Unraveling The Effects of DICER1 Overexpression on Immune-Related Genes Expression in Mesenchymal Stromal/Stem Cells: Insights for Therapeutic Applications

Document Type : Original Article

Authors

1 Immunology Research Centre, Division of Inflammation and Inflammatory Diseases, Mashhad University of Medical Sciences, Mashhad, Iran

2 Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture, and Research (ACECR) - Khorasan Razavi, Iran

3 Division of Oncology, Laboratory of Cellular Therapy, Department of Medical and Surgical Sciences for Children and Adults, University Hospital of Modena and Reggio Emilia, Modena, Italy

4 Blood Borne Infections Research Center, Academic Center for Education, Culture, and Research (ACECR) - Khorasan Razavi, Iran

5 Basic Sciences Research Institute, Mashhad University of Medical Sciences, Mashhad, Iran

6 Cancer Research Center, Mashhad University of Medical Sciences, Mashhad, Iran

7 Department of Infectious Disease, Faculty of Medicine, Imperial College London, London, United Kingdom

Abstract

Objective: The immunoregulatory properties of mesenchymal stromal/stem cells (MSCs) bring a promise for the
treatment of inflammatory diseases. However, their ability to suppress the immune system is unstable. To enhance their
effectiveness against immune responses, it may be necessary to manipulate MSCs. Although some dsRNA transcripts
come from invading viruses, the majority of dsRNA has an endogenous origin and is known as endo-siRNA. DICER1 is
a ribonuclease protein that can generate small RNAs to modulate gene expression at the post-transcriptional level. We
aimed to evaluate the expression of several immune-related genes at mRNA and protein levels in MSCs overexpressing
DICER1 exogenously.
Materials and Methods: In this comparative transcriptomic experimental study, the adipose-derived MSCs (Ad-MSCs) were
transfected using the pCAGGS-Flag-hsDicer vector for the DICER1 overexpression. Following the RNA extraction,
mRNA expression level of DICER1 and several inflammatory cytokines were examined. We performed a relative
real-time polymerase chain reaction (PCR) assay and transcriptome analysis between two groups including DICER1-
transfected MSCs and control MSCs. Moreover, media from the transfected MSCs were evaluated for various interferon
response factors by ELISA.
Results: The overexpression of DICER1 is associated with a significant increase in the mRNA expression level of
COX-2, DDX-58, IFIH1, MYD88, RNase L, TLR3/4, and TDO2 genes and a downregulation of the TSG-6 gene in
MSCs. Moreover, the expression levels of IL-1, 6, 8, 17, 18, CCL2, INF-γ, TGF-β, and TNF-α were higher in the
DICER1-transfected MSCs group.
Conclusion: It seems that the ectopic expression of DICER1 in Ad-MSCs is linked to alterations in the expression level
of immune-related genes. It is suggested that the manipulation of immune-related pathways in MSCs via the Dicer1
overexpression could facilitate the development of MSCs with distinct immunoregulatory phenotypes.

Keywords

Main Subjects

References

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