Figure 1.

Ponatinib inhibits venous malformation (VM) formation and expansion. A, Immunoblot analysis of nontransfected human umbilical vein endothelial cell (HUVEC-NT), HUVEC transfected with wild-type TIE2 (WT) or with mutant TIE2 (L914F) probed with indicated antibodies. B, Immunoblot analysis of HUVEC-NT, -TIE2-WT, and -TIE2-L914F probed with indicated antibodies. Cells were treated with dimethyl sulfoxide (control) or 500 nmol/L ponatinib for 1 h. C, Treatment scheme: HUVEC-TIE2-L914F injected mice were treated daily with vehicle or ponatinib (30 mg/kg per day, oral gavage) for 14 d. Lesion size measured by caliper every 2 d; Data expressed as mean±SD, linear mixed-effect model (n=5–6 mice with 2 lesions/group). D, Representative images of lesions; scale bar: 1 cm (left). Representative images of Hematoxylin and Eosin (H&E) stained lesion sections; scale bar: 500 μm (right). E, Quantification of vascular area (%). Data expressed as average value for 2 lesions on each mouse, mean shown by horizontal bars, Student t test (n=5–6 mice). F, Relative mRNA levels of c-ABL and ARG in sh scramble and sh c-ABL/ARG (target 1) HUVEC-TIE2-L914F measured 48 h after shRNA induction. G, Immunoblot of HUVEC-TIE2-L914F sh Scramble and sh c-ABL/ARG (target 1) at 48 h. H, Proliferation rates of HUVEC-TIE2-L914F sh Scramble and sh c-ABL/ARG (target 1). Data expressed as mean±SEM, linear mixed-effect model. I, HUVEC-TIE2-L914F sh Scramble and sh c-ABL/ARG (target 1) cells were injected into nude mice and lesion size measured by caliper every 2 days; Data expressed as mean±SEM, linear mixed-effect model (n=8–9 mice with 2 lesions/group). J, Representative images of lesions (left) and sections stained for H&E and UEA (Ulex europaeus agglutinin I; right). Scale bar: 1 cm (lesions) and 100 μm (H&E and UEA staining). K, Quantification of vascular area. Data expressed as average value for 2 lesions on each mouse, mean shown by horizontal bars, Student t test (n=8–9 mice).