![Figure 2. RUNX1-inhibition potently inhibits the homing of AML cells to the vascular niche. (A) Cumulative result of the E-selectin static adhesion assay. MLL-ENL+ mouse leukemic cells were seeded to the E-selectin–coated plate at the indicated concentrations, and adhesive cells were counted using a fluorometer (n = 4). (B) Schematic representation of the treatment and analysis schedule in mice. C57BL/6 mice were treated with andrographolide (25 mg/kg body weight, 3 times/week, IP), A 205804 (10 mg/kg body weight, 3 times/week, orally), or Chb-M′ (320 μg/kg body weight, 3 times/week, IV) for 2 weeks. Twenty-four hours after the last treatment, mice were transplanted with 1 × 106 MLL-ENL+ leukemic cells marked with GFP fluorescence. After another 24 hours, when all mice were sacrificed for flow cytometric analysis, the frequency of GFP+ cells in the femur and spleen was determined. (C) Frequency of GFP+ leukemic cells in the femur and the spleen in mice treated with control DMSO or andrographolide (n = 5). (D) Frequency of GFP+ leukemic cells in the femur and spleen in mice treated with control DMSO or A 205804 (n = 5). (E) Frequency of GFP+ leukemic cells in the femur and spleen in mice treated with control DMSO or Chb-M′ (n = 5). (F) Schematic representation of the treatment and analysis schedule in mice. NOG mice were preconditioned with Chb-M′ (320 μg/kg body weight, 3 times/week, IV) or control DMSO for 2 weeks. Twenty-four hours after the last administration, mice were transplanted with 1 × 106 MV4-11 cells. Seven days later, when treatment with Chb-M′ (320 μg/kg body weight, twice per week, IV) or control DMSO began, we looked for signs of leukemia development. (G) Overall survival of mice transplanted with MV4-11 cells followed by treatment with Chb-M′ or control DMSO as in panel A (DMSO, DMSO [DD]: n = 11; Chb-M′, DMSO [M′D]: n = 9; DMSO, Chb-M′ [DM′]: n = 11; Chb-M′, Chb-M′ [M′M′]: n = 11). Data represent mean ± SEM. P values were determined using a log-rank (Mantel-Cox) test. *P < .05, **P < .01 (2-tailed Student t test). N.S., not significant.](https://ash.silverchair-cdn.com/ash/content_public/journal/bloodadvances/2/5/10.1182_bloodadvances.2017009324/3/advances009324f2.jpeg?Expires=1735535966&Signature=E0frxnIjmBaW1vkfwNtvkU6jcg~-4Um0Y~Wa53yKGIb8NmenkQLabom8WfrLwJy~ZU~Wx4pNHeM~NTUh3vNwkuhNtStmol~JmuIP5xkQDx77Gmszj6een1wKcp7pXoJy40sd3YMrobeH2YlrigBKDtJga9MW1hS7Q3u1LMKS6JWA-wmkVkcrBG75nDdkqHgNKumvg1mKm~uZw6t5XSHiIjyaVI45iuavVvJAdNHQxUj0UjU2qWxdgJ-ETzeS1f6picsglsuZQlT~sEYP6E8gnQunqstCOP9pgUOmV6MmooHYsDTEm~T~E6hRSWBcHEwXwuKJc9auRsNxi~uYTH~Nbw__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
RUNX1-inhibition potently inhibits the homing of AML cells to the vascular niche. (A) Cumulative result of the E-selectin static adhesion assay. MLL-ENL+ mouse leukemic cells were seeded to the E-selectin–coated plate at the indicated concentrations, and adhesive cells were counted using a fluorometer (n = 4). (B) Schematic representation of the treatment and analysis schedule in mice. C57BL/6 mice were treated with andrographolide (25 mg/kg body weight, 3 times/week, IP), A 205804 (10 mg/kg body weight, 3 times/week, orally), or Chb-M′ (320 μg/kg body weight, 3 times/week, IV) for 2 weeks. Twenty-four hours after the last treatment, mice were transplanted with 1 × 106 MLL-ENL+ leukemic cells marked with GFP fluorescence. After another 24 hours, when all mice were sacrificed for flow cytometric analysis, the frequency of GFP+ cells in the femur and spleen was determined. (C) Frequency of GFP+ leukemic cells in the femur and the spleen in mice treated with control DMSO or andrographolide (n = 5). (D) Frequency of GFP+ leukemic cells in the femur and spleen in mice treated with control DMSO or A 205804 (n = 5). (E) Frequency of GFP+ leukemic cells in the femur and spleen in mice treated with control DMSO or Chb-M′ (n = 5). (F) Schematic representation of the treatment and analysis schedule in mice. NOG mice were preconditioned with Chb-M′ (320 μg/kg body weight, 3 times/week, IV) or control DMSO for 2 weeks. Twenty-four hours after the last administration, mice were transplanted with 1 × 106 MV4-11 cells. Seven days later, when treatment with Chb-M′ (320 μg/kg body weight, twice per week, IV) or control DMSO began, we looked for signs of leukemia development. (G) Overall survival of mice transplanted with MV4-11 cells followed by treatment with Chb-M′ or control DMSO as in panel A (DMSO, DMSO [DD]: n = 11; Chb-M′, DMSO [M′D]: n = 9; DMSO, Chb-M′ [DM′]: n = 11; Chb-M′, Chb-M′ [M′M′]: n = 11). Data represent mean ± SEM. P values were determined using a log-rank (Mantel-Cox) test. *P < .05, **P < .01 (2-tailed Student t test). N.S., not significant.
