Our long-term objective is to develop safe and efficient methods for liver-directed gene therapy for genetic disorders of hepatic metabolism, using inherited jaundice due to bilirubin-uridinediphospho-glucuronate glucuronosyltransferase (bilirubin-UGT) deficiency as a model target. Absence of hepatic bilirubin-UGT activity leads to accumulation of the bilirubin in plasma and consequent brain damage, resulting in the potentially lethal Crigler-Najjar syndrome type I (CN-I). Currently, liver transplantation is the only definitive therapy. Gunn rats, the animal model for CN-I, will be used for these studies. During the last four years, both viral and non-viral vehicles for delivery of normal human bilirubin-UGT genes to Gunn rat liver were devised and tested. In this continuation application, two strategies for correcting hepatic bilirubin-UGT deficiency in vivo will be pursued. In the first approach, recombinant viral vectors will be used to substitute hepatic bilirubin-UGT by delivering a normal gene. Recombinant adenoviruses are very efficient in transferring genes into the liver in vivo, but the transgene expression is transient because the virus is episomal and the host immune response precludes its repeated administration. Three methods for specific tolerization of the host to adenoviral antigens, developed in our laboratory, show promise for long-term adenovirus- mediated gene therapy without immunosuppression. To refine these methods for future clinical application, the mechanisms by which they induce specific tolerance will be elucidated. In addition, a much less immunogenic vector, based on the SV40 virus, will be developed and tested. The second approach will utilize RNA/DNA chimeric molecules to repair the genetic lesion in Gunn rats, using our highly efficient vectors for liver-specific nucleic acid delivery by receptor-mediated endocytosis. The gene transfer efficiency will be assessed by molecular, enzymatic and metabolic studies. Successful completion of these studies will provide a basis for gene therapies for CN-I and other inherited metabolic disorders, such as deficiency of alpha1-antitrypsin and urea cycle enzymes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK046057-08
Application #
6176194
Study Section
General Medicine A Subcommittee 2 (GMA)
Program Officer
Doo, Edward
Project Start
1993-08-01
Project End
2003-07-31
Budget Start
2000-08-01
Budget End
2001-07-31
Support Year
8
Fiscal Year
2000
Total Cost
$284,232
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
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Yannam, Govardhana Rao; Han, Bing; Setoyama, Kentaro et al. (2014) A nonhuman primate model of human radiation-induced venocclusive liver disease and hepatocyte injury. Int J Radiat Oncol Biol Phys 88:404-411
Zhou, Hongchao; Dong, Xinyuan; Kabarriti, Rafi et al. (2012) Single liver lobe repopulation with wildtype hepatocytes using regional hepatic irradiation cures jaundice in Gunn rats. PLoS One 7:e46775
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Yamanouchi, Kosho; Zhou, Hongchao; Roy-Chowdhury, Namita et al. (2009) Hepatic irradiation augments engraftment of donor cells following hepatocyte transplantation. Hepatology 49:258-67
Wang, Xia; Sarkar, Debi P; Mani, Prashant et al. (2009) Long-term reduction of jaundice in Gunn rats by nonviral liver-targeted delivery of Sleeping Beauty transposon. Hepatology 50:815-24
Jiang, Jinlan; Salido, Eduardo C; Guha, Chandan et al. (2008) Correction of hyperoxaluria by liver repopulation with hepatocytes in a mouse model of primary hyperoxaluria type-1. Transplantation 85:1253-60
Kawashita, Yujo; Guha, Chandan; Moitra, Rituparna et al. (2008) Hepatic repopulation with stably transduced conditionally immortalized hepatocytes in the Gunn rat. J Hepatol 49:99-106
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Salido, Eduardo C; Li, Xiao M; Lu, Yang et al. (2006) Alanine-glyoxylate aminotransferase-deficient mice, a model for primary hyperoxaluria that responds to adenoviral gene transfer. Proc Natl Acad Sci U S A 103:18249-54

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