Abstract 19021: Metabolomic Characterization of Responses to Angiotensin Peptides in Human Brain Microvascular Endothelial Cells

Abstract

Introduction and hypothesis: Metabolomics offers a robust platform for dissecting the intricate molecular responses of biological models under distinct stimulation scenarios. The mechanism by which Angiotensin 1-7 modulates Angiotensin II signaling in endothelial cells is unclear and may involve events at the cellular membrane or downstream signaling pathways. We hypothesize that metabolomics can help us identify shared and distinct metabolic responses induced by these two molecules. Methods: hCMEC/D3 cells were cultured and then subjected to stimulation with Ang-II and Ang 1-7 during their logarithmic growth phase. Subsequent metabolomic profiling incorporated both RP-LC-QTOF-MS and GC-QTOF-MS analytical techniques. Our data analysis strategy was multilayered, implementing quality control alongside various multivariate statistical methods, such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and hierarchical clustering. Results: Our results underscore a discernible pattern of decreased and increased metabolite expression in response to angiotensin treatments, with noteworthy variations observed in the expression of glycerophospholipids, sterol lipids, and sphingolipids amongst the Ang-II, Ang 1-7, and control groups. Intriguingly, an "ON/OFF" pattern for numerous metabolites emerged across the experimental groups, hinting at shared biological activity provoked by both angiotensin treatments. The compounds Erucamide, Dihydroxy-chol-en-oic Acid, and LPE O-19:0;O significantly contributed to the discrimination between Ang-II and Ang 1-7 treatment groups when compared with controls. Conclusions: These findings offer a novel perspective on the distinct metabolic responses induced by Ang-II and Ang 1-7 treatments in hCMEC/D3 cells, and may shed light on the cellular mechanisms that underlie the impact of these angiotensins, and other vasoactive peptides, on the blood-brain barrier. Overall, our study reaffirms the value of metabolomics as a powerful tool for gaining new insights into the molecular effects of vasoactive peptides on endothelial cells.