Chromosomal and telomeric reprogramming following ES-somatic cell fusion

Sümer, Hüseyin; Nicholls, Craig; Pinto, Alexander R.; Indraharan, Dinesh; Li, Jun; Lim, Mei Ling; Liu, Junping; Verma, Paul J.

doi:10.1007/s00412-009-0245-1

Cited by 14 publications

(9 citation statements)

References 38 publications

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“…This is due to the repression of telomerase during differentiation which coincides with telomere shortening, which can ultimately lead to replicative senescence (Osterhage and Friedman, 2009). It has previously been shown that telomerase activity and telomere lengthening occurs following reprogramming by SCNT (Tian et al, 2000;Wakayama et al, 2000), the induction of pluripotency by transcription factors (Marion et al, 2009;Takahashi and Yamanaka, 2006) and ES-somatic cell fusion (Cowan et al, 2005;Sumer et al, 2010b). In the present study we report that the length of telomeres and levels of telomerase activity appeared to be restored to pluripotent stem cell levels in both ES-somatic and iPS-somatic cell hybrids.…”

Section: Rt-pcrsupporting

confidence: 63%

“…2A). Furthermore, the cell cycle profiles of the two hybrid lines were shown to be similar to that of ES and iPS ous data (Sumer et al, 2010b), increased telomere fluorescence intensity was observed in ES cells compared to MEFs, with ES cell telomeres an average of 2.3 times longer than the MEF cell line telomeres. Similarly the induction of pluripotency of the QS MEFs resulted in iPS cells with a mean telomere fluorescence 2.5 fold longer following reprogramming (Fig.…”

Section: Characterization Of Cell Hybridsmentioning

confidence: 59%

“…It has previously been shown that both mouse iPS (Maherali et al, 2007;Sumer et al, 2010a) and SCNT derived ntES cells somatic cell hybrids is high and similar to those of ES cells (Cowan et al, 2005;Sumer et al, 2010b), and may be the underlying mechanism for the lengthening of the somatic cell derived chromosomes telomeres in hybrids (Sumer et al, 2010b). Telomere repeat amplification protocol (TRAP) assay was conducted on cell lysates from equal number of cells in order to determine the levels of telomerase activity of the various cell lines (Fig.…”

Section: Discussionmentioning

confidence: 99%

“…The resulting ES-somatic cell hybrids have most properties of pluripotency in vitro, such as gene expression profiles and epigenetic regulation, and are able to differentiate into cell types of the three germ lineages in teratoma assays when injected into immuno-compromised mice (Cowan et al, 2005;Tada et al, 2001;Yu et al, 2006). Furthermore, telomerase activity in ES-somatic hybrids is observed to be high and similar to those of ES cells (Hanna et al, 2007), resulting in the lengthening of the somatic cell derived chromosomes telomeres (Sumer et al, 2010b).…”

Section: Introductionmentioning

confidence: 97%

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Comparison of reprogramming ability of mouse ES and iPS cells measured by somatic cell fusion

Sümer¹,

Nicholls²,

Li³

et al. 2010

Int. J. Dev. Biol.

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The ectopic expression of the key transcription factors Oct4, Sox2, c-Myc, and Klf-4 have been shown to reprogram somatic cells to a pluripotent state. In turn these induced pluripotent stem (iPS) cells, like embryonic stem (ES) cells, have been shown to be able to reprogram somatic cells by cell fusion. In this study we compare the differences and similarities between ES and iPS cells measured by somatic cell fusion to somatic cells harboring an Oct4-GFP transgene. We found that iPS cells were just as potent as ES cells at reprogramming the somatic genome as measured by Oct4-GFP reactivation. The resulting ES-somatic and iPS-somatic cell hybrids were characterized for expression of key pluripotency genes, immunostaining for Oct4, SSEA-1, and the ability to differentiate into cell types representative of the three germ layers. In addition to restoring pluripotency to the somatic genome following cell fusion, the telomere maintenance mechanisms of both the ES and iPS cells were found to be dominant in the resulting ES-somatic and iPS-somatic cell hybrids, resulting in the lengthening of the somatic telomeres following cellular reprogramming. Therefore this study supports the view that iPS cells can be virtually indistinguishable from ES cells, even with regard to their reprogramming ability.

