Figure 2.

AML Engraftment Increases Vascular Permeability in the BM

(A) Schematic of the experiment. Human AML cells engineered to express GFP and luciferase were injected intravenously into NSG mice and engraftment was monitored via bioluminescence and BM aspirate. Once engraftment was confirmed, 3 mg of 65–85 kDa TRITC-dextran was injected intravenously and 15 min later, calvaria were imaged via 2P microscopy.

(B) Representative z stack of the TRITC-dextran-labeled BM vasculature in the calvarium of non-transplanted control or from mice transplanted with GFP-HL60 cells. Left: TRITC-dextran signal alone. Right: merge of TRITC-dextran and GFP signals. Data are representative of triplicates. Scale bars represent 30 μm.

(C) Vascular leakiness quantified in vivo at different time points after dextran injection in non-transplanted mice (ctrl) and U937 or AML6 patient cells-engrafted mice.

(D) Vascular leakiness quantification in the BM of non-transplanted mice or mice transplanted with different human AML cell lines and patient-derived samples, as depicted, and imaged 10 min after administration of vascular dyes. Ctrl, n = 11; CB, n = 19; HL60, n = 12; ML1, n = 7; U937, n = 4; AML9, n = 6; AML6, n = 14; AML5, n = 4; AML1, n = 12; AML3, n = 4; AML2, n = 3; AML8, n = 7; AML7, n = 6. Data are shown as whiskers minimum-to-maximum plots, the line inside the box representing the mean, the top and the bottom lines representing the 75% and 25% percentiles, respectively, and the lines above and below the box representing the SD.

(E) Representative z stacks of BM vasculature in the calvarium of non-transplanted mice (ctrl) or mice engrafted with HL60 or AML9 cells, as depicted. Scale bars represent 80 μm.

ns, not significant; ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001. See also Figure S2.