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

Abstract

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, 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^PRL-1. We analyzed the protein levels of adipogenesis markers and their signaling pathways in OFs from GO patients.

Results: The characteristics of PD-MSCs^PRL-1 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^PRL-1 compared to naïve cells. The mRNA and protein expression of adipogenic markers was decreased in PD-MSCs^PRL-1. Insulin-like growth factor-binding proteins (IGFBPs) secreting PD-MSCs^PRL-1 downregulated the phosphorylated PI3K/AKT/mTOR expression in OFs from GO patients. Interestingly, IGFBP2, - 4, - 6, and - 7 expression in PD-MSCs^PRL-1, 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^PRL-1 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.

Keywords: Adipogenesis; Gene modification; Graves’ ophthalmopathy; Phosphatase of regenerating liver-1; Placenta-derived mesenchymal stem cells.