title: Single tyrosine mutation in AAV8 vector capsid enhances gene lung delivery and does not alter lung morphofunction in mice.
abstract: OBJECTIVE
  Vectors derived from adeno-associated viruses (AAVs) are important gene delivery tools for treating pulmonary diseases. Phosphorylation of surface-exposed tyrosine residues from AAV2 capsid targets the viral particles for ubiquitination and proteasome-mediated degradation, and mutations of these tyrosine residues lead to highly efficient vector transduction in vitro and in vivo in different organs. We evaluated the pulmonary transduction efficiency of AAV8 vectors containing point mutations in surface-exposed capsid tyrosine residues.
  
  
  METHODS
  Male C57BL/6 mice (20-25 g, n=24) were randomly assigned into three groups: control group animals received intratracheal (i.t.) instillation of saline (50 μl), wild-type AAV8 group, and capsid mutant Y733F AAV8 group, which received (i.t.) AAV8 vectors containing the DNA sequence of enhanced green fluorescence protein (eGFP). Four weeks after instillation, lung mechanics and morphometry, vector transduction (immunohistochemistry and mRNA expression of eGFP), and inflammatory cytokines and growth factor expression were analyzed.
  
  
  RESULTS
  Tyrosine-mutant AAV8 vectors displayed significantly increased transduction efficiency in the lung compared with their wild-type counterparts. No significant differences were observed in lung mechanics and morphometry between experimental groups. There was no evidence of inflammatory response in any group.
  
  
  CONCLUSIONS
  AAV8 vectors may be useful for new therapeutic strategies for the treatment of pulmonary diseases.
authors.affiliation: Laboratory of Cellular and Molecular Physiology, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Sabrina V
authors.initials: SV
authors.lastname: Martini
authors.forename: Adriana L
authors.initials: AL
authors.lastname: da Silva
authors.forename: Debora
authors.initials: D
authors.lastname: Ferreira
authors.forename: Karina
authors.initials: K
authors.lastname: Gomes
authors.forename: Felipe M
authors.initials: FM
authors.lastname: Ornellas
authors.forename: Miquéias
authors.initials: M
authors.lastname: Lopes-Pacheco
authors.forename: Emilia
authors.initials: E
authors.lastname: Zin
authors.forename: Hilda
authors.initials: H
authors.lastname: Petrs-Silva
authors.forename: Patricia R M
authors.initials: PR
authors.lastname: Rocco
authors.forename: Marcelo M
authors.initials: MM
authors.lastname: Morales
mesh_terms: D000818:Animals; D001483:Base Sequence; D002213:Capsid; D016207:Cytokines / Q000235:genetics; D017931:DNA Primers; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D008168:Lung / Q000378:metabolism / Q000503:physiopathology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016297:Mutagenesis, Site-Directed; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D014443:Tyrosine / Q000235:genetics*
chemical_list: D016207:Cytokines; D017931:DNA Primers; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D014443:Tyrosine