title: Safety and Biodistribution Evaluation of rAAV2tYF-CB-hRS1, a Recombinant Adeno-Associated Virus Vector Expressing Retinoschisin, in RS1-Deficient Mice.
abstract: Applied Genetic Technologies Corporation is developing a recombinant adeno-associated virus (rAAV) vector for treatment of X-linked retinoschisis (XLRS), an inherited retinal disease characterized by splitting (schisis) of the layers of the retina, which causes poor vision. We report here results of a study evaluating the safety and biodistribution of rAAV2tYF-CB-hRS1 in RS1-deficient mice. Three groups of male RS1-deficient mice received an intravitreal injection in one eye of either vehicle, or rAAV2tYF-CB-hRS1 at one of two dose levels (1 × 10(9) or 4 × 10(9) vg/eye) and were sacrificed 30 or 90 days later. The intravitreal injection procedure was well tolerated in all groups, with no test article-related changes in ophthalmic examinations. Two low-dose vector-treated animals had minimally to mildly higher white blood cell counts at day 90. There were no other intergroup differences in hematology or clinical chemistry analyses and no test article-related gross necropsy observations. Microscopic pathology results demonstrated minimal to slight mononuclear cell infiltrates in 80% of vector-injected eyes at day 30 and 20% of vector-injected eyes at day 90. Immunohistochemistry studies showed RS1 labeling of the retina in all vector-treated eyes. At the day 90 sacrifice, there was a decrease in the severity of splitting/disorganization of the inner nuclear layer of the retina in high-dose vector-treated eyes. Biodistribution studies demonstrated vector DNA in vector-injected eyes but not in any nonocular tissue. These results support the use of rAAV2tYF-CB-hRS1 in clinical studies in patients with XLRS.
authors.affiliation: 1 Applied Genetic Technologies Corporation, Alachua, Florida .
authors.forename: Guo-Jie
authors.initials: GJ
authors.lastname: Ye
authors.affiliation: 2 Department of Pediatrics, University of Florida Powell Gene Therapy Center , Gainesville, Florida .
authors.forename: Thomas
authors.initials: T
authors.lastname: Conlon
authors.affiliation: 2 Department of Pediatrics, University of Florida Powell Gene Therapy Center , Gainesville, Florida .
authors.forename: Kirsten
authors.initials: K
authors.lastname: Erger
authors.affiliation: 3 Covance Laboratories, Inc., Madison, Wisconsin.
authors.forename: Peter
authors.initials: P
authors.lastname: Sonnentag
authors.affiliation: 3 Covance Laboratories, Inc., Madison, Wisconsin.
authors.forename: Alok K
authors.initials: AK
authors.lastname: Sharma
authors.affiliation: 4 Laboratory Corporation of America® Holdings, Seattle, Washington.
authors.forename: Kellie
authors.initials: K
authors.lastname: Howard
authors.affiliation: 1 Applied Genetic Technologies Corporation, Alachua, Florida .
authors.forename: David R
authors.initials: DR
authors.lastname: Knop
authors.affiliation: 1 Applied Genetic Technologies Corporation, Alachua, Florida .
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Chulay
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D002460:Cell Line; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D005136:Eye Proteins / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects / Q000235:genetics* / Q000493:pharmacokinetics*; D006801:Humans; D007150:Immunohistochemistry; D058449:Intravitreal Injections; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D012160:Retina / Q000378:metabolism / Q000473:pathology; D041441:Retinoschisis / Q000235:genetics* / Q000628:therapy*; D013601:T-Lymphocytes / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D014018:Tissue Distribution; D018675:Toxicity Tests; D019076:Transgenes
chemical_list: D000914:Antibodies, Viral; D005136:Eye Proteins; C108726:RS1 protein, human