Fig. 3: Azacitidine treatment supports the increased anti-leukemic effect of anti-CD123 CAR T cells in MOLM-13 xenograft mice.
From: Demethylating therapy increases anti-CD123 CAR T cell cytotoxicity against acute myeloid leukemia
a Schematic diagram summarizing the experimental plan for the MOLM-13Luc AML and AZA xenograft model using Rag2−/−Il2rγ−/− recipients. Scatter plots showing the expression of CD123 at D17 in the b BM and c spleen, represented as a fold change of MFI of AMLMOLM-13-bearing mice treated with 2.5 mg/kg AZA (n = 5) with respect to MFI of mice treated with PBS (n = 5). d Scatter plots showing the expression of CD123 at D17 in the PB, represented as raw MFI values of AMLMOLM-13-bearing mice treated with AZA (n = 5) compared to mice treated with PBS (n = 5). e Scatter plots depicting the absolute cell count (cells/μL) of hCD45 in AMLMOLM-13-bearing mice treated with AZA (n = 5) compared with AMLMOLM-13-bearing mice treated with PBS (n = 5). Mice in b–e were pair-matched for similar hCD45 engraftment levels and CD123 MFI. f Kaplan–Meier analysis of percentage survival for each treatment group (PBS n = 15; NTD Tc n = 15; anti-CD123 CAR Tc n = 16; AZA n = 21; NTD Tc + AZA n = 18; anti-CD123 CAR Tc + AZA n = 20). Attrition of mice: as described above. All p-values were calculated with respect to the anti-CD123 CAR Tc + AZA group. g Quantification of the BLI signal for each treatment group over time (PBS n = 10; NTD Tc n = 12; anti-CD123 CAR Tc n = 12; AZA n = 16; NTD Tc + AZA n = 15; anti-CD123 CAR Tc + AZA n = 18). On day 35, at least 5 mice per group were still alive and included in the analysis. h Representative serial BLI from depicting leukemia burden in MOLM-13 engrafted mice treated with PBS, NTD Tc, anti-CD123 CAR Tc, AZA only, AZA with NTD Tc, and anti-CD123 CAR Tc + AZA. Data are represented colorimetrically (photons s−1 cm−1) with the scale bars indicating upper (max) and lower (min) BLI thresholds at each analysis time point. All graphed data are represented as mean ± SEM and pooled from two (g, h) or three (b–e) independent experiments. p-values were calculated using f two-sided Mantel–Cox test (log-rank) and b–e two-sided paired parametric student’s t-test.
