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Nrf-2 overexpression in mesenchymal stem cells reduces oxidative stress-induced apoptosis and cytotoxicity
Mohammad Mohammadzadeh1, Raheleh Halabian, Ahmad Gharehbaghian
1Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, Tehran, Iran.
Abstract:
The most prominent capabilities of mesenchymal stem cells (MCSs) which make them promising for therapeutic applications are their capacity to endure and implant in the target tissue. However, the therapeutic applications of these cells are limited due to their early death within the first few days following transplantation. Therefore, to improve cell therapy efficacy, it is necessary to manipulate MSCs to resist severe stresses imposed by microenvironment. In this study, we manipulated MSCs to express a cytoprotective factor, nuclear factor erythroid-2 related factor 2 (Nrf2) to address this issue. Full-length human Nrf2 cDNA was isolated and TOPO cloned into TOPO cloning vector and then transferred to gateway adapted adenovirus expression vector by LR recombination reaction. Afterwards, the Nrf2 bearing recombinant virus was prepared in appropriate mammalian cell line and used to infect MSCs. The viability and apoptosis of the Nrf2 expressing MSCs were evaluated following hypoxic and oxidative stress conditions. Transient expression of Nrf2 by MSCs protected them against cell death and the apoptosis triggered by hypoxic and oxidative stress conditions. Nrf2 also enhanced the activity of SOD and HO-1. These findings could be used as a strategy for prevention of graft cell death in MSC-based cell therapy. It also indicates that management of cellular stress responses can be used for practical applications.
Insights
Mesenchymal stem cells (MSCs) show promise for therapy but often die post-transplant. Enhancing MSCs to express nuclear factor erythroid-2 related factor 2 (Nrf2) improves their survival against stress, boosting cell therapy potential.
Area of Science:
- Stem cell biology
- Regenerative medicine
- Molecular biology
Background:
- Mesenchymal stem cells (MSCs) possess therapeutic potential due to their engraftment capacity.
- Limited survival of transplanted MSCs under stress hinders effective cell therapy.
- Strategies to enhance MSC resilience are crucial for improving therapeutic outcomes.
Purpose of the Study:
- To engineer Mesenchymal stem cells (MSCs) to express nuclear factor erythroid-2 related factor 2 (Nrf2).
- To evaluate the cytoprotective effects of Nrf2 expression in MSCs under stress conditions.
- To investigate the impact of Nrf2 on MSC viability and apoptosis.
Main Methods:
- Human Nrf2 cDNA was cloned into an adenovirus expression vector.
- Recombinant adenovirus was used to infect MSCs, inducing Nrf2 expression.
- MSC viability and apoptosis were assessed under hypoxic and oxidative stress.
Main Results:
- Transient Nrf2 expression in MSCs significantly protected against hypoxic and oxidative stress-induced cell death.
- Nrf2 enhanced the activity of antioxidant enzymes superoxide dismutase (SOD) and heme oxygenase-1 (HO-1).
- Engineered MSCs demonstrated improved resistance to apoptosis.
Conclusions:
- Overexpression of Nrf2 is a viable strategy to enhance MSC survival in cell therapy.
- Managing cellular stress responses through genetic manipulation can improve graft survival.
- This approach holds potential for advancing MSC-based therapeutic applications.
