ABSTRACT
Pregnant and lactating people remain therapeutic orphans as they are excluded from the vast majority of
clinical drug development and therapeutic trials. Moreover, current practices in drug evaluation in pregnancy
have been hindered by the lack of effective biomarkers and innovative study designs. There is a need to
develop novel placental-specific biomarkers in order to assess placental function and response to therapeutics,
as a way to inform on their safety and efficacy. An example of these novel biomarkers is placental (fetal)
specific extracellular vesicles (EVs). Recent advances in characterizing the cargo content of these EVs
demonstrated their potential to be used as placental biomarkers. We have shown using funding support from
the MPRINT that fourteen proteins found in EVs significantly correlated with aspirin use (FDR<0.1) in at-risk
people, but that more power is needed to confidently assess the relationship with aspirin dosage and
pregnancy outcomes. In addition, prior studies showed that aspirin affects endothelial and trophoblast cells,
thus potentially modulating exosome derived from these cells and their cargo contents. Leveraging our prior
proof of principle success, previously collected, and ongoing collection of maternal plasma from people at-risk
of preeclampsia (PE) receiving 81mg or 162mg of aspirin daily and low-risk people at the Ohio State
University, we are submitting this application with the overarching goal to develop a novel platform to augment
the MPRINT resources using exosome profiling, as novel biomarkers, to monitor placental mediated adverse
pregnancy outcomes and response to therapeutics. For this, we will use PE as a hallmark of these outcomes
and aspirin as the therapeutic agent to show case the utility of the platform. In this study, we will test the
hypothesis that differential expression of EVs proteome cargo is associated with placental and pregnancy
health in response to aspirin treatment at different gestational periods. We will test the following specific aims:
1) Validate our preliminary proteome data and model using larger cohort from additional biobanked samples of
at-risk people receiving 81mg or 162 mg aspirin, characterize fetal specific EVs (placental alkaline
phosphatase [PLAP] +ve) isolated from maternal plasma, and determine the differences in between the
maternal vs. fetal EV cargo difference with aspirin treatment; and 2) Determine the changes associated with
EV proteome profile (maternal and fetal) in at-risk people receiving aspirin prospectively and correlate with
clinical outcomes (development of PE) and angiogenic and inflammatory biomarkers associated with PE. This
project has the potential to play a seminal role in the development of a novel platform for therapeutics research
in pregnancy. This translational research platform will substantially add to the MPRINT Network repertoire and
be available for scientific community through the already established relationship of the team with other
networks such as the Maternal Fetal Medicine Units Network, MPRINT, and the Foundation of the NIH.
