功能增强的胎盘间充质干细胞抑制格雷夫斯眼病眼眶成纤维细胞的脂肪生成。

Functionally enhanced placenta-derived mesenchymal stem cells inhibit adipogenesis in orbital fibroblasts with Graves' ophthalmopathy.

机构信息

Department of Biomedical Science, CHA University, Seongnam, 13488, Republic of Korea.

Center for Non-Clinical Development, CHA Advanced Research Institute CHA University, Seongnam, 13488, Republic of Korea.

出版信息

Stem Cell Res Ther. 2020 Nov 5;11(1):469. doi: 10.1186/s13287-020-01982-3.

BACKGROUND

Placenta-derived mesenchymal stem cells (PD-MSCs) have unique immunomodulatory properties. Phosphatase of regenerating liver-1 (PRL-1) regulates the self-renewal ability of stem cells and promotes proliferation. Graves' ophthalmopathy (GO) is an autoimmune inflammatory disease of the orbit and is characterized by increased orbital levels of adipose tissue. Here, we evaluated the therapeutic mechanism for regulation of adipogenesis by PRL-1-overexpressing PD-MSCs (PD-MSCs, PRL-1+) in orbital fibroblast (OF) with GO patients.

METHODS

PD-MSCs isolated from human placenta were transfected with the PRL-1 gene using nonviral transfection method. Primary OFs were isolated from orbital adipose tissue specimens from GO patients. After maturation as adipogenic differentiation, normal and GO-derived OFs were cocultured with naïve and PD-MSCs. We analyzed the protein levels of adipogenesis markers and their signaling pathways in OFs from GO patients.

RESULTS

The characteristics of PD-MSCs were similar to those of naïve cells. OFs from GO patients induced adipocyte differentiation and had significantly decreased a lipid accumulation after coculture with PD-MSCs compared to naïve cells. The mRNA and protein expression of adipogenic markers was decreased in PD-MSCs. Insulin-like growth factor-binding proteins (IGFBPs) secreting PD-MSCs downregulated the phosphorylated PI3K/AKT/mTOR expression in OFs from GO patients. Interestingly, IGFBP2, - 4, - 6, and - 7 expression in PD-MSCs, which was mediated by integrin alpha 4 (ITGA4) and beta 7 (ITGB7), was higher than that in naïve cells and upregulated phosphorylated FAK downstream factor.

CONCLUSION

In summary, IGFBPs secreting PD-MSC inhibit adipogenesis in OFs from GO patients by upregulating phosphorylated FAK and downregulating PI3K/AKT/mTOR signaling pathway. The functional enhancement of PD-MSCs by nonviral gene modification provides a novel therapeutic strategy for the treatment of degenerative diseases.

背景

胎盘来源的间充质干细胞（PD-MSCs）具有独特的免疫调节特性。肝再生磷酸酶-1（PRL-1）调节干细胞的自我更新能力，并促进增殖。格雷夫斯眼病（GO）是一种眼眶的自身免疫性炎症性疾病，其特征是眼眶脂肪组织水平升高。在这里，我们评估了过表达 PRL-1 的 PD-MSCs（PD-MSCs，PRL-1+）对 GO 患者眼眶成纤维细胞（OF）脂肪生成的调节作用。

方法

使用非病毒转染方法将 PRL-1 基因转染到 PD-MSCs 中。从 GO 患者的眼眶脂肪组织标本中分离出原代 OF。作为成脂分化成熟后，将正常和 GO 来源的 OF 与幼稚和成纤维细胞共培养。我们分析了 GO 患者来源的 OF 中脂肪生成标志物及其信号通路的蛋白水平。

结果

PD-MSCs 的特征与幼稚细胞相似。与幼稚细胞相比，GO 患者来源的 OF 诱导脂肪细胞分化，与 PD-MSCs 共培养后脂滴积累明显减少。PD-MSCs 中脂肪生成标志物的 mRNA 和蛋白表达降低。分泌胰岛素样生长因子结合蛋白（IGFBPs）的 PD-MSCs 下调了 GO 患者来源的 OF 中磷酸化 PI3K/AKT/mTOR 的表达。有趣的是，PD-MSCs 中 IGFBP2、-4、-6 和-7 的表达，由整合素 alpha 4（ITGA4）和 beta 7（ITGB7）介导，高于幼稚细胞，并上调磷酸化 FAK 下游因子。