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Published on: March 29, 2018
Tracking expression of virally mediated BMP-2 in gene therapy for bone repair
Seth C Gamradt1, Nobuhiro Abe, Matthew E Bahamonde
1Department 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
Abstract:
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.
