Maturation, not initiation, is the major roadblock during reprogramming toward pluripotency from human fibroblasts

Tanabe, Koji; Nakamura, Michiko; Narita, Megumi; Takahashi, Kazutoshi; Yamanaka, Shinya

doi:10.1073/pnas.1310291110

Cited by 119 publications

(137 citation statements)

References 26 publications

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“…Among these cells, KLF4 particularly plays important roles for the activation of HERVHs. Our data also revealed that NANOG as a replacer of KLF4 in iPSC generation can induce less HERV-H activity during reprogramming (11). Therefore, the reason why the significant difference of reprogramming activity between KLF4 and NANOG can be explained with our data.…”

Section: Discussionsupporting

confidence: 74%

“…To this end, we examined LTR7 activities during the course of iPSC generation. We sorted TRA-1-60-positive (+) reprogrammed cells on various days after retroviral transduction of OCT3/4, SOX2, KLF4, and myelocytomatosis oncogene (c-MYC) (subsequently referred to as OSKM) and analyzed their global gene expression by microarrays (11,12). Using principle component analyses (PCA) with the 144 DD-iPSC markers, we found similarities between DD-iPSC subclones and TRA-1-60 (+) intermediate reprogrammed cells (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Dynamic regulation of human endogenous retroviruses mediates factor-induced reprogramming and differentiation potential

Ohnuki

Tanabe

Sutou

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Significance In this study, we found that human endogenous retoriviruses type-H (HERV-Hs) are transiently hyperactivated during reprogramming toward induced pluripotent stem cells (iPSCs) and play important roles in this process. However, when reprogramming is complete and cells acquire full pluripotency, HERV-H activity should decrease to levels comparable with those in embryonic stem cells because failure to resilence this activity leads to the differentiation-defective phenotype in neural lineage. We also found that during reprogramming, reprogramming factors, including POU class 5 homeobox 1 (OCT3/4), sex determining region Y-box 2 (SOX2), and Krüppel-like factor 4 (KLF4) (OSK) bind to and activate long-terminal repeats of HERV-Hs. KLF4 possibly precludes Tripartite motif containing 28 and recruits not only OCT3/4 and SOX2, but also E1A binding protein p300 (p300) histone acethyltransferase on HERV-H loci. Therefore, OKSM-induced HERV-H activation constitutes an unanticipated and critical mechanism for iPSC formation.

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Section: Discussionsupporting

confidence: 74%

Section: Resultsmentioning

confidence: 99%

Dynamic regulation of human endogenous retroviruses mediates factor-induced reprogramming and differentiation potential

Ohnuki

Tanabe

Sutou

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

245

282

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“…Tanabe et al [72] have recently identified the maturation stage of iPS cell reprogramming as being a major bottleneck in the process, which is likely to account for the low efficiency of the process generally. They demonstrate that LIN28, but not NANOG, shp53 or CYCLIN D1, promotes maturation of iPS cells.…”

Section: Maturationmentioning

confidence: 99%

“…Only around 1% of cells that initiate reprogramming make it to the stabilisation stage [72] . This can be explained by the observation made by Golipour et al [81] that not all cells are "stabilisation competent".…”

