Figure 4.
Figure 4. Evaluation of the efficacy of pacritinib in CMML patient samples in vitro and in vivo. (A) Clinical and genetic characteristics of CMML patient samples used for pacritinib treatment in vitro and in vivo. (B) BM samples from CMML patients were cultured with pacritinib for 48 hours, and the half maximal inhibitory concentration was calculated for each patient sample based on the cell viability after drug exposure (n = 10). (C) Percentages of viable, early-apoptotic, late-apoptotic, and necrotic cells based on 4′,6-diamidino-2-phenylindole and Annexin V flow cytometric analysis of CMML BM MNCs treated with vehicle or pacritinib at the indicated doses in vitro (shown are mean percentages ± standard deviation across 10 patient samples). (D) Heatmap representation of phospho-STAT5 levels in CMML BM MNCs by intracellular flow cytometry after treatment with pacritinib at the indicated doses for 1 hour (top; median data across 10 patients is shown). Cells were also prestimulated with GM-CSF at 10 ng/mL for 15 minutes after incubation with pacritinib (bottom; n = 10). Levels of phospho-STAT5 in cells treated with pacritinib were evaluated considering the level of phospho-STAT5 in cells treated with dimethyl sulfoxide (DMSO) as 100% in each patient sample. (E) Colony numbers after 14 days of CMML BM MNCs seeded in methylcellulose with 1 μM pacritinib or dimethyl sulfoxide (CFU-GM, colony forming unit-granulocyte macrophage; median value is shown represented by the line inside the box ± standard deviation represented by lines above and below the box; n = 7 patient samples; *P < .05). (F) Log2 ratio of human CD45 (hCD45)+ cells in the BM of pacritinib- vs vehicle-treated PDX models from 5 CMML patients (n = 2 or 4 mice per patient). Genetic information for each patient is shown in supplemental Figure 3B. (G) Representative flow cytometric analysis of hCD45 in NSGS recipients xenografted with CMML BM MNCs following 7 days of oral administration of vehicle or 150 mg/kg/d pacritinib (mCD45.1: mouse CD45.1). (H) Comparison of hCD45 chimerism in the BM and spleen NSGS-recipient mice treated with either vehicle or pacritinib (mean value ± standard deviation and number of individual mice are represented in the figure).

Evaluation of the efficacy of pacritinib in CMML patient samples in vitro and in vivo. (A) Clinical and genetic characteristics of CMML patient samples used for pacritinib treatment in vitro and in vivo. (B) BM samples from CMML patients were cultured with pacritinib for 48 hours, and the half maximal inhibitory concentration was calculated for each patient sample based on the cell viability after drug exposure (n = 10). (C) Percentages of viable, early-apoptotic, late-apoptotic, and necrotic cells based on 4′,6-diamidino-2-phenylindole and Annexin V flow cytometric analysis of CMML BM MNCs treated with vehicle or pacritinib at the indicated doses in vitro (shown are mean percentages ± standard deviation across 10 patient samples). (D) Heatmap representation of phospho-STAT5 levels in CMML BM MNCs by intracellular flow cytometry after treatment with pacritinib at the indicated doses for 1 hour (top; median data across 10 patients is shown). Cells were also prestimulated with GM-CSF at 10 ng/mL for 15 minutes after incubation with pacritinib (bottom; n = 10). Levels of phospho-STAT5 in cells treated with pacritinib were evaluated considering the level of phospho-STAT5 in cells treated with dimethyl sulfoxide (DMSO) as 100% in each patient sample. (E) Colony numbers after 14 days of CMML BM MNCs seeded in methylcellulose with 1 μM pacritinib or dimethyl sulfoxide (CFU-GM, colony forming unit-granulocyte macrophage; median value is shown represented by the line inside the box ± standard deviation represented by lines above and below the box; n = 7 patient samples; *P < .05). (F) Log2 ratio of human CD45 (hCD45)+ cells in the BM of pacritinib- vs vehicle-treated PDX models from 5 CMML patients (n = 2 or 4 mice per patient). Genetic information for each patient is shown in supplemental Figure 3B. (G) Representative flow cytometric analysis of hCD45 in NSGS recipients xenografted with CMML BM MNCs following 7 days of oral administration of vehicle or 150 mg/kg/d pacritinib (mCD45.1: mouse CD45.1). (H) Comparison of hCD45 chimerism in the BM and spleen NSGS-recipient mice treated with either vehicle or pacritinib (mean value ± standard deviation and number of individual mice are represented in the figure).

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal