Investigation of TGFβ1-Induced Long Noncoding RNAs in Endothelial Cells.

Krishna K Singh,Pratiek N Matkar,Adrian Quan,Subodh Verma,Mohammed Al-Omran,Laura-Eve Mantella,Hwee Teoh

doi:10.1155/2016/2459687

Abstract

Objective. To evaluate the relationship between TGFβ signaling and endothelial lncRNA expression. Methods. Human umbilical vein endothelial cell (HUVECs) lncRNAs and mRNAs were profiled with the Arraystar Human lncRNA Expression Microarray V3.0 after 24 hours of exposure to TGFβ1 (10 ng/mL). Results. Of the 30,584 lncRNAs screened, 2,051 were significantly upregulated and 2,393 were appreciably downregulated (P < 0.05) in response to TGFβ. In the same HUVEC samples, 2,148 of the 26,106 mRNAs screened were upregulated and 1,290 were downregulated. Of these 2,051 differentially expressed upregulated lncRNAs, MALAT1, which is known to be induced by TGFβ in endothelial cells, was the most (~220-fold) upregulated lncRNA. Bioinformatics analyses indicated that the differentially expressed upregulated mRNAs are primarily enriched in hippo signaling, Wnt signaling, focal adhesion, neuroactive ligand-receptor interaction, and pathways in cancer. The most downregulated are notably involved in olfactory transduction, PI3-Akt signaling, Ras signaling, neuroactive ligand-receptor interaction, and apoptosis. Conclusions. This is the first lncRNA and mRNA transcriptome profile of TGFβ-mediated changes in human endothelial cells. These observations may reveal potential new targets of TGFβ in endothelial cells and novel therapeutic avenues for cardiovascular disease-associated endothelial dysfunction.

Highlights

Transforming growth factor-β (TGFβ) belongs to a large superfamily of linked proteins, comprising activins, bone morphogenetic proteins (BMPs), growth/differentiation factors, and anti-Mullerian hormone [1] that regulates proliferation, differentiation, migration, and survival in diverse cell populations depending on the cell type [2]
Box-andWhisker plots constructed to visualize the distribution of the fluorescent intensities revealed very similar normalized log 2 ratios for both long noncoding RNAs (lncRNAs) and mRNA and comparable quality of the array data, across the board (Supplementary Figure 1)
Scatter plots provided a profile of HUVEC lncRNAs (Figure 1(a)) and mRNAs (Figure 1(b)) that were upregulated, downregulated, or unaffected by exposure to TGFβ1 treatment

Summary

Introduction

Transforming growth factor-β (TGFβ) belongs to a large superfamily of linked proteins, comprising activins, bone morphogenetic proteins (BMPs), growth/differentiation factors, and anti-Mullerian hormone [1] that regulates proliferation, differentiation, migration, and survival in diverse cell populations depending on the cell type [2]. TGFβ1, TGFβ2, and TGFβ3 are the most common of the isoforms that are involved in these functions [3]. Prior to binding to its specific type I and type II serine/threonine kinase receptors, the latent form of TGFβ is activated by proteases or thrombospondin. It is well documented that TGFβ signaling involves one TGFβ type II receptor and two distinct TGFβ type I receptors, that is, the endothelium limited activin receptor-like kinase (ALK1) and the largely expressed ALK5. Activated ALK5 after ligand binding transduces signals from the membrane to the nucleus via phosphorylation of a specific subset of intracellular effectors termed Smads [3, 4].

Methods

Results

Conclusion