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Section: Rt-pcrsupporting

confidence: 63%

Section: Characterization Of Cell Hybridsmentioning

confidence: 59%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 97%

See 2 more Smart Citations

Comparison of reprogramming ability of mouse ES and iPS cells measured by somatic cell fusion

Sümer¹,

Nicholls²,

Li³

et al. 2010

Int. J. Dev. Biol.

Self Cite

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show abstract

“…Telomere repeat amplification protocol assays and telomere length analysis were performed as previously described (44) with minor modifications (45,46).…”

Section: Methodsmentioning

confidence: 99%

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induces cancer cell senescence by interacting with telomerase RNA component

Nicholls

Pinto

et al. 2012

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

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Oxidative stress regulates telomere homeostasis and cellular aging by unclear mechanisms. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) is a key mediator of many oxidative stress responses, involving GAPDH nuclear translocation and induction of cell death. We report here that GAPDH interacts with the telomerase RNA component (TERC), inhibits telomerase activity, and induces telomere shortening and breast cancer cell senescence. The Rossmann fold containing NAD + binding region on GAPDH is responsible for the interaction with TERC, whereas a lysine residue in the GAPDH catalytic domain is required for inhibiting telomerase activity and disrupting telomere maintenance. Furthermore, the GAPDH substrate glyceraldehyde-3-phosphate (G3P) and the nitric oxide donor S-nitrosoglutathione (GSNO) both negatively regulate GAPDH inhibition of telomerase activity. Thus, we demonstrate that GAPDH is regulated to target the telomerase complex, resulting in an arrest of telomere maintenance and cancer cell proliferation. and apoptosis. Although production of daughter cells costs energy, it remains largely elusive how energy metabolism regulates normal cell aging and lifespan. Considerable evidence suggests that metabolic waste causes premature senescence and apoptosis, shortening cellular lifespan. Accumulation of reactive oxygen species (ROS) damages cellular enzymes, organelle membranes, and chromosomal DNA including telomeres (the ends of chromosomes) (1, 2). Whereas enzymes that reduce the production of metabolic hazards may operate to extend cellular lifespan, recent studies demonstrate that the energy producing enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) that limits ROS production undergoes structural modifications and travel into the nucleus (3-7) to interact with telomeres directly and to potentially alleviate stress-induced cell aging (8, 9).Human telomeres are composed of arrays of a few thousand tandem DNA repeats of TTAGGG and various binding proteins. Telomeres undergo progressive shortening as somatic cells divide, due to the inability of cells to replicate the extreme ends of chromosomes. When telomeres are critically short, cells exit the cell cycle and undergo cell senescence (10-13). As a measure to maintain telomeres for continuous renewal of germ lines, development of embryonic stem cells, clonal expansion of lymphocytes, and immortalization of neoplastic cells, telomerase is activated to synthesize and maintain telomeres by reverse transcription (10-13).Telomerase is a large ribonucleoprotein complex containing the catalytic subunit telomerase reverse transcriptase (TERT) and an RNA template (reviewed in refs. 13 and 14). Human telomerase reverse transcriptase (hTERT) comprises 1,132 amino acid residues and human telomerase RNA (hTERC) is composed of 451 nucleotides (15-18). Reconstitution of telomerase activity in telomerase-negative cells can be achieved by expression of hTERT, demonstrating that hTERT is the rate-limiting component of the enzyme complex (19,20).GAPDH has long been recognize...

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From Pluripotency to Differentiation: The Role of mtDNA in Stem Cell Models of Mitochondrial Diseases

Facucho-Oliveira

Kulkarni

Machado-Oliveira

et al. 2012

Mitochondrial DNA, Mitochondria, Disease and Stem Cells

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Chromosomal and telomeric reprogramming following ES-somatic cell fusion

Cited by 14 publications

References 38 publications

Comparison of reprogramming ability of mouse ES and iPS cells measured by somatic cell fusion

Comparison of reprogramming ability of mouse ES and iPS cells measured by somatic cell fusion

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induces cancer cell senescence by interacting with telomerase RNA component

From Pluripotency to Differentiation: The Role of mtDNA in Stem Cell Models of Mitochondrial Diseases

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