Figure 7.
Targeting Vascular Permeability Potentiates HSPC Function
(A) Cellular ROS levels in BM-derived HSC of non-transplanted mice or mice engrafted with AML6 patient-derived cells. Data are representative of triplicates.
(B) Percentage of murine HSC among total murine CD45+ cells in the blood of non-transplanted mice and mice engrafted with AML6 patient-derived cells. ctrl, n = 5; AML6, n = 3. Data are shown as mean ± SEM.
(C) Percentage of murine HSC among total murine CD45+ cells in the BM of non-transplanted mice of mice engrafted with AML patient-derived samples (AML6, 9). ctrl, n = 5; AML patients, n = 12. Data are shown as mean ± SEM.
(D) Absolute number of murine HSC in the BM of mice transplanted with AML patient-derived cells, as depicted, and treated with AraC alone or in combination with NOS inhibitors. Each dot represents a mouse. Error bars represent mean.
(E–G) Percentage in blood (E) and absolute number in BM (F) and spleen (G) of donor CD45.1, retrieved from NSG mice transplanted with AML6 patient-derived cells and treated with AraC alone or in combination with NOS inhibitors, injected in secondary CD45.2 Rag2-KO/Gammac-KO mice. AraC, n = 4; A + N, n = 3; A + C, n = 3. Data are shown as mean ± SEM.
(H) Schematic of the experiment. Mice engrafted with human CD34+ CB cells were treated with NOS inhibitors (NG-monomethyl-L-arginine [NMMA] 20 mg/kg/day intraperitoneally or cavtratin 2 mg/kg/day intraperitoneally) for 2 weeks, 5 days/week. Mice were euthanized and BMC harvested for subsequent analysis and secondary transplantation.
(I) Percentage of HSPCs in the peripheral blood of mice engrafted with CB-derived HSPCs and treated or not with NMMA or cavtratin. Data are shown as mean ± SEM.
(J) Absolute number of HSPC in the BM of mice engrafted with CB-derived HSPC cells and treated or not with NMMA or cavtratin. Data are shown as mean ± SEM.
(K) Absolute number of human CD45+ cells in the BM (left) and spleen (right) of secondary NSG mice transplanted with primary xenografts described in (H). Data are shown as mean ± SEM.
(L) Schematic of the experiment. Non-irradiated NBSGW mice were simultaneously transplanted with tomato+ HSPCs (red) and non-transduced AML6 (black) patient-derived cells, and 2 weeks after transplantation control solvent or NMMA were administered for 6 weeks, 5 days/week. Two days after the last treatment, mice were euthanized and BM was analyzed.
(M) Percentage of tomato+ (normal, red) and tomato− (leukemic, black) cells among hCD45+ in the BM of mice described in (L); n = 4. Data are shown as mean ± SEM.
(N) Absolute number of tomato+ hCD45+ (normal) cells in the BM of mice described in (L). Data are shown as mean ± SEM.
ns, not significant; ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. See also Figure S7.