miRNA-126-3p carried by human umbilical cord mesenchymal stem cell enhances endothelial function through exosome-mediated mechanisms in vitro and attenuates vein graft neointimal formation in vivo

Abstract

Background The aim of this study was to determine whether the combination of MSC implantation with miRNA-126-3p overexpression would further improve the surgical results after vein grafting. Methods human umbilical cord MSCs (hucMSCs) and human umbilical vein endothelial cells (HUVECs) were isolated from human umbilical cords and characterized by a series of experiments. Lentivirus vector encoding miRNA-126-3p was transfected into hucMSCs and verified by PCR. We analyzed the miRNA-126-3p-hucMSC function in vascular endothelial cells by using a series of co-culture experiments. miRNA-126-3p-hucMSCs-exosomes were separated from cell culture supernatants and identified by WB and TEM. We validated the role of miRNA-126-3p-hucMSCs-exosomes on HUVECs proliferative and migratory and angiogenic activities by using a series of function experiments. We further performed co-culture experiments to detect downstream target genes and signaling pathways of miRNA-126-3p-hucMSCs in HUVECs. We established a rat vein grafting model, CM-Dil-labeled hucMSCs were injected intravenously into rats, and the transplanted cells homing to the vein grafts were detected by fluorescent microscopy. We performed historical and immunohistochemical experiments to exam miRNA-126-3p-hucMSC transplantation on vein graft neointimal formation and reendothelialization in vitro. Results We successfully isolated and identified primary hucMSCs and HUVECs. Primary hucMSCs were transfected with lentiviral vectors carrying miRNA-126-3p at a MOI 75. Co-culture studies indicated that overexpression of miRNA-126-3p in hucMSCs enhanced HUVECs proliferation, migration, and tube formation in vivo. We successfully separated hucMSCs-exosomes and found that miRNA-126-3p-hucMSCs-exosomes can strengthen the proliferative, migratory, and tube formation capacities of HUVECs. Further PCR and WB analysis indicated that, SPRED-1/PIK3R2/AKT/ERK1/2 pathways are involved in this process. In the rat vein arterialization model, reendothelialization analysis showed that transplantation with hucMSCs modified with miRNA-126-3p had a higher reendothelialization of the vein grafts. The subsequent historical and immunohistochemical examination revealed that delivery with miRNA-126-3p overexpressed hucMSCs significantly reduced vein graft intimal hyperplasia in rats. Conclusion These results suggest hucMSC-based miRNA-126-3p gene therapy may be a novel option for the treatment of vein graft disease after CABG.

miR-126-3p-loaded small extracellular vesicles secreted by urine-derived stem cells released from a phototriggered imine crosslink hydrogel could enhance vaginal epithelization after vaginoplasty

“…Among the predicted target genes, Spred1 and PIK3R2 have been reported to be related to migration and differentiation. Many studies have demonstrated that Spred1 and PIK3R2 are direct target genes of miR-126-3p and can be suppressed by miR-126-3p [ 23 – 25 ]. As shown in Fig.…”

miR-126-3p containing exosomes derived from human umbilical cord mesenchymal stem cells promote angiogenesis and attenuate ovarian granulosa cell apoptosis in a preclinical rat model of premature ovarian failure

“…The experimental procedures strictly followed the guidelines of the Ethics Committee of Qilu Hospital of Shandong University. Human umbilical cords were obtained from the informed, consenting delivery women, and the isolation and culture of hucMSCs were conducted based on the standard procedure as we have previously described [ 10 , 11 ]. The hucMSCs were maintained in α-MEM (SH30265.01B, Hyclone, USA) containing 10% FBS (16140-071, Gibco, USA) and passaged by trypsin (25200-056, Gibco, USA).…”

“…To construct the cell lines for stable and high expression of miR-126-3p, we selected lentivirus as overexpression vector in this study. The hucMSCs were transfected with miR-126-3p and control sequences by lentiviral vectors (Biolink, China) as we have reported [ 11 ]. The transfection efficiency of vectors was verified by GFP fluorescence signal and PCR.…”

“…Recently, we demonstrated that hucMSC-exosomes can be up-taken by endothelial cell and promote its angiogenic activities in vitro [ 10 ]. Moreover, we obtained data that miR-126-3p can be packed into hucMSCs-exosomes and overexpression of miR-126-3p in hucMSCs can further enhance its vascular protective effects by exosomes mechanism [ 11 ]. However, the effect of miR-126-3p-overexpressed hucMSCs-exosomes in ovarian function and structure remains unclear.…”

Mechanisms of Action of MiRNAs and LncRNAs in Extracellular Vesicle in Atherosclerosis

“…EVs are involved in the exchange of information between MSCs and other cells, including ECs, VSMCs, and macrophages, regulating ECs angiogenesis and proliferation, VSMCs proliferation and migration, and macrophages polarization and infiltration. MiRNA-30b and miRNA-126-3P of EVs from MSCs delivered into recipient HUVECs increase cell tube-like structure formation and migration, and regulate angiogenesis (59,102). Similarly, MSCs-derived EVs containing miRNA-125a, which enhances endothelial tip cell formation (61).…”