Nonviral gene transfer into liver and muscle for treatment of hyperbilirubinemia in the gunn rat.

We evaluated naked plasmid DNA (pDNA)-mediated expression of human hepatic bilirubin UDP-glucuronosyltransferase (hUGT1A1) in skeletal muscle to correct hyperbilirubinemia in the UGT1A1-deficient Gunn rat, an animal model of Crigler-Najjar syndrome type I (CN-I). After delivery of pDNA encoding hUGT1A1 via hepatic vein or femoral artery, in vitro bilirubin glucuronidation activity was detectable in Gunn rat liver and muscle extracts. Expression of hUGT1A1 in Gunn rat liver or muscle resulted in excretion of bilirubin glucuronides in bile. Total biliary bilirubin concentrations increased from a pretreatment average of 10.5 +/- 2.1 microM to 29.2 +/- 4.2 microM after gene transfer into the liver, and to 28.6 +/- 3.8 microM after gene transfer into muscle. Total serum bilirubin decreased by up to 31.2 +/- 6.9 and 29.2 +/- 3.7% and remained significantly lower for at least 1 and 2 weeks, respectively. Tissue damage associated with the procedure was minimal and reversible. Our results demonstrate that muscle can be genetically modified to glucuronidate bilirubin, leading to elimination in bile. A 30% decrease in serum bilirubin, if sustained, would provide meaningful clinical benefit for CN-I patients. However, to be clinically useful, this method needs further optimization and stable gene expression must be achieved.