title: Rapid cohort generation and analysis of disease spectrum of large animal model of cone dystrophy.
abstract: Large animal models are an important resource for the understanding of human disease and for evaluating the applicability of new therapies to human patients. For many diseases, such as cone dystrophy, research effort is hampered by the lack of such models. Lentiviral transgenesis is a methodology broadly applicable to animals from many different species. When conjugated to the expression of a dominant mutant protein, this technology offers an attractive approach to generate new large animal models in a heterogeneous background. We adopted this strategy to mimic the phenotype diversity encounter in humans and generate a cohort of pigs for cone dystrophy by expressing a dominant mutant allele of the guanylate cyclase 2D (GUCY2D) gene. Sixty percent of the piglets were transgenic, with mutant GUCY2D mRNA detected in the retina of all animals tested. Functional impairment of vision was observed among the transgenic pigs at 3 months of age, with a follow-up at 1 year indicating a subsequent slower progression of phenotype. Abnormal retina morphology, notably among the cone photoreceptor cell population, was observed exclusively amongst the transgenic animals. Of particular note, these transgenic animals were characterized by a range in the severity of the phenotype, reflecting the human clinical situation. We demonstrate that a transgenic approach using lentiviral vectors offers a powerful tool for large animal model development. Not only is the efficiency of transgenesis higher than conventional transgenic methodology but this technique also produces a heterogeneous cohort of transgenic animals that mimics the genetic variation encountered in human patients.
authors: Unit of Gene Therapy and Stem Cell Biology, Jules-Gonin Eye Hospital, University of Lausanne, Lausanne, Switzerland.
authors: Corinne
authors: C
authors: Kostic
authors: Simon Geoffrey
authors: SG
authors: Lillico
authors: Sylvain Vincent
authors: SV
authors: Crippa
authors: Nicolas
authors: N
authors: Grandchamp
authors: Héloïse
authors: H
authors: Pilet
authors: Stéphanie
authors: S
authors: Philippe
authors: Zen
authors: Z
authors: Lu
authors: Tim James
authors: TJ
authors: King
authors: Jacques
authors: J
authors: Mallet
authors: Chamsy
authors: C
authors: Sarkis
authors: Yvan
authors: Y
authors: Arsenijevic
authors: Christopher Bruce Alexander
authors: CB
authors: Whitelaw
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D030801:Animals, Genetically Modified; D004195:Disease Models, Animal; D004596:Electroretinography; D005799:Genes, Dominant; D018740:Genetic Heterogeneity; D005822:Genetic Vectors; D006162:Guanylate Cyclase / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D017949:Retinal Cone Photoreceptor Cells / Q000201:enzymology / Q000473:pathology*; D058499:Retinal Dystrophies / Q000235:genetics* / Q000473:pathology; D017386:Sequence Homology, Amino Acid; D012720:Severity of Illness Index; D013552:Swine / Q000235:genetics; D019076:Transgenes; D014792:Visual Acuity
chemical_list: D007527:Isoenzymes; D006162:Guanylate Cyclase
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