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Homo sapiens

ABSTRACT: Transcriptome analysis of human pulmonary endothelial cells exposed to laminar shear stress

PROVIDER: PRJDB9991 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	DRR232680.fastq.gz	Fastqsanger.gz
	DRR232681.fastq.gz	Fastqsanger.gz
	DRR232682.fastq.gz	Fastqsanger.gz
	DRR232683.fastq.gz	Fastqsanger.gz
	DRR232684.fastq.gz	Fastqsanger.gz

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HUVECs cultured under normal or high laminar shear stress

Project description:Many studies have characterised the effect of normal laminar shear stress (LSS) on endothelial responses, however elevated shear stress, as would be experienced overlying a stenotic plaque, has not been studied in depth. Therefore we used transcriptomics and related functional analyses to compare cells exposed to laminar shear stress at 15 or 75 dynes/cm2 for 24 hours (LSS15-normal or LSS75-high shear stress). Human umbilical vein endothelial cells (HUVEC n=4 per flow condition from different batches of pooled donor HUVEC p2) were cultured for 24 hours on gelatin coated slides under laminar shear stress of either 15 dynes/cm2 or 75 dynes/cm2 (LSS15 or LSS75) to assess the effect of high shear stress on endothelial cells. Total RNA was obtained using Qiagen kit and array analysis performed by Service XS (Leiden, Netherlands).

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Project description:Atherosclerosis is an important pathological factor in the development of cardiovascular diseases. In addition to increased plasma lipid concentrations, irregular/oscillatory shear stress and inflammatory processes trigger pathophysiological changes. Inhibitors of the transcription modulatory bromo- and extra-terminal domain (BET) protein family (BETi) could offer a possible therapeutic approach due to their anti-inflammatory properties. In this study, the influence of laminar shear stress, inflammation and BETi on human endothelial cells in an atherosclerosis in vitro model was investigated using global protein expression profiling. For this purpose, human umbilical cord derived vascular endothelial cells (HUVEC) were treated with TNFα to mimic the inflammatory condition and were exposed to 24h laminar shear stress in the presence or absence of a BRD4 inhibitor, JQ1. Data-independent acquisition mass spectrometry (DIA-MS) alloqed us to quantify 3316 proteins for further statistical analysis. Differentially regulated proteins indicate a clear influence of inflammation and shear stress on human endothelial cells. Overall, application of JQ1 is led to significant changes in the proteome, including a strong anti-inflammatory response as well as a potentially negative impact on atherosclerosis formation. To our knowledge, this is the first proteomics study on HUVEC which investigates the influence of shear stress and BET inhibition in TNFα inflammatory endothelial cell culture model.

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