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Potentiation of TNF‐induced inflammatory transcriptional regulation by SFK activation (278.3)
Author(s) -
Adam Alejandro,
Lu Yao Wei,
Alsaffar Hiba,
Martino Nina,
Lowery Anthony,
Schwarz John,
Vincent Peter
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.278.3
Subject(s) - tumor necrosis factor alpha , lymphotoxin , chemistry , microbiology and biotechnology , messenger rna , transcriptional regulation , kinase , biology , gene expression , endocrinology , gene , biochemistry
TNF induces dramatic changes in the transcriptional profile of endothelial cells to promote a pro‐inflammatory phenotype that allows leukocyte transendothelial migration and loss of barrier function. We have previously shown that treatment with 50 pg/ml TNF increases endothelial permeability after 4‐6 h in monolayers of human dermal microvascular endothelial cells expressing DN‐Csk to activate endogenous Src Family Kinases (SFKs) but not in control (LacZ) monolayers. We thus reasoned that this delay in increasing permeability could be due to a requirement in transcriptional changes. Here, we demonstrate that activation of SFKs potentiates the transcriptional effects induced by TNF. After 6 h, TNF elevated ICAM1, E‐Selectin, HSPA1A/B and complement C3 mRNA levels 41‐, 76‐, 12‐ and 24‐fold respectively. In the presence of DN‐Csk, the same treatment induced these genes 149‐, 310‐, 38‐ and 202‐fold, respectively. Similar effects were observed in the mRNA of TNF itself (46 vs 193 fold) and lymphotoxin A (5 vs 21 fold), but not lymphotoxin B (86 vs 83 fold). ATF3 was induced 7 and 20‐fold respectively. EGR2 mRNA was induced 5‐fold by DN‐Csk and 9‐fold by the dual treatment, while Fos mRNA was induced 4‐fold in DN‐Csk‐expressing cells after 6h TNF and 9‐fold after 18 h. While these treatments did not lead to changes in the expression of many junctional genes, ZO‐1 and VE‐cadherin localization was drastically altered as determined by immunofluorescence. A pro‐inflammatory transcriptional profile, together with local posttranslational changes at the cell‐cell junctions, could explain the synergistic loss of barrier function observed with dual TNF signaling and endogenous SFK activation. Grant Funding Source : Supported by an AHA SDG grant 13SDG17100110 to APA.