Fig. 7
Knockdown of DNMT3A in a PDX-AML model leads to significantly increased survival. A Schematic overview about in vivo treatments after i.v. injection of 1 × 107 blasts of DNMT3A-mut/FLT3-ITD-mut patient #751 AML into NSG mice. Mice were treated by i.p. injection with PBS, 4 mg/kg αCD33-mAB-P/P-control (scrambled: “scr”)-siRNA, αCD33-mAB-P/P-DNMT3A-siRNA or 4 mg/kg αCD33-mAB-P/P-FLT3-siRNA three times weekly (n = 5 in each group). B Kaplan–Meier survival curves: Treatment with αCD33-mAB-P/P-DNMT3A-siRNA led to survival benefit for all mice within this setting, αCD33-mAB-P/P-FLT3-siRNA showed a trend. C αCD33-mAB-P/P-DNMT3A treatment significantly reduces hCD45-positive circulating leukmia cells in mouse blood. Day 12 was chosen, because control mice were still alive. FLT3-siRNA treated mice showed the same trend. D-U In the same PDX model, engrafted animals were treated with αCD33-mAB-P/P-DNMT3A nanocarriers with Cy5-labeled siRNA (lower row) or non-labeled (upper row) as well as PBS control (middle row). D, E, F. Bright field photographs of Cy5-FRI in G, H, I Mice treated with αCD33-mAB-P/P-nanocarriers loaded with Cy5-labeled siRNA showed Cy5 signals in human leukemia engrafted bone marrow (I, L) as well as excretion by kidney (I), while no or very weak signals were detected in non-labeled siRNA treated (G) and PBS controls (H). M-U Organ sections immunostained for human IgG showed distinct signals in αCD33-mAB-P/P -nanocarrier-treated, but not control mice in bone marrow (αCD33-mAB signal) and kidney (excretion), but not in irrelevant organs such as heart. α, anti; D3A, DNMT3A; FRI, Fluorescence reflectance imaging