TY - JOUR
T1 - Long-term reduction of serum bilirubin levels in gunn rats by retroviral gene transfer in vivo
AU - Tada, K.
AU - Chowdhury, N. R.
AU - Neufeld, David S.
AU - Bosma, P. J.
AU - Heard, M.
AU - Prasad, V. R.
AU - Chowdhury, J. R.
PY - 1998
Y1 - 1998
N2 - Conjugation with glucuronic acid, mediated by bilirubin- uridinediphosphoglucuronate glucuronosyltransferase (bilirubin-UGT), is essential for efficient billary excretion of bilirubin. Inherited absence of this enzyme activity results in the potentially lethal Crigler-Najjar syndrome type I in humans and lifelong hyperbilirubinemia in Gunn rats. To develop a gene therapy for bilirubin-UGT deficiency, we constructed a high- titer replication-deficient amphotropic recombinant retrovirus (MFG-S hB- UGT1) capable of transferring the gene encoding bilirubin-UGT1, the principal bilirubin-UGT isoform in human liver. To stimulate hepatocyte proliferation, Gunn rats were subjected to 66% hepatectomy. After 24 hours, the portal vein, the hepatic artery, and the inferior vena cave above and below the hepatic vein were clamped, and the portal vein and the isolated segment of the yens cave were cannulated. The liver was perfused with the MFG-S hB-UGT1 preparation through the portal vein at 5 ml/min for 10 minutes, then circulation was restored. Control rat livers were perfused with a recombinant retrovirus expressing Escherichia coli β-galactosidase. In MFG-S hB-UGT1-perfused rats, but not in controls, expression of human bilirubin-UGT1 was shown by immunotransblotting, bilirubin-UGT assay of liver homogenates, and biliary excretion of bilirubin diglucuronide and monoglucuronide. Mean serum bilirubin levels decreased by 20% to 25% in 3 weeks and remained at that level throughout the study period (18 months). This is the first report of long-term amelioration of inherited jaundice by retrovirus-directed gene therapy in an animal model for Crigler-Najjar syndrome.
AB - Conjugation with glucuronic acid, mediated by bilirubin- uridinediphosphoglucuronate glucuronosyltransferase (bilirubin-UGT), is essential for efficient billary excretion of bilirubin. Inherited absence of this enzyme activity results in the potentially lethal Crigler-Najjar syndrome type I in humans and lifelong hyperbilirubinemia in Gunn rats. To develop a gene therapy for bilirubin-UGT deficiency, we constructed a high- titer replication-deficient amphotropic recombinant retrovirus (MFG-S hB- UGT1) capable of transferring the gene encoding bilirubin-UGT1, the principal bilirubin-UGT isoform in human liver. To stimulate hepatocyte proliferation, Gunn rats were subjected to 66% hepatectomy. After 24 hours, the portal vein, the hepatic artery, and the inferior vena cave above and below the hepatic vein were clamped, and the portal vein and the isolated segment of the yens cave were cannulated. The liver was perfused with the MFG-S hB-UGT1 preparation through the portal vein at 5 ml/min for 10 minutes, then circulation was restored. Control rat livers were perfused with a recombinant retrovirus expressing Escherichia coli β-galactosidase. In MFG-S hB-UGT1-perfused rats, but not in controls, expression of human bilirubin-UGT1 was shown by immunotransblotting, bilirubin-UGT assay of liver homogenates, and biliary excretion of bilirubin diglucuronide and monoglucuronide. Mean serum bilirubin levels decreased by 20% to 25% in 3 weeks and remained at that level throughout the study period (18 months). This is the first report of long-term amelioration of inherited jaundice by retrovirus-directed gene therapy in an animal model for Crigler-Najjar syndrome.
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U2 - 10.1002/lt.500040111
DO - 10.1002/lt.500040111
M3 - Article
C2 - 9457971
AN - SCOPUS:0031973035
SN - 1074-3022
VL - 4
SP - 78
EP - 88
JO - Liver Transplantation and Surgery
JF - Liver Transplantation and Surgery
IS - 1
ER -