Single cell 'omic profiles of human aortic endothelial cells in vitro and human atherosclerotic lesions ex vivo reveals heterogeneity of endothelial subtype and response to activating perturbations.

Autor:	Adelus ML; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA.; The Clinical Translational Sciences Graduate Program, The University of Arizona, Tucson, AZ, 85721, USA., Ding J; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Tran BT; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Conklin AC; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Golebiewski AK; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Stolze LK; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Whalen MB; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA., Cusanovich DA; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA.; Asthma and Airway Disease Research Center, The University of Arizona, Tucson, AZ, 85721, USA., Romanoski CE; The Department of Cellular and Molecular Medicine, The University of Arizona, Tucson, AZ 85721, USA.; The Clinical Translational Sciences Graduate Program, The University of Arizona, Tucson, AZ, 85721, USA.; Asthma and Airway Disease Research Center, The University of Arizona, Tucson, AZ, 85721, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Jan 17. Date of Electronic Publication: 2024 Jan 17.
DOI:	10.1101/2023.04.03.535495
Abstrakt:	Objective: Endothelial cells (ECs), macrophages, and vascular smooth muscle cells (VSMCs) are major cell types in atherosclerosis progression, and heterogeneity in EC sub-phenotypes are becoming increasingly appreciated. Still, studies quantifying EC heterogeneity across whole transcriptomes and epigenomes in both in vitro and in vivo models are lacking. Approach and Results: To create an in vitro dataset to study human EC heterogeneity, multiomic profiling concurrently measuring transcriptomes and accessible chromatin in the same single cells was performed on six distinct primary cultures of human aortic ECs (HAECs). To model pro-inflammatory and activating environments characteristic of the atherosclerotic microenvironment in vitro , HAECs from at least three donors were exposed to three distinct perturbations with their respective controls: transforming growth factor beta-2 (TGFB2), interleukin-1 beta (IL1B), and siRNA-mediated knock-down of the endothelial transcription factor ERG (siERG). To form a comprehensive in vivo/ex vivo dataset of human atherosclerotic cell types, meta-analysis of single cell transcriptomes across 17 human arterial specimens was performed. Two computational approaches quantitatively evaluated the similarity in molecular profiles between heterogeneous in vitro and in vivo cell profiles. HAEC cultures were reproducibly populated by 4 major clusters with distinct pathway enrichment profiles: EC1-angiogenic, EC2-proliferative, EC3-activated/mesenchymal-like, and EC4-mesenchymal. Exposure to siERG, IL1B or TGFB2 elicited mostly distinct transcriptional and accessible chromatin responses. EC1 and EC2, the most canonically 'healthy' EC populations, were affected predominantly by siERG; the activated cluster EC3 was most responsive to IL1B; and the mesenchymal population EC4 was most affected by TGFB2. Quantitative comparisons between in vitro and in vivo transcriptomes confirmed EC1 and EC2 as most canonically EC-like, and EC4 as most mesenchymal with minimal effects elicited by siERG and IL1B. Lastly, accessible chromatin regions unique to EC2 and EC4 were most enriched for coronary artery disease (CAD)-associated SNPs from GWAS, suggesting these cell phenotypes harbor CAD-modulating mechanisms. Conclusion: Primary EC cultures contain markedly heterogeneous cell subtypes defined by their molecular profiles. Surprisingly, the perturbations used here, which have been reported by others to be involved in the pathogenesis of atherosclerosis as well as induce endothelial-to-mesenchymal transition (EndMT), only modestly shifted cells between subpopulations, suggesting relatively stable molecular phenotypes in culture. Identifying consistently heterogeneous EC subpopulations between in vitro and in vivo models should pave the way for improving in vitro systems while enabling the mechanisms governing heterogeneous cell state decisions. Competing Interests: CONFLICT OF INTEREST STATEMENT The authors declare that there is no conflict of interest.
Databáze:	MEDLINE
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