The overall goal of this P01 is to develop and evaluate approaches for liver-directed gene therapy for ornithine transcarbamylase deficiency (OTCD) that are based on treatment of the acute hyperammonemic crisis by transient expression of OTC, and long-term prevention of hyperammonemia by stable correction of OTCD. OTCD is well suited for the development of gene therapy: Current treatment has failed to avert a high mortality or morbidity rate; Given the relative frequency of OTCD, an adequate population is available for study; The OTC gene has been sequenced and preliminary studies of gene therapy in animal models have been promising; The OTC gene is not rate-limiting for ureagenesis, so excessive activity should not have adverse consequences; and Restoration of enzyme activity in the liver, should suffice to normalize metabolism without the need to introduce the gene into the central nervous system. The grant consists of three projects and four cores. Project I seeks to define the molecular basis for the current limitations of in vivo gene therapy using first generation recombinant adenoviruses in liver, which include transient expression of the transduced gene and the development of inflammation at the site of transgene expression; to develop second generation recombinant adenoviruses expressing OTC in which essential genes other than E1 have been inactivated; to analyze these improved recombinant adenoviruses in vitro and in vivo; and to develop methods for efficiently transducing functional OTC genes into primary cultures of human heepatocytes using recombinant retroviruses. Project II focuses on two murine models of OTCD, the Spf and Spf/ash mouse.
The specific aims are to study the efficacy of in vivo gene therapy in correcting the metabolic and neurochemical and behavioral consequences of OTCD in the adult and neonatal animal. Project III focuses on using recombinant adenoviruses and retroviruses to treat OTCD in humans. We hypothesize that a recombinant adenovirus containing the OTC gene will be effective and safe for the short-term treatment of neonates and older children with OTCD who are in hyperammonemic crisis. We further hypothesize that long-term correction of OTCD can be achieved either by ex vivo or in vivo gene therapy using a recombinant retrovirus or a second generation recombinant adenovirus. Finally, we hypothesize that a cellular immune response following gene therapy may affect safety, efficacy and duration of gene therapy. The cores that support these projects include an Administrative Core, a Recombinant Viruses Core (containing a Vector development laboratory and a Human Applications Laboratory), a Toxicology Core (containing an Animal Models program and a Cell Morphology Laboratory), and a Mass Spectroscopy Core to perform stable isotope studies of urea turnover.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Research Program Projects (P01)
Project #
3P01HD032649-05S1
Application #
6126434
Study Section
Special Emphasis Panel (SRC (GT))
Program Officer
Hanson, James W
Project Start
1994-12-15
Project End
2000-11-30
Budget Start
1999-07-16
Budget End
2000-11-30
Support Year
5
Fiscal Year
1999
Total Cost
Indirect Cost
Name
Children's Hospital of Philadelphia
Department
Type
DUNS #
073757627
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Raper, Steven E; Chirmule, Narendra; Lee, Frank S et al. (2003) Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. Mol Genet Metab 80:148-58
Raper, Steven E; Yudkoff, Marc; Chirmule, Narendra et al. (2002) A pilot study of in vivo liver-directed gene transfer with an adenoviral vector in partial ornithine transcarbamylase deficiency. Hum Gene Ther 13:163-75
Ye, X; Zimmer, K P; Brown, R et al. (2001) Differences in the human and mouse amino-terminal leader peptides of ornithine transcarbamylase affect mitochondrial import and efficacy of adenoviral vectors. Hum Gene Ther 12:1035-46
Chen, S J; Tazelaar, J; Wilson, J M (2001) Selective repopulation of normal mouse liver by hepatocytes transduced in vivo with recombinant adeno-associated virus. Hum Gene Ther 12:45-50
Chen, S J; Tazelaar, J; Moscioni, A D et al. (2000) In vivo selection of hepatocytes transduced with adeno-associated viral vectors. Mol Ther 1:414-22
Ye, X; Whiteman, B; Jerebtsova, M et al. (2000) Correction of argininosuccinate synthetase (AS) deficiency in a murine model of citrullinemia with recombinant adenovirus carrying human AS cDNA. Gene Ther 7:1777-82
Xiao, W; Chirmule, N; Schnell, M A et al. (2000) Route of administration determines induction of T-cell-independent humoral responses to adeno-associated virus vectors. Mol Ther 1:323-9
Ye, X; Robinson, M B; Pabin, C et al. (2000) Transient depletion of CD4 lymphocyte improves efficacy of repeated administration of recombinant adenovirus in the ornithine transcarbamylase deficient sparse fur mouse. Gene Ther 7:1761-7
Mitchell, M; Jerebtsova, M; Batshaw, M L et al. (2000) Long-term gene transfer to mouse fetuses with recombinant adenovirus and adeno-associated virus (AAV) vectors. Gene Ther 7:1986-92
Chirmule, N; Moscioni, A D; Qian, Y et al. (1999) Fas-Fas ligand interactions play a major role in effector functions of cytotoxic T lymphocytes after adenovirus vector-mediated gene transfer. Hum Gene Ther 10:259-69

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