Adenovirus-mediated expression of the HO-1 protein within MSCs decreased cytotoxicity and inhibited apoptosis induced by oxidative stresses

Abstract

The capacity of mesenchymal stem cells (MSCs) to survive and engraft in the target tissue may lead to promising therapeutic effects. However, the fact that the majority of MSCs die during the first few days following transplantation complicates cell therapy. Hence, it is necessary to strengthen the stem cells to withstand the rigors of the microenvironment to improve the efficacy of cell therapy. In this study, we manipulated MSCs to express a cytoprotective factor, heme oxygenase-1 (HO-1), to address this issue. Full-length cDNA of human HO-1 was isolated and cloned into TOPO vector by TOPO cloning reaction. Then, the construct was ligated to gateway adapted adenovirus expression vector by LR recombination reaction. Afterwards, the recombinant virus expressing HO-1 was produced in appropriate mammalian cell line and used to infect MSCs. The HO-1 engineered MSCs were exposed to hypoxic and oxidative stress conditions followed by evaluation of the cells' viability and apoptosis. Transient expression of HO-1 was detected within MSCs. It was observed that HO-1 expression could protect MSCs against cell death and the apoptosis triggered by hypoxic and oxidative stress conditions. The MSCs-HO-1 retained their ability to differentiate into adipogenic, chondrogenic, or osteogenic lineages. These findings could be applied as a strategy for prevention of graft cell death in MSCs-based cell therapy and is a good demonstration of how an understanding of cellular stress responses can be used for practical applications.

Heme oxygenase-1-transduced bone marrow mesenchymal stem cells in reducing acute rejection and improving small bowel transplantation outcomes in rats

“…Cellular morphology and proliferation, in-vitro differentiation, and surface molecule characterization of our results all demonstrated that Ad/HO-1 did not alter the biological properties of BMMSCs, which was in agreement with previous studies [9, 43, 45]. Immunocytochemical staining revealed that the red fluorescence intensity in the cytoplasm was higher, while western blot analysis showed more HO-1 expression in Ad/HO-1/BMMSCs than in BMMSCs and in Ad/BMMSCs, indicating that the HO-1 gene was successfully overexpressed in the cytoplasm of Ad/HO-1/BMMSCs.…”

Mesenchymal Stem/Stromal Cells in Liver Fibrosis: Recent Findings, Old/New Caveats and Future Perspectives

“…Despite the lack of enough in vivo tracking evidence to support any of those views, some articles showed new strategies aiming at prolonging the survival of MSCs in an adverse environment. P2 hBM-MSCs genetically-modified to transiently express cytoprotective factors (i.e., heme oxygenase-1 or nuclear factor erythroid-2 related factor 2), by using adenoviral vectors, were able to survive better in hypoxic and oxidative-stress conditions [86,87].…”

Heme oxygenase-1, a novel target for the treatment of diabetic complications: focus on diabetic peripheral neuropathy

“…Stem cells genetically combined with HO-1 using Adenovirus vector improved the success of stem cell transplantation. HO-1 has shown cytoprotective effects in mesenchymal stem cells by preventing hypoxia and oxidative stress damage besides retention of the differentiation properties [56].Furthermore ischemia/reperfusion induced renal injury during transplantation was prevented by adenovirus mediated HO-1 delivery [57]. Diabetic mice treated with adenoviral vectors containing HO-1 showed increase in wound healing and neovascularization [58].Recombinant adeno-associated virus (rAAV) offers safe, effective and long term therapy with minimum deleterious effect on host cell and non-immunogenicity making the delivery of HO-1 more effective.…”