Epigenetic Regulation of Filaggrin Gene Expression in Human Epidermal Keratinocytes
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Background: Loss-of-function mutations in the filaggrin gene (FLG), which encodes an epidermal protein crucial for the formation of a functional skin barrier, have been identified as a major predisposing factor in the etiopathogenesis of atopic dermatitis (AD). Recent reports of relatively low frequencies of FLG-null mutations among specific ethnic groups with AD necessitated analysis of the epigenetic regulation which may control FLG expression without altering its DNA sequence. Objective: The study aimed to identify DNA methylation- dependent regulation of FLG expression. Methods: Quantitative polymerase chain reaction was performed to determine the restoration of FLG mRNA expression in normal human epidermal keratinocyte (NHEK) cells after treatment with epigenetic modulating agents. Bisulfite genomic sequencing and pyrosequencing analyses of the FLG promoter region were conducted to identify the citical CpG sites relevant to FLG expression. We performed small-scale pilot study for epidermal tissues obtained from Korean patients with severe AD. Results: We here show that DNA methylation in the FLG with non-CpG island promoter is responsible for the transcriptional regulation of FLG in undifferentiated NHEK cells. The methylation frequencies in a single CpG site of the FLG promoter were significantly higher in lesional epidermis than those in matched nonlesional epidermis of subjects with severe AD. Conclusion: Our in vitro and clinical studies point to this unique CpG site as a potential DNA methylation marker of FLG, which can be a promising therapeutic target in the complications of filaggrin-related skin barrier dysfunction as well as in AD.
Abstract
Background: Loss-of-function mutations in the filaggrin gene (FLG), which encodes an epidermal protein crucial for the formation of a functional skin barrier, have been identified as a major predisposing factor in the etiopathogenesis of atopic dermatitis (AD). Recent reports of relatively low frequencies of FLG-null mutations among specific ethnic groups with AD necessitated analysis of the epigenetic regulation which may control FLG expression without altering its DNA sequence. Objective: The study aimed to identify DNA methylation- dependent regulation of FLG expression. Methods: Quantitative polymerase chain reaction was performed to determine the restoration of FLG mRNA expression in normal human epidermal keratinocyte (NHEK) cells after treatment with epigenetic modulating agents. Bisulfite genomic sequencing and pyrosequencing analyses of the FLG promoter region were conducted to identify the citical CpG sites relevant to FLG expression. We performed small-scale pilot study for epidermal tissues obtained from Korean patients with severe AD. Results: We here show that DNA methylation in the FLG with non-CpG island promoter is responsible for the transcriptional regulation of FLG in undifferentiated NHEK cells. The methylation frequencies in a single CpG site of the FLG promoter were significantly higher in lesional epidermis than those in matched nonlesional epidermis of subjects with severe AD. Conclusion: Our in vitro and clinical studies point to this unique CpG site as a potential DNA methylation marker of FLG, which can be a promising therapeutic target in the complications of filaggrin-related skin barrier dysfunction as well as in AD.
참고문헌 (40)
- Brown SJ, "One remarkable molecule : filaggrin", J Invest Dermatol, 132(3 Pt 2), pp.751~762 (2012)
- Elias PM, ""Outside-to-inside"(and now back to"outside")pathogenic mechanisms in atopic dermatitis", J Invest Dermatol, 128, pp.1067~1070 (2008)
- Smith FJ, "Loss-of-function mutations in the gene encoding filaggrin cause ichthyosis vulgaris", Nat Genet, 38, pp.337~342 (2006)
- Palmer CN, "Common loss-of-function variants of the epidermal barrier protein filaggrin are a major predisposing factor for atopic dermatitis", Nat Genet, 38, pp.441~446 (2006)
- Sandilands A, "Comprehensive analysis of the gene encoding filaggrin uncovers prevalent and rare mutations in ichthyosis vulgaris and atopic eczema", Nat Genet, 39, pp.650~654 (2007)
- Nomura T, "Unique mutations in the filaggrin gene in Japanese patients with ichthyosis vulgaris and atopic dermatitis", J Allergy Clin Immunol, 119, pp.434~440 (2007)
