Advertisement

Comparative gene expression profiling reveals key pathways and genes different in skin epidermal stem cells and corneal epithelial cells

  • Yanjie Guo
  • Weini Wu
  • Xiya Ma
  • Mingyan Shi
  • Xueyi YangEmail author
Research Article
  • 78 Downloads

Abstract

Background

Corneal epithelial cells (CECs) are required for corneal transparency and visual function, and corneal injuries may cause corneal blindness. Skin epidermal stem cells (SESCs), which share the same origin with CECs and have the potential of multi-directional differentiation are ideal seed cells for tissue engineered corneal construction to treat corneal blindness.

Objective

This study aims to investigate critical genes and pathways that may modulate the transdifferentiation from SESCs to CECs.

Methods

Isolated SESCs and CECs were used for gene expression analysis by microarray. GO and KEGG pathway of differently expressed genes (DEGs) were enriched using DAVID. The protein–protein interaction (PPI) network were then constructed using Cytoscape and highly interconnected module was subsequently isolated from the network by Molecular Complex Detection. Expression of the hub genes and other selected genes were then verified by qRT-PCR.

Results

We found 112 upregulated and 105 downregulated genes in CECs compared with SESCs. These DEGs were significantly enriched in focal adhesion, PI3K–Akt and TNF signaling pathway. Highly interconnected module of PPI network contains ten genes. Further regulatory network of these genes showed that ESR1 and SLC2A4 were hub genes.

Conclusion

Our study identified gene expression in SESCs and CECs and suggested that several crucial genes and pathways may play critical roles in transdifferentiation from SESCs to CECs. It may help uncover molecular mechanisms and offer a foundation for promoting tissue-engineered cornea into clinical application.

Keywords

Corneal epithelia cells Skin epidermal stem cells Microarray Transdifferentiation Genes expression profiling 

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (nos. 31240089 and 31701121) and the Programs for Science and Technology Development of Henan Province (172102310550). We thank LetPub (http://www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Compliance with ethical standards

Conflict of interest

Yanjie Guo, Weini Wu, Xiya Ma, Mingyan Shi and Xueyi Yang declare that they have no conflict of interest.

Ethical approval

This study had been approved by the Animal Care and Welfare Committee of Luoyang Normal University (number 12009158).

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

13258_2019_814_MOESM1_ESM.xlsx (20 kb)
Supplementary material 1 (XLSX 19 kb)
13258_2019_814_MOESM2_ESM.xls (630 kb)
Supplementary material 2 (XLS 630 kb)
13258_2019_814_MOESM3_ESM.xls (32 kb)
Supplementary material 3 (XLS 32 kb)
13258_2019_814_MOESM4_ESM.xls (142 kb)
Supplementary material 4 (XLS 142 kb)
13258_2019_814_MOESM5_ESM.xls (22 kb)
Supplementary material 5 (XLS 21 kb)
13258_2019_814_MOESM6_ESM.xlsx (38 kb)
Supplementary material 6 (XLSX 38 kb)

