Int J Stem Cells.  2014 Nov;7(2):108-117. 10.15283/ijsc.2014.7.2.108.

Transcriptional Profiles of Imprinted Genes in Human Embryonic Stem Cells During In vitro Differentiation

Affiliations
  • 1Department of Biological Sciences and Center for Stem Cell Differentiation, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Korea. ymhan@kaist.ac.kr
  • 2Department of Biochemistry and Molecular Biology, College of Medicine, Hanyang University, Seoul, Korea.

Abstract

BACKGROUND AND OBJECTIVES
Genomic imprinting is an inheritance phenomenon by which a subset of genes are expressed from one allele of two homologous chromosomes in a parent of origin-specific manner. Even though fine-tuned regulation of genomic imprinting process is essential for normal development, no other means are available to study genomic imprinting in human during embryonic development. In relation with this bottleneck, differentiation of human embryonic stem cells (hESCs) into specialized lineages may be considered as an alternative to mimic human development.
METHODS AND RESULTS
In this study, hESCs were differentiated into three lineage cell types to analyze temporal and spatial expression of imprinted genes. Of 19 imprinted genes examined, 15 imprinted genes showed similar transcriptional level among two hESC lines and two human induced pluripotent stem cell (hiPSC) lines. Expressional patterns of most imprinted genes were varied in progenitors and fully differentiated cells which were derived from hESCs. Also, no consistence was observed in the expression pattern of imprinted genes within an imprinting domain during in vitro differentiation of hESCs into three lineage cell types.
CONCLUSIONS
Transcriptional expression of imprinted genes is regulated in a cell type-specific manner in hESCs during in vitro differentiation.

Keyword

Human embryonic stem cells; Genomic imprinting; In vitro differentiation; Transcriptional expression

MeSH Terms

Alleles
Embryonic Development
Embryonic Stem Cells*
Female
Genomic Imprinting
Human Development
Humans
Parents
Pluripotent Stem Cells
Pregnancy
Wills

Figure

  • Fig. 1. Transcriptional expression of imprinted genes between four hPSC lines. Transcriptional levels of the imprinted genes that are included in IGF2-H19 (A), DIO3-DLK1 (B), KCNQ1 (C), PWS/AS (D), and PEG10 (E) domains were examined in undifferentiated hPSCs by qRT-PCR. The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p <0.01). Abbreviations: CRL_iPSCs; hiPSCs derived from CRL foreskin fibroblasts, HDF_iPSCs; hiPSCs derived from human dermal fibro-blasts.

  • Fig. 2. Relative expression levels of imprinted genes in IGF2/H19 domain in hESCs during in vitro differentiation. (A) A schematic diagram of IGF2/H19 domain (Ch 11p15). Relative expression level of IGF and H19 imprinted genes in hESCs during in vitro differentiation to DA neurons (B), osteoblasts (C), and hepatocytes (D). The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p<0.01, ***p<0.001). Abbreviations, ES: H9-hESCs; NP: neuronal progenitors; DA: dopaminergic neurons; MP: mesenchymal progenitors; OB: osteoblasts; DE: definitive endoderm; HEP: hepatocytes.

  • Fig. 3. Relative expression levels of imprinted genes in KCNQ1 domain in hESCs during in vitro differentiation. (A) A schematic diagram of KCNQ1 domain (Ch 11p15). This KCNQ1 domain contains several imprinted genes, including OSBPL5, PHLDA2, CDKN1C, KCNQ1, and KCNQ1OT1. Relative expression level of several imprinted genes in hESCs during in vitro differentiation to DA neurons (B), osteoblasts (C), and hepatocytes (D). The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p<0.01, ***p<0.001).

  • Fig. 4. Relative expression levels of imprinted genes in PWS/AS domain in hESCs during in vitro differentiation. (A) A schematic diagram of PWS/AS domain (Ch. 15q11-q13). This domain contains several imprinted genes, including MAGEL2, NDN, SNRPN, IPW, UBE3A, and ATP10A. Relative expression level of several imprinted genes in hESCs during in vitro differentiation to DA neurons (B), osteoblasts (C), and hepatocytes (D). The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p<0.01, ***p<0.001).

  • Fig. 5. Relative expression levels of imprinted genes in PEG10 domain in hESCs during in vitro differentiation. (A) A schematic diagram of PEG10 domain (Ch. 7q21.3). This domain contains three imprinted genes such as SGCE, PEG10 and PPP1R9A. Relative expression level of these imprinted genes in hESCs during in vitro differentiation to DA neurons (B), osteoblasts (C), and hepatocytes (D). The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p<0.01, ***p<0.001).

  • Fig. 6. Relative expression levels of imprinted genes in DIO3-DLK1 domain in hESCs during in vitro differentiation. (A) A schematic diagram of DIO3-DLK1 domain (Ch. 14q32.2). This domain contains four imprinted genes such as DLK1, MEG3, DIO3 and RTL1. Relative expression level of DIO3 and RTL1 imprinted genes in hESCs during in vitro differentiation to DA neurons (B), osteoblasts (C), and hepatocytes (D). The error bars indicate±SD for three independent experiments (n=3, *p<0.05, **p<0.01, ***p<0.001).


