title: Thixotropic solutions enhance viral-mediated gene transfer to airway epithelia.
abstract: Adenovirus-mediated gene transfer to airway epithelia is inefficient in part because its receptor is absent on the apical surface of the airways. Targeting adenovirus to other receptors, increasing the viral concentration, and even prolonging the incubation time with adenovirus vectors can partially overcome the lack of receptors and facilitate gene transfer. Unfortunately, mucociliary clearance would prevent prolonged incubation time in vivo. Thixotropic solutions (TS) are gels that upon a vigorous shearing force reversibly become liquid. We hypothesized that formulating recombinant adenoviruses in TS would decrease virus clearance and thus enhance gene transfer to the airway epithelia. We found that clearance of virus-sized fluorescent beads by human airway epithelia in vitro and by monkey trachea in vivo were markedly decreased when the beads were formulated in TS compared with phosphate-buffered saline (PBS). Adenovirus formulated in TS significantly increased adenovirus-mediated gene transfer of a reporter gene in human airway epithelia in vitro and in murine airway epithelia in vivo. Furthermore, an adenovirus encoding the cystic fibrosis transmembrane regulator (CFTR) gene (AdCFTR) formulated in TS was more efficient in correcting the chloride transport defect in cystic fibrosis airway epithelia than AdCFTR formulated in PBS. These data indicate a novel strategy to augment the efficiency of gene transfer to the airways that may be applicable to a number of different gene transfer vectors and could be of value in gene transfer to cystic fibrosis (CF) airway epithelia in vivo.
authors.affiliation: Department of Internal Medicine, Division of Pharmaceutics College of Pharmacy, and Howard Hughes Medical Institute, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Seiler
authors.forename: Paul
authors.initials: P
authors.lastname: Luner
authors.forename: Thomas O
authors.initials: TO
authors.lastname: Moninger
authors.forename: Philip H
authors.initials: PH
authors.lastname: Karp
authors.forename: Shaf
authors.initials: S
authors.lastname: Keshavjee
authors.forename: Joseph
authors.initials: J
authors.lastname: Zabner
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D002266:Carboxymethylcellulose Sodium / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004847:Epithelial Cells / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D005782:Gels / Q000737:chemistry; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008810:Mice, Inbred C57BL; D008863:Microspheres; D020545:Respiratory Mucosa / Q000166:cytology / Q000502:physiology; D012996:Solutions / Q000737:chemistry
chemical_list: C505032:CFTR protein, human; D005782:Gels; D012996:Solutions; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D002266:Carboxymethylcellulose Sodium