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Proteomic alterations in response to Hypoxia Inducible Factor 2# in normoxic Neuroblastoma cells Flora Cimmino, Lucia Pezone, Marianna Avitabile, Luca Persano, Monica Vitale, Mauro Sassi, Silvia Bresolin, Valentina Serafin, Nicola Zambrano, Andrea Scaloni, Giuseppe Basso, Achille Iolascon, and Mario Capasso J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.6b00457 • Publication Date (Web): 05 Sep 2016 Downloaded from http://pubs.acs.org on September 6, 2016
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Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
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Journal of Proteome Research
Proteomic alterations in response to Hypoxia Inducible Factor 2α in normoxic Neuroblastoma cells
Flora Cimmino1,2,*, Lucia Pezone2-3, Marianna Avitabile1,2, Luca Persano4, Monica Vitale1,2, Mauro Sassi5, Silvia Bresolin6, Valentina Serafin6, Nicola Zambrano1,2, Andrea Scaloni5, Giuseppe Basso6, Achille Iolascon1,2, Mario Capasso1,2
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Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli “Federico II”, Naples, Italy
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CEINGE Biotecnologie Avanzate, Naples, Italy
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Scuola di Medicina e Chirurgia, Università degli Studi di Verona, Verona, Italy
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Istituto di Ricerca Pediatrica Città della Speranza - IRP, Padua, Italy
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Proteomics and Mass Spectrometry Laboratory, ISPAAM, National Research Council, Naples, Italy
6
Dipartimento di Salute della Donna e del Bambino, Università degli Studi di Padova, Padua, Italy
Keywords: Hypoxia inducible factor 2α, Neuroblastoma, Proteomics, Patient stratification
Corresponding Author: Cimmino Flora, PhD CEINGE – Biotecnologie Avanzate Via Gaetano Salvatore 486 80145 Napoli, Italy Email:
[email protected] Phone: +39 081 3737 73
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Summary Hypoxia Inducible Factor (HIF)-2α protein expression in solid tumors promotes stem-like phenotype in cancer stem cells and increases tumorigenic potential in non-stem cancer cells. Recently, we have shown HIF-1/2α gene expression is correlated to neuroblastoma (NB) poor survival and to undifferentiated tumor state; HIF-2α protein was demonstrated to enhance aggressive features of the disease. In this study, we used proteomic experiments on NB cells to investigate HIF-2α downstream-regulated proteins and/or pathways with the aim of providing novel therapeutic targets and/or bad prognosis markers. We verified that pathways mostly altered by HIF-2α perturbation are involved in tumor progression. In particular, HIF-2α induces alteration of central metabolism and splicing control pathways. Simultaneously, WNT, RAS/MAPK and PI3K/AKT activity and/or expression are affected and may impact on the sensitivity and the intensity of HIF-2α-regulated pathways. Furthermore, genes coding the identified HIF-2α-related markers built a signature able to stratify NB patients with unfavorable outcome. Taken together, our findings underline the relevance of dissecting the downstream effects of a poor survival marker in developing targeted therapy and improving patient stratification. Future prospective studies are needed to translate the use of these data into the clinical practice.
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Journal of Proteome Research
Introduction Hypoxia-inducible factors (HIF-1α, HIF-2α) are key mediators of the cellular response to low oxygen concentrations (hypoxia). In the absence of oxygen, they are stabilized against degradation and thereby coordinate hypoxia adaptation through the expression of several target genes involved in neo-angiogenesis, glucose transport and metabolism, which are required for tumor growth and metastasis formation 1. HIF-1α participates in the initial response to hypoxia being most active during short periods (2–24 h) of intense hypoxia or anoxia (
