title: Corrective GUSB transfer to the canine mucopolysaccharidosis VII cornea using a helper-dependent canine adenovirus vector.
abstract: Corneal transparency is maintained, in part, by specialized fibroblasts called keratocytes, which reside in the fibrous lamellae of the stroma. Corneal clouding, a condition that impairs visual acuity, is associated with numerous diseases, including mucopolysaccharidosis (MPS) type VII. MPS VII is due to deficiency in β-glucuronidase (β-glu) enzymatic activity, which leads to accumulation of glycosaminoglycans (GAGs), and secondary accumulation of gangliosides. Here, we tested the efficacy of canine adenovirus type 2 (CAV-2) vectors to transduce keratocyte in vivo in mice and nonhuman primates, and ex vivo in dog and human corneal explants. Following efficacy studies, we asked if we could treat corneal clouding by the injection a helper-dependent (HD) CAV-2 vector (HD-RIGIE) harboring the human cDNA coding for β-glu (GUSB) in the canine MPS VII cornea. β-Glu activity, GAG content, and lysosome morphology and physiopathology were analyzed. We found that HD-RIGIE injections efficiently transduced coxsackievirus adenovirus receptor-expressing keratocytes in the four species and, compared to mock-injected controls, improved the pathology in the canine MPS VII cornea. The key criterion to corrective therapy was the steady controlled release of β-glu and its diffusion throughout the collagen-dense stroma. These data support the continued evaluation of HD CAV-2 vectors to treat diseases affecting corneal keratocytes.
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Serratrice
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Aurelie
authors.initials: A
authors.lastname: Cubizolle
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Sandy
authors.initials: S
authors.lastname: Ibanes
authors.affiliation: Université Montpellier 2, Montpellier, France; Inserm U710, Montpellier, France; Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Nadine
authors.initials: N
authors.lastname: Mestre-Francés
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Neus
authors.initials: N
authors.lastname: Bayo-Puxan
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Creyssels
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Aurelie
authors.initials: A
authors.lastname: Gennetier
authors.affiliation: Institut Régional du Cancer Montpellier, Inserm U896, Montpellier, France.
authors.forename: Florence
authors.initials: F
authors.lastname: Bernex
authors.affiliation: Université Montpellier 2, Montpellier, France; Inserm U710, Montpellier, France; Ecole Pratique des Hautes Etudes, Paris, France.
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Verdier
authors.affiliation: Department of Pathobiology, School of Veterinary Medicine, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Haskins
authors.affiliation: Tissue Bank, Centre Hospitalier Régional Universitaire de Montpellier, Montpellier, France.
authors.forename: Guilhem
authors.initials: G
authors.lastname: Couderc
authors.affiliation: Inserm U563, Toulouse, France; Departement d'Ophtalmologie, Hôpital Purpan, Toulouse, France.
authors.forename: Francois
authors.initials: F
authors.lastname: Malecaze
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Kalatzis
authors.affiliation: Institut de Génétique Moléculaire de Montpellier, CNRS 5535, Montpellier, France; Université de Montpellier I, Montpellier, France; Université Montpellier 2, Montpellier, France. Electronic address: eric.kremer@igmm.cnrs.fr.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Kremer
mesh_terms: D006516:Adenoviruses, Canine / Q000235:genetics*; D000260:Adenoviruses, Human / Q000235:genetics; D000818:Animals; D016565:Cheirogaleidae; D003318:Corneal Opacity / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D003319:Corneal Stroma / Q000201:enzymology* / Q000473:pathology / Q000648:ultrastructure; D004195:Disease Models, Animal; D004285:Dogs; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D005966:Glucuronidase / Q000235:genetics*; D006025:Glycosaminoglycans / Q000378:metabolism; D006378:Helper Viruses; D006801:Humans; D066298:In Vitro Techniques; D008247:Lysosomes / Q000201:enzymology; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D046529:Microscopy, Electron, Transmission; D016538:Mucopolysaccharidosis VII / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D013045:Species Specificity
chemical_list: D006025:Glycosaminoglycans; D005966:Glucuronidase
keywords: Clouding; Cornea; Keratocytes; Lysosomal storage disorders; Mucopolysaccharidosis; Primates; Tissue engineering; β-Glucuronidase