Section: Stabilisationmentioning

confidence: 99%

Cell signalling pathways underlying induced pluripotent stem cell reprogramming

Hawkins

Joy

McKay

2014

WJSC

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Induced pluripotent stem (iPS) cells, somatic cells reprogrammed to the pluripotent state by forced expression of defined factors, represent a uniquely valuable resource for research and regenerative medicine. However, this methodology remains inefficient due to incomplete mechanistic understanding of the reprogramming process. In recent years, various groups have endeavoured to interrogate the cell signalling that governs the reprogramming process, including LIF/STAT3, BMP, PI3K, FGF2, Wnt, TGFβ and MAPK pathways, with the aim of increasing our understanding and identifying new mechanisms of improving safety, reproducibility and efficiency. This has led to a unified model of reprogramming that consists of 3 stages: initiation, maturation and stabilisation. Initiation of reprogramming occurs in almost all cells that receive the reprogramming transgenes; most commonly Oct4, Sox2, Klf4 and cMyc , and involves a phenotypic mesenchymal-to-epithelial transition. The initiation stage is also characterised by increased proliferation and a metabolic switch from oxidative phosphorylation to glycolysis. The maturation stage is considered the major bottleneck within the process, resulting in very few "stabilisation competent" cells progressing to the final stabilisation phase. To reach this stage in both mouse and human cells, pre-iPS cells must activate endogenous expression of the core circuitry of pluripotency, comprising Oct4, Sox2, and Nanog , and thus reach a state of transgene independence. By the stabilisation stage, iPS cells generally use the same signalling networks that govern pluripotency in embryonic stem cells. These pathways differ between mouse and human cells although recent work has demonstrated that this is context dependent. As iPS cell generation technologies move forward, tools are being developed to interrogate the process in more detail, thus allowing a greater understanding of this intriguing biological phenomenon.© 2014 Baishideng Publishing Group Inc. All rights reserved.Key words: Pluripotency; Reprogramming; Induced pluripotent stem; Cell signalling; Embryonic stem Core tip: Induced pluripotent stem (iPS) cells present great promise, both to research and to medicine. However, we know very little regarding the mechanisms that occur throughout the iPS cell reprogramming process and thus the process remains inefficient. In this review, we discuss the 3 stages of reprogramming, initiation, maturation and stabilisation, and clarify the signalling pathways underlying each phase. We draw together the current knowledge to propose a model for the interactions between the key pathways in iPS cell reprogramming with the aim of illuminating this complex yet fascinating process.Hawkins K, Joy S, McKay T. Cell signalling pathways underlying induced pluripotent stem cell reprogramming. World J Stem Cells

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“…Mouse intermediate reprogrammed cells can be labelled by stagespecific embryonic antigen (SSEA)-1 that is recognized as a specific marker of mouse embryonic stem cells (ESCs)/iPSCs [5][6][7] . We demonstrated that human cells positive ( þ ) for tumorrelated antigen (TRA)-1-60 8,9 induced by OSKM are intermediate reprogrammed cells to being iPSCs 10 . Behaviour of TRA-1-60 ( þ ) intermediate reprogrammed cells suggested that maturation, but not the initiation step, is a bottleneck of cell fate conversion towards human iPSCs.…”

mentioning

confidence: 99%

Induction of pluripotency in human somatic cells via a transient state resembling primitive streak-like mesendoderm

et al. 2014

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During mammalian embryonic development, the primitive streak initiates the differentiation of pluripotent epiblast cells into germ layers. Pluripotency can be reacquired in committed somatic cells using a combination of a handful of transcription factors, such as OCT3/4, SOX2, KLF4 and c-MYC (hereafter referred to as OSKM), albeit with low efficiency. Here we show that during OSKM-induced reprogramming towards pluripotency in human cells, intermediate cells transiently show gene expression profiles resembling mesendoderm, which is a major component of the primitive streak. Based on these findings, we discover that forkhead box H1 (FOXH1), a transcription factor required for anterior primitive streak specification during early development, significantly enhances the reprogramming efficiency of human fibroblasts by promoting their maturation, including mesenchymal to epithelial transition and the activation of late pluripotency markers. These results demonstrate that during the reprogramming process, human somatic cells go through a transient state that resembles mesendoderm.

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Maturation, not initiation, is the major roadblock during reprogramming toward pluripotency from human fibroblasts

Cited by 119 publications

References 26 publications

Dynamic regulation of human endogenous retroviruses mediates factor-induced reprogramming and differentiation potential

Dynamic regulation of human endogenous retroviruses mediates factor-induced reprogramming and differentiation potential

Cell signalling pathways underlying induced pluripotent stem cell reprogramming

Induction of pluripotency in human somatic cells via a transient state resembling primitive streak-like mesendoderm

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