Comparison of upstream regulators in human ex vivo cultured cornea limbal epithelial stem cells and differentiated corneal epithelial cells

Zoltán Veréb, Réka Albert, Szilárd Póliska, Ole K. Olstad, Saeed Akhtar, Morten C. Moe, G. Petrovski

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background: The surface of the human eye is covered by corneal epithelial cells (CECs) which regenerate from a small population of limbal epithelial stem cells (LESCs). Cell therapy with LESCs is a non-penetrating treatment for preventing blindness due to LESC deficiency or dysfunction. Our aim was to identify new putative molecular markers and upstream regulators in the LESCs and associated molecular pathways.Results: Genome-wide microarray transcriptional profiling was used to compare LESCs to differentiated human CECs. Ingenuity-based pathway analysis was applied to identify upstream regulators and pathways specific to LESCs. ELISA and flow cytometry were used to measure secreted and surface expressed proteins, respectively. More than 2 fold increase and decrease in expression could be found in 1830 genes between the two cell types. A number of molecules functioning in cellular movement (381), proliferation (567), development (552), death and survival (520), and cell-to-cell signaling (290) were detected having top biological functions in LESCs and several of these were confirmed by flow cytometric surface protein analysis. Custom-selected gene groups related to stemness, differentiation, cell adhesion, cytokines and growth factors as well as angiogenesis could be analyzed. The results show that LESCs play a key role not only in epithelial differentiation and tissue repair, but also in controlling angiogenesis and extracellular matrix integrity. Some pro-inflammatory cytokines were found to be important in stemness-, differentiation- and angiogenesis-related biological functions: IL-6 and IL-8 participated in most of these biological pathways as validated by their secretion from LESC cultures.Conclusions: The gene and molecular pathways may provide a more specific understanding of the signaling molecules associated with LESCs, therefore, help better identify and use these cells in the treatment of ocular surface diseases.

Original languageEnglish
Article number900
JournalBMC Genomics
Volume14
Issue number1
DOIs
Publication statusPublished - Dec 17 2013

Fingerprint

Cornea
Stem Cells
Epithelial Cells
Membrane Proteins
Cytokines
Genes
Eye Diseases
Blindness
Cell- and Tissue-Based Therapy
Interleukin-8
Extracellular Matrix
Interleukin-6
Cell Survival
Intercellular Signaling Peptides and Proteins
Flow Cytometry
Epithelium
Cell Culture Techniques
Enzyme-Linked Immunosorbent Assay
Genome

Keywords

  • Angiogenesis
  • Cell adhesion
  • Corneal epithelial cells
  • Cytokines
  • Gene array
  • IL-6
  • IL-8
  • Limbal epithelial stem cells
  • Upstream regulators

ASJC Scopus subject areas

  • Biotechnology
  • Genetics

Cite this

Comparison of upstream regulators in human ex vivo cultured cornea limbal epithelial stem cells and differentiated corneal epithelial cells. / Veréb, Zoltán; Albert, Réka; Póliska, Szilárd; Olstad, Ole K.; Akhtar, Saeed; Moe, Morten C.; Petrovski, G.

In: BMC Genomics, Vol. 14, No. 1, 900, 17.12.2013.

Research output: Contribution to journalArticle

Veréb, Zoltán ; Albert, Réka ; Póliska, Szilárd ; Olstad, Ole K. ; Akhtar, Saeed ; Moe, Morten C. ; Petrovski, G. / Comparison of upstream regulators in human ex vivo cultured cornea limbal epithelial stem cells and differentiated corneal epithelial cells. In: BMC Genomics. 2013 ; Vol. 14, No. 1.
@article{84be32635d474cb3b51a59475819246f,
title = "Comparison of upstream regulators in human ex vivo cultured cornea limbal epithelial stem cells and differentiated corneal epithelial cells",
abstract = "Background: The surface of the human eye is covered by corneal epithelial cells (CECs) which regenerate from a small population of limbal epithelial stem cells (LESCs). Cell therapy with LESCs is a non-penetrating treatment for preventing blindness due to LESC deficiency or dysfunction. Our aim was to identify new putative molecular markers and upstream regulators in the LESCs and associated molecular pathways.Results: Genome-wide microarray transcriptional profiling was used to compare LESCs to differentiated human CECs. Ingenuity-based pathway analysis was applied to identify upstream regulators and pathways specific to LESCs. ELISA and flow cytometry were used to measure secreted and surface expressed proteins, respectively. More than 2 fold increase and decrease in expression could be found in 1830 genes between the two cell types. A number of molecules functioning in cellular movement (381), proliferation (567), development (552), death and survival (520), and cell-to-cell signaling (290) were detected having top biological functions in LESCs and several of these were confirmed by flow cytometric surface protein analysis. Custom-selected gene groups related to stemness, differentiation, cell adhesion, cytokines and growth factors as well as angiogenesis could be analyzed. The results show that LESCs play a key role not only in epithelial differentiation and tissue repair, but also in controlling angiogenesis and extracellular matrix integrity. Some pro-inflammatory cytokines were found to be important in stemness-, differentiation- and angiogenesis-related biological functions: IL-6 and IL-8 participated in most of these biological pathways as validated by their secretion from LESC cultures.Conclusions: The gene and molecular pathways may provide a more specific understanding of the signaling molecules associated with LESCs, therefore, help better identify and use these cells in the treatment of ocular surface diseases.",
keywords = "Angiogenesis, Cell adhesion, Corneal epithelial cells, Cytokines, Gene array, IL-6, IL-8, Limbal epithelial stem cells, Upstream regulators",
author = "Zolt{\'a}n Ver{\'e}b and R{\'e}ka Albert and Szil{\'a}rd P{\'o}liska and Olstad, {Ole K.} and Saeed Akhtar and Moe, {Morten C.} and G. Petrovski",
year = "2013",
month = "12",
day = "17",
doi = "10.1186/1471-2164-14-900",
language = "English",
volume = "14",
journal = "BMC Genomics",
issn = "1471-2164",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Comparison of upstream regulators in human ex vivo cultured cornea limbal epithelial stem cells and differentiated corneal epithelial cells

