title: A translationally optimized AAV-UGT1A1 vector drives safe and long-lasting correction of Crigler-Najjar syndrome.
abstract: Crigler-Najjar syndrome is a severe metabolic disease of the liver due to a reduced activity of the UDP Glucuronosyltransferase 1A1 (UGT1A1) enzyme. In an effort to translate to the clinic an adeno-associated virus vector mediated liver gene transfer approach to treat Crigler-Najjar syndrome, we developed and optimized a vector expressing the UGT1A1 transgene. For this purpose, we designed and testedin vitroandin vivomultiple codon-optimized UGT1A1 transgene cDNAs. We also optimized noncoding sequences in the transgene expression cassette. Our results indicate that transgene codon-optimization is a strategy that can improve efficacy of gene transfer but needs to be carefully testedin vitroandin vivo. Additionally, while inclusion of introns can enhance gene expression, optimization of these introns, and in particular removal of cryptic ATGs and splice sites, is an important maneuver to enhance safety and efficacy of gene transfer. Finally, using a translationally optimized adeno-associated virus vector expressing the UGT1A1 transgene, we demonstrated rescue of the phenotype of Crigler-Najjar syndrome in two animal models of the disease, Gunn rats andUgt1a1-/-mice. We also showed long-term (>1 year) correction of the disease in Gunn rats. These results support further translation of the approach to humans.
authors.affiliation: Genethon , Evry, France.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Ronzitti
authors.affiliation: International Centre for Genetic Engineering and Biotechnology , Trieste, Italy.
authors.forename: Giulia
authors.initials: G
authors.lastname: Bortolussi
authors.affiliation: Tytgat Institute for Liver and Intestinal Research, Academic Medical Center , Amsterdam, the Netherlands.
authors.forename: Remco
authors.initials: R
authors.lastname: van Dijk
authors.affiliation: Genethon , Evry, France.
authors.forename: Fanny
authors.initials: F
authors.lastname: Collaud
authors.affiliation: Genethon , Evry, France.
authors.forename: Severine
authors.initials: S
authors.lastname: Charles
authors.affiliation: Genethon , Evry, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Leborgne
authors.affiliation: Genethon, Evry, France; Universite' Pierre et Marie Curie - Paris 6, Paris, France.
authors.forename: Patrice
authors.initials: P
authors.lastname: Vidal
authors.affiliation: Genethon , Evry, France.
authors.forename: Samia
authors.initials: S
authors.lastname: Martin
authors.affiliation: Genethon , Evry, France.
authors.forename: Bernard
authors.initials: B
authors.lastname: Gjata
authors.affiliation: Genethon , Evry, France.
authors.forename: Marcelo Simon
authors.initials: MS
authors.lastname: Sola
authors.affiliation: Genethon , Evry, France.
authors.forename: Laetitia
authors.initials: L
authors.lastname: van Wittenberghe
authors.affiliation: Genethon , Evry, France.
authors.forename: Alban
authors.initials: A
authors.lastname: Vignaud
authors.affiliation: Genethon , Evry, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Veron
authors.affiliation: Tytgat Institute for Liver and Intestinal Research, Academic Medical Center , Amsterdam, the Netherlands.
authors.forename: Piter J
authors.initials: PJ
authors.lastname: Bosma
authors.affiliation: International Centre for Genetic Engineering and Biotechnology , Trieste, Italy.
authors.forename: Andres F
authors.initials: AF
authors.lastname: Muro
authors.affiliation: Genethon, Evry, France; Universite' Pierre et Marie Curie - Paris 6, Paris, France; INSERM U951, Evry, France.
authors.forename: Federico
authors.initials: F
authors.lastname: Mingozzi