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JNK/SAPK signalling is essential for efficient reprogramming of human fibroblasts to induced pluripotent stem cells

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 Lako

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).


Abstract

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.


Publication metadata

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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