Liver-targeted transcutaneous UMGD of liver-specific FVIII plasmid into dogs. (A) The plasmid construct pHP-hF8-X10, which carries an hFVIII variant with 10 residues in the A1 domain of hFVIII replaced with porcine residues (hFVIII-X10), is driven by a liver-specific enhancer-promoter (hepatic locus control region [HCR]-hAAT). (B) A representative diagnostic ultrasound image outlines the dispersion of MBs within the target liver lobe (circled in black, top). Angiography with x-ray contrast (Visipaque) was performed to ensure that the catheter was correctly placed into the hepatic vein and to show widespread agent distribution in the lobe. The bottom of the panel shows placement of the balloon catheter to guide infusion and occlude blood flow from the hepatic vein for LLL treatment. (C) Once the treatment area was identified, pHP-hF8-X10 (0.67 mg/kg) and MBs (0.1 mL/kg) were infused through the catheter as the ultrasound was applied to the surface of the abdomen. bpA, bovine growth hormone polyadenylation signal; hAAT (p), human α1-antitrypsin promoter; hFIXIntA, human FIX first intron.