CAR-dependent and CAR-independent pathways of adenovirus vector-mediated gene transfer and expression in human fibroblasts. Academic Article uri icon

Overview

MeSH

  • Capsid
  • Cells, Cultured
  • Fibroblasts
  • Gene Expression
  • Humans
  • Transgenes
  • Up-Regulation

MeSH Major

  • Adenoviruses, Human
  • Capsid Proteins
  • Gene Transfer Techniques
  • Genetic Vectors
  • Receptors, Virus

abstract

  • Primary fibroblasts are not efficiently transduced by subgroup C adenovirus (Ad) vectors because they express low levels of the high-affinity Coxsackie virus and adenovirus receptor (CAR). In the present study, we have used primary human dermal fibroblasts as a model to explore strategies by which Ad vectors can be designed to enter cells deficient in CAR. Using an Ad vector expressing the human CAR cDNA (AdCAR) at high multiplicity of infection, primary fibroblasts were converted from being CAR deficient to CAR sufficient. Efficiency of subsequent gene transfer by standard Ad5-based vectors and Ad5-based vectors with alterations in penton and fiber was evaluated. Marked enhancement of binding and transgene expression by standard Ad5 vectors was achieved in CAR-sufficient fibroblasts. Expression by AdDeltaRGDbetagal, an Ad5-based vector lacking the arginine-glycine-aspartate (RGD) alphaV integrin recognition site from its penton base, was achieved in CAR-sufficient, but not CAR-deficient, cells. Fiber-altered Ad5-based vectors, including (a) AdF(pK7)betagal (bearing seven lysines on the end of fiber) (b) AdF(RGD)betagal (bearing a high-affinity RGD sequence on the end of fiber), and (c) AdF9sK betagal (bearing a short fiber and Ad9 knob), demonstrated enhanced gene transfer in CAR-deficient fibroblasts, with no further enhancement in CAR-sufficient fibroblasts. Together, these observations demonstrate that CAR deficiency on Ad targets can be circumvented either by supplying CAR or by modifying the Ad fiber to bind to other cell-surface receptors.

publication date

  • February 1999

has subject area

  • Adenoviruses, Human
  • Capsid
  • Capsid Proteins
  • Cells, Cultured
  • Fibroblasts
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors
  • Humans
  • Receptors, Virus
  • Transgenes
  • Up-Regulation

Research

keywords

  • Journal Article

Identity

Language

  • eng

PubMed Central ID

  • PMC408101

Digital Object Identifier (DOI)

  • 10.1172/JCI5309

PubMed ID

  • 10021467

Additional Document Info

start page

  • 579

end page

  • 587

volume

  • 103

number

  • 4