 4 mice per group for case16, 17,18. n = 5 mice per group for case8, 10, 19. n = 6 mice per group for case20). Two-sided unpaired Student’s t test. d After transplantation of FLT3 ITD⁺ AML cells (from case #2), mice were treated with CPZ or normal saline. Histopathological analysis was performed on BM samples isolated from these mice 9 weeks after transplantation. Representative images are HE staining (upper panel) and immunohistochemical staining to detect engrafted hCD34⁺ and hCD45⁺ cells (lower panel). Images are representative of three independent experiments. Scale bars, 100 µm at 200 × 20 µm at 1000×. e PCR analyses were performed using genomic DNA isolated from BMMNCs before transplantation and from BM samples isolated from CPZ- or normal saline (as a control)-treated mice 9 weeks after transplantation. Images are representative of three independent experiments.