MMP-2 and MMP-14 Silencing Inhibits VEGFR2 Cleavage and Induces the Differentiation of Porcine Adipose-Derived Mesenchymal Stem Cells to Endothelial Cells

Abstract

The molecular mechanisms that control the ability of adipose‐derived mesenchymal stem cells (AMSCs) to remodel three‐dimensional extracellular matrix barriers during differentiation are not clearly understood. Herein, we studied the expression of matrix metalloproteinases (MMPs) during the differentiation of AMSCs to endothelial cells (ECs) in vitro. MSCs were isolated from porcine abdominal adipose tissue, and characterized by immunopositivity to CD44, CD90, CD105, and immunonegativity to CD14 and CD45. Plasticity of AMSCs was confirmed by multilineage differentiation. The mRNA transcripts for MMPs and Tissue Inhibitor of Metalloproteinases (TIMPs), and protein expression of EC markers were analyzed. The enzyme activity and protein expression were analyzed by gelatin zymography, enzyme‐linked immunosorbent assay (ELISA), and Western blot. The differentiation of AMSCs to ECs was confirmed by mRNA and protein expressions of the endothelial markers. The mRNA transcripts for MMP‐2 and MMP‐14 were significantly increased during the differentiation of MSCs into ECs. Findings revealed an elevated MMP‐14 and MMP‐2 expression, and MMP2 enzyme activity. Silencing of MMP‐2 and MMP‐14 significantly increased the expression of EC markers, formation of capillary tubes, and acetylated‐low‐density lipoprotein uptake, and decreased the cleavage of vascular endothelial growth factor receptor type 2 (VEGFR2). Inhibition of VEGFR2 significantly decreased the expression of EC markers. These novel findings demonstrate that the upregulation of MMP2 and MMP14 has an inhibitory effect on the differentiation of AMSCs to ECs, and silencing these MMPs inhibit the cleavage of VEGFR2 and stimulate the differentiation of AMSCs to ECs. These findings provide a potential mechanism for the regulatory role of MMP‐2 and MMP‐14 in the re‐endothelialization of coronary arteries following intervention. Stem Cells Translational Medicine 2017;6:1385–1398

Mesenchymal stromal cell therapy for liver diseases

“…This observation was in comparison with MSCs treated with endothelial growth medium only. 143 In other work, the expression level of heme oxgenase 1 (HO-1 [HMOX1]) was genetically modified in MSCs and shown to increase MSCs resis-tance to cell death under oxidative stress conditions and enhance their anti-apoptotic properties. 144 Moreover, more MSCs overexpressing HO-1 survived following exposure to H 2 O 2 and hypoxia, indicating that HO-1 may shape the stress responsive and cytoprotective properties of MSCs.…”

Role of the CXCR4-SDF1-HMGB1 pathway in the directional migration of cells and regeneration of affected organs

“…MSCs are capable of differentiating into several mesoderm lineages, including adipogenic, osteogenic, chondrogenic and myogenic lineages, depending on the multitude of stimuli and inhibitors present in the tissue microenvironment[ 50 ]. The micro-environment plays an important role in the activation or downregulation of transcription factors that regulate the expression of genes responsible for the induction and progression of tissue-specific differentiation[ 51 ]. MSCs can also generate neural cells in the ectodermal layer, and hepatic cells and pancreatic cells in the endodermal layer[ 52 ].…”

Behavior of Metalloproteinases in Adipose Tissue, Liver and Arterial Wall: An Update of Extracellular Matrix Remodeling

“…However, the molecular mechanism is still not completely described. Almalki et al [29] recently studied in vitro the expression of MMPs during the differentiation of AMSCs isolated from porcine abdominal AT to endothelial cells [29]. The authors demonstrated that the up-regulation of MMP-2 and MMP-14 has an inhibitory effect on the differentiation of AMSCs to endothelial cells; the silencing these MMPs inhibits the cleavage of the VEGF-receptor and stimulates the differentiation of AMSCs to endothelial cells and consequently the formation of endothelial tubes.…”