Abstract 13269: Effects of Nicotine-Containing Product Challenges on Interleukin-6 Pathway Expression in Human Pluripotent-Derived Arterial Endothelial Cells: The Clues Study

Abstract

Background: Electronic nicotine delivery devices (ENDs) induce interleukin-6 (IL-6) pathway expression in airway epithelial cells, but their systemic effects on arterial endothelial cells (AECs) are unknown. We hypothesized differential expression (DE) of IL-6 pathway genes by human pluripotent-derived AECs exposed in vitro to participant serum drawn before and after use of ENDS by chronic vapers, use of combustible cigarettes by smokers, and no product use by controls. Methods: For this pilot study, we randomly selected 30 participants from the CLUES Study: 10 exclusive vapers (exhaled carbon monoxide [CO] <5 ppm, + urine NicCheck I), 10 exclusive smokers (CO >5 ppm, + urine), and 10 non-vaping/non-smoking controls (CO <5 ppm, negative urine). We generated purified populations of AECs (H9 CPTC C13 derivative human embryonic stem cell line) and exposed them for 6 hours to 50% concentrated participant serum drawn before and 15’ after a product use challenge. Total RNA was sequenced on a NextSeq 2000. The DESeq2 suite characterized DE with an a priori focus on genes in the IL-6 biosynthetic pathway gene ontology set. Z scores of normalized log2 gene expression values were determined and expressed as heat maps (Figure). Gene Set Variational Analysis (GSVA) was used to interrogate for enriched gene sets. P values were not adjusted for multiple hypothesis testing and are exploratory. Results: Vapers were a mean (SD) 28.9 (8.1) years old (30% female), smokers were 38.3 (11.6) years old (40% female), and controls were 33.6 (17.7) years old (50% female). Compared to controls, vapers had increased expression of the IL-6 pathway at baseline (p=0.004), but smokers did not (p=0.38). Following product use, vapers (p=0.0009) but not smokers (p=0.79) had increased expression of genes in the IL-6 biosynthetic pathway (Figure). Conclusion: Chronic vapers, but not smokers, may have increased expression of genes in the IL-6 pathway prior to product use with further increases afterwards.