Abstract
We proposed a novel combined gene therapy of human vascular endothelial growth factor 165 gene (hVEGF165) and human bone morphogenetic protein 2 gene (hBMP2) for bone regeneration by lentivirus-mediated co-transfection of both genes into rat bone marrow-derived mesenchymal stromal cells (MSCs). Both genes were successfully co-expressed in MSCs confirmed by real-time PCR and ELISA. And the alkaline phosphatase activity of MSCs was significantly augmented by the co-transfection with both genes than any single gene transfection (P < 0.01). These results demonstrated the feasibility of the combined gene therapy by using MSCs lentivirally co-transfected with hVEGF165 and hBMP2 for bone regeneration.
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Cheng H, Jiang W, Phillips FM, Haydon RC, Peng Y, Zhou L, Luu HH, An N, Breyer B, Vanichakarn P, Szatkowski JP, Park JY, He TC (2003) Osteogenic activity of the fourteen types of human bone morphogenetic proteins (BMPs). J Bone Joint Surg Am 85:1544–1552
Deckers MM, Karperien M, van der Bent C, Yamashita T, Papapoulos SE, Lowik CW (2000) Expression of vascular endothelial growth factors and their receptors during osteoblast differentiation. Endocrinology 141:1667–1674
Hsu WK, Sugiyama O, Park SH, Conduah A, Feeley BT, Liu NQ, Krenek L, Virk MS, An DS, Chen IS, Lieberman JR (2007) Lentiviral-mediated BMP-2 gene transfer enhances healing of segmental femoral defects in rats. Bone 40:931–938
Jiang XQ, Chen JG, Gittens S, Chen CJ, Zhang XL, Zhang ZY (2005) The ectopic study of tissue-engineered bone with hBMP-4 gene modified bone marrow stromal cells in rabbits. Chin Med J 118:281–288
Jung HS, Kwon BS, Lee SW (2005) Tumor-specific gene delivery using RNA-targeting Tetrahymena group I intron. Biotechnol Lett 27:567–574
Lieberman JR, Ghivizzani SC, Evans CH (2002) Gene transfer approaches to the healing of bone and cartilage. Mol Ther 6:141–147
Lu W, Zhou X, Hong B, Liu J, Yue Z (2004) Suppression of invasion in human U87 glioma cells by adenovirus-mediated co-transfer of TIMP-2 and PTEN gene. Cancer Lett 214:205–213
McMahon JM, Conroy S, Lyons M, Greiser U, O’shea C, Strappe P, Howard L, Murphy M, Barry F, O’Brien T (2006) Gene transfer into rat mesenchymal stem cells: a comparative study of viral and nonviral vectors. Stem Cells Develop 15:87–96
Midy V, Plouet J (1994) Vasculotropin/vascular endothelial growth factor induces differentiation in cultured osteoblasts. Biochem Biophys Res Commun 199:380–386
Naldini L, Blomer U, Gage FH, Trono D, Verma IM (1996) Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc Natl Acad Sci USA 93:11382–11388
Ojima T, Iwahashi M, Nakamura M, Matsuda K, Naka T, Nakamori M, Ueda K, Ishida K, Yamaue H (2006) The boosting effect of co-transduction with cytokine genes on cancer vaccine therapy using genetically modified dendritic cells expressing tumor-associated antigen. Int J Oncol 28:947–953
Spinella-Jaegle S, Roman-Roman S, Faucheu C, Dunn FW, Kawai S, Gallea S, Stiot V, Blanchet AM, Courtois B, Baron R, Rawadi G (2001) Opposite effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on osteoblast differentiation. Bone 29:323–330
Street J, Bao M, deGuzman L, Bunting S, Peale FV Jr, Ferrara N, Steinmetz H, Hoeffel J, Cleland JL, Daugherty A, van Bruggen N, Redmond HP, Carano RA, Filvaroff EH (2002) Vascular endothelial growth factor stimulates bone repair by promoting angiogenesis and bone turnover. Proc Natl Acad Sci USA 99:9656–9661
Sugiyama O, An DS, Kung SP, Feeley BT, Gamradt S, Liu NQ, Chen IS, Lieberman JR (2005) Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo. Mol Ther 11:390–398
Tsuda H, Wada T, Yamashita T, Hamada H (2005) Enhanced osteoinduction by mesenchymal stem cells transfected with a fiber-mutant adenoviral BMP2 gene. J Gene Med 7:1322–1334
Wang Y, Zeng B, Li X (2006) Expression of human calcitonin by microencapsulated recombinant myoblasts. Biotechnol Lett 28:1453–1458
Yeh LC, Lee JC (2006) Co-transfection with the osteogenic protein (OP)-1 gene and the insulin-like growth factor (IGF)-I gene enhanced osteoblastic cell differentiation. Biochim Biophys Acta 1763:57–63
Zelzer E, McLean W, Ng YS, Fukai N, Reginato AM, Lovejoy S, D’Amore PA, Olsen BR (2002) Skeletal defects in VEGF(120/120) mice reveal multiple roles for VEGF in skeletogenesis. Development 129:1893–1904
Acknowledgements
This work was supported by Key Program of Shanghai Science and Technology Committee (054119520). Hereby, we would like to express our great appreciation to the following people from Shanghai Tissue Engineering Research and Development Center for their kindness and support in this study: Lei Cui, Yu-ling Li, Heng Zhou, Guang-peng Liu, Jian Sun.
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Jiang, J., Fan, Cy. & Zeng, Bf. Osteogenic differentiation effects on rat bone marrow-derived mesenchymal stromal cells by lentivirus-mediated co-transfection of human BMP2 gene and VEGF165 gene. Biotechnol Lett 30, 197–203 (2008). https://doi.org/10.1007/s10529-007-9532-1
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DOI: https://doi.org/10.1007/s10529-007-9532-1