Project description:Transcriptional profiling of HMEC-1 cells comparing untreated control cells with TNF-alpha-treated cells. Goal was to include more evidence by adding another research method. Keywords: Inflammatory stimulation
Project description:CUT&RUN on Spen-degron TX1072 mESCs was performed in 2 biological replicates. Cells were treated with doxycycline (1ug/mL) for 0h, 4h, 8h, 24h and 8h doxycycline and auxin (500uM).
Project description:TNFa stimulated human cultured podocytes compared with unstimulated (vehicle control (VC)) podocytes. Incubation of podocytes with high concentrations of TNF alpha led to an overexpression of ICAM1 in particular. We studied this system as a comparator for organoids also stimulated with the same concentration of TNFa. Four conditions (6 replicates each) 1) treatment (5 ng/mL TNFa) for 24 hours 2) control (PBS) for 24h 3) treatment (5 ng/mL TNFa) for 48 hours 4) control (PBS) for 48h
Project description:TNFa stimulated human cultured podocytes compared with unstimulated (vehicle control (VC)) podocytes. Incubation of podocytes with high concentrations of TNF alpha led to an overexpression of ICAM1 in particular. We studied this system as a comparator for organoids also stimulated with the same concentration of TNFa. Four conditions (6 replicates each) 1) treatment (5 ng/mL TNFa) for 24 hours 2) control (PBS) for 24h 3) treatment (5 ng/mL TNFa) for 48 hours 4) control (PBS) for 48h
Project description:Glucocorticoids used in pharmacological doses for the treatment of a variety of medical conditions, and endogenous glucocorticoid excess – Cushing’s syndrome, may result in several adverse effects, but currently there is no clinically useful biomarker of glucocorticoid activity. A three-phase protein biomarker discovery strategy was used. Proteomic biomarker discovery and qualification was conducted on the secretome of ex vivo-stimulated peripheral blood mononuclear cells (PBMC) isolated from 6 volunteers, incubated ± dexamethasone 100 ng/mL for 4h and 24h. Untargeted proteomics with label-free quantification (LFQ) was conducted to discover candidate proteins which were quantified using targeted proteomics by a custom multiple reaction monitoring mass spectrometry (MRM-MS) assay. Five proteins were selected for serum measurement by immunoassay in 20 healthy volunteers, with blood drawn at baseline and 12h after 4 mg oral dexamethasone. Paired analysis of the discovery proteomics data (576 and 280 proteins for the 4h and 24h secretomes, respectively) generated a shortlist of candidates which were qualified using MRM-MS to obtain protein level intensity data for 39 proteins. In the validation cohort, 3/5 proteins were dexamethasone-responsive, two significantly decreased: lysozyme C (mean±SEM) – 101±5.5 vs 67±4.4 ng/mL, (P<0.0001); nucleophosmin-1 (median (interquartile range)) – 16.6 (14.4-18.4) vs 14.2 (11.1-17.4) ng/mL, (P<0.01), while high mobility group box 2 (mean±SEM) significantly increased – 819±34 vs 984±60 pg/mL (P<0.01). Using an ex vivo proteomic approach in PBMC, we have identified and verified circulating glucocorticoid-responsive proteins which may have potential as serum biomarkers.
Project description:HMEC cultures were left untreated or stimulated for 5h with 2 ng/ml TNF. Comparison of the gene expression profiles revealed the TNF-mediated gene expression changes. Keywords: parallel sample
Project description:Endothelial-mesenchymal-transition (EndMT) is an important source of cancer-associated fibroblasts (CAFs), which are known to facilitate tumor progression. We have previously shown that EndMT is present in pancreatic tumors and that deficiency of the Tie1 receptor induces EndMT in human endothelial cells. Pancreatic tumors are characterized by the presence of tumor necrosis factor-α (TNF-α). We now show that TNF-α strongly induces human endothelial cells to undergo EndMT. In order to know the secretory feature of cells which undergo EndMT by TNF-α, we conducted a comparative analysis of HMVEC secretome treated or not for 24h and 48h with TNF-α. Secretome study shows that cells treated with TNF-α have an important fibroblast-like secretory capacity, and a proinflamatory signature. Moreover, Ingenuity Pathway Analysis (IPA) shows that pathways implicated in migration, inflammation and fibrosis are predicted to be activated and that necrosis and apoptosis pathways are inhibited. Accordingly cell survival, viability and cycle progression are activated. We show that TNF-α- treated cells secrete proteins related to 16 protumoral pathways, confirming their fibroblastic characteristic. Finally, among the predicted upstream regulators activated, IPA analysis shows that, TNFSF12 and its receptor are present at hight levels in PDAC patients. Altogether these results show the fibroblastic characteristic of treated cells and demonstrate that TNF-α induces CAFs.