Small RNA Analysis within Human Trophoblast-Derived Extracellular Vesicles and Effect of These Structures on the Transcriptome Profile of Human Neural Progenitor Cells [small RNAseq]
The fetal brain is thought to be dependent upon the placenta for factors that play a specific role in its early development. The tight linkage between the two organs has given rise to the term, the placenta-brain axis. The mouse placenta produces a unique signature pattern of miRNAs target transcripts associated with early fetal brain development, including ones regulating neurogenesis. We hypothesize that placenta-derived extracellular vesicles (EV) transport their molecular cargo, including small RNA, from the placenta to the brain, whereupon their contents can induce pleiotropic effects. We isolated EV from human trophoblast (TB) differentiated in vitro and the parental induced pluripotent stem cells (iPSC) from which the TB was derived, and small RNAseq was done to characterize small RNA profiles within these EV. Human neural progenitor cells (NPC) generated from the same iPSC were exposed to EV derived from TB or iPSC or only treated with control media. Results indicate EV derived from human TB cells contain unique profiles of miRNA and long non-coding RNA (lncRNA) relative to EV isolated from parental iPS cells. Human TB-derived EV readily internalized by human NPC. EV from TB cells induce transcripts in NPC associated with forebrain formation and neurogenesis, suggesting that such EV might program early fetal brain development. Results shed light on the inner workings of the placenta-brain axis. Understanding how contents of TB-derived EV, namely small RNA, affect NPC might yield new insights and potential treatment strategies for neurobehavioral disorders that originate in utero, such as autism spectrum disorders (ASD).  The fetal brain is thought to be dependent upon the placenta for factors that play a specific role in its early development. The tight linkage between the two organs has given rise to the term, the placenta-brain axis. The mouse placenta produces a unique signature pattern of miRNAs target transcripts associated with early fetal brain development, including ones regulating neurogenesis. We hypothesize that placenta-derived extracellular vesicles (EV) transport their molecular cargo, including small RNA, from the placenta to the brain, whereupon their contents can induce pleiotropic effects. We isolated EV from human trophoblast (TB) differentiated in vitro and the parental induced pluripotent stem cells (iPSC) from which the TB was derived, and small RNAseq was done to characterize small RNA profiles within these EV. Human neural progenitor cells (NPC) generated from the same iPSC were exposed to EV derived from TB or iPSC or only treated with control media. Results indicate EV derived from human TB cells contain unique profiles of miRNA and long non-coding RNA (lncRNA) relative to EV isolated from parental iPS cells. Human TB-derived EV readily internalized by human NPC. EV from TB cells induce transcripts in NPC associated with forebrain formation and neurogenesis, suggesting that such EV might program early fetal brain development. Results shed light on the inner workings of the placenta-brain axis. Understanding how contents of TB-derived EV, namely small RNA, affect NPC might yield new insights and potential treatment strategies for neurobehavioral disorders that originate in utero, such as autism spectrum disorders (ASD). Overall design: 3 Treatment groups of neural progenitor cells--one group which received EVs from day 6 iPS cells for 24 hours; one group which received EVs from day 6 TB cells for 24 hours; and one control group which did not receive any EVs for 24 hours; each group has four replicates