Single hepatic venous injection of liver-specific naked plasmid vector expressing human UGT1A1 leads to long-term correction of hyperbilirubinemia and prevention of chronic bilirubin toxicity in Gunn rats.

Previously, we have demonstrated that hepatic venous injection of pcDNA3hUGT1A1 expressing human bilirubin glucuronosyl transferase 1A1 (hUGT1A1) under the control of the cytomegalovirus promoter results in excretion of bilirubin glucuronides in bile and significant decrease in serum bilirubin for at least 2 weeks in the Gunn rat, an animal model of Crigler-Najjar syndrome type I. In this study we compared repeat delivery of pcDNA3hUGT1A1 with single injection of pBShUGT1A1 expressing hUGT1A1 under liver-specific regulatory control, for treatment of hyperbilirubinemia in the Gunn rat. Although repeat injections of pcDNA3hUGT1A1 consistently reduced serum bilirubin levels, the effect did not exceed 2 weeks; hUGT1A1 was detectable in livers only for 2 weeks, despite the presence of vector and transcript for at least 1 month. In contrast, injection of pBShUGT1A1 resulted in persistence of vector, transcript, and recombinant protein and sustained correction of hyperbilirubinemia for at least 8 months; furthermore, renal tubular damage, the principal manifestation of chronic bilirubin toxicity in the Gunn rat, was prevented. Sera from animals treated with pBShUGT1A1 consistently contained anti-hUGT1A1 antibodies, but a significant increase in the number of hepatic CD4(+) and CD8(+) cells was seen only in the pcDNA3hUGT1A1 group; thus liver-specific expression of hUGT1A1 may attenuate immune response. Our results provide further evidence of the feasibility of long-term correction of hepatic enzyme deficiencies with plasmid vectors optimized for expression in the liver.