Fig. 8: Anti-miR-126 therapy by miRisten effectively inhibits CM-AML burden and abrogates LSC activity.

a Schematic of experimental design. CM/tdTomato⁺ AML cells (1 × 10⁶) were transplanted into cohorts of WTwild-type syngenic mice. After 8 weeks, mice were treated with SCR control or miRisten (20 mg/kg/dose, daily for 3 weeks), or a “5 + 3” chemotherapy regimen consisted of Ara-C (50 mg/kg/day, 5 days) plus DNR (1.5 mg/kg/day, 3 days), or miRisten and “5 + 3” chemotherapy combined. AML engraftment was analyzed 7 days after the last dose of treatment and BM cells were transplanted into second recipients, which were analyzed for engraftment at 7 weeks or monitored for leukemia onset and survival. b Representative images of spleens from recipients of SCR control (Ctrl), miRisten, A + D, or miRisten/A + D group. c Spleen weight from treated recipients of SCR Ctrl (black; n = 10), miRisten (red; n = 8; miRisten vs. Ctrl p < 0.0001), A + D (blue; n = 8; A + D vs. Ctrl p < 0.0001), or miRisten/A + D (purple; n = 8; miRisten/A + D vs. Ctrl p < 0.0001; miRisten/A + D vs. miRisten p = 0.0004; miRisten/A + D vs. A + D p = 0.0337). d Representative FACS plots showing gating strategy and frequency of dTomato⁺ AML cells in the BM of SCR control, miRisten, A + D, or miRisten/A + D group. e Frequency of dTomato⁺ AML cells in the PB of mice treated with SCR control (black; n = 10), miRisten (red; n = 8; miRisten vs. Ctrl p = 0.0324), A + D (blue; n = 8; A + D vs. Ctrl p = 0.0012; A + D vs. miRisten p = 0.0047), or miRisten/A + D (purple; n = 8; miRisten/A + D vs. Ctrl p = 0.0018; miRisten/A + D vs. miRisten p = 0.014). f Frequency of dTomato⁺ AML cells in the SP of mice treated with SCR control (black; n = 10), miRisten (red; n = 8; miRisten vs. Ctrl p = 0.0103), A + D (blue; n = 8; A + D vs. Ctrl p < 0.0001), or miRisten/A + D (purple; n = 8; miRisten/A + D vs. Ctrl p < 0.0001; miRisten/A + D vs. miRisten p = 0.0004; miRisten/A + D vs. A + D p = 0.0071). g Frequency of dTomato⁺ AML cells in the BM of mice treated with SCR control (black; n = 10), miRisten (red; n = 8; miRisten vs. Ctrl p = 0.0038), A + D (blue; n = 8; A + D vs. Ctrl p < 0.0001), or miRisten/A + D (purple; n = 8; miRisten/A + D vs. Ctrl p < 0.0001; miRisten/A + D vs. miRisten p < 0.0001; miRisten/A + D vs. A + D p = 0.0291). h White blood cell (WBC) counts of second transplant recipients of SCR control (black; n = 10), miRisten (red; n = 10; miRisten vs. Ctrl p = 0.0225), A + D (blue; n = 10; A + D vs. Ctrl p = 0.0005), or miRisten/A + D (purple; n = 11; miRisten/A + D vs. Ctrl p = 0.0002) treated mice at 7 weeks. i Frequency of dTomato⁺ AML cells in the PB of second transplant recipients treated with SCR Ctrl (black; n = 10), miRisten (red; n = 10; miRisten vs. Ctrl p = 0.0358), A + D (blue; n = 10; A + D vs. Ctrl p = 0.0098), or miRisten/A + D (purple; n = 11; miRisten/A + D vs. Ctrl p < 0.0001; miRisten/A + D vs. A + D p = 0.0049) at 7 weeks. j The Kaplan–Meier survival curve of second transplant recipients treated with SCR Ctrl (black line; n = 10; median survival 84.5 days), miRisten (red line; n = 10; median survival 95.5 days; miRisten vs. Ctrl p = 0.0004), A + D (blue line; n = 10; median survival 84 days), or miRisten/A + D (purple line; n = 11; median survival 102 days; miRisten/A + D vs. Ctrl p = 0.0002; miRisten/A + D vs. miRisten p = 0.0386). The statistical significance was determined using log-rank (Mantel–Cox) test. Each dot in c, e, f, g, h, i represents data from an individual mouse and presented as the mean ± SEM; statistics for all comparisons shown were determined using two-tailed T-tests (*p < 0.05; **p < 0.01; ***p < 0.001).