Direct cell reprogramming is a stochastic process amenable to acceleration

By J. Hanna, Krishanu Saha, Bernardo F. Pando, Jeroen S. van Zon, C. Lengner, M. Creyghton, Alexander van Oudenaarden, R. Jaenisch. 2009

1,070 citations recorded by the source corpus.

Direct reprogramming of somatic cells into induced pluripotent stem (iPS) cells can be achieved by overexpression of Oct4, Sox2, Klf4 and c-Myc transcription factors, but only a minority of donor somatic cells can be reprogrammed to pluripotency. Here we demonstrate that reprogramming by these transcription factors is a continuous stochastic process where almost all mouse donor cells eventually give rise to iPS cells on continued growth and transcription factor expression. Additional inhibition of the p53/p21 pathway or overexpression of Lin28 increased the cell division rate and resulted in an accelerated kinetics of iPS cell formation that was directly proportional to the increase in cell proliferation. In contrast, Nanog overexpression accelerated reprogramming in a predominantly cell-division-rate-independent manner. Quantitative analyses define distinct cell-division-rate-dependent and -independent modes for accelerating the stochastic course of reprogramming, and suggest that the number of cell divisions is a key parameter driving epigenetic reprogramming to pluripotency.

Subfields: Molecular Biology

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In Molecular Biology

1. K. Hochedlinger
2. S. Yamanaka
3. M. Stadtfeld
4. R. Jaenisch
5. Kathrin Plath
6. G. Daley
7. K. Zaret
8. Helen M. Blau

Citations in the corpus

• Induced pluripotency: history, mechanisms, and applications. (2010, background): “These data suggest, for example, that Oct4, Nanog, Sox2, and Klf4 are part of a core pluripotency network that serves two main purposes: the repression of genes associated with differentiation, and the activation of ESC-specific targets (Boyer et al. 2005; Loh et al. 2006).”
• Cellular Senescence: Aging, Cancer, and Injury. (2019, background): “LIN28 expression improves reprogramming efficiency in mouse fibroblasts, whereas NANOG expression only increases reprogramming kinetics (148).”
• Senescence impairs successful reprogramming to pluripotent stem cells. (2009, background): “reprogramming kinetics rather than the overall efficiency of reprogramming (Hanna et al., 2009).”
• Incomplete DNA methylation underlies a transcriptional memory of the somatic cell in human iPS cells (2011, neutral): “The proximity of multiple genes being repressed may synergize in recruiting the silencing machinery, whereas silencing may be inefficient or delayed in more isolated regions, where stochastic events thought to underlie the reprogramming proces”
• Single-cell expression analyses during cellular reprogramming reveal an early stochastic and a late hierarchic phase. (2012, background): “Given these data, a stochastic model has emerged to explain how forced expression of the transcription factors initiates the process that eventually leads to the pluripotent state in only a small fraction of the transduced cells (Hanna et al., 2009; Yamanaka, 2009).”
• Dedifferentiation, transdifferentiation and reprogramming: three routes to regeneration (2011, background): “For instance, it takes a substantial period of time to induce pluripotency and, within any given population, individual cells will become pluripotent at different times despite expressing similar levels of exogenous gene”
• Nuclear reprogramming to a pluripotent state by three approaches (2010, background): “On the basis of these findings, many researchers now think that reprogramming during iPS-cell generation entails stochastic (that is, random) event”
• Conversion of mouse fibroblasts into cardiomyocytes using a direct reprogramming strategy (2011, background): “and activator of transcription), PI(3)K (phosphatidylinositol-3-kinase) or Wnt signalling during the early stages of the protocol might boost the effectiveness of cardiomyogenesis by preventing even a small number of iPSC intermediates from forming.”
• Highly efficient reprogramming to pluripotency and directed differentiation of human cells using synthetic modified mRNA (2010, methodology): “…employed a five-factor cocktail including a modified RNA encoding LIN28 (KMOSL), which has been shown to facilitate reprogramming (Yu et al., 2007; Hanna et al., 2009), and we supplemented the media with valproic acid (VPA), a histone deacetylase inhibitor that has been reported to increase…”
• Identification and classification of chromosomal aberrations in human induced pluripotent stem cells. (2010, result): “This is congruent with recent findings that cell division rate is a crucial parameter in the success and efficiency of the reprogramming process (Hanna et al., 2009).”