title: Adenovirus gene transfer to amelogenesis imperfecta ameloblast-like cells.
abstract: To explore gene therapy strategies for amelogenesis imperfecta (AI), a human ameloblast-like cell population was established from third molars of an AI-affected patient. These cells were characterized by expression of cytokeratin 14, major enamel proteins and alkaline phosphatase staining. Suboptimal transduction of the ameloblast-like cells by an adenovirus type 5 (Ad5) vector was consistent with lower levels of the coxsackie-and-adenovirus receptor (CAR) on those cells relative to CAR-positive A549 cells. To overcome CAR -deficiency, we evaluated capsid-modified Ad5 vectors with various genetic capsid modifications including "pK7" and/or "RGD" motif-containing short peptides incorporated in the capsid protein fiber as well as fiber chimera with the Ad serotype 3 (Ad3) fiber "knob" domain. All fiber modifications provided an augmented transduction of AI-ameloblasts, revealed following vector dose normalization in A549 cells with a superior effect (up to 404-fold) of pK7/RGD double modification. This robust infectivity enhancement occurred through vector binding to both α(v)β3/α(v)β5 integrins and heparan sulfate proteoglycans (HSPGs) highly expressed by AI-ameloblasts as revealed by gene transfer blocking experiments. This work thus not only pioneers establishment of human AI ameloblast-like cell population as a model for in vitro studies but also reveals an optimal infectivity-enhancement strategy for a potential Ad5 vector-mediated gene therapy for AI.
authors: Department of Periodontics, University of Alabama at Birmingham School of Dentistry, Birmingham, Alabama, United States of America.
authors: Anton V
authors: AV
authors: Borovjagin
authors: Juan
authors: J
authors: Dong
authors: Michael J
authors: MJ
authors: Passineau
authors: Changchun
authors: C
authors: Ren
authors: Ejvis
authors: E
authors: Lamani
authors: Olga A
authors: OA
authors: Mamaeva
authors: Hongju
authors: H
authors: Wu
authors: Enid
authors: E
authors: Keyser
authors: Miho
authors: M
authors: Murakami
authors: Shuo
authors: S
authors: Chen
authors: Mary
authors: M
authors: MacDougall
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D000293:Adolescent; D000565:Ameloblasts / Q000378:metabolism*; D000567:Amelogenesis Imperfecta / Q000473:pathology*; D002213:Capsid / Q000378:metabolism; D045744:Cell Line, Tumor; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D011991:Receptors, Virus / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods*; D056189:Viral Tropism
chemical_list: C568771:CLMP protein, human; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D011991:Receptors, Virus