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Basic Science

Poster Session 4

(1137) Pregnancy hormones effects on coronary artery cellular gene regulation and functional correlates of SCAD

Wednesday, February 14, 2024
1:00 PM – 2:30 PM  
Location: Prince George's Exhibit Hall CDE, Lower Atrium Level
Objective: Spontaneous coronary artery dissection (SCAD) occurs during hormonal-mediated events such as pregnancy. The objective of this study was to determine the effect of pregnancy-level estradiol and progesterone on coronary artery smooth muscle cells (huCASMCs) and cellular behavior relevant to arterial destabilization such as in SCAD.

Study Design: Human CASMCs were propagated and incubated with either control media or 100nM estradiol plus 1000nM progesterone (pregnancy) for 48 hours. Experiments were performed in triplicate. To query transcriptional activation by hormone exposure, RNA sequencing (RNASeq) was analyzed to detect differentially expressed genes and processes. To validate transcriptomic findings on functional models, traction force microscopy (TFM) was performed on the hormone and control cells using a polydimethylsiloxane (PDMS)  layer with a predefined elasticity modulus and fluorescent nanoparticles with deflections measured to quantify force generation. A wound healing assay was performed to assess cellular migration. Bromodeoxyuridine (BrdU) incorporation was measured to determine cellular proliferation. Analyses were performed using DESeq2 in R, Friedman tests, and Kruskal-Wallis tests where appropriate.

Results: Over 100 transcripts were differentially expressed between the control and pregnancy conditions (Fig.1). Significant biologic processes identified by gene ontology analysis included cell proliferation (p=0.01), cell adhesion (p=0.02) and cell movement (p=0.01). TFM showed increased force in the pregnancy condition when induced with KCl (263±11 N/m2 versus 181±8 N/m2, p=0.01). Although the pregnancy cells were less proliferative (24 hours of BrdU incorporation, p=0.03), they showed increased migration in the wound healing assay (p=0.01).

Conclusion: Results from this in vitro study highlight candidate transcriptional and cellular behavior of the vascular smooth muscle induced by estradiol and progesterone that support specific mechanistic hypotheses for the regulation coronary artery smooth muscle cells that may have relevance to pregnancy-related cardiovascular events.