title: Safety and Biodistribution Evaluation in CNGB3-Deficient Mice of rAAV2tYF-PR1.7-hCNGB3, a Recombinant AAV Vector for Treatment of Achromatopsia.
abstract: Applied Genetic Technologies Corporation (AGTC) is developing rAAV2tYF-PR1.7-hCNGB3, a recombinant adeno-associated virus (rAAV) vector expressing the human CNGB3 gene, for treatment of achromatopsia, an inherited retinal disorder characterized by markedly reduced visual acuity, extreme light sensitivity, and absence of color discrimination. We report here results of a study evaluating safety and biodistribution of rAAV2tYF-PR1.7-hCNGB3 in CNGB3-deficient mice. Three groups of animals (n = 35 males and 35 females per group) received a subretinal injection in one eye of 1 μl containing either vehicle or rAAV2tYF-PR1.7-hCNGB3 at one of two dose concentrations (1 × 10(12) or 4.2 × 10(12) vg/ml) and were euthanized 4 or 13 weeks later. There were no test-article-related changes in clinical observations, body weights, food consumption, ocular examinations, clinical pathology parameters, organ weights, or macroscopic observations at necropsy. Cone-mediated electroretinography (ERG) responses were detected after vector administration in the treated eyes in 90% of animals in the higher dose group and 31% of animals in the lower dose group. Rod-mediated ERG responses were reduced in the treated eye for all groups, with the greatest reduction in males given the higher dose of vector, but returned to normal by the end of the study. Microscopic pathology results demonstrated minimal mononuclear cell infiltrates in the retina and vitreous of some animals at the interim euthanasia and in the vitreous of some animals at the terminal euthanasia. Serum anti-AAV antibodies developed in most vector-injected animals. No animals developed antibodies to hCNGB3. Biodistribution studies demonstrated high levels of vector DNA in vector-injected eyes but little or no vector DNA in nonocular tissue. These results support the use of rAAV2tYF-PR1.7-hCNGB3 in clinical studies in patients with achromatopsia caused by CNGB3 mutations.
authors.affiliation: 1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
authors.forename: Guo-jie
authors.initials: GJ
authors.lastname: Ye
authors.affiliation: 2 Covance Laboratories Inc. , Madison, Wisconsin.
authors.forename: Ewa
authors.initials: E
authors.lastname: Budzynski
authors.affiliation: 2 Covance Laboratories Inc. , Madison, Wisconsin.
authors.forename: Peter
authors.initials: P
authors.lastname: Sonnentag
authors.affiliation: 3 OSOD , Madison, Wisconsin.
authors.forename: T Michael
authors.initials: TM
authors.lastname: Nork
authors.affiliation: 3 OSOD , Madison, Wisconsin.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Miller
authors.affiliation: 2 Covance Laboratories Inc. , Madison, Wisconsin.
authors.forename: Leslie
authors.initials: L
authors.lastname: McPherson
authors.affiliation: 3 OSOD , Madison, Wisconsin.
authors.forename: James N
authors.initials: JN
authors.lastname: Ver Hoeve
authors.affiliation: 4 Laboratory Corporation of America Holdings , Seattle, Washington.
authors.forename: Leia M
authors.initials: LM
authors.lastname: Smith
authors.affiliation: 2 Covance Laboratories Inc. , Madison, Wisconsin.
authors.forename: Tara
authors.initials: T
authors.lastname: Arndt
authors.affiliation: 1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
authors.forename: Savitri
authors.initials: S
authors.lastname: Mandapati
authors.affiliation: 1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
authors.forename: Paulette M
authors.initials: PM
authors.lastname: Robinson
authors.affiliation: 5 University of Pennsylvania , Philadelphia, Pennsylvania.
authors.forename: Roberto
authors.initials: R
authors.lastname: Calcedo
authors.affiliation: 1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
authors.forename: David R
authors.initials: DR
authors.lastname: Knop
authors.affiliation: 6 University of Florida , Gainesville, Florida.
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
authors.affiliation: 1 Applied Genetic Technologies Corporation (AGTC) , Alachua, Florida.
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Chulay
mesh_terms: D000818:Animals; D003117:Color Vision Defects / Q000235:genetics / Q000628:therapy*; D054815:Cyclic Nucleotide-Gated Cation Channels / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000009:adverse effects*; D000229:Dependovirus / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects*; D006801:Humans; D056965:Injections, Intraocular; D008297:Male; D051379:Mice; D012160:Retina / Q000378:metabolism
chemical_list: C415904:CNGB3 protein, human; C532622:CNGB3 protein, mouse; D054815:Cyclic Nucleotide-Gated Cation Channels; D004274:DNA, Recombinant