Differential Gene Transcription of Extracellular Matrix Components in Response to In Vivo Corneal Crosslinking (CXL) in Rabbit Corneas.
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State: Public
Version: Final published version
License: CC BY-NC-ND 4.0
State: Public
Version: Final published version
License: CC BY-NC-ND 4.0
Serval ID
serval:BIB_52326A38C07C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Differential Gene Transcription of Extracellular Matrix Components in Response to In Vivo Corneal Crosslinking (CXL) in Rabbit Corneas.
Journal
Translational vision science & technology
ISSN
2164-2591 (Print)
ISSN-L
2164-2591
Publication state
Published
Issued date
12/2017
Peer-reviewed
Oui
Volume
6
Number
6
Pages
8
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Publication Status: epublish
Abstract
We studied changes in gene transcription after corneal crosslinking (CXL) in the rabbit cornea in vivo and identified potential molecular signaling pathways.
A total of 15 corneas of eight male New-Zealand-White rabbits were de-epithelialized and equally divided into five groups. Group 1 served as an untreated control. Groups 2 to 5 were soaked with 0.1% riboflavin for 20 minutes, which in Groups 3 to 5 was followed by UV-A irradiation at a fluence of 5.4 J/cm <sup>2</sup> . Ultraviolet A (UVA) irradiation was delivered at 3 mW/cm <sup>2</sup> for 30 minutes (Group 3, standard CXL protocol), 9 mW/cm <sup>2</sup> for 10 minutes (Group 4, accelerated), and 18 mW/cm <sup>2</sup> for 5 minutes (Group 5, accelerated). At 1 week after treatment, corneal buttons were obtained; mRNA was extracted and subjected to cDNA sequencing (RNA-seq).
A total of 297 differentially transcribed genes were identified after CXL treatment. CXL downregulated extracellular matrix components (collagen types 1A1, 1A2, 6A2, 11A1, keratocan, fibromodulin) and upregulated glycan biosynthesis and proteoglycan glycosylation (GALNT 3, 7, and 8, B3GALT2). Also, CXL activated pathways related to protein crosslinking (transglutaminase 2 and 6). In 9.1% of the significantly different genes, CXL at 3 mW/cm <sup>2</sup> (Group 1) induced a more distinct change in gene transcription than the accelerated CXL protocols, which induced a lower biomechanical stiffening effect.
Several target genes have been identified that might be related to the biomechanical stability and shape of the cornea. Stiffening-dependent differential gene transcription suggests the activation of mechano-sensitive pathways.
A better understanding of the molecular mechanisms behind CXL will permit an optimization and individualization of the clinical treatment protocol.
A total of 15 corneas of eight male New-Zealand-White rabbits were de-epithelialized and equally divided into five groups. Group 1 served as an untreated control. Groups 2 to 5 were soaked with 0.1% riboflavin for 20 minutes, which in Groups 3 to 5 was followed by UV-A irradiation at a fluence of 5.4 J/cm <sup>2</sup> . Ultraviolet A (UVA) irradiation was delivered at 3 mW/cm <sup>2</sup> for 30 minutes (Group 3, standard CXL protocol), 9 mW/cm <sup>2</sup> for 10 minutes (Group 4, accelerated), and 18 mW/cm <sup>2</sup> for 5 minutes (Group 5, accelerated). At 1 week after treatment, corneal buttons were obtained; mRNA was extracted and subjected to cDNA sequencing (RNA-seq).
A total of 297 differentially transcribed genes were identified after CXL treatment. CXL downregulated extracellular matrix components (collagen types 1A1, 1A2, 6A2, 11A1, keratocan, fibromodulin) and upregulated glycan biosynthesis and proteoglycan glycosylation (GALNT 3, 7, and 8, B3GALT2). Also, CXL activated pathways related to protein crosslinking (transglutaminase 2 and 6). In 9.1% of the significantly different genes, CXL at 3 mW/cm <sup>2</sup> (Group 1) induced a more distinct change in gene transcription than the accelerated CXL protocols, which induced a lower biomechanical stiffening effect.
Several target genes have been identified that might be related to the biomechanical stability and shape of the cornea. Stiffening-dependent differential gene transcription suggests the activation of mechano-sensitive pathways.
A better understanding of the molecular mechanisms behind CXL will permit an optimization and individualization of the clinical treatment protocol.
Keywords
corneal biomechanics, corneal crosslinking, differential transcription, extracellular matrix, glycosylation
Pubmed
Web of science
Open Access
Yes
Create date
11/01/2018 18:02
Last modification date
09/08/2024 14:59