RNA-seq identifies a role for the PPAR?/? inverse agonist GSK0660 in the regulation of TNF?-induced cytokine signaling in retinal endothelial cells.

ABSTRACT: The peroxisome proliferator-activated receptor beta/delta (PPAR?/?) is a transcription factor with roles in metabolism, angiogenesis, and inflammation. It has yet undefined roles in retinal inflammation and diabetic retinopathy (DR). We used RNA-seq to better understand the role of the antagonist and inverse agonist of PPAR?/?, GSK0660, in TNF?-induced inflammation. Understanding the underlying mechanisms of vascular inflammation could lead to new treatments for DR.RNA was isolated from human retinal microvascular endothelial cells treated with a vehicle, TNF?, or TNF? plus GSK0660. RNA-seq was performed with a 50 bp single read protocol. The differential expression was determined using edgeR and gene ontology, and a pathway analysis was performed using DAVID. RNA-seq validation was performed using qRT-PCR using the primers for ANGPTL4, CCL8, NOV, CXCL10, and PDPK1.TNF? differentially regulated 1,830 transcripts, many of which are involved in the cytokine-cytokine receptor interaction, chemokine signaling, and inflammatory response. Additionally, TNF? highly upregulated genes involved in leukocyte recruitment, including CCL5, CX3CL1, and CXCL10. GSK0660 differentially regulated 273 transcripts in TNF?-treated cells compared to TNF? alone. A pathway analysis revealed the enrichment of cytokine-cytokine receptor signaling. In particular, GSK0660 blocks the TNF?-induced upregulation of CCL8, a chemokine involved in leukocyte recruitment.TNF? regulates several genes related to retinal leukostasis in retinal endothelial cells. GSK0660 blocks the effect of TNF? on the expressions of cytokines involved in leukocyte recruitment, including CCL8, CCL17, and CXCL10 and it may therefore block TNF?-induced retinal leukostasis.

SUBMITTER: Savage SR 

PROVIDER: S-EPMC4443583 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature