Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector.
Strategies using mesenchymal stem cell (MSC)-mediated gene therapy have been developed to improve bone healing. However, transduction efficiency into MSCs by each vector is not always high. To overcome this problem, we used a modified adenoviral vector (Adv-F/RGD) with an RGD-containing peptide in the HI loop of the fiber knob domain of adenovirus type 5 (Ad5). Transduction efficiency into bone marrow-derived MSCs with Adv-F/RGD increased 12-fold compared with a vector containing the wild-type fiber (Adv-F/wt) by beta-galactosidase chemiluminescent assay. As a next step, we constructed AxCAhBMP2-F/RGD and AxCAhBMP2-F/wt carrying human bone morphogenetic protein 2 (BMP2). At the same multiplicity of infection, MSCs infected with AxCAhBMP2-F/RGD produced higher amounts of BMP2 than cells infected with AxCAhBMP2-F/wt, and also differentiated towards the osteogenic lineage more efficiently in vitro. Furthermore, using ex vivo gene transduction, we evaluated the potential for ectopic bone formation by the transduced MSCs in vivo. Transduction with AxCAhBMP2-F/RGD exhibited greatly enhanced new bone formation. These data suggest that Adv-F/RGD is useful for introducing foreign genes into MSCs and that it will be a powerful gene therapy tool for bone regeneration and other tissue engineering.
Tsuda
Hajime
H
Department of Molecular Medicine, Sapporo Medical University, Japan.
Wada
Takuro
T
Ito
Yoshinori
Y
Uchida
Hiroaki
H
Dehari
Hironari
H
Nakamura
Kiminori
K
Sasaki
Katsunori
K
Kobune
Masayoshi
M
Yamashita
Toshihiko
T
Hamada
Hirofumi
H
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001846:Bone Development / Q000502:physiology*; D001853:Bone Marrow; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016023:Integrins / Q000235:genetics / Q000378:metabolism; D008648:Mesoderm / Q000378:metabolism*; D023421:Models, Animal; D009842:Oligopeptides / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta; D019076:Transgenes / Q000502:physiology
C525718:BMP2 protein, human; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; C568771:CLMP protein, human; C568773:Clmp protein, rat; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D016023:Integrins; D009842:Oligopeptides; D012333:RNA, Messenger; D011991:Receptors, Virus; D016212:Transforming Growth Factor beta; C047981:arginyl-glycyl-aspartic acid