PMID:22003382

Citation	Borovjagin, AV, Dong, J, Passineau, MJ, Ren, C, Lamani, E, Mamaeva, OA, Wu, H, Keyser, E, Murakami, M, Chen, S and MacDougall, M (2011) Adenovirus gene transfer to amelogenesis imperfecta ameloblast-like cells. PLoS ONE 6:e24281
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.
Links	PubMed PMC3189176 Online version:10.1371/journal.pone.0024281
Keywords	Adenoviridae/genetics; Adenoviridae/physiology; Adolescent; Ameloblasts/metabolism; Amelogenesis Imperfecta/pathology; Capsid/metabolism; Cell Line, Tumor; Coxsackie and Adenovirus Receptor-Like Membrane Protein; Genetic Vectors/genetics; Humans; Receptors, Virus/genetics; Transduction, Genetic/methods; Viral Tropism
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
HUMAN:CXAR		GO:0005737: cytoplasm	ECO:0000314:		C		Figure 2E shows CAR expression in cytoplasm (white arrows) in two different cell types	complete CACAO 6767
HUMAN:CXAR		GO:0005923: tight junction	ECO:0000314:		C		Figure 2E shows CAR protein is expressed at tight junctions (Red Arrows) in specific cell type	complete CACAO 6768
HUMAN:CXAR	part_of	GO:0005737: cytoplasm	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete
HUMAN:CXAR	part_of	GO:0005923: bicellular tight junction	ECO:0000314: direct assay evidence used in manual assertion		C		Seeded From UniProt	complete

References

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