References

  1. Ahmad S, Stewart R, Yung S, Kolli S, Armstrong L, Stojkovic M, Figueiredo F, Lako M (2007) Differentiation of human embryonic stem cells into corneal epithelial-like cells by in vitro replication of the corneal epithelial stem cell niche. Stem Cells 25:1145–1155CrossRefGoogle Scholar
  2. Blazejewska EA, Schlotzer-Schrehardt U, Zenkel M, Bachmann B, Chankiewitz E, Jacobi C, Kruse FE (2009) Corneal limbal microenvironment can induce transdifferentiation of hair follicle stem cells into corneal epithelial-like cells. Stem Cells 27:642–652CrossRefGoogle Scholar
  3. Bose A, Teh MT, Mackenzie IC, Waseem A (2013) Keratin k15 as a biomarker of epidermal stem cells. Int J Mol Sci 14:19385–19398CrossRefGoogle Scholar
  4. Chen HC, Yeh LK, Tsai YJ, Lai CH, Chen CC, Lai JY, Sun CC, Chang G, Hwang TL, Chen JK, Ma DH (2012) Expression of angiogenesis-related factors in human corneas after cultivated oral mucosal epithelial transplantation. Investig Ophthalmol Vis Sci 53:5615–5623CrossRefGoogle Scholar
  5. Datta SR, Brunet A, Greenberg ME (1999) Cellular survival: a play in three Akts. Genes Dev 13:2905–2927CrossRefGoogle Scholar
  6. Dyce PW, Wen L, Li J (2006) In vitro germline potential of stem cells derived from fetal porcine skin. Nat Cell Biol 8:384–390CrossRefGoogle Scholar
  7. Forni MF, Trombetta-Lima M, Sogayar MC (2012) Stem cells in embryonic skin development. Biol Res 45:215–222CrossRefGoogle Scholar
  8. Friend J, Snip RC, Kiorpes TC, Thoft RA (1980) Insulin sensitivity and sorbitol production of the normal rabbit corneal epithelium in vitro. Investig Ophthalmol Vis Sci 19:913–919Google Scholar
  9. Ghadially R (2012) 25 years of epidermal stem cell research. J Investig Dermatol 132:797–810CrossRefGoogle Scholar
  10. Gomes JA, Geraldes Monteiro B, Melo GB, Smith RL, Pereira Cavenaghi, da Silva M, Lizier NF, Kerkis A, Cerruti H, Kerkis I (2010) Corneal reconstruction with tissue-engineered cell sheets composed of human immature dental pulp stem cells. Investig Ophthalmol Vis Sci 51:1408–1414CrossRefGoogle Scholar
  11. Gu S, Xing C, Han J, Tso MO, Hong J (2009) Differentiation of rabbit bone marrow mesenchymal stem cells into corneal epithelial cells in vivo and ex vivo. Mol Vis 15:99–107Google Scholar
  12. Hollenstein K, Dawson RJ, Locher KP (2007) Structure and mechanism of ABC transporter proteins. Curr Opin Struct Biol 17:412–418CrossRefGoogle Scholar
  13. Jones JC (2011) Isolation and culture of bovine corneal epithelial cells. Cold Spring Harb Protoc 2011:1012–1013CrossRefGoogle Scholar
  14. Katikireddy KR, Dana R, Jurkunas UV (2014) Differentiation potential of limbal fibroblasts and bone marrow mesenchymal stem cells to corneal epithelial cells. Stem Cells 32:717–729CrossRefGoogle Scholar
  15. Ksander BR, Kolovou PE, Wilson BJ, Saab KR, Guo Q, Ma J, McGuire SP, Gregory MS, Vincent WJ, Perez VL et al (2014) ABCB5 is a limbal stem cell gene required for corneal development and repair. Nature 511:353–357CrossRefGoogle Scholar
  16. Kuipers DP, Scripture JP, Gunnink SM, Salie MJ, Schotanus MP, Ubels JL, Louters LL (2013) Differential regulation of GLUT1 activity in human corneal limbal epithelial cells and fibroblasts. Biochimie 95:258–263CrossRefGoogle Scholar
  17. Lavker RM, Sun TT (2000) Epidermal stem cells: properties, markers, and location. Proc Natl Acad Sci USA 97:13473–13475CrossRefGoogle Scholar
  18. Leto D, Saltiel AR (2012) Regulation of glucose transport by insulin: traffic control of GLUT4. Nat Rev Mol Cell Biol 13:383–396CrossRefGoogle Scholar
  19. Madhira SL, Vemuganti G, Bhaduri A, Gaddipati S, Sangwan VS, Ghanekar Y (2008) Culture and characterization of oral mucosal epithelial cells on human amniotic membrane for ocular surface reconstruction. Mol Vis 14:189–196Google Scholar
  20. Murayama K, Kimura T, Tarutani M, Tomooka M, Hayashi R, Okabe M, Nishida K, Itami S, Katayama I, Nakano T (2007) Akt activation induces epidermal hyperplasia and proliferation of epidermal progenitors. Oncogene 26:4882–4888CrossRefGoogle Scholar
  21. Nakamura T, Kinoshita S (2003) Ocular surface reconstruction using cultivated mucosal epithelial stem cells. Cornea 22:S75–S80CrossRefGoogle Scholar
  22. Nishida K, Yamato M, Hayashida Y, Watanabe K, Yamamoto K, Adachi E, Nagai S, Kikuchi A, Maeda N, Watanabe H et al (2004) Corneal reconstruction with tissue-engineered cell sheets composed of autologous oral mucosal epithelium. N Engl J Med 351:1187–1196CrossRefGoogle Scholar
