Abstract: Bone marrow-derived mesenchymal stem cells (MSCs) are being explored for clinical applications, and genetic engineering represents a
useful strategy for boosting the therapeutic potency of MSCs. Vascular endothelial growth factor (VEGF)-based gene therapy protocols
have been used to treat tissue ischemia, and a combined VEGF/MSC therapeutics is appealing due to their synergistic paracrine actions.
However, multiple VEGF splice variants exhibit differences in their mitogenicity, chemotactic efficacy, receptor interaction, and tissue
distribution, and the differential regulatory effects of multiple VEGF isoforms on the function of MSCs have not been characterized. We
expressed three rat VEGF-A splice variants VEGF120, 164, and 188 in MSCs using adenoviral vectors, and analyzed their effects on MSC
proliferation, differentiation, survival, and trophic factor production. The three VEGF splice variants exert common and differential effects
on MSCs. All three expressed VEGFs are potent in promoting MSC proliferation. VEGF120 and 188 are more effective in amplifying
expression of multiple growth factor and cytokine genes. VEGF164 on the other hand is more potent in promoting expression of genes
associated with MSC remodeling and endothelial differentiation. The longer isoform VEGF188, which is preferentially retained by
proteoglycans, facilitates bone morphogenetic protein-7 (BMP7)-mediated MSC osteogenesis. Under serum starvation condition, virally
expressed VEGF188 preferentially enhances serum withdrawal-mediated cell death involving nitric oxide production. This work indicates
that seeking the best possible match of an optimal VEGF isoform to a given disease setting can generate maximum therapeutic benefits and
minimize unwanted side effects in combined stem cell and gene therapy.