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Enhancement of Immunoregulatory Function of Modified Bone Marrow Mesenchymal Stem Cells by Targeting SOCS1

Xiaoming Zhang¹, Fei Hua², Ziying Yang²

¹Department of Cardiothoracic Surgery, Suzhou Municipal Hospital, Suzhou, China.

Biomed Research International

|June 2, 2018

Abstract

Objective:

The study aim to investigate the role of microRNA-155 (miR-155) on the immunoregulatory function of bone marrow mesenchymal stem cells (MSCs).

Methods:

MSCs were isolated from 2-week-old Sprague-Dawley rats and identified by flow cytometry using anti-CD29, anti-CD44, anti-CD34, and anti-CD45 antibodies. MSCs were transfected with miR155-mimics, miR155-inhibitor, and control oligos, respectively, and then cocultured with spleen mononuclear cells (SMCs). The mRNA levels of Th1, Th2, Th17, and Treg cell-specific transcription factors (Tbx21, Gata3, Rorc, and Foxp3, resp.) and the miR-155 target gene SOCS1 were detected by quantitative real-time PCR (qPCR) in SMCs. The proportion of CD4⁺ FOXP3⁺ Treg cells was detected by flow cytometry. In addition, the effects of MSCs transfected with miR-155 on the migration of rat SMCs were investigated by transwell chamber.

Results:

CD29 and CD44 were expressed in MSCs, while CD34 and CD45 were negative. The percentage of CD4⁺ FOXP3⁺ Treg cells in the SMC population was significantly higher compared with that noted in SMCs control group (p < 0.001) following 72 hours of coculture with miR155-mimics-transfected SMCs. In contrast, the percentage of CD4⁺ FOXP3⁺ Treg cells in the SMCs cocultured with miR155-inhibitor-transfected MSCs was significantly lower compared with that noted in SMCs control group (p < 0.001). MiR155-mimics-transfected MSCs inhibited the expression of Tbx21, Rorc, and SOCS1, while the expression of Gata3 and Foxp3 was increased. In contrast to the downregulation of the aforementioned genes, miR155-inhibitor-transfected MSCs resulted in upregulation of Tbx21, Rorc, and SOCS1 expression levels and inhibition of Gata3 and Foxp3. In the transwell assay, miR155-mimics-transfected MSCs exhibited lower levels of SMCs migration, while the miR155-inhibitor-transfected MSCs demonstrated significantly higher levels of migration, compared with the blank control group (p < 0.01, resp.).

Conclusion:

miR-155 favors the differentiation of T cells into Th2 and Treg cells in MSCs, while it inhibits the differentiation to Th1 and Th17 cells.

Insights

MicroRNA-155 (miR-155) influences bone marrow mesenchymal stem cells (MSCs) to promote T helper 2 (Th2) and regulatory T (Treg) cell differentiation while suppressing T helper 1 (Th1) and T helper 17 (Th17) cell development.

Area of Science:

Immunology
Stem Cell Biology
Molecular Biology

Background:

Bone marrow mesenchymal stem cells (MSCs) possess immunomodulatory properties.
MicroRNAs (miRNAs) play critical roles in regulating cellular functions, including immune responses.
The specific role of microRNA-155 (miR-155) in MSC-mediated immunomodulation requires further elucidation.

Purpose of the Study:

To investigate the impact of miR-155 on the immunoregulatory capacity of MSCs.
To determine how manipulating miR-155 levels in MSCs affects T cell differentiation.
To assess the influence of miR-155 on MSC-mediated suppression of T cell migration.

Main Methods:

MSCs were isolated from Sprague-Dawley rats and characterized using flow cytometry.
MSCs were transfected with miR-155 mimics or inhibitors and co-cultured with rat spleen mononuclear cells (SMCs).
T cell differentiation (Th1, Th2, Th17, Treg) and gene expression (SOCS1, Tbx21, Gata3, Rorc, Foxp3) were analyzed via qPCR and flow cytometry; SMC migration was assessed using Transwell assays.

Main Results:

Overexpression of miR-155 in MSCs significantly increased the proportion of CD4+FOXP3+ Treg cells and enhanced Gata3 and Foxp3 expression, while inhibiting Tbx21 and Rorc.
Inhibition of miR-155 in MSCs led to a significant decrease in Treg cells and reversed the expression patterns of T cell transcription factors.
MSCs with miR-155 overexpression reduced SMC migration, whereas miR-155 inhibition increased SMC migration.

Conclusions:

miR-155 plays a crucial role in modulating MSC immunoregulatory functions.
miR-155 promotes Th2 and Treg cell differentiation and suppresses Th1 and Th17 cell differentiation.
Modulating miR-155 in MSCs offers a potential strategy for controlling immune responses.

Read full analysis

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