Figure 1.

AML-Derived Alteration of the BM Vascular Architecture and Function

(A) Quantification of CD31⁺ ECs in the BM (shown as percentage of CD45^-Ter119^- BM cells) of non-transplanted mice (ctrl) and mice transplanted with HSPCs (CB) or AML patient-derived samples, as depicted. Each dot represents an individual mouse. Ctrl, n = 22; CB, n = 29; AML patients (AML1, 2, 3, 5, 6, 7, 8, 9), n = 30. Data are shown as mean ± SEM.

(B) Absolute number of CD31⁺ ECs in the BM (2 femurs, 2 tibias, and 2 iliac crests) of non-transplanted mice (ctrl) and mice transplanted with CB-derived HSPCs (CB) or AML patient-derived samples, as depicted. Each dot represents an individual mouse. Ctrl, n = 17; CB, n = 26; AML patients (AML1, 2, 6, 7, 8, 9), n = 23. Data are shown as mean ± SEM.

(C) Frequency of Sca-1^high and Sca-1^low CD31⁺ cells in the BM of non-transplanted mice (ctrl) and mice transplanted with AML patient-derived samples. Ctrl, n = 19; AML patients (AML 2, 3, 6, 8, 9), n = 16. Data are shown as mean ± SEM.

(D) Representative 3D reconstruction of BM vasculature of the calvarium of non-transplanted mice (ctrl) and mice transplanted with CB-derived HSPCs (CB) or AML patient-derived samples, as depicted, imaged via 2P microscopy 1 min after injection of 655-conjugated NT-Qtracker as vessel-pooling agent. White arrows pointing at sinusoids. Data are representative of triplicates in 4 independent experiments. Scale bars represent 70 μm.

(E) Quantification of vascular mean diameter in the calvarium BM of non-transplanted mice (ctrl) and mice transplanted with CB-derived HSPCs (CB) or human AML-derived samples, as depicted, using IMARIS filament tool. Dots represent the diameter of vascular fragments in the z stack of the calvarium of at least 3 mice per group. Ctrl, n = 235; CB, n = 232; HL60, n = 91; ML1, n = 148; U937, n = 85; AML6, n = 130; AML8, n = 94. Red lines represent the mean ± SEM.

(F) Representative 3D reconstruction of BM hypoxia imaged via intravital microscopy using the HypoxiSense probe together with vasculature (dextran) and Nestin⁺ cells in non-transplanted mice or mice transplanted with AML6 patient-derived cells, as depicted. Scale bars represent 50 μm.

(G) Distribution and relative frequency of vessel distances to hypoxic areas in the BM of non-transplanted mice (ctrl) or mice transplanted with AML6 patient-derived cells, as depicted.

ns, not significant; ^∗p < 0.05; ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001. See also Figure S1.