Bidkhori Hamid Reza, Farshchian Moein, Hasanzadeh Halimeh, Jafarzadeh Esfehani Reza, Alsadat Mahmoudian Reihaneh, Moradi Marjaneh Mahdi, Rafatpanah Houshang
Immunology Research Centre, Division of Inflammation and Inflammatory Diseases, Mashhad University of Medical Sciences, Mashhad, Iran.
Stem Cells and Regenerative Medicine Department, Academic Center for Education, Culture, and Research (ACECR) - Khorasan Razavi, Iran.
Cell J. 2023 Oct 1;25(10):696-705. doi: 10.22074/cellj.2023.1988987.1221.
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.
In this comparative transcriptomic experimental study, the adipose-derived MSCs (Ad-MSCs) were transfected using the pCAGGS-Flag-hsDicer vector for the overexpression. Following the RNA extraction, mRNA expression level of 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.
The overexpression of is associated with a significant increase in the mRNA expression level of and genes and a downregulation of the TSG-6 gene in MSCs. Moreover, the expression levels of and were higher in the DICER1-transfected MSCs group.
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.
间充质基质/干细胞（MSCs）的免疫调节特性为炎症性疾病的治疗带来了希望。然而，它们抑制免疫系统的能力并不稳定。为了提高其对抗免疫反应的有效性，可能有必要对MSCs进行调控。虽然一些双链RNA转录本来自入侵病毒，但大多数双链RNA具有内源性起源，被称为内源性小干扰RNA（endo-siRNA）。DICER1是一种核糖核酸酶蛋白，可产生小RNA以在转录后水平调节基因表达。我们旨在评估外源性过表达DICER1的MSCs中几种免疫相关基因在mRNA和蛋白质水平的表达。
在这项比较转录组学实验研究中，使用pCAGGS-Flag-hsDicer载体转染脂肪来源的MSCs（Ad-MSCs）以进行过表达。RNA提取后，检测 以及几种炎性细胞因子的mRNA表达水平。我们对包括DICER1转染的MSCs和对照MSCs的两组进行了相对实时聚合酶链反应（PCR）分析和转录组分析。此外，通过酶联免疫吸附测定（ELISA）评估转染的MSCs培养基中的各种干扰素反应因子。
的过表达与MSCs中 和 基因的mRNA表达水平显著增加以及TSG-6基因的下调有关。此外，DICER1转染的MSCs组中 和 的表达水平更高。
似乎Ad-MSCs中DICER1的异位表达与免疫相关基因表达水平的改变有关。建议通过Dicer1过表达对MSCs中的免疫相关途径进行调控，可能有助于开发具有独特免疫调节表型的MSCs。
