Extracellular vesicles alter trophoblast function in pregnancies complicated by COVID-19

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and resulting coronavirus disease (COVID-19) causes placental dysfunction, which increases the risk of adverse pregnancy outcomes. While abnormal placental pathology resulting from COVID-19 is common, direct infection of the placenta is rare. This suggests that pathophysiology associated with maternal COVID-19, rather than direct placental infection, is responsible for placental dysfunction and alteration of the placental transcriptome. We hypothesized that maternal circulating extracellular vesicles (EVs), altered by COVID-19 during pregnancy, contribute to placental dysfunction. To examine this hypothesis, we characterized maternal circulating EVs from pregnancies complicated by COVID-19 and tested their effects on trophoblast cell physiology in vitro. We found that the gestational timing of COVID-19 is a major determinant of circulating EV function and cargo. In vitro trophoblast exposure to EVs isolated from patients with an active infection at the time of delivery, but not EVs isolated from Controls, altered key trophoblast functions including hormone production and invasion. Thus, circulating EVs from participants with an active infection, both symptomatic and asymptomatic cases, can disrupt vital trophoblast functions. EV cargo differed between participants with COVID-19 and Controls, which may contribute to the disruption of the placental transcriptome and morphology. Our findings show that COVID-19 can have effects throughout pregnancy on circulating EVs and circulating EVs are likely to participate in placental dysfunction induced by COVID-19. Overall design: There are 3 sets of RNAseq; 1) placenta biopsy RNAseq, 2) BeWo cell RNAseq, 3) extracellular vesicle (EV) RNAseq. The RNA-related methods for placenta and Bewo cell RNAseq are the same. The RNA-related methods for EV RNAseq are different. The transcriptome was analyzed to measure the effect of COVID-19 during pregnancy on the placenta and circulating EVs. Placenta biopsies were collected at delivery from controls (no known SARS-CoV-2 infection (C), resolved infection in the first (R1), second (R2), and third (R3) trimester, and active infection (AI) at the time of delivery. RNA sequencing was also performed to analyze the transcriptome of BeWo cells exposed in vitro to plasma EVs isolated from subjects with an active infection (BCD) or controls (BCC). Finally, the transcriptome of plasma-derived EVs from controls (C), resolved infection in the first (R1), second (R2), and third (R3) trimester, and active infection (AI) at the time of delivery.