Macrophage migration inhibitory factor  facilitates the therapeutic efficacy of mesenchymal stem cells derived exosomes in acute myocardial infarction through upregulating miR-133a-3p

Abstract

Background Exosome transplantation is a promising cell-free therapeutic approach for the treatment of ischemic heart disease. The purpose of this study was to explore whether exosomes derived from Macrophage migration inhibitory factor (MIF) engineered umbilical cord MSCs (ucMSCs) exhibit superior cardioprotective effects in a rat model of AMI and reveal the mechanisms underlying it. Results Exosomes isolated from ucMSCs (MSC-Exo), MIF engineered ucMSCs (MIF-Exo) and MIF downregulated ucMSCs (siMIF-Exo) were used to investigate cellular protective function in human umbilical vein endothelial cells (HUVECs) and H9C2 cardiomyocytes under hypoxia and serum deprivation (H/SD) and infarcted hearts in rats. Compared with MSC-Exo and siMIF-Exo, MIF-Exo significantly enhanced proliferation, migration, and angiogenesis of HUVECs and inhibited H9C2 cardiomyocyte apoptosis under H/SD in vitro. MIF-Exo also significantly inhibited cardiomyocyte apoptosis, reduced fibrotic area, and improved cardiac function as measured by echocardiography in infarcted rats in vivo. Exosomal miRNAs sequencing and qRT-PCR confirmed miRNA-133a-3p significantly increased in MIF-Exo. The biological effects of HUVECs and H9C2 cardiomyocytes were attenuated with incubation of MIF-Exo and miR-133a-3p inhibitors. These effects were accentuated with incubation of siMIF-Exo and miR-133a-3p mimics that increased the phosphorylation of AKT protein in these cells. Conclusion MIF-Exo can provide cardioprotective effects by promoting angiogenesis, inhibiting apoptosis, reducing fibrosis, and preserving heart function in vitro and in vivo. The mechanism in the biological activities of MIF-Exo involves miR-133a-3p and the downstream AKT signaling pathway.

The Application Potential and Advance of Mesenchymal Stem Cell-Derived Exosomes in Myocardial Infarction

“…Additionally, the exosomes from SIRT1-overexpressing ADSCs can help to restore the function of cell migration and tube formation and recruitment of EPCs to the repair area through the Nrf2/CXCL12/CXCR7 pathway [84]. Macrophage migration inhibitory factor engineered hUC-MSC-produced exosomes also significantly enhanced proliferation, migration, and angiogenesis by delivering miR-133a-3p to HUVEC [85]. TIMP2-modified hUC-MSC-derived exosomes can promote the secretion level of Sfrp2 which contributed to HUVEC proliferation, migration, and tube formation in vitro and angiogenesis in a rat MI model [86].…”

Exosomes Derived From miR-214-3p Overexpressing Mesenchymal Stem Cells Promote Myocardial Repair

“…Isolation of exosome was proceed as described previously 9 . Brie y, huMSCs with 80% con uence were washed with PBS and then cocultured in exosome free medium for 48 h. The supernatant were centrifuged at 1500g for 30 minutes and incubated with Exosome Isolation Reagent (C10130-2, RiboBio, China) for 12 hours at a temperature of 4°C, and centrifuged at 2000 g for half an hour.…”

“…Masson trichrome staining and Hematoxylin-Eosin staining Heart tissues were collected, xed and sliced to 5μm. Mason's trichrome staining as described above 9 . The proportion of infarct area is the sum of infarct area of each section/sum of LV area of each section × 100%.…”

“…Western blotting was performed with a standard protocol as previously described 9 . Antibodies were used as follows: GAPDH (1:1000; 5174S; Cell Signaling Technology), Bcl S1.…”

“…MiRNAs controls gene-expressing by matching with 3 'untranslated region (UTR) of targeted mRNA, leading to mRNA degradation or translation inhibition 8 . More and more studies have shown that miRNAs take part in the regulation of myocardial angiogenesis, brosis and apoptosis after myocardial infarction, and become potential drug candidates for the treatment of myocardial infarction [9][10][11] . However, the poor stability of miRNAs in vivo and poor cellular uptake limit their clinical application 12 .…”