JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells

Irina Neganova, Evgenija Shmeleva, Jennifer Munkley, Valeria Chichagova, George Anyfantis, Rhys Anderson, Joao Passos, David J. Elliott, Lyle Armstrong, Majlinda Lako

Research output: Contribution to journalArticle

8 Citations (Scopus)

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. c-Jun N-terminal kinase (JNK)/stress-activated protein kinase (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 (MET) 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 signaling 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 signaling 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 signaling resulted in reduced cell proliferation, disruption of MET and loss of the pluripotent phenotype, which either singly or in combination prevented establishment of pluripotent colonies. Together these data provide new evidence for an indispensable role for JNK/SAPK signaling to overcome the well-established molecular barriers in human somatic cell induced reprogramming.

Original languageEnglish (US)
Pages (from-to)1198-1212
Number of pages15
JournalStem Cells
Volume34
Issue number5
DOIs
StatePublished - May 1 2016
Externally publishedYes

Fingerprint

Induced Pluripotent Stem Cells
JNK Mitogen-Activated Protein Kinases
Heat-Shock Proteins
Protein Kinases
Fibroblasts
Cell Proliferation
Epithelial-Mesenchymal Transition
Mitogen-Activated Protein Kinase Kinases
RNA Interference
Cues
Cell Cycle
Epithelial Cells
Phenotype
Gene Expression

Keywords

  • hESC
  • hiPSC
  • JNKs
  • MKK4
  • MKK7
  • SAPK

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this

Neganova, I., Shmeleva, E., Munkley, J., Chichagova, V., Anyfantis, G., Anderson, R., ... Lako, M. (2016). JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells. Stem Cells, 34(5), 1198-1212. https://doi.org/10.1002/stem.2327

JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells. / Neganova, Irina; Shmeleva, Evgenija; Munkley, Jennifer; Chichagova, Valeria; Anyfantis, George; Anderson, Rhys; Passos, Joao; Elliott, David J.; Armstrong, Lyle; Lako, Majlinda.

In: Stem Cells, Vol. 34, No. 5, 01.05.2016, p. 1198-1212.

Research output: Contribution to journalArticle

Neganova, I, Shmeleva, E, Munkley, J, Chichagova, V, Anyfantis, G, Anderson, R, Passos, J, Elliott, DJ, Armstrong, L & Lako, M 2016, 'JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells', Stem Cells, vol. 34, no. 5, pp. 1198-1212. https://doi.org/10.1002/stem.2327
Neganova I, Shmeleva E, Munkley J, Chichagova V, Anyfantis G, Anderson R et al. JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells. Stem Cells. 2016 May 1;34(5):1198-1212. https://doi.org/10.1002/stem.2327
Neganova, Irina ; Shmeleva, Evgenija ; Munkley, Jennifer ; Chichagova, Valeria ; Anyfantis, George ; Anderson, Rhys ; Passos, Joao ; Elliott, David J. ; Armstrong, Lyle ; Lako, Majlinda. / JNK/SAPK Signaling Is Essential for Efficient Reprogramming of Human Fibroblasts to Induced Pluripotent Stem Cells. In: Stem Cells. 2016 ; Vol. 34, No. 5. pp. 1198-1212.
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