Figure 3.

Persistence of Increased Vascular Permeability after Chemotherapy

(A) Schematic of the experiment. Patient-derived AML cells were injected intravenously into NSG mice and engraftment was monitored via BM aspirate. Once BM engraftment (>40%) assessed, 10 mg/kg/day of AraC or solvent was administrated subcutaneously for one or two alternate weeks, depending on the patient sample. Seven days after the last administration of AraC, vessel-pooling agents were administrated intravenously. Mice were euthanized 10 min later and calvaria imaged via 2P microscopy.

(B) Representative 3D reconstruction of the BM vasculature in the calvarium of mice engrafted with different patient-derived samples treated with solvent (top) or AraC (bottom). Scale bars represent 70 μm.

(C) Vascular leakiness in the BM of mice transplanted with different human AML patient-derived samples, treated with solvent or AraC, as depicted. Three or more replicates were used for each condition. 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.

(D) Absolute number of CD31⁺ ECs in the BM (n = 2 femurs) of mice transplanted with AML6 (top) or AML9 (bottom) patient-derived cells, treated with solvent or AraC, as depicted; n = 3 in each condition. Data are shown as mean ± SEM.

(E) Hypoxia represented as HypoxiSense signal intensity/voxel in the calvaria BM of non-transplanted mice (ctrl) and mice transplanted with CB-derived HSPCs (CB) or AML patient-derived samples, treated or not with AraC, as depicted. Ctrl, n = 9; CB UT, n = 7; CB AraC, n = 8; AML patients (AML6 and 9) UT, n = 16; AraC, n = 16. Data are shown as whiskers minimum-to-maximum plots, the line inside the box representing the mean, the top and the bottom line representing the 75% and 25% percentiles, respectively, and the lines above and below the box representing the SD.

(F) Representative 3D reconstruction of BM hypoxia imaged via intravital microscopy using the HypoxiSense probe together with vasculature (dextran) and Nestin⁺ cells in mice transplanted with AML6 patient-derived cells and treated or not with AraC, as depicted. Scale bars represent 50 μm.

(G) Distribution and relative frequency of vessel distances to hypoxic areas in the BM of mice transplanted with AML6 patient-derived cells, and treated or not with AraC, as depicted.

ns, not significant; ^∗∗p < 0.01. See also Figure S3; Tables S1 and S2.