PMID- 24586617 OWN - NLM STAT- MEDLINE DCOM- 20141014 LR - 20220310 IS - 1932-6203 (Electronic) IS - 1932-6203 (Linking) VI - 9 IP - 2 DP - 2014 TI - The redox function of APE1 is involved in the differentiation process of stem cells toward a neuronal cell fate. PG - e89232 LID - 10.1371/journal.pone.0089232 [doi] LID - e89232 AB - Low-to-moderate levels of reactive oxygen species (ROS) govern different steps of neurogenesis via molecular pathways that have been decrypted only partially. Although it has been postulated that redox-sensitive molecules are involved in neuronal differentiation, the molecular bases for this process have not been elucidated yet. The aim of this work was therefore to study the role played by the redox-sensitive, multifunctional protein APE1/Ref-1 (APE1) in the differentiation process of human adipose tissue-derived multipotent adult stem cells (hAT-MASC) and embryonic carcinoma stem cells (EC) towards a neuronal phenotype. METHODS AND RESULTS: Applying a definite protocol, hAT-MASC can adopt a neural fate. During this maturation process, differentiating cells significantly increase their intracellular Reactive Oxygen Species (ROS) levels and increase the APE1 nuclear fraction bound to chromatin. This latter event is paralleled by the increase of nuclear NF-kappaB, a transcription factor regulated by APE1 in a redox-dependent fashion. Importantly, the addition of the antioxidant N-acetyl cysteine (NAC) to the differentiation medium partially prevents the nuclear accumulation of APE1, increasing the neuronal differentiation of hAT-MASC. To investigate the involvement of APE1 in the differentiation process, we employed E3330, a specific inhibitor of the APE1 redox function. The addition of E3330, either to the neurogenic embryonic carcinoma cell line NT2-D1or to hAT-MASC, increases the differentiation of stem cells towards a neural phenotype, biasing the differentiation towards specific subtypes, such as dopaminergic cells. In conclusion, during the differentiation process of stem cells towards a neuroectodermic phenotype, APE1 is recruited, in a ROS-dependent manner, to the chromatin. This event is associated with an inhibitory effect of APE1 on neurogenesis that may be reversed by E3330. Therefore, E3330 may be employed both to boost neural differentiation and to bias the differentiation potential of stem cells towards specific neuronal subtypes. These findings provide a molecular basis for the redox-mediated hypothesis of neuronal differentiation program. FAU - Domenis, Rossana AU - Domenis R AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Bergamin, Natascha AU - Bergamin N AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Gianfranceschi, Giuseppe AU - Gianfranceschi G AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Vascotto, Carlo AU - Vascotto C AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Romanello, Milena AU - Romanello M AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Rigo, Silvia AU - Rigo S AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Vagnarelli, Giovanna AU - Vagnarelli G AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Faggiani, Massimo AU - Faggiani M AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Parodi, Piercamillo AU - Parodi P AD - Department of Experimental and Clinical Medical Sciences, University of Udine, Udine, Italy. FAU - Kelley, Mark R AU - Kelley MR AD - Department of Pediatrics (Section of Hematology/Oncology), Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, Indiana, United States of America. FAU - Beltrami, Carlo Alberto AU - Beltrami CA AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Cesselli, Daniela AU - Cesselli D AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Tell, Gianluca AU - Tell G AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. FAU - Beltrami, Antonio Paolo AU - Beltrami AP AD - Department of Medical and Biological Sciences, University of Udine, Udine, Italy. LA - eng GR - R01 CA167291/CA/NCI NIH HHS/United States GR - CA121168/CA/NCI NIH HHS/United States GR - CA167291/CA/NCI NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, Non-U.S. Gov't DEP - 20140219 PL - United States TA - PLoS One JT - PloS one JID - 101285081 RN - 0 (Benzoquinones) RN - 0 (Chromatin) RN - 0 (NF-kappa B) RN - 0 (Propionates) RN - 0 (Reactive Oxygen Species) RN - 136164-66-4 (E 3330) RN - EC 4.2.99.18 (APEX1 protein, human) RN - EC 4.2.99.18 (DNA-(Apurinic or Apyrimidinic Site) Lyase) SB - IM MH - Adipose Tissue/*cytology MH - Adult MH - Adult Stem Cells/metabolism/*physiology MH - Benzoquinones MH - Blotting, Western MH - Cell Differentiation/*physiology MH - Chromatin/metabolism MH - DNA-(Apurinic or Apyrimidinic Site) Lyase/*metabolism MH - Flow Cytometry MH - Humans MH - Microscopy, Fluorescence MH - Multipotent Stem Cells/metabolism/*physiology MH - NF-kappa B/metabolism MH - Neurogenesis/*physiology MH - Oxidation-Reduction MH - Propionates MH - Reactive Oxygen Species/metabolism MH - Real-Time Polymerase Chain Reaction MH - Reverse Transcriptase Polymerase Chain Reaction MH - Statistics, Nonparametric PMC - PMC3929656 COIS- Competing Interests: The authors have read the journal's policy and declare that they have no conflict of interest, with the exception of Dr. Mark R. Kelley, who is the Chief Scientific Founder and consultant for ApeX Therapeutics, a company that has licensed IP from his work. Dr. Beltrami is a PLoS ONE Editorial Board member, but this does not alter the authors' adherence to all the PLOS ONE policies on sharing data and materials. EDAT- 2014/03/04 06:00 MHDA- 2014/10/15 06:00 PMCR- 2014/02/19 CRDT- 2014/03/04 06:00 PHST- 2013/10/18 00:00 [received] PHST- 2014/01/16 00:00 [accepted] PHST- 2014/03/04 06:00 [entrez] PHST- 2014/03/04 06:00 [pubmed] PHST- 2014/10/15 06:00 [medline] PHST- 2014/02/19 00:00 [pmc-release] AID - PONE-D-13-42917 [pii] AID - 10.1371/journal.pone.0089232 [doi] PST - epublish SO - PLoS One. 2014 Feb 19;9(2):e89232. doi: 10.1371/journal.pone.0089232. eCollection 2014.