title: AAV Gene Therapy for MPS1-associated Corneal Blindness.
abstract: Although cord blood transplantation has significantly extended the lifespan of mucopolysaccharidosis type 1 (MPS1) patients, over 95% manifest cornea clouding with about 50% progressing to blindness. As corneal transplants are met with high rejection rates in MPS1 children, there remains no treatment to prevent blindness or restore vision in MPS1 children. Since MPS1 is caused by mutations in idua, which encodes alpha-L-iduronidase, a gene addition strategy to prevent, and potentially reverse, MPS1-associated corneal blindness was investigated. Initially, a codon optimized idua cDNA expression cassette (opt-IDUA) was validated for IDUA production and function following adeno-associated virus (AAV) vector transduction of MPS1 patient fibroblasts. Then, an AAV serotype evaluation in human cornea explants identified an AAV8 and 9 chimeric capsid (8G9) as most efficient for transduction. AAV8G9-opt-IDUA administered to human corneas via intrastromal injection demonstrated widespread transduction, which included cells that naturally produce IDUA, and resulted in a >10-fold supraphysiological increase in IDUA activity. No significant apoptosis related to AAV vectors or IDUA was observed under any conditions in both human corneas and MPS1 patient fibroblasts. The collective preclinical data demonstrate safe and efficient IDUA delivery to human corneas, which may prevent and potentially reverse MPS1-associated cornea blindness.
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.
authors.forename: Melisa
authors.initials: M
authors.lastname: Vance
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Ophthalmology, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Telmo
authors.initials: T
authors.lastname: Llanga
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Ophthalmology, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Will
authors.initials: W
authors.lastname: Bennett
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.
authors.forename: Kenton
authors.initials: K
authors.lastname: Woodard
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Genetics, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Giridhar
authors.initials: G
authors.lastname: Murlidharan
authors.affiliation: Department of Ophthalmology, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Neil
authors.initials: N
authors.lastname: Chungfat
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Genetics, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Aravind
authors.initials: A
authors.lastname: Asokan
authors.affiliation: College of Veterinary Medicine, NCSU-CVM, Clinical Sciences, Raleigh, NC, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Gilger
authors.affiliation: Department of Pediatrics, Duke University, Durham, NC, 27710, USA.
authors.forename: Joanne
authors.initials: J
authors.lastname: Kurtzberg
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Pharmacology, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: R Jude
authors.initials: RJ
authors.lastname: Samulski
authors.affiliation: Gene Therapy Center, University of North Carolina at Chapel Hill, NC, 27599, USA.|Department of Ophthalmology, University of North Carolina, Chapel Hill, NC, 27599, USA.
authors.forename: Matthew L
authors.initials: ML
authors.lastname: Hirsch
mesh_terms: D017209:Apoptosis / Q000235:genetics; D001766:Blindness / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D015153:Blotting, Western; D002478:Cells, Cultured; D003315:Cornea / Q000378:metabolism / Q000473:pathology; D003316:Corneal Diseases / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007068:Iduronidase / Q000235:genetics* / Q000378:metabolism; D018613:Microscopy, Confocal; D008059:Mucopolysaccharidosis I / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D009924:Organ Culture Techniques; D014162:Transfection / Q000379:methods
chemical_list: D049452:Green Fluorescent Proteins; D007068:Iduronidase