The Expression of the Cancer-Associated lncRNA Snhg15 Is Modulated by EphrinA5-Induced Signaling.

Daniel Pensold,Asa Walberg,Georg Pitschelatow,Kai Braunsteffer,Angelika Lampert,Julia Reichard,Amin Ravaei,Julia Gehrmann,Jenice Linde,Geraldine Zimmer-Bensch,Ivan G Costa

doi:10.3390/ijms22031332

Abstract

The Eph receptor tyrosine kinases and their respective ephrin-ligands are an important family of membrane receptors, being involved in developmental processes such as proliferation, migration, and in the formation of brain cancer such as glioma. Intracellular signaling pathways, which are activated by Eph receptor signaling, are well characterized. In contrast, it is unknown so far whether ephrins modulate the expression of lncRNAs, which would enable the transduction of environmental stimuli into our genome through a great gene regulatory spectrum. Applying a combination of functional in vitro assays, RNA sequencing, and qPCR analysis, we found that the proliferation and migration promoting stimulation of mouse cerebellar granule cells (CB) with ephrinA5 diminishes the expression of the cancer-related lncRNA Snhg15. In a human medulloblastoma cell line (DAOY) ephrinA5 stimulation similarly reduced SNHG15 expression. Computational analysis identified triple-helix-mediated DNA-binding sites of Snhg15 in promoters of genes found up-regulated upon ephrinA5 stimulation and known to be involved in tumorigenic processes. Our findings propose a crucial role of Snhg15 downstream of ephrinA5-induced signaling in regulating gene transcription in the nucleus. These findings could be potentially relevant for the regulation of tumorigenic processes in the context of glioma.

Highlights

Eph receptors represent the largest subfamily of receptor tyrosine kinases (RTKs), initially isolated from an erythropoietin-producing human hepatocellular carcinoma line [1]
We investigated whether ephrinA5 modulates the expression of long non-coding RNAs (lncRNAs) in immortalized cerebellar granule (CB) cells, which are often used as a medulloblastoma model based on the observation that medulloblastoma can arise from CB progenitor cells [24,25]
To test whether ephrinA5 triggers changes in lncRNA expression, we stimulated immortalized CB cells with recombinant ephrinA5-Fc, or Fc as control. Both were clustered with an Alexa488-labelled anti-human IgG antibody to achieve Eph receptor activation according to previous studies, and at the same time allowing for visualization of ephrinA5Fc-binding sites [2,3]

Summary

Introduction

Eph receptors represent the largest subfamily of receptor tyrosine kinases (RTKs), initially isolated from an erythropoietin-producing human hepatocellular carcinoma line [1]. By interacting with their cognate ligands, the ephrins (Eph receptor-interacting ligands), Eph receptors mediate important aspects of embryogenesis and the development of numerous tissues, including the brain [1]; such as proliferation, cell adhesion, axon guidance, cell migration, and others [1,2,3,4,5,6,7]. The ligands and receptors can promote and inhibit tumorigenicity depending on the downstream forward and reverse signaling generated To dissect their potential as therapeutic targets, downstream signaling, and effects on gene expression have to be better understood

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