Immunomodulatory Properties of Mouse Mesenchymal Stromal/Stem Cells Upon Ectopic Expression of Immunoregulator Nanos2

Background Mesenchymal stromal/stem cells (MSCs) are known for their involvement in modulating the immune system of mammals. This potency could be enhanced by different strategies, including regulation of key proteins, in order to meet desirable therapeutic properties. Nanos2, encoding an RNA-binding protein involved in regulation of key spermatogonial signaling pathways, has been demonstrated to downregulate a range of immune related genes in mouse embryonic fibroblasts (MEFs). Accordingly, it was hypothesized that Nanos2 functions as a potent immunosuppressing factor. This study was aimed to measure the expression profile of the immune-related genes in mouse mesenchymal stromal/stem cells (mMSCs) and assess their functional properties after Nanos2 ectopic expression. Methods As inflammatory mediators, interferon (IFN-gamma) and poly(I:C) were used to provoke an immune response. The interactions between the control and engineered mMSCs overexpressing Nanos2, with mouse peripheral blood mononuclear cells (mPBMCs) were then compared. The sensitivity of these cells to an inflammatory environment was assessed by using a conditioned medium containing high levels of inflammatory cytokines. Finally, the functional properties of the cells were investigated both in vivo and in vitro in presence of tumor and immune cells. Results Deep transcriptome analysis indicated that numerous genes were downregulated as a result of higher Nanos2 expression. Most of the genes subjected to gene expression alteration, were responsible for controlling responses to external stimuli, cell-cell adhesion, and wound healing. In comparison to the control cells, Nanos2-overexpressing cells showed lower expression of several immune-related genes after pretreatment with IFN-gamma and poly(I:C). They also exhibited inhibitory effects against mPBMCs proliferation. Tumor growth rate, in B16-F0 administered mice was obviously increased upon their treatment with the Nanos2-mMSCs, while no tumor or very small ones were developed in the control group. In addition, the cytotoxic environment had no significant effects on Nanos2-mMSCs. Conclusions According to the literature, MSCs are believed to be tuned very precisely by their internal and external conditions to act as either pro-inflammatory or anti-inflammatory agents. We show here that Nanos2 plays a significant role in promoting anti-inflammatory properties when expressed at higher levels by MSCs. This approach could be adopted for controlling the excessive inflammatory conditions in clinical programs, however more experiments are required to confirm it.

Automated summary

Nanos2 expression in mouse mesenchymal stromal/stem cells downregulates immune-related genes and exhibits anti-inflammatory effects. This approach may be useful in controlling clinical inflammatory conditions, although further research is needed.