Downregulated miR-199a-3p in Preeclamptic Placenta-derived Exosomes from Cord Blood Hinders VEGF-induced Fetal Glomerular Dysplasia Through Inhibiting Akt1(S473) Phosphorylation via Targeting PHLPP2.

BACKGROUND

Preeclampsia is amultisystem disorder involving in inflammatory responses and metabolic dysfunction of maternal-fetal circulation. Recently, researchers found it threatens renal health of offspring in adulthood. Growing evidence indicated chronic kidney disease is associated with glomeruli deficiencies during intrauterine development. Our previous study showed placenta-derived exosomes from cord plasma with preeclampsia impede fetal glomerular vascularization, during which we postulate microRNAs may function as epigenetic switches for gene silencing of human glomerular endothelial cells. However, the specific miRNAs in placenta-derived exosomes engaged in glomerular vascularization remain unclear.

METHODS

Small RNA sequencing of placental-derived exosomes and bioinformatics analysis were applied to identify differentially expressed miRNAs, followed by real-time polymerase chain reaction for verification. Transient expression and inhibition of candidate miRNA were performed by transfection with chemically synthesized miRNA oligonucleotides. Functional assays of HGECs including cell proliferation assays, EDU assays, migration assays, tube formation assays and monolayer cell barrier permeability assays were performed after transfection. Further, dual luciferase assay was used to explore the target genes of candidate miRNA, followed by RT-qPCR, western blot and rescue assays. Antagomirs transfection of C57BL/6 J fetal mice via Amniotic cavity injection in vivo and C57BL/6 J fetal mice kidney explants culture in vitro were performed to evaluate number of glomeruli, renal development.

RESULTS

Preeclampsia downregulates miR-199a-3p in placenta-derived exosomes from cord plasma. Suppression of endogenous miR-199a-3p in HGECs inhibits angiogenesis, proliferation, migration and permeability. A dual luciferase assay and rescue assays confirmed that miR-199a-3p targets PH domain leucine-rich repeat-containing protein phosphatase 2, regulating the phosphorylation of Akt serine/threonine kinase 1 (S473). C57BL/6 J fetal mice with miR-199a-3p downregulation have low glomerulus counts and relative growth rate.

CONCLUSIONS

miR-199a-3p in placenta-derived exosomes from cord plasma controls VEGF-induced glomerular angiogenesis. Moreover, it provides a distinct perspective for the mechanisms underlying the increased risk for renal disease in the offspring of preeclampsia patients via placenta-derived exosomal miRNAs.