MiR-101-containing extracellular vesicles bind to BRD4 and enhance proliferation and migration of trophoblasts in preeclampsia

Abstract

Background: Preeclampsia (PE) is a frequently occurring pregnancy disorder in the placenta, which results in various maternal and fetal complications. The current study aims to evaluate the role of extracellular vesicles (EVs)encapsulated microRNA (miR)-101 in biological processes of trophoblasts in PE and its underlying mechanism. Methods: Human umbilical cord mesenchymal stem cell (HUCMSC) and HUCMSC-derived EVs were isolated and cultured, after which EV characterization was carried out using PKH67 staining. In silico analyses were adopted to predict the downstream target genes of miR-101, and dual luciferase reporter gene assay was applied to validate the binding affinity. Furthermore, loss-and gain-of-function approaches were adopted to determine the role of miR-101 and bromodomain-containing protein 4 (BRD4) in trophoblast proliferation and invasion using EDU staining and transwell assay. In addition, a rat model of PE was established to verify the function of EV-encapsulated miR-101 in vivo. Results: Placental tissues obtained from PE patients presented with downregulated miR-101 expression and upregulated BRD4 and CXCL11 expression. EV-encapsulated miR-101 from HUCMSCs could be delivered into the trophoblast HTR-8/SVneo cells, thus enhancing proliferation and migration of trophoblasts. Mechanically, miR-101 targeted and negatively regulated BRD4 expression. BRD4 knockdown promoted the proliferation and migration of trophoblasts by suppressing NF-κB/CXCL11 axis. EV-encapsulated miR-101 from HUCMSCs also reduced blood pressure and 24 h urine protein in vivo, thereby ameliorating PE. Conclusion: In summary, EV-encapsulated miR-101 promoted proliferation and migration of placental trophoblasts through the inhibition of BRD4 expression via NF-κB/CXCL11 inactivation.

Association of placenta‐derived extracellular vesicles with pre‐eclampsia and associated hypercoagulability: a clinical observational study

“…Although anionic phospholipid‐expressing pcEVs accounted for less than 1% of total placental EVs, they were nearly three times higher in PE patients than in NP women, and predicted PE with higher confidence. These findings are consistent with recent reports that syncytiotrophoblast‐derived EVs contribute to the pathogenesis of PE by EV‐bound metalloprotease neprilysin, 38 which cleaves several peptides that regulate vascular tension, 39 and microRNA‐101, which regulates trophoblast proliferation 40 . A recent report further shows that the while pcEVs express both PLAP and syncytin‐1, levels of syncytin‐1 + pcEVs were significantly reduced in PE patients, as compared to NP women 41 .…”

Downregulated ribosomal protein L39 inhibits trophoblast cell migration and invasion by targeting E‐cadherin in the placenta of patients with preeclampsia

“…26 The abnormal migration and invasion of trophoblasts impairs the function of the placenta and is one of the main mechanisms underlying PE pathogenesis. 27,28 In the present study, the expression of RPL39 was decreased in placental tissues of early-onset PE patients. Moreover, RPL39 knockdown suppressed trophoblast cells proliferation, migration, and invasion by upregulating E-cadherin expression.…”

“…PE is a pregnancy‐related complication, and its etiology is primarily associated with pathological placentation 26 . The abnormal migration and invasion of trophoblasts impairs the function of the placenta and is one of the main mechanisms underlying PE pathogenesis 27,28 . In the present study, the expression of RPL39 was decreased in placental tissues of early‐onset PE patients.…”

Roles of noncoding RNAs in preeclampsia

“… [ 26 ] miR-195 placenta downregulated ActRIIB/ActRIIA miR-195 could promote cell invasion via directly targeting ActRIIB/ActRIIA in human trophoblast cells [ 27 ] miR-16 placenta/mesenchymal stem cell (MSC) upregulated CCNE1 /VEGF-A/Notch2 over-expressed miR-16 inhibited the proliferation and migration of decidua-derived mesenchymal stem cells /BeWo and JEG-3 cells, and induced cell-cycle arrest by targeting cyclin E1 [ 28 , 29 ] miRNA-376c placenta/plasma/exosome downregulated ALK5/ALK7/25-OH-VD miR-376c inhibits both ALK5 and ALK7 expression to impair transforming growth factor-β/Nodal signaling, leading to increases in cell proliferation and invasion [ 30 , 31 ] miR-29b decidua-derived mesenchymal stem cell (dMSC)/placenta upregulated MMP2/MCL1/ VEGFA/ ITGB1/HDAC4 miR-29b induced apoptosis and inhibited invasion and angiogenesis of trophoblast cells. [ 32 ] miR-101 placenta downregulated ERp44/BRD4/CXCL6 miR-101 could promote apoptosis and inhibite the proliferation and migration of trophoblasts [ 10 , 33 , 34 ] miR-18a placenta/plasma downregulated Smad2/ER1/ESRα miR-18a could promote trophoblast cell invasion and suppress apoptosis of human trophoblast cells [ 35 – 37 ] miR-20a placenta upregulated Foxa1/VEGF the upregulated miR-20a in human preeclampsia tissue can inhibit the proliferative and invasive activities of trophoblast cells [ 23 , 38 ] miR-125b-1-3p placenta upregulated S1PR1 miR-125b-1-3p inhibited the invasiveness of human...…”