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Lookup NU author(s): Dr Irina Neganova, Dr Evgeniya Shmeleva, Dr Jennifer Munkley, Dr Valeria Chichagova, Dr George Anyfantis, Rhys Anderson, Dr Joao Passos, Professor David Elliott, Professor Lyle Armstrong, Professor Majlinda LakoORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Reprogramming of somatic cells to the phenotypic state termed “induced pluripotency” is thought to occur through three consecutive stages: initiation, maturation and stabilisation. The initiation phase is stochastic but nevertheless very important as it sets the gene expression pattern that permits completion of reprogramming; hence a better understanding of this phase and how this is regulated may provide the molecular cues for improving the reprogramming process. JNK/SAPKs are stress activated MAPK kinases that play an essential role in several processes known to be important for successful completion of the initiation phase such as cellular proliferation, mesenchymal to epithelial transition and cell cycle regulation. In view of this we postulated that manipulation of this pathway would have significant impacts on reprogramming of human fibroblasts to induced pluripotent stem cells. Accordingly, we found that key components of the JNK/SAPK signalling pathway increase expression as early as day 3 of the reprogramming process and continue to rise in reprogrammed cells throughout the initiation and maturation stages. Using both chemical inhibitors and RNA interference of MKK4, MKK7 and JNK1, we tested the role of JNK/SAPK signalling during the initiation stage of neonatal and adult fibroblast reprogramming. These resulted in complete abrogation of fully reprogrammed colonies and the emergence of partially reprogrammed colonies which disaggregated and were lost from culture during the maturation stage. Inhibition of JNK/SAPK signalling results in reduced cell proliferation, disruption of mesenchymal to epithelial transition and loss of the pluripotent phenotype, which either singly or in combination prevent establishment of pluripotent colonies. Together these data provide new evidence for an indispensable role for JNK/SAPK signalling to overcome the well-established molecular barriers in human somatic cell induced reprogramming. This article is protected by copyright. All rights reserved.
Author(s): Neganova A, Shmeleva E, Munkley J, Chichagova V, Anyfantis G, Anderson R, Passos AJ, Elliott DJ, Armstrong L, Lako M
Publication type: Article
Publication status: Published
Journal: Stem Cells
Year: 2016
Volume: 34
Issue: 5
Pages: 1198-1212
Print publication date: 01/05/2016
Online publication date: 11/02/2016
Acceptance date: 12/01/2016
Date deposited: 08/03/2016
ISSN (print): 1066-5099
ISSN (electronic): 1549-4918
Publisher: AlphaMed Press
URL: http://dx.doi.org/10.1002/stem.2327
DOI: 10.1002/stem.2327
PubMed id: 26867034
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