Regulation of C-Myc Expression by Ahnak Promotes Induced Pluripotent Stem Cell Generation

Overview

Journal J Biol Chem

Specialty Biochemistry

Date 2015 Nov 25

PMID 26598518

Citations 7

Authors

Hee Jung Lim

Jusong Kim

Chang-Hwan Park

Sang A Lee

Man Ryul Lee

Kye-Seong Kim

Jaesang Kim

Yun Soo Bae

Affiliations

Soon will be listed here.

Abstract

We have previously reported that Ahnak-mediated TGFβ signaling leads to down-regulation of c-Myc expression. Here, we show that inhibition of Ahnak can promote generation of induced pluripotent stem cells (iPSC) via up-regulation of endogenous c-Myc. Consistent with the c-Myc inhibitory role of Ahnak, mouse embryonic fibroblasts from Ahnak-deficient mouse (Ahnak(-/-) MEF) show an increased level of c-Myc expression compared with wild type MEF. Generation of iPSC with just three of the four Yamanaka factors, Oct4, Sox2, and Klf4 (hereafter 3F), was significantly enhanced in Ahnak(-/-) MEF. Similar results were obtained when Ahnak-specific shRNA was applied to wild type MEF. Of note, expressionof Ahnak was significantly induced during the formation of embryoid bodies from embryonic stem cells, suggesting that Ahnak-mediated c-Myc inhibition is involved in embryoid body formation and the initial differentiation of pluripotent stem cells. The iPSC from 3F-infected Ahnak(-/-) MEF cells (Ahnak(-/-)-iPSC-3F) showed expression of all stem cell markers examined and the capability to form three primary germ layers. Moreover, injection of Ahnak(-/-)-iPSC-3F into athymic nude mice led to development of teratoma containing tissues from all three primary germ layers, indicating that iPSC from Ahnak(-/-) MEF are bona fide pluripotent stem cells. Taken together, these data provide evidence for a new role for Ahnak in cell fate determination during development and suggest that manipulation of Ahnak and the associated signaling pathway may provide a means to regulate iPSC generation.

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References

Avilion A, Nicolis S, Pevny L, Perez L, Vivian N, Lovell-Badge R . Multipotent cell lineages in early mouse development depend on SOX2 function. Genes Dev. 2003; 17(1):126-40. PMC: 195970. DOI: 10.1101/gad.224503. View

Theunissen T, Jaenisch R . Molecular control of induced pluripotency. Cell Stem Cell. 2014; 14(6):720-34. PMC: 4112734. DOI: 10.1016/j.stem.2014.05.002. View

Lee I, You J, Ha K, Bae D, Suh P, Rhee S . AHNAK-mediated activation of phospholipase C-gamma1 through protein kinase C. J Biol Chem. 2004; 279(25):26645-53. DOI: 10.1074/jbc.M311525200. View

Rosner M, Vigano M, Ozato K, Timmons P, Poirier F, Rigby P . A POU-domain transcription factor in early stem cells and germ cells of the mammalian embryo. Nature. 1990; 345(6277):686-92. DOI: 10.1038/345686a0. View

Sekiya F, Bae Y, Jhon D, Hwang S, Rhee S . AHNAK, a protein that binds and activates phospholipase C-gamma1 in the presence of arachidonic acid. J Biol Chem. 1999; 274(20):13900-7. DOI: 10.1074/jbc.274.20.13900. View

Cartwright P, McLean C, Sheppard A, Rivett D, Jones K, Dalton S . LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism. Development. 2005; 132(5):885-96. DOI: 10.1242/dev.01670. View

Adhikary S, Eilers M . Transcriptional regulation and transformation by Myc proteins. Nat Rev Mol Cell Biol. 2005; 6(8):635-45. DOI: 10.1038/nrm1703. View

Takahashi K, Yamanaka S . Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006; 126(4):663-76. DOI: 10.1016/j.cell.2006.07.024. View

Wernig M, Meissner A, Foreman R, Brambrink T, Ku M, Hochedlinger K . In vitro reprogramming of fibroblasts into a pluripotent ES-cell-like state. Nature. 2007; 448(7151):318-24. DOI: 10.1038/nature05944. View

10.

Okita K, Ichisaka T, Yamanaka S . Generation of germline-competent induced pluripotent stem cells. Nature. 2007; 448(7151):313-7. DOI: 10.1038/nature05934. View

11.

Yu J, Vodyanik M, Smuga-Otto K, Antosiewicz-Bourget J, Frane J, Tian S . Induced pluripotent stem cell lines derived from human somatic cells. Science. 2007; 318(5858):1917-20. DOI: 10.1126/science.1151526. View

12.

Lee I, Sohn M, Lim H, Yoon S, Oh H, Shin S . Ahnak functions as a tumor suppressor via modulation of TGFβ/Smad signaling pathway. Oncogene. 2014; 33(38):4675-84. PMC: 4180639. DOI: 10.1038/onc.2014.69. View

13.

Dai P, Harada Y, Miyachi H, Tanaka H, Kitano S, Adachi T . Combining TGF-β signal inhibition and connexin43 silencing for iPSC induction from mouse cardiomyocytes. Sci Rep. 2014; 4:7323. PMC: 4255192. DOI: 10.1038/srep07323. View

14.

Tan F, Qian C, Tang K, Abd-Allah S, Jing N . Inhibition of transforming growth factor β (TGF-β) signaling can substitute for Oct4 protein in reprogramming and maintain pluripotency. J Biol Chem. 2014; 290(7):4500-11. PMC: 4326853. DOI: 10.1074/jbc.M114.609016. View

15.

Vrbsky J, Tereh T, Kyrylenko S, Dvorak P, Krejci L . MEK and TGF-beta Inhibition Promotes Reprogramming without the Use of Transcription Factor. PLoS One. 2015; 10(6):e0127739. PMC: 4454598. DOI: 10.1371/journal.pone.0127739. View

16.

Nandan M, Yang V . The role of Krüppel-like factors in the reprogramming of somatic cells to induced pluripotent stem cells. Histol Histopathol. 2009; 24(10):1343-55. PMC: 2753264. DOI: 10.14670/HH-24.1343. View

17.

Nakagawa M, Koyanagi M, Tanabe K, Takahashi K, Ichisaka T, Aoi T . Generation of induced pluripotent stem cells without Myc from mouse and human fibroblasts. Nat Biotechnol. 2007; 26(1):101-6. DOI: 10.1038/nbt1374. View

18.

Park I, Zhao R, West J, Yabuuchi A, Huo H, Ince T . Reprogramming of human somatic cells to pluripotency with defined factors. Nature. 2007; 451(7175):141-6. DOI: 10.1038/nature06534. View

19.

Lowry W, Richter L, Yachechko R, Pyle A, Tchieu J, Sridharan R . Generation of human induced pluripotent stem cells from dermal fibroblasts. Proc Natl Acad Sci U S A. 2008; 105(8):2883-8. PMC: 2268554. DOI: 10.1073/pnas.0711983105. View

20.

Lee I, Lim H, Yoon S, Seong J, Bae D, Rhee S . Ahnak protein activates protein kinase C (PKC) through dissociation of the PKC-protein phosphatase 2A complex. J Biol Chem. 2008; 283(10):6312-20. DOI: 10.1074/jbc.M706878200. View