Project description:Effect of TNF-alpha on microRNAs levels in Human Umbilical Endothelial Cells (HUVECs). HUVEC that were treated or not for 2 or 24 hours with TNF (10 ng/ml). Duplicate samples (1 or 2) of two different isolations of HUVEC (A or B)
Project description:To assess if HUVEC and HUAEC differ in suscptibility to inflammation and inflammation resolution, the cells were stimulated for 24 hrs with TNF followed by RNA isolation. Two additional groups were first stimulated with TNF (24 hr) followed by TNF removal (12 and 24 hrs). Expression profiles were compared to basal conditions, i.e. cells cultured in normal medium We used micro-arrays to assess if Gene-xpression profiles between HUVEC and HUAEC differ under inflammatory conditions and under conditions of inflammation resolution
Project description:HUVEC were left untreated or stimulated for 5h with 2 ng/ml TNF. Comparsion of the gene profiles revealed TNF-mediated gene expression changes in HUVEC. Keywords: parallel sample
Project description:Anti-TNF therapies are a core anti-inflammatory approach for chronic diseases such as rheumatoid arthritis and Crohn’s Disease. Previously, we and others found that TNF blocks the emergence and function of alternatively-activated or M2 macrophages involved in wound healing and tissue-reparative functions. Conceivably, anti-TNF drugs could mediate their protective effects in part by an altered balanced of macrophage activity. To understand the mechanistic basis of how TNF regulates tissue-reparative macrophages we used RNAseq, scRNAseq, ATACseq, time-resolved phospho-proteomics, gene-specific approaches, metabolic analysis and signaling pathway deconvolution. Our findings reveal that TNF controls tissue-reparative macrophage gene expression in a highly gene-specific way dependent on JNK signaling. We uncover principles of the selectively inhibition by TNF via the type 1 TNF receptor on specific populations of alternative activated macrophages.
Project description:Here, we have compared the gene expression repertoire of CD40L (CD154) and TNF stimulated HUVEC and report that unexpectedly, apart from a stronger response to TNF, no major qualitative differences could be observed. This applies for the period of up to six hours, a time where the alternative pathway has already been activated. Analysis of the early events after receptor engagement revealed that both TNF and CD40L activate the classical NF-kappaB pathway, and confirm activation of the alternative by the latter. Furthermore, using genetic and pharmacological inhibition of the classical pathway we show that activation of the alternative occurs independently of the former. This reveals novel insights into NF- kappaB signaling by CD40L and TNF in endothelial cells
Project description:To investigate sex differences in endothelial cell response to AngII, male and female human umbilical vein endothelial cells (HUVEC) were treated with AngII for 24 hours. RNA seq was run, and demonstrated that female HUVEC had elevated genes associated with oxidative stress and inflammatory responses compared to male HUVEC following AngII treatment.
Project description:HUVEC were left untreated or stimulated for 5h with 2 ng/ml TNF. Comparsion of the gene profiles revealed TNF-mediated gene expression changes in HUVEC.
Project description:The response of endothelial cells to diabetic-simulating stimuli is poorly understood. To evaluate the changes of enhancer activation status in HUVEC, genome-wide enrichment of H3K27ac (marker of enhancer and open chromatin) was examined in HUVECs treated with combined high glucose and TNF-alpha. HUVECs incubated in 25 mM mannitol were used as osmolarity control.