Placental homing peptide guides HIF1α‑silenced exosomes conjugates for targeted enhancement of invasion of trophoblast cells

Abstract

There is a current lack of availability of therapeutics to treat Preeclampsia (PE), primarily due to the risk of harm to the fetus. Hypoxia-inducible factor-1α (HIF1α) is highly expressed in trophoblast cells and suppresses their invasive ability. Extensive studies have confirmed the positive effects of mesenchymal stem cell (MSC)-derived exosomes on PE. The aim of the present study was to develop a method for targeted delivery of HIF1α-silenced exosomes to the placenta. HIF1α was overexpressed in JEG-3 cells. Then, the glucose uptake, lactate production, proliferation and invasion of HIF1α-elevated JEG-3 cells were detected. Exosomal membrane protein lysosome-associated membrane glycoprotein 2b and placental homing peptide CCGKRK gene sequence amplified by PCR were conjugated using short hairpin RNA-HIF1α (sh-HIF1α) sequence (exo-pep-sh-HIF1α), which were then transfected into MSCs cultured in vitro . Exosomes were isolated from the supernatant of the aforementioned MSCs and identified by determining the size and exosomal markers. Finally, the invasion ability of MSCs-derived exosomes treated JEG-3 cells were detected using Transwell assays. HIF1α was demonstrated to remarkably promote the uptake of glucose and the production of lactate in JEG-3 cells. In addition, high levels of HIF1α facilitated the proliferation of JEG-3 cells, while suppressing their invasion ability. Bone marrow derived MSCs were cultured in vitro and exosomes were successfully isolated from these cells. Exo-pep-sh-HIF1α significantly reduced placental HIF1α expression, and induced significant enhancement of placental invasion. Overall, placental homing peptide-guided HIF1α-silenced exosomes effectively facilitated the invasion of placental trophoblasts, which could be used for the targeted delivery of payloads to the placenta and serve as a novel placenta-specific therapeutic approach.