Long-term reduction of serum bilirubin levels in gunn rats by retroviral gene transfer in vivo

K. Tada, N. R. Chowdhury, David S. Neufeld, P. J. Bosma, M. Heard, Vinayaka R. Prasad, Jayanta Roy-Chowdhury

Research output: Contribution to journalArticle

41 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)78-88
Number of pages11
JournalLiver Transplantation and Surgery
Volume4
Issue number1
StatePublished - 1998

Fingerprint

Gunn Rats
Bilirubin
Serum
Genes
Crigler-Najjar Syndrome
Retroviridae
Portal Vein
Liver
Genetic Therapy
Galactosidases
Glucuronosyltransferase
Glucuronic Acid
Hyperbilirubinemia
Hepatic Veins
Hepatic Artery
Hepatectomy
Jaundice
Hepatocytes
Protein Isoforms
Animal Models

ASJC Scopus subject areas

  • Hepatology
  • Surgery

Cite this

Long-term reduction of serum bilirubin levels in gunn rats by retroviral gene transfer in vivo. / Tada, K.; Chowdhury, N. R.; Neufeld, David S.; Bosma, P. J.; Heard, M.; Prasad, Vinayaka R.; Roy-Chowdhury, Jayanta.

In: Liver Transplantation and Surgery, Vol. 4, No. 1, 1998, p. 78-88.

Research output: Contribution to journalArticle

@article{6da27cf460554c18974f7c269fa3bd4f,
title = "Long-term reduction of serum bilirubin levels in gunn rats by retroviral gene transfer in vivo",
abstract = "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.",
author = "K. Tada and Chowdhury, {N. R.} and Neufeld, {David S.} and Bosma, {P. J.} and M. Heard and Prasad, {Vinayaka R.} and Jayanta Roy-Chowdhury",
year = "1998",
language = "English (US)",
volume = "4",
pages = "78--88",
journal = "Liver Transplantation",
issn = "1527-6465",
publisher = "John Wiley and Sons Ltd",
number = "1",

}

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, Vinayaka R.

AU - Roy-Chowdhury, Jayanta

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.

UR - http://www.scopus.com/inward/record.url?scp=0031973035&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031973035&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 78

EP - 88

JO - Liver Transplantation

JF - Liver Transplantation

SN - 1527-6465

IS - 1

ER -