Tracking expression of virally mediated BMP-2 in gene therapy for bone repair.
Ex vivo gene therapy using stem cells transduced with viral vectors is a useful method for delivering a therapeutic protein to augment bone repair in animal models. However, the duration of cell-mediated protein production and the fate of the transduced cells are unknown. We constructed an adenoviral vector encoding Myc epitope tagged bone morphogenetic protein (BMP)-2 gene (AdBMP-2). Rat bone marrow cells transduced with this vector produced biologically active BMP-2 protein, which was confirmed by Western blot analysis and alkaline phosphatase assay. Implantation of bone marrow cells infected ex vivo with AdBMP-2 caused orthotopic bone formation in mouse hindlimbs and bony union of critical-sized mouse radial defects. Immunohistochemical analysis revealed that rBMCs expressed Myc epitope-tagged BMP-2 protein for 14 days in vivo and became incorporated in the endochondral fracture callus. This novel adenovirus encoding for epitope-tagged BMP-2 can be used for immunohistochemical tracking of transduced cells in ex vivo gene therapy for bone repair.
Gamradt
Seth C
SC
Department of Orthopaedic Surgery, School of Medicine, University of California-Los Angeles Medical Center, 10833 Le Conte Avenue, Los Angeles, CA 90095, USA. sgamradt@mednet.ucla.edu
Abe
Nobuhiro
N
Bahamonde
Matthew E
ME
Lee
Yu-Po
YP
Nelson
Scott D
SD
Lyons
Karen M
KM
Lieberman
Jay R
JR
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D016259:Genes, myc; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D007150:Immunohistochemistry; D051379:Mice; D016513:Mice, SCID; D011885:Radius Fractures / Q000378:metabolism* / Q000601:surgery; D051381:Rats; D011917:Rats, Inbred Lew; D014161:Transduction, Genetic; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
C525719:Bmp2 protein, mouse; C525720:Bmp2 protein, rat; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D016212:Transforming Growth Factor beta