Microtubular disruption prolongs the expression of human bilirubin-uridinediphosphoglucuronate-glucuronosyltransferase-1 gene transferred into gunn rat livers

Namita Roy Chowdhury, Richard M. Hays, Vasudeva R. Bommineni, Nicholas Franki, Jayanta Roy-Chowdhury, Catherine H. Wu, George Y. Wu

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

DNA delivered to the liver by asialoglycoprotein receptor-mediated endocytosis is degraded in lysosomes within 48 h. To test the hypothesis that microtubular disruption should promote transgene persistence by interrupting endosomal translocation to lysosomes, plasmids containing bacterial chloramphenicol acetyltransferase (pSV2-CAT) or human bilirubin-UDP-glucuronosyltransferase-1 (pSVKS-hBUGT1) genes were complexed with asialoglycoprotein-polylysine conjugates, and 1 mg of the complexed DNA was injected intravenously into bilirubin-UDP-glucuronosyltransferase-deficient Gunn rats. 30 min before DNA injection, one group received 0.75 mg of colchicine/kg of body weight intraperitoneally, which was shown by immununofluoreseent confocal microscopy to disrupt the microtubular network. Control rats received normal saline. In colchicine-pretreated rats receiving pSV2-CAT, hepatic chloramphenicol acetyltransferase activity persisted for 9-14 weeks, whereas in the saline-pretreated group the activity was detectable for 48 h only. In colchicine-pretreated Gunn rats receiving pSVK3-hBUGT1, the DNA persisted in liver for 10 weeks, bilirubin glucuronides were excreted in bile, and serum bilirubin levels declined by 25-35% in 2-4 weeks and remained reduced for 8 weeks. Without colchicine pretreatment, the DNA was detectable in liver for 2 days only, and serum bilirubin levels were not reduced. Thus, microtubular disruption provides a noninvasive method for prolonging the effect of liver-targeted gene therapy.

Original languageEnglish (US)
Pages (from-to)2341-2346
Number of pages6
JournalJournal of Biological Chemistry
Volume271
Issue number4
StatePublished - Jan 26 1996

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Gunn Rats
Glucuronosyltransferase
Bilirubin
Liver
bilirubin glucuronoside glucuronosyltransferase
Colchicine
Rats
Genes
DNA
Chloramphenicol O-Acetyltransferase
Lysosomes
Rat control
Asialoglycoprotein Receptor
Gene therapy
Confocal microscopy
Endocytosis
Serum
Transgenes
Confocal Microscopy
Bile

ASJC Scopus subject areas

  • Biochemistry

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Microtubular disruption prolongs the expression of human bilirubin-uridinediphosphoglucuronate-glucuronosyltransferase-1 gene transferred into gunn rat livers. / Chowdhury, Namita Roy; Hays, Richard M.; Bommineni, Vasudeva R.; Franki, Nicholas; Roy-Chowdhury, Jayanta; Wu, Catherine H.; Wu, George Y.

In: Journal of Biological Chemistry, Vol. 271, No. 4, 26.01.1996, p. 2341-2346.

Research output: Contribution to journalArticle

Chowdhury, Namita Roy ; Hays, Richard M. ; Bommineni, Vasudeva R. ; Franki, Nicholas ; Roy-Chowdhury, Jayanta ; Wu, Catherine H. ; Wu, George Y. / Microtubular disruption prolongs the expression of human bilirubin-uridinediphosphoglucuronate-glucuronosyltransferase-1 gene transferred into gunn rat livers. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 4. pp. 2341-2346.
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abstract = "DNA delivered to the liver by asialoglycoprotein receptor-mediated endocytosis is degraded in lysosomes within 48 h. To test the hypothesis that microtubular disruption should promote transgene persistence by interrupting endosomal translocation to lysosomes, plasmids containing bacterial chloramphenicol acetyltransferase (pSV2-CAT) or human bilirubin-UDP-glucuronosyltransferase-1 (pSVKS-hBUGT1) genes were complexed with asialoglycoprotein-polylysine conjugates, and 1 mg of the complexed DNA was injected intravenously into bilirubin-UDP-glucuronosyltransferase-deficient Gunn rats. 30 min before DNA injection, one group received 0.75 mg of colchicine/kg of body weight intraperitoneally, which was shown by immununofluoreseent confocal microscopy to disrupt the microtubular network. Control rats received normal saline. In colchicine-pretreated rats receiving pSV2-CAT, hepatic chloramphenicol acetyltransferase activity persisted for 9-14 weeks, whereas in the saline-pretreated group the activity was detectable for 48 h only. In colchicine-pretreated Gunn rats receiving pSVK3-hBUGT1, the DNA persisted in liver for 10 weeks, bilirubin glucuronides were excreted in bile, and serum bilirubin levels declined by 25-35{\%} in 2-4 weeks and remained reduced for 8 weeks. Without colchicine pretreatment, the DNA was detectable in liver for 2 days only, and serum bilirubin levels were not reduced. Thus, microtubular disruption provides a noninvasive method for prolonging the effect of liver-targeted gene therapy.",
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