title: Targeted gene delivery to the enteric nervous system using AAV: a comparison across serotypes and capsid mutants.
abstract: Recombinant adeno-associated virus (AAV) vectors are one of the most widely used gene transfer systems in research and clinical trials. AAV can transduce a wide range of biological tissues, however to date, there has been no investigation on targeted AAV transduction of the enteric nervous system (ENS). Here, we examined the efficiency, tropism, spread, and immunogenicity of AAV transduction in the ENS. Rats received direct injections of various AAV serotypes expressing green fluorescent protein (GFP) into the descending colon. AAV serotypes tested included; AAV 1, 2, 5, 6, 8, or 9 and the AAV2 and AAV8 capsid mutants, AAV2-Y444F, AAV2-tripleY-F, AAV2-tripleY-F+T-V, AAV8-Y733F, and AAV8-doubeY-F+T-V. Transduction, as determined by GFP-positive cells, occurred in neurons and enteric glia within the myenteric and submucosal plexuses of the ENS. AAV6 and AAV9 showed the highest levels of transduction within the ENS. Transduction efficiency scaled with titer and time, was translated to the murine ENS, and produced no vector-related immune response. A single injection of AAV into the colon covered an area of ~47 mm(2). AAV9 primarily transduced neurons, while AAV6 transduced enteric glia and neurons. This is the first report on targeted AAV transduction of neurons and glia in the ENS.
authors.affiliation: 1] Department of Translational Science and Molecular Medicine, College of Human Medicine, Michigan State University, Grand Rapids, Michigan, USA [2] Neuroscience Program, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Benskey
authors.affiliation: Department of Translational Science and Molecular Medicine, College of Human Medicine, Michigan State University, Grand Rapids, Michigan, USA.
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Kuhn
authors.affiliation: 1] Neuroscience Program, Michigan State University, East Lansing, Michigan, USA [2] Department of Pharmacology and Toxicology, Michigan State University, East Lansing, Michigan, USA.
authors.forename: James J
authors.initials: JJ
authors.lastname: Galligan
authors.affiliation: Department of Translational Science and Molecular Medicine, College of Human Medicine, Michigan State University, Grand Rapids, Michigan, USA.
authors.forename: Joanna
authors.initials: J
authors.lastname: Garcia
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, USA.
authors.forename: Shannon E
authors.initials: SE
authors.lastname: Boye
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, USA.
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
authors.affiliation: Department of Pediatrics, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
authors.forename: Christian
authors.initials: C
authors.lastname: Mueller
authors.affiliation: Department of Ophthalmology, University of Florida, Gainesville, Florida, USA.
authors.forename: Sanford L
authors.initials: SL
authors.lastname: Boye
authors.affiliation: 1] Department of Translational Science and Molecular Medicine, College of Human Medicine, Michigan State University, Grand Rapids, Michigan, USA [2] Neuroscience Program, Michigan State University, East Lansing, Michigan, USA.
authors.forename: Fredric P
authors.initials: FP
authors.lastname: Manfredsson
mesh_terms: D000818:Animals; D002213:Capsid / Q000737:chemistry / Q000378:metabolism; D044683:Colon, Descending / Q000166:cytology* / Q000294:innervation; D000229:Dependovirus / Q000145:classification / Q000235:genetics*; D017615:Enteric Nervous System / Q000166:cytology*; D015870:Gene Expression; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000627:therapeutic use; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009154:Mutation; D009457:Neuroglia / Q000166:cytology*; D009474:Neurons / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D065288:Serogroup; D052416:Targeted Gene Repair / Q000379:methods; D014161:Transduction, Genetic / Q000379:methods*; D056189:Viral Tropism / Q000235:genetics
chemical_list: D049452:Green Fluorescent Proteins