  23. Ouyang H, Xue Y, Lin Y, Zhang X, Xi L, Patel S, Cai H, Luo J, Zhang M, Zhang M et al (2014) WNT7A and PAX6 define corneal epithelium homeostasis and pathogenesis. Nature 511:358–361CrossRefGoogle Scholar
  24. Pellegrini G, De Luca M (2014) Eyes on the prize: limbal stem cells and corneal restoration. Cell Stem Cell 15:121–122CrossRefGoogle Scholar
  25. Pellegrini G, Traverso CE, Franzi AT, Zingirian M, Cancedda RD, De Luca M (1997) Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium. Lancet 349:990–993CrossRefGoogle Scholar
  26. Rajala MS, Rajala RV, Astley RA, Butt AL, Chodosh J (2005) Corneal cell survival in adenovirus type 19 infection requires phosphoinositide 3-kinase/Akt activation. J Virol 79:12332–12341CrossRefGoogle Scholar
  27. Ramachandran C, Basu S, Sangwan VS, Balasubramanian D (2014) Concise review: the coming of age of stem cell treatment for corneal surface damage. Stem Cells Transl Med 3:1160–1168CrossRefGoogle Scholar
  28. Saichanma S, Bunyaratvej A, Sila-Asna M (2012) In vitro transdifferentiation of corneal epithelial-like cells from human skin-derived precursor cells. Int J Ophthalmol 5:158–163Google Scholar
  29. Shen S, Wertheimer E, Sampson SR, Tennenbaum T (2000) Characterization of glucose transport system in keratinocytes: insulin and IGF-1 differentially affect specific transporters. J Investig Dermatol 115:949–954CrossRefGoogle Scholar
  30. Spravchikov N, Sizyakov G, Gartsbein M, Accili D, Tennenbaum T, Wertheimer E (2001) Glucose effects on skin keratinocytes: implications for diabetes skin complications. Diabetes 50:1627–1635CrossRefGoogle Scholar
  31. Takacs L, Toth E, Berta A, Vereb G (2009) Stem cells of the adult cornea: from cytometric markers to therapeutic applications. Cytometry A 75:54–66CrossRefGoogle Scholar
  32. Takahashi H, Kaminski AE, Zieske JD (1996) Glucose transporter 1 expression is enhanced during corneal epithelial wound repair. Exp Eye Res 63:649–659CrossRefGoogle Scholar
  33. Toma JG, Akhavan M, Fernandes KJ, Barnabe-Heider F, Sadikot A, Kaplan DR, Miller FD (2001) Isolation of multipotent adult stem cells from the dermis of mammalian skin. Nat Cell Biol 3:778–784CrossRefGoogle Scholar
  34. Utheim TP, Utheim OA, Khan QE, Sehic A (2016) Culture of oral mucosal epithelial cells for the purpose of treating limbal stem cell deficiency. J Funct Biomater 7(1):5CrossRefGoogle Scholar
  35. Xie W, Chow LT, Paterson AJ, Chin EK, Kudlow JE (1999) Conditional expression of the ErbB2 oncogene elicits reversible hyperplasia in stratified epithelia and up-regulation of TGFalpha expression in transgenic mice. Oncogene 18:3593–3607CrossRefGoogle Scholar
  36. Xie Q, Yang Y, Huang J, Ninkovic J, Walcher T, Wolf L, Vitenzon A, Zheng D, Gotz M, Beebe DC et al (2013) Pax6 interactions with chromatin and identification of its novel direct target genes in lens and forebrain. PLoS One 8:e54507CrossRefGoogle Scholar
  37. Yamaoka T, Yan F, Cao H, Hobbs SS, Dise RS, Tong W, Polk DB (2008) Transactivation of EGF receptor and ErbB2 protects intestinal epithelial cells from TNF-induced apoptosis. Proc Natl Acad Sci USA 105:11772–11777CrossRefGoogle Scholar
  38. Yang X, Qu L, Wang X, Zhao M, Li W, Hua J, Shi M, Moldovan N, Wang H, Dou Z (2007) Plasticity of epidermal adult stem cells derived from adult goat ear skin. Mol Reprod Dev 74:386–396CrossRefGoogle Scholar
  39. Yang X, Moldovan NI, Zhao Q, Mi S, Zhou Z, Chen D, Gao Z, Tong D, Dou Z (2008) Reconstruction of damaged cornea by autologous transplantation of epidermal adult stem cells. Mol Vis 14:1064–1070Google Scholar
  40. Yang H, Wang Z, Capo-Aponte JE, Zhang F, Pan Z, Reinach PS (2010) Epidermal growth factor receptor transactivation by the cannabinoid receptor (CB1) and transient receptor potential vanilloid 1 (TRPV1) induces differential responses in corneal epithelial cells. Exp Eye Res 91:462–471CrossRefGoogle Scholar
  41. Yao R, Cooper GM (1995) Requirement for phosphatidylinositol-3 kinase in the prevention of apoptosis by nerve growth factor. Science 267:2003–2006CrossRefGoogle Scholar

Copyright information

© The Genetics Society of Korea 2019

Authors and Affiliations

  • Yanjie Guo
    • 1
  • Weini Wu
    • 1
  • Xiya Ma
    • 1
  • Mingyan Shi
    • 1
  • Xueyi Yang
    • 1
    Email author
  1. 1.Life Science CollegeLuoyang Normal UniversityLuoyangChina

Personalised recommendations