AU - Veréb, Zoltán

AU - Albert, Réka

AU - Póliska, Szilárd

AU - Olstad, Ole K.

AU - Akhtar, Saeed

AU - Moe, Morten C.

AU - Petrovski, G.

PY - 2013/12/17

Y1 - 2013/12/17

N2 - Background: The surface of the human eye is covered by corneal epithelial cells (CECs) which regenerate from a small population of limbal epithelial stem cells (LESCs). Cell therapy with LESCs is a non-penetrating treatment for preventing blindness due to LESC deficiency or dysfunction. Our aim was to identify new putative molecular markers and upstream regulators in the LESCs and associated molecular pathways.Results: Genome-wide microarray transcriptional profiling was used to compare LESCs to differentiated human CECs. Ingenuity-based pathway analysis was applied to identify upstream regulators and pathways specific to LESCs. ELISA and flow cytometry were used to measure secreted and surface expressed proteins, respectively. More than 2 fold increase and decrease in expression could be found in 1830 genes between the two cell types. A number of molecules functioning in cellular movement (381), proliferation (567), development (552), death and survival (520), and cell-to-cell signaling (290) were detected having top biological functions in LESCs and several of these were confirmed by flow cytometric surface protein analysis. Custom-selected gene groups related to stemness, differentiation, cell adhesion, cytokines and growth factors as well as angiogenesis could be analyzed. The results show that LESCs play a key role not only in epithelial differentiation and tissue repair, but also in controlling angiogenesis and extracellular matrix integrity. Some pro-inflammatory cytokines were found to be important in stemness-, differentiation- and angiogenesis-related biological functions: IL-6 and IL-8 participated in most of these biological pathways as validated by their secretion from LESC cultures.Conclusions: The gene and molecular pathways may provide a more specific understanding of the signaling molecules associated with LESCs, therefore, help better identify and use these cells in the treatment of ocular surface diseases.

AB - Background: The surface of the human eye is covered by corneal epithelial cells (CECs) which regenerate from a small population of limbal epithelial stem cells (LESCs). Cell therapy with LESCs is a non-penetrating treatment for preventing blindness due to LESC deficiency or dysfunction. Our aim was to identify new putative molecular markers and upstream regulators in the LESCs and associated molecular pathways.Results: Genome-wide microarray transcriptional profiling was used to compare LESCs to differentiated human CECs. Ingenuity-based pathway analysis was applied to identify upstream regulators and pathways specific to LESCs. ELISA and flow cytometry were used to measure secreted and surface expressed proteins, respectively. More than 2 fold increase and decrease in expression could be found in 1830 genes between the two cell types. A number of molecules functioning in cellular movement (381), proliferation (567), development (552), death and survival (520), and cell-to-cell signaling (290) were detected having top biological functions in LESCs and several of these were confirmed by flow cytometric surface protein analysis. Custom-selected gene groups related to stemness, differentiation, cell adhesion, cytokines and growth factors as well as angiogenesis could be analyzed. The results show that LESCs play a key role not only in epithelial differentiation and tissue repair, but also in controlling angiogenesis and extracellular matrix integrity. Some pro-inflammatory cytokines were found to be important in stemness-, differentiation- and angiogenesis-related biological functions: IL-6 and IL-8 participated in most of these biological pathways as validated by their secretion from LESC cultures.Conclusions: The gene and molecular pathways may provide a more specific understanding of the signaling molecules associated with LESCs, therefore, help better identify and use these cells in the treatment of ocular surface diseases.

KW - Angiogenesis

KW - Cell adhesion

KW - Corneal epithelial cells

KW - Cytokines

KW - Gene array

KW - IL-6

KW - IL-8

KW - Limbal epithelial stem cells

KW - Upstream regulators

UR - http://www.scopus.com/inward/record.url?scp=84890289273&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84890289273&partnerID=8YFLogxK

U2 - 10.1186/1471-2164-14-900

DO - 10.1186/1471-2164-14-900

M3 - Article

C2 - 24344983

AN - SCOPUS:84890289273

VL - 14

JO - BMC Genomics

JF - BMC Genomics

SN - 1471-2164

IS - 1

M1 - 900

ER -