Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector

Abstract

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.

Antitumor effect of genetically engineered mesenchymal stem cells in a rat glioma model

“…28,29 Human IL-2-transduced MSCs (MSCs-IL2) were prepared by infection with modified adenoviral vector as previously described. 15 This fiber-mutant vector was employed because rat primary MSCs are relatively resistant to wild-type adenoviral infection due to their low level of expression of the adenoviral receptor, coxsackie-adenovirus receptor Gene-modified mesenchymal stem cells against glioma K Nakamura et al (CAR). In agreement with our previous observations, high levels of hIL-2 were detected in the supernatant of MSCs infected with relatively low concentrations of AxCAhIL2-F/RGD (8.670.5 and 24.071.7 ng/ml/10 4 cells/72 h at 300 and 1000 particle units/cell, respectively).…”

“…Among bone marrow stem cells, much attention has been paid to mesenchymal stem cells (MSCs, also referred to as bone marrow stromal cells) because of their plasticity for differentiation into classical mesenchymal lineages, such as adipocytes, chondrocytes, and osteocytes, [15][16][17][18] and also neuronal lineages. [19][20][21][22] This neurogenic potential of MSCs was revealed under special culture conditions or following intracranial implantation.…”

Genetically Engineered Mesenchymal Stem Cells: The Ongoing Research for Bone Tissue Engineering

“…BMP-2, approved by Food and Drug Administration (FDA) for clinical practice, is the most potent member in promoting bone and cartilage development and therefore wins a popular choice for MSCs-based BTE (Lieberman et al, 1998;Lieberman et al, 1999;Lou et al, 1999;Turgeman et al, 2001;Olmsted-Davis et al, 2002;Blum et al, 2003;Gugala et al, 2003;Park et al, 2003;Riew et al, 2003;Tsuda et al, 2003;Kumar et al, 2004;Hasharoni et al, 2005;Egermann et al, 2006;Feeley et al, 2006). The BMP-2-modified MSCs are proven to increase the alkaline phosphatase (ALP) activity, mineralization, and cell proliferation in vitro and induce ectopic bone formation, heal critical size bone defect, repair fracture, and trigger spinal fusion in vivo (Lou et al, 1999;Moutsatsos et al, 2001;Turgeman et al, 2001;Blum et al, 2003;Park et al, 2003;Riew et al, 2003;Tsuda et al, 2003;Hasharoni et al, 2005;Egermann et al, 2006;Feeley et al, 2006). BMP-7 plays a key role in osteoblast differentiation, and there is only one study in which MSCs are engineered with BMP-7.…”

“…Adenovirus vector is one of the most popular vectors for MSC gene therapy aimed at BTE Dumont et al, 2002;Olmsted-Davis et al, 2002;Blum et al, 2003;Dayoub et al, 2003;Gugala et al, 2003;Li et al, 2003;Park et al, 2003;Riew et al, 2003;Sugiyama et al, 2003;Tsuda et al, 2003;Yang et al, 2003;Kumar et al, 2004;Zheng et al, 2004;Zhao et al, 2005;Egermann et al, 2006;Feeley et al, 2006;Zachos et al, 2006). Adenovirus vector is characterized by efficient transduction of quiescent cells and a relatively high capacity for transgene insertion.…”

“…An optimal scaffold should provide genetically modified MSCs with the appropriate environment for attachment, proliferation, and differentiation, preventing them from migrating into other places of the body. The scaffold may be soluble for direct injection or solid for surgical implantation (Peng et al, 2001;Turgeman et al, 2001;Blum et al, 2003;Park et al, 2003;Rose et al, 2003;Tsuda et al, 2003;Feeley et al, 2006). Genetically engineered MSCs have the ability to induce bone regeneration not only through paracrine secretion of growth factors to adjacent cells but also by an autocrine effect on MSCs themselves to promote their osteogenic differentiation (Gazit et al, 1999).…”