Research Paper Volume 9, Issue 12 pp 2559—2586

Central role of the p53 pathway in the noncoding-RNA response to oxidative stress

Paola Fuschi 1, , Matteo Carrara 1, , Christine Voellenkle 1, , Jose Manuel Garcia-Manteiga 2, , Paolo Righini 3, , Biagina Maimone 1, , Elena Sangalli 4, , Francesco Villa 4, , Claudia Specchia 4, 5, , Mario Picozza 6, , Giovanni Nano 3, 7, , Carlo Gaetano 8, , Gaia Spinetti 4, , Annibale A. Puca 4, 9, , Alessandra Magenta 6, , Fabio Martelli 1, ,

  • 1 Molecular Cardiology Laboratory, IRCCS Policlinico S. Donato, 20097, San Donato Milanese, Milan, Italy
  • 2 Center for Translational Genomics and BioInformatics, IRCCS San Raffaele Scientific Institute, 20132, Milan, Italy
  • 3 Operative Unit of Vascular Surgery I, IRCCS Policlinico S. Donato, 20097, San Donato Milanese, Milan, Italy
  • 4 Cardiovascular Research Unit, IRCCS MultiMedica, 20138, Milan, Italy
  • 5 Department of Molecular and Traslational Medicine, University of Brescia, 25123, Brescia, Italy
  • 6 Vascular Pathology Laboratory, Istituto Dermopatico dell’Immacolata-IRCCS, FLMM, 00167, Rome, Italy
  • 7 University of Milan, 20133, Milan, Italy
  • 8 Division of Cardiovascular Epigenetics, Department of Cardiology, Goethe University, Frankfurt am Main, 60596, Germany
  • 9 Department of Medicine and Surgery, University of Salerno, 84084, Salerno, Italy

received: October 12, 2017 ; accepted: December 1, 2017 ; published: December 12, 2017 ;

https://doi.org/10.18632/aging.101341
How to Cite

Copyright: Fuschi et al. This is an open‐access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Abstract

Oxidative stress plays a fundamental role in many conditions. Specifically, redox imbalance inhibits endothelial cell (EC) growth, inducing cell death and senescence. We used global transcriptome profiling to investigate the involvement of noncoding-RNAs in these phenotypes. By RNA-sequencing, transcriptome changes were analyzed in human ECs exposed to H2O2, highlighting a pivotal role of p53-signaling. Bioinformatic analysis and validation in p53-silenced ECs, identified several p53-targets among both mRNAs and long noncoding-RNAs (lncRNAs), including MALAT1 and NEAT1. Among microRNAs (miRNAs), miR-192-5p was the most induced by H2O2 treatment, in a p53-dependent manner. Down-modulated mRNA-targets of miR-192-5p were involved in cell cycle, DNA repair and stress response. Accordingly, miR-192-5p overexpression significantly decreased EC proliferation, inducing cell death. A central role of the p53-pathway was also confirmed by the analysis of differential exon usage: Upon H2O2 treatment, the expression of p53-dependent 5’-isoforms of MDM2 and PVT1 increased selectively. The transcriptomic alterations identified in H2O2-treated ECs were also observed in other physiological and pathological conditions where redox control plays a fundamental role, such as ECs undergoing replicative senescence, skeletal muscles of critical limb-ischemia patients and the peripheral-blood mononuclear cells of long-living individuals. Collectively, these findings indicate a prominent role of noncoding-RNAs in oxidative stress response.

Abbreviations

BrdU: bromodeoxyuridine; ChIP-seq: ChIP-sequencing; CLI: critical limb ischemia; EC: endothelial cell; FDR: False Discovery Rate; LLIs: long-living individuals; lncRNAs: long noncoding-RNAs; miRNAs: microRNAs; ncRNAs: noncoding-RNAs; PBMCs: peripheral blood mononuclear cells; SEM: standard error of the mean.