Use of a chimeric adenovirus vector enhances BMP2 production and bone formation

Hum Gene Ther. 2002 Jul 20;13(11):1337-47. doi: 10.1089/104303402760128568.

Abstract

Recombinant adenoviral vectors have potential for the treatment of a variety of musculoskeletal defects and such gene therapy systems have been a recent research focus in orthopedic surgery. In studies reported here, two different adenovirus vectors have been compared for their ability to transduce human bone marrow mesenchymal stem cells (hBM-MSCs) and elicit bone formation in vivo. Vectors consisted either of standard adenovirus type 5 (Ad5) vector or a chimeric adenovirus type 5 vector that contains an adenovirus type 35 fiber (Ad5F35), which has been recently demonstrated to bestow a different cellular tropism, and a complete cDNA encoding human bone morphogenetic 2 (BMP2). Studies were also conducted to compare the transduction efficiency of these vectors using enhanced green fluorescent protein (GFP). hBM-MSCs transduced with Ad5F35 vectors had higher levels of transgene expression than those transduced with Ad5 vectors. The results also demonstrate that hBM-MSCs lack the coxsackie-adenovirus receptor (CAR), which is responsible for cellular adsorption of Ad5. Therefore, the data suggest that Ad5 virus adsorption to hBM-MSCs is inefficient. Ad5BMP2- or Ad5F35BMP2-transduced hBM-MSCs were also compared in an in vivo heterotopic bone formation assay. Mineralized bone was radiologically identified only in muscle that received the Ad5F35BMP2 transduced hBM-MSCs. In summary, Ad5F35BMP2 can efficiently transduce hBM-MSCs leading to enhanced bone formation in vivo.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics*
  • Animals
  • Bone Marrow Cells / cytology
  • Bone Marrow Cells / drug effects
  • Bone Marrow Cells / metabolism
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins / biosynthesis
  • Bone Morphogenetic Proteins / genetics*
  • Calcification, Physiologic
  • Carcinoma / pathology
  • Cell Line
  • Cell Transformation, Viral
  • Cells, Cultured
  • Coxsackie and Adenovirus Receptor-Like Membrane Protein
  • DNA, Recombinant / genetics
  • Gene Expression
  • Gene Transfer Techniques
  • Genetic Vectors* / administration & dosage
  • Green Fluorescent Proteins
  • Humans
  • Injections, Intramuscular
  • Luminescent Proteins / metabolism
  • Lung Neoplasms / pathology
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Osteogenesis*
  • Receptors, Virus / metabolism
  • Recombinant Fusion Proteins / pharmacology
  • Recombinant Proteins / metabolism
  • Stem Cells / cytology
  • Stem Cells / drug effects
  • Stem Cells / metabolism
  • Stromal Cells / cytology
  • Stromal Cells / drug effects
  • Stromal Cells / metabolism
  • Transduction, Genetic
  • Transforming Growth Factor beta*
  • Transgenes
  • Tumor Cells, Cultured

Substances

  • BMP2 protein, human
  • Bmp2 protein, mouse
  • Bone Morphogenetic Protein 2
  • Bone Morphogenetic Proteins
  • CLMP protein, human
  • CLMP protein, mouse
  • Coxsackie and Adenovirus Receptor-Like Membrane Protein
  • DNA, Recombinant
  • Luminescent Proteins
  • Receptors, Virus
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • Green Fluorescent Proteins