Project description:Endothelial cells of blood vessel playing a key role in the metastasis of tumor cells particularly the brain microvascular endothelial cells in the brain-tropic metastasis of lung tumor cells. Currently, studies towards the influence contributed by tumor cells to the alteration of endothelial cells have been extensively conducted by contrast studies for the alteration of gene-expression signature of tumor cells under the influence of endothelial cells remains poorly characterized, thus in this study the transcriptome of the human small-cell lung cancer cell NCI-H209 was sequenced via RNA-Seq after conditioned by human brain microvascular endothelial cell (HBMEC) for characterizing the alteration of gene-expression signature.
Project description:The purpose is to obtain samples for mRNA analysis in primary human lung microvascular endothelial cells treated with universal interferon alpha beta or interferon gamma.
Project description:To evaluate the impact of mTOR activation by Tsc2KD in MVPC on the microvascular endothelial cell population we utilized single cell transcriptomic analysis of fractionated lung tissue. Mouse lung samples were pooled and cell sorting was 2 days and 10 weeks post tamoxifen induction. Following annotation of the original 28 lung subclusters, capillary MVEC were localized to 7 subclusters. Temporal comparison of the MVEC populations demonstrated that in WT the population was transcriptionally stable from 2 days to 10 weeks postinduction, as would be expected during angiostasis. In contrast lung MVEC from the mice with mTOR+ Tsc2KD MVPC demonstrated significant increases in gene expression at 10 weeks relative to day 2 post-induction. Increased gene expression was related to mesenchymal stem cell differentiation, microvascular endothelial differentiation, mTOR activation, autophagy and apoptosis. These detailed complementary analyses identify mechanistic consequences of Tsc2KD and mTOR dysregulation in MVPC.
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:We explored the mechanistic involvement of the growth arrest and DNA damageinducible gene, GADD45a, in LPS- and ventilator-induced inflammatory lung injury (VILI). Multiple biochemical and genomic parameters of inflammatory lung injury indicated GADD45a-/- mice to be modestly susceptible to intratracheal LPS-induced lung injury and profoundly susceptible to high tidal volume ventilation-induced lung injury (VILI) with increases in microvascular permeability and levels of inflammatory cytokines in bronchoalveolar lavage. Expression profiling of lung tissues from GADD45a-/- mice revealed strong dysregulation in the B cell receptor signaling pathway suggesting involvement of PI3 kinase/Akt signaling components while the wild type controls depicted no observable changes. Western blot analyses of lung homogenates confirmed ~50% reduction in Akt protein levels in GADD45a-/- mice accompanied by marked increases in Akt ubiquitination. Electrical resistance measurements across human lung endothelial cell monolayers with either reduced GADD45a or Akt expression (siRNAs) revealed significant potentiation of LPS-induced human lung endothelial barrier dysfunction which was attenuated by overexpression of a constitutively active Akt1 transgene. These studies validate GADD45a as a novel candidate gene in inflammatory lung injury and a significant participant in vascular barrier regulation via effects on Akt-mediated endothelial signaling
