ETS-Related Gene Activation Preserves Adherens Junctions and Permeability in Microvascular Endothelial Cells

Abstract

ERG (ETS-related gene) is a member of the ETS (Erythroblast-transformation specific) family of transcription factors abundantly present in vascular endothelial cells. Recent studies demonstrate that ERG has important roles in blood vessel stability and angiogenesis. However, it is unclear how ERG is potentially involved in microvascular barrier functions and permeability. A wide variety of diseases and clinical conditions including trauma-hemorrhagic shock and burn injury are associated with microvascular dysfunctions, which causes excessive microvascular permeability, tissue edema and eventually, multiple organ dysfunction and death. The main purpose of this study was to determine the specific role of ERG in regulating microvascular permeability in human lung microvascular endothelial cells (HLMEC) and to evaluate if exogenous ERG will protect the barrier. The HLMECs were grown on Transwell inserts as monolayers and were transfected with ERG CRISPR/cas9 knockdown plasmid, ERG CRISPR activation plasmid, recombinant ERG protein or their respective controls. Recombinant vascular endothelial growth factor (VEGF) was used as an inducer of permeability for evaluating the effect of ERG activation on permeability. Changes in barrier integrity and permeability were studied using monolayer permeability assay and immunofluorescence of adherens junction proteins (VE-cadherin and b-catenin) respectively. CRISPR/cas9-based ERG knockdown as well as VEGF treatment induced monolayer hyperpermeability, VE-cadherin, and b-catenin junctional relocation and cytoskeletal F-actin stress fiber formation. CRISPR based ERG activation and recombinant ERG transfection attenuated VEGF-induced monolayer hyperpermeability. ERG activation preserved the adherens junctions and cytoskeleton. These results demonstrate that ERG is a potent regulator of barrier integrity and permeability in human lung microvascular endothelial cells and endogenously or exogenously enhancing ERG provides protection against barrier dysfunction and hyperpermeability.

Dynamics of Local Temperature in the Fingertips After the Cuff Occlusion Test: Infrared Diagnosis of Adaptation Reserves to Hypoxia and Assessment of Survivability of Victims at Massive Blood Loss

“…A probable mechanism for the disappearance of reactive local hyperthermia in the fingertips of the patients studied with blood loss may be the development of microvascular endothelial dysfunction [ 27 , 28 ]. Endothelial cells are known to regulate vasoreactivity and vascular permeability through the synthesis of nitric oxide, a powerful vasodilator [ 29 ].…”

Erythroblast transformation‐specific‐related gene promotes metastasis of oral squamous cell carcinoma by transcriptionally upregulating peroxiredoxin 1

“…4 Numerous studies have reported on ERG's regulatory roles in cell growth, proliferation, differentiation, and apoptosis. 5,6 Studies have confirmed the association between ERG expression and the development of different cancers. In prostate cancer cells, for instance, the fusion of the ERG and TMPRSS2 genes can facilitate cancer cell proliferation, invasion, angiogenesis, invasiveness, and activate downstream oncogenes, 7,8 thereby enhancing their transcriptional activity.…”

“…Erythroblast transformation‐specific‐related gene ( ERG ), a member of ETS transcription factor family, is a transcription factor that is widely expressed and can be induced by various growth and differentiation stimulators in many cell types 4 . Numerous studies have reported on ERG's regulatory roles in cell growth, proliferation, differentiation, and apoptosis 5,6 . Studies have confirmed the association between ERG expression and the development of different cancers.…”

Research Progress on Natural Plant Molecules in Regulating the Blood–Brain Barrier in Alzheimer’s Disease

“…Despite limited evidence regarding histone modification, DNA methylation, and histone deacetylase activity in BMECs, recent studies [90,91] have demonstrated that dysregulation of microRNAs and long non-coding RNAs disrupts the integrity of the BBB. Specifically, the dysregulation of miR124, miR-107, and LINC00662 alters the transcription of ZO-1, claudin-5, and occludin in BMECs of AD patients through the modification of members belonging to the erythroid transformation-specific (ETS) transcription factor family and ETS-related genes [92].…”