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

Stem Cell Research & Therapy

Chunmei Zhang¹  Weidong Bing²  Yanwen Bi²  Limei Wang³  Qingxi Qu³  Xuanxuan Jing⁴  Linghong Liu⁵ 

[1] Department of Cardiology, Qilu Hospital of Shandong University, 250012, Jinan, Shandong, People’s Republic of China;Department of Cardiovascular Surgery, Qilu Hospital of Shandong University, 250012, Jinan, People’s Republic of China;Department of Obstetrics and Gynecology, Qilu Hospital of Shandong University, 250012, Jinan, People’s Republic of China;Department of Ultrasound, Qilu Hospital of Shandong University, 250012, Jinan, Shandong, People’s Republic of China;Laboratory of Cryomedicine, Qilu Hospital of Shandong University, 250012, Jinan, Shandong, People’s Republic of China;Research Center of Stem Cell and Regenerative Medicine, Shandong University, 250012, Jinan, Shandong, People’s Republic of China;

关键词: miRNA-126-3p;    Exosomes;    Mesenchymal stem cell;    Vein graft;    Reendothelialization;    Neointimal hyperplasia;   

DOI  :  10.1186/s13287-020-01978-z

来源: Springer

【 摘 要 】

BackgroundThe 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.Methodshuman 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.ResultsWe 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.ConclusionThese results suggest hucMSC-based miRNA-126-3p gene therapy may be a novel option for the treatment of vein graft disease after CABG.

【 授权许可】

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