title: Transport across a polarized monolayer of Caco-2 cells by transferrin receptor-mediated adenovirus transcytosis.
abstract: Adenoviral vectors have a poor record of transgene delivery efficiency through physical barriers such as the epithelium or endothelium. We report here the construction of an adenoviral vector that has the capability to be transported across polarized epithelial monolayers of Caco-2 cells (a colon carcinoma cell line) by transcytosis. This transcytosis is transferrin receptor (TfR)-mediated with use of a bifunctional adaptor, soluble coxsackie adenovirus receptor (sCAR)-Tf, and is both temperature and iron dependent. Under experimental conditions, the adenoviral transcytosis was inhibited by pretreatment of Caco-2 cells with colchicine, an inhibitor of transcytosis, and was not enhanced by pretreatment with Brefeldin A (BFA), an enhancer of transcytosis. In these Caco-2 cells, the transcytosis rate was 0.3 +/- 1.3% (SD). The transcytosed adenoviruses remain biologically functional. These data suggest the potential clinical benefit under conditions where drug delivery is a challenge, such as within the airway epithelium, at the bladder lumen urothelial cell interface, and across the blood-brain barrier for clinical treatment of lung, urogenital, and brain disorders, respectively, by adenoviral transcytosis of transgene delivery.
authors.affiliation: Division of Human Gene Therapy, Department of Medicine, Pathology, and The Gene Therapy Center, University of Alabama at Birmingham, Birmingham, AL 35291, USA.
authors.forename: Zeng B
authors.initials: ZB
authors.lastname: Zhu
authors.forename: Sharmila K
authors.initials: SK
authors.lastname: Makhija
authors.forename: Baogen
authors.initials: B
authors.lastname: Lu
authors.forename: Minghui
authors.initials: M
authors.lastname: Wang
authors.forename: Angel A
authors.initials: AA
authors.lastname: Rivera
authors.forename: Meredith
authors.initials: M
authors.lastname: Preuss
authors.forename: Fen
authors.initials: F
authors.lastname: Zhou
authors.forename: Gene P
authors.initials: GP
authors.lastname: Siegal
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Alvarez
authors.forename: David T
authors.initials: DT
authors.lastname: Curiel
mesh_terms: D000074183:ATPases Associated with Diverse Cellular Activities; D000256:Adenoviridae / Q000502:physiology*; D001692:Biological Transport; D020126:Brefeldin A / Q000494:pharmacology; D018938:Caco-2 Cells; D015815:Cell Adhesion Molecules / Q000502:physiology; D016764:Cell Polarity; D004705:Endocytosis; D005822:Genetic Vectors; D006801:Humans; D007501:Iron / Q000502:physiology; D008666:Metalloendopeptidases; D011990:Receptors, Transferrin / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000502:physiology
chemical_list: D015815:Cell Adhesion Molecules; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins; D020126:Brefeldin A; D007501:Iron; D008666:Metalloendopeptidases; C112889:SPG7 protein, human; D000074183:ATPases Associated with Diverse Cellular Activities