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AICAR activates the pluripotency transcriptional network in embryonic stem cells and induces KLF4 and KLF2 expression in fibroblasts.

Adamo L, Zhang Y, García-Cardeña G - BMC Pharmacol. (2009)

Bottom Line: Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4.AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells.AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Laboratory for Systems Biology, Center for Excellence in Vascular Biology, Harvard Medical School and Brigham and Women's Hospital, Boston, MA, USA. ladamo@hms.harvard.edu

ABSTRACT

Background: Pluripotency, the property of a cell to differentiate into all cellular types of a given organism, is central to the development of stem cell-based therapies and regenerative medicine. Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4. Recently, it has been shown that pluripotency can be induced in somatic cells by viral-mediated expression of the transcription factors Oct3/4, Sox2, Klf4, and c-Myc.

Results: Here we show that 5-Aminoimidazole-4-carboxamide-1-b-riboside (AICAR) is able to activate the molecular circuitry of pluripotency in mouse embryonic stem cells (mESC) and maintain Nanog and Oct4 expression in mESC exposed to the differentiating agent retinoic acid. We also show that AICAR is able to induce Klf4, Klf2 and Myc expression in both mESC and murine fibroblasts.

Conclusion: AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells. AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency.

Show MeSH
AICAR activates the pluripotency transcriptional network in mESC. Murine embryonic stem cells were exposed to 1 mM AICAR for 72 h. AICAR induced upregulation of the master regulators of pluripotency Nanog and Oct4 and of the pluripotency related transcription factors Klf4, Klf2, Myc and Sox2. Taqman real time quantitative PCR. N = 3. Graphs represent average +/- SEM. *** = P < 0.0001, ** = P < 0.001.
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Figure 1: AICAR activates the pluripotency transcriptional network in mESC. Murine embryonic stem cells were exposed to 1 mM AICAR for 72 h. AICAR induced upregulation of the master regulators of pluripotency Nanog and Oct4 and of the pluripotency related transcription factors Klf4, Klf2, Myc and Sox2. Taqman real time quantitative PCR. N = 3. Graphs represent average +/- SEM. *** = P < 0.0001, ** = P < 0.001.

Mentions: The J1 mESC line is a well characterized pluripotent cell line derived from the 129 Sv/J mouse strain. This cell line was used to assess the effects of AICAR on gene expression in pluripotent cells. ES cells were plated at 5000/cm2 and exposed to 1 mM AICAR for 72 h. Gene expression was measured by total RNA extraction followed by RNA reverse transcription and real-time Taqman quantitative PCR. As seen in Figure 1, AICAR induced a 9 fold upregulation of Klf4 and a 2.7 fold upregulation of Klf2. Concomitantly, AICAR treatment induced the activation of the pluripotency transcription network as demonstrated by the upregulation of the pluripotency keeper transcription factors Nanog [12,13], Oct4 [14], Myc [15] and Sox2 [16]


AICAR activates the pluripotency transcriptional network in embryonic stem cells and induces KLF4 and KLF2 expression in fibroblasts.

Adamo L, Zhang Y, García-Cardeña G - BMC Pharmacol. (2009)

AICAR activates the pluripotency transcriptional network in mESC. Murine embryonic stem cells were exposed to 1 mM AICAR for 72 h. AICAR induced upregulation of the master regulators of pluripotency Nanog and Oct4 and of the pluripotency related transcription factors Klf4, Klf2, Myc and Sox2. Taqman real time quantitative PCR. N = 3. Graphs represent average +/- SEM. *** = P < 0.0001, ** = P < 0.001.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2651871&req=5

Figure 1: AICAR activates the pluripotency transcriptional network in mESC. Murine embryonic stem cells were exposed to 1 mM AICAR for 72 h. AICAR induced upregulation of the master regulators of pluripotency Nanog and Oct4 and of the pluripotency related transcription factors Klf4, Klf2, Myc and Sox2. Taqman real time quantitative PCR. N = 3. Graphs represent average +/- SEM. *** = P < 0.0001, ** = P < 0.001.
Mentions: The J1 mESC line is a well characterized pluripotent cell line derived from the 129 Sv/J mouse strain. This cell line was used to assess the effects of AICAR on gene expression in pluripotent cells. ES cells were plated at 5000/cm2 and exposed to 1 mM AICAR for 72 h. Gene expression was measured by total RNA extraction followed by RNA reverse transcription and real-time Taqman quantitative PCR. As seen in Figure 1, AICAR induced a 9 fold upregulation of Klf4 and a 2.7 fold upregulation of Klf2. Concomitantly, AICAR treatment induced the activation of the pluripotency transcription network as demonstrated by the upregulation of the pluripotency keeper transcription factors Nanog [12,13], Oct4 [14], Myc [15] and Sox2 [16]

Bottom Line: Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4.AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells.AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pathology, Laboratory for Systems Biology, Center for Excellence in Vascular Biology, Harvard Medical School and Brigham and Women's Hospital, Boston, MA, USA. ladamo@hms.harvard.edu

ABSTRACT

Background: Pluripotency, the property of a cell to differentiate into all cellular types of a given organism, is central to the development of stem cell-based therapies and regenerative medicine. Stem cell pluripotency is the result of the orchestrated activation of a complex transcriptional network characterized by the expression of a set of transcription factors including the master regulators of pluripotency Nanog and Oct4. Recently, it has been shown that pluripotency can be induced in somatic cells by viral-mediated expression of the transcription factors Oct3/4, Sox2, Klf4, and c-Myc.

Results: Here we show that 5-Aminoimidazole-4-carboxamide-1-b-riboside (AICAR) is able to activate the molecular circuitry of pluripotency in mouse embryonic stem cells (mESC) and maintain Nanog and Oct4 expression in mESC exposed to the differentiating agent retinoic acid. We also show that AICAR is able to induce Klf4, Klf2 and Myc expression in both mESC and murine fibroblasts.

Conclusion: AICAR is able to activate the molecular circuitry of pluripotency in mESC and to induce the expression of several key regulators of pluripotency in somatic cells. AICAR is therefore a useful pharmacological entity for studying small molecule mediated induction of pluripotency.

Show MeSH