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Adeno‐associated virus gene repair corrects a mouse model of hereditary tyrosinemia in vivo
Author(s) -
Paulk Nicole K.,
Wursthorn Karsten,
Wang Zhongya,
Finegold Milton J.,
Kay Mark A.,
Grompe Markus
Publication year - 2010
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.23481
Subject(s) - biology , homologous recombination , tyrosinemia , genetic enhancement , gene , transgene , gene targeting , adeno associated virus , in vivo , dna repair , gene delivery , genetics , cancer research , tyrosine , vector (molecular biology) , recombinant dna , biochemistry
Adeno‐associated virus (AAV) vectors are ideal for performing gene repair due to their ability to target multiple different genomic loci, low immunogenicity, capability to achieve targeted and stable expression through integration, and low mutagenic and oncogenic potential. However, many handicaps to gene repair therapy remain. Most notable is the low frequency of correction in vivo . To date, this frequency is too low to be of therapeutic value for any disease. To address this, a point‐mutation–based mouse model of the metabolic disease hereditary tyrosinemia type I was used to test whether targeted AAV integration by homologous recombination could achieve high‐level stable gene repair in vivo . Both neonatal and adult mice were treated with AAV serotypes 2 and 8 carrying a wild‐type genomic sequence for repairing the mutated Fah (fumarylacetoacetate hydrolase) gene. Hepatic gene repair was quantified by immunohistochemistry and supported with reverse transcription polymerase chain reaction and serology for functional correction parameters. Successful gene repair was observed with both serotypes but was more efficient with AAV8. Correction frequencies of up to 10 −3 were achieved and highly reproducible within typical dose ranges. In this model, repaired hepatocytes have a selective growth advantage and are thus able to proliferate to efficiently repopulate mutant livers and cure the underlying metabolic disease. Conclusion: AAV‐mediated gene repair is feasible in vivo and can functionally correct an appropriate selection‐based metabolic liver disease in both adults and neonates. (H EPATOLOGY 2010.)