Reference

References

1. Edwards CA, Ferguson-Smith AC. Mechanisms regulating imprinted genes in clusters. Curr Opin Cell Biol. 2007. 19:281–289.
Article
2. Verona RI, Mann MR, Bartolomei MS. Genomic imprinting: intricacies of epigenetic regulation in clusters. Annu Rev Cell Dev Biol. 2003. 19:237–259.
Article
3. Reik W, Dean W, Walter J. Epigenetic reprogramming in mammalian development. Science. 2001. 293:1089–1093.
Article
4. Ideraabdullah FY, Vigneau S, Bartolomei MS. Genomic imprinting mechanisms in mammals. Mutat Res. 2008. 647:77–85.
Article
5. Frost JM, Moore GE. The importance of imprinting in the human placenta. PLoS Genet. 2010. 6:e1001015.
Article
6. Xie W, Barr CL, Kim A, Yue F, Lee AY, Eubanks J, Dempster EL, Ren B. Base-resolution analyses of sequence and parent-of-origin dependent DNA methylation in the mouse genome. Cell. 2012. 148:816–831.
Article
7. Peters J. The role of genomic imprinting in biology and disease: an expanding view. Nat Rev Genet. 2014. 15:517–530.
Article
8. Morison IM, Ramsay JP, Spencer HG. A census of mammalian imprinting. Trends Genet. 2005. 21:457–465.
Article
9. Ko JY, Park CH, Koh HC, Cho YH, Kyhm JH, Kim YS, Lee I, Lee YS, Lee SH. Human embryonic stem cell-derived neural precursors as a continuous, stable, and on-demand source for human dopamine neurons. J Neurochem. 2007. 103:1417–1429.
Article
10. Mahmood A, Harkness L, Schrøder HD, Abdallah BM, Kassem M. Enhanced differentiation of human embryonic stem cells to mesenchymal progenitors by inhibition of TGF-beta/activin/nodal signaling using SB-431542. J Bone Miner Res. 2010. 25:1216–1233.
Article
11. Cai J, Zhao Y, Liu Y, Ye F, Song Z, Qin H, Meng S, Chen Y, Zhou R, Song X, Guo Y, Ding M, Deng H. Directed differentiation of human embryonic stem cells into functional hepatic cells. Hepatology. 2007. 45:1229–1239.
Article
12. Park SW, Kim J, Park JL, Ko JY, Im I, Do HS, Kim H, Tran NT, Lee SH, Kim YS, Cho YS, Lee DR, Han YM. Variable allelic expression of imprinted genes in human pluripotent stem cells during differentiation into specialized cell types in vitro. Biochem Biophys Res Commun. 2014. 446:493–498.
Article
13. Bell AC, Felsenfeld G. Methylation of a CTCF-dependent boundary controls imprinted expression of the Igf2 gene. Nature. 2000. 405:482–485.
Article
14. Ono R, Nakamura K, Inoue K, Naruse M, Usami T, Wakisaka-Saito N, Hino T, Suzuki-Migishima R, Ogonuki N, Miki H, Kohda T, Ogura A, Yokoyama M, Kaneko-Ishino T, Ishino F. Deletion of Peg10, an imprinted gene acquired from a retrotransposon, causes early embryonic lethality. Nat Genet. 2006. 38:101–106.
Article
15. Berg JS, Lin KK, Sonnet C, Boles NC, Weksberg DC, Nguyen H, Holt LJ, Rickwood D, Daly RJ, Goodell MA. Imprinted genes that regulate early mammalian growth are coexpressed in somatic stem cells. PLoS One. 2011. 6:e26410.
Article
16. Sun BW, Yang AC, Feng Y, Sun YJ, Zhu Yf, Zhang Y, Jiang H, Li CL, Gao FR, Zhang ZH, Wang WC, Kong XY, Jin G, Fu SJ, Jin Y. Temporal and parental-specific expression of imprinted genes in a newly derived Chinese human embryonic stem cell line and embryoid bodies. Hum Mol Genet. 2006. 15:65–75.
Article
17. Onyango P, Jiang S, Uejima H, Shamblott MJ, Gearhart JD, Cui H, Feinberg AP. Monoallelic expression and methylation of imprinted genes in human and mouse embryonic germ cell lineages. Proc Natl Acad Sci U S A. 2002. 99:10599–10604.
Article
18. Freed WJ, Chen J, Bäckman CM, Schwartz CM, Vazin T, Cai J, Spivak CE, Lupica CR, Rao MS, Zeng X. Gene expression profile of neuronal progenitor cells derived from hESCs: activation of chromosome 11p15.5 and comparison to human dopaminergic neurons. PLoS One. 2008. 3:e1422.
Article
Full Text Links
  • IJSC
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr