Project description:Using RNA-seq we report changes in gene expression, over a 24 hour time course, in human lung microvascular endothelial cells subjected to 10 Gy X-irradiation.
Project description:The vascular endothelium may play a role in the response to infectious agents and in the pathophysiology of disease processes resulting in capillary leak such as septic shock and acute respiratory distress syndrome. In order to study the effect of endotoxin on endothelial cell function, human lung microvascular endothelial cells in culture were exposed to lipopolysaccharide (LPS), 10 ng, for 4, 8, or 24 hours and changes in mRNA expression were studied using Affymetrix HG U133plus2 gene arrays. A principal components analysis revealed that LPS treatment was the primary source of variability in the data. LPS treatment of endothelial cells for 4, 8, or 24 hours resulted in the upregulation by two-fold or greater of 275, 260 and 141 genes respectively. LPS treatment resulted in the down regulation by 50% or greater of 176, 263 and 79 genes at 4, 8, or 24 hours respectively. Up regulated genes at 4 or 8 hours were enriched in those encoding for cytokines, secreted proteins, cell membrane proteins and proteins controlling signal transduction and transcriptional regulation. Down regulated genes at each of the time points included those coding for cell membrane proteins, transcriptional regulation and metabolism. At each time point, a significant proportion of the genes identified as changed were unique to that time point. Experiment Overall Design: In order to study the effect of endotoxin on endothelial cell function, human lung microvascular endothelial cells in culture were exposed to lipopolysaccharide (LPS), 10 ng, for 4, 8, or 24 hours and changes in mRNA expression were studied using Affymetrix HG U133plus2 gene arrays. Controls for each time point with no LPS exposure were also run. An N of 4 for each time point and treatment was used.
Project description:Human microvascular endothelial cells were stimulated to either proliferate or undergo tubulogenesis over an 8 hour time period. Keywords: Time Series
Project description:Our understanding on mechanisms of ischemia-reperfusion induced lung injury during lung preservation and transplantation is based on clinical observations and animal studies. Herein, we used cell and systems biology approaches to explore these mechanisms at transcriptomics levels, especially by focusing on the differences between human lung endothelial and epithelial cells, which are crucial for maintaining essential lung structure and function. Human pulmonary microvascular endothelial cells and human lung epithelial cells were cultured to confluent, subjected to different cold ischemic time (CIT) to mimic static cold storage with preservation solution, and then subjected to warm reperfusion with serum containing culture medium to similar lung transplantation. Cell morphology, viability and transcriptomic profiles were studied. Ischemia-reperfusion induced a CIT time-dependent cell death, which was associated with dramatic changes in gene expression. Under normal control conditions, endothelial cells showed gene clusters enriched in vascular process and inflammation, while epithelial cells showed gene clusters enriched in protein biosynthesis and metabolism. CIT 6 h alone or after reperfusion had little effects on these phenotypic characteristics. After CIT 18 h, protein biosynthesis related gene clusters disappeared in epithelial cells; after reperfusion, metabolism-related gene cluster in epithelial cells and multiple gene clusters in the endothelial cells also disappeared. Human pulmonary endothelial and epithelial cells have distinct phenotypic transcriptomic signatures. Severe cellular injury reduces these gene expression signatures in a cell type dependent manner. Therapeutics that preserve these transcriptomic signatures may represent new treatment to prevent acute lung injury during lung transplantation.
Project description:Tert-immortalized microvascular endothelial (TIME) cells are infected with human herpesvirus (HHV-8) or mocked infected to determine expression changes upon infection.
Project description:Here, we investigated the time-course changes in the pattern of microRNA (miRNA) expression of TNFα and IFNγ-stimulated and unstimulated hCMEC/D3 cells, an immortalized human cerebral microvascular endothelial cell line. In order to investigate pro-inflammatory cytokine-induced changes in miRNA levels in hCMEC/D3 cells, we challenged brain endothelial cells with TNFα and IFNγ (100 ng/ml) for 2 h, 6 h and 24 h and determined microRNA expression in cytokine-stimulated and unstimulated cells
Project description:We developed a novel substrate-selective inhibitor of p38 MAPK, UM101, and compared its effects on TNF-induced gene expression by human lung microvascular endothelial cells (HMVECLs) with the prototypical p38 catalytic inhibitor, SB203580
