3 years ago

Promoterless gene targeting without nucleases rescues lethality of a Crigler-Najjar syndrome mouse model

Promoterless gene targeting without nucleases rescues lethality of a Crigler-Najjar syndrome mouse model
Fabiola Porro, Giulia Bortolussi, Lorena Zentilin, Andrés F Muro, Alessia De Caneva, Mark A Kay, Simone Vodret, Adi Barzel, Alessandra Iaconcig
Crigler-Najjar syndrome type I (CNSI) is a rare monogenic disease characterized by severe neonatal unconjugated hyperbilirubinemia with a lifelong risk of neurological damage and death. Liver transplantation is the only curative option, which has several limitations and risks. We applied an in vivo gene targeting approach based on the insertion, without the use of nucleases, of a promoterless therapeutic cDNA into the albumin locus of a mouse model reproducing all major features of CNSI. Neonatal transduction with the donor vector resulted in the complete rescue from neonatal lethality, with a therapeutic reduction in plasma bilirubin lasting for at least 12 months, the latest time point analyzed. Mutant mice, which expressed about 5–6% of WT Ugt1a1 levels, showed normal liver histology and motor-coordination abilities, suggesting no functional liver or brain abnormalities. These results proved that the promoterless gene therapy is applicable for CNSI, providing therapeutic levels of an intracellular ER membrane-bound enzyme responsible for a lethal liver metabolic disease. Gene targeting into the albumin locus without using nucleases rescues lethality in a severe genetic liver disease in mice, thereby avoiding the loss of therapeutic DNA by hepatocyte duplication, a critical issue in AAV-mediated gene replacement therapy.

Publisher URL: http://onlinelibrary.wiley.com/resolve/doi

DOI: 10.15252/emmm.201707601

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