Abstract | BACKGROUND: METHODS: Wild-type (n = 3), GCD2-heterozygous (n = 1), and GCD2-homozygous (n = 3) primary human corneal fibroblasts were harvested from human donors and patients prepared. Microarray and gene-expression profiling, Chromatin immunoprecipitation microarray analysis, and Methylated DNA isolation assay-assisted CpG microarrays was performed in Wild-type and GCD2-homozygous human cells. RESULTS: Transcription and extracellular-secretion levels of TGFBIp were high in normal cells compared with those in GCD2-derived cells and were related to H3K4me3 levels but not to DNA methylation over the TGFBI locus. TGFβ1 increased the expression of TGFBIp and the ECM-associated genes connective tissue growth factor, collagen-α2[Ι], and plasminogen activator inhibitor-1 in normal corneal fibroblasts. Increased levels of gene-activating markers (H3K4me1/3) and decreased levels of repressive markers (H3K27me3) at the promoters of those gene accompanied the changes in expression. TGFβ1 also increased recruitment of the H3K4 methyltransferase MLL1 and of SET7/9 and also the binding of Smad3 to the promoters. Knockdown of both MLL1 and SET7/9 significantly blocked the TGFβ1-induced gene expression and inhibited TGFβ1-induced changes in promoter H3K4me1/3 levels. Those effects were very weak, however, in GCD2-derived corneal fibroblasts. CONCLUSIONS: Taken together, the results show the functional role of H3K4me in TGFβ1-mediated TGFBIp and ECM gene expression in corneal fibroblasts. Pharmacologic and other therapies that regulate these modifications could have potential cornea-protective effects for granular corneal dystrophy.
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Authors | Yong-Sun Maeng, Ga-Hyun Lee, Seung-Il Choi, Kyu Seo Kim, Eung Kweon Kim |
Journal | BMC medical genomics
(BMC Med Genomics)
Vol. 8
Pg. 74
(Nov 09 2015)
ISSN: 1755-8794 [Electronic] England |
PMID | 26553048
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- Extracellular Matrix Proteins
- Histones
- KMT2A protein, human
- Smad3 Protein
- Transforming Growth Factor beta
- Transforming Growth Factor beta1
- betaIG-H3 protein
- Myeloid-Lymphoid Leukemia Protein
- Histone Methyltransferases
- Histone-Lysine N-Methyltransferase
- SETD7 protein, human
- Lysine
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Topics |
- Cornea
(cytology, pathology)
- Corneal Dystrophies, Hereditary
(genetics, metabolism, pathology)
- Extracellular Matrix
(metabolism)
- Extracellular Matrix Proteins
(genetics, metabolism)
- Fibroblasts
(cytology, pathology)
- Gene Expression Regulation
- Gene Knockdown Techniques
- Histone Methyltransferases
- Histone-Lysine N-Methyltransferase
(deficiency, genetics, metabolism)
- Histones
(chemistry, metabolism)
- Homozygote
- Humans
- Lysine
(metabolism)
- Methylation
- Myeloid-Lymphoid Leukemia Protein
(deficiency, genetics, metabolism)
- Protein Transport
- Smad3 Protein
(metabolism)
- Transcription, Genetic
- Transforming Growth Factor beta
(genetics, metabolism)
- Transforming Growth Factor beta1
(metabolism)
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