A critical role for p38MAPK signalling pathway during reprogramming of human fibroblasts to iPSCs.
Reprogramming of somatic cells to induced pluripotent stem cells (iPSCs) holds enormous promise for regenerative medicine. Reprogramming is a stepwise process with well-defined stages of initiation, maturation and stabilisation which are critically dependent on interactions between key pluripotency transcription factors, epigenetic regulators and signalling pathways. In this manuscript we have investigated the role of p38 MAPK signalling pathway and have shown a subpopulation- and phase-specific pattern of activation occurring during the initiation and maturation stage of reprogramming in partially and fully reprogrammed cells respectively. Downregulation of p38 MAPK activity via RNA interference or small molecule inhibitor led to cell accumulation in G1 phase of the cell cycle and reduced expression of cell cycle regulators during the initiation stage of reprogramming. This was associated with a significant downregulation of key pluripotency marker expression, disruption of mesenchymal to epithelial transition (MET), increased expression of differentiation markers and presence of partially reprogrammed cells which retained a typical gene expression profile of mesendodermal cells and were unable to progress to fully reprogrammed phenotype. Together our data indicate an important role for p38 MAPK activity in proliferation, MET progression and establishment of pluripotent phenotype, which are necessary steps for the development of human iPSCs.
Institute of Genetic Medicine, Newcastle University, International Centre for Life, NE1 3BZ, UK.
Irina
I
Neganova
Institute of Genetic Medicine, Newcastle University, International Centre for Life, NE1 3BZ, UK.
Valeria
V
Chichagova
Institute of Genetic Medicine, Newcastle University, International Centre for Life, NE1 3BZ, UK.
Lyle
L
Armstrong
Institute of Genetic Medicine, Newcastle University, International Centre for Life, NE1 3BZ, UK.
Majlinda
M
Lako
D015415:Biomarkers; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D020935:MAP Kinase Signaling System / Q000187:drug effects; D047428:Protein Kinase Inhibitors / Q000494:pharmacology
D015415:Biomarkers; D047428:Protein Kinase Inhibitors