- Hamada T, "De novo occurrence of the filaggrin mutation p.R501X with prevalent mutation c.3321delA in a Japanese family with ichthyosis vulgaris complicated by atopic dermatitis", J Invest Dermatol, 128, pp.1323~1325 (2008)
- Zhang X, "Novel and recurrent mutations in the filaggrin gene in Chinese patients with ichthyosis vulgaris", Br J Dermatol, 163, pp.63~69 (2010)
- Zhang H, "Mutations in the filaggrin gene in Han Chinese patients with atopic dermatitis", Allergy, 66, pp.420~427 (2011)
- Chen H, "Wide spectrum of filaggrin-null mutations in atopic dermatitis highlights differences between Singaporean Chinese and European populations", Br J Dermatol, 165, pp.106~114 (2011)
- Hsu CK, "Analysis of Taiwanese ichthyosis vulgaris families further demonstrates differences in FLG mutations between European and Asian populations", Br J Dermatol, 161, pp.448~451 (2009)
- Eun Joo Kim, "Genetic Polymorphism of FLG in Korean Ichthyosis Vulgaris Patients", Annals of Dermatology, 23(2), pp.170~176 (2011), 대한피부과학회
- Park J, "Novel FLG null mutations in Korean patients with atopic dermatitis and comparison of the mutational spectra in Asian populations", J Dermatol, 42, pp.867~873 (2015)
- Winge MC, "Novel filaggrin mutation but no other loss-of-function variants found in Ethiopian patients with atopic dermatitis", Br J Dermatol, 165, pp.1074~1080 (2011)
- Margolis DJ, "Exome sequencing of filaggrin and related genes in African-American children with atopic dermatitis", J Invest Dermatol, 134, pp.2272~2274 (2014)
- Polcari I, "Filaggrin gene mutations in African Americans with both ichthyosis vulgaris and atopic dermatitis", Pediatr Dermatol, 31, pp.489~492 (2014)
- Thawer-Esmail F, "South African amaXhosa patients with atopic dermatitis have decreased levels of filaggrin breakdown products but no loss-of-function mutations in filaggrin", J Allergy Clin Immunol, 133, pp.280~282 (2014)
- Berger SL, "An operational definition of epigenetics", Genes Dev, 23, pp.781~783 (2009)
- Jaenisch R, "Epigenetic regulation of gene expression : how the genome integrates intrinsic and environmental signals", Nat Genet, 33(Suppl), pp.245~254 (2003)
- Botchkarev VA, "Epigenetic regulation of gene expression in keratinocytes", J Invest Dermatol, 132, pp.2505~2521 (2012)
- Schübeler D, "Function and information content of DNA methylation", Nature, 517, pp.321~326 (2015)
- Jones PA, "Functions of DNA methylation: islands, start sites, gene bodies and beyond", Nat Rev Genet, 13, pp.484~492 (2012)
- Li Y, "Aberrant DNA methylation in skin diseases", J Dermatol Sci, 54, pp.143~149 (2009)
- Ziyab AH, "DNA methylation of the filaggrin gene adds to the risk of eczema associated with loss-of-function variants", J Eur Acad Dermatol Venereol, 27, pp.e420~e423 (2013)
- Lee J, "DNA methylation-mediated downregulation of DEFB1 in prostate cancer cells", PLoS One, 11, pp.e0166664~ (2016)
- Alaskhar Alhamwe B, "Histone modifications and their role in epigenetics of atopy and allergic diseases", Allergy Asthma Clin Immunol, 14, pp.39~ (2018)
- Bhardwaj N, "MicroRNAs in atopic dermatitis : a review", J Transl Genet Genom, 1, pp.15~22 (2017)
- Gardiner-Garden M, "CpG islands in vertebrate genomes", J Mol Biol, 196, pp.261~282 (1987)
- Stadler MB, "DNA-binding factors shape the mouse methylome at distal regulatory regions", Nature, 480, pp.490~495 (2011)
- Ziller MJ, "Charting a dynamic DNA methylation landscape of the human genome", Nature, 500, pp.477~481 (2013)
- Rodríguez E, "An integrated epigenetic and transcriptomic analysis reveals distinct tissue-specific patterns of DNA methylation associated with atopic dermatitis", J Invest Dermatol, 134, pp.1873~1883 (2014)
- Tan HT, "Methylation of the filaggrin gene promoter does not affect gene expression and allergy", Pediatr Allergy Immunol, 25, pp.608~610 (2014)
- Weber M, "Distribution, silencing potential and evolutionary impact of promoter DNA methylation in the human genome", Nat Genet, 39, pp.457~466 (2007)
- Meissner A, "Genome-scale DNA methylation maps of pluripotent and differentiated cells", Nature, 454, pp.766~770 (2008)
- Kezic S, "Levels of filaggrin degradation products are influenced by both filaggrin genotype and atopic dermatitis severity", Allergy, 66, pp.934~940 (2011)
- Bataille V, "The use of the twin model to investigate the genetics and epigenetics of skin diseases with genomic, transcriptomic and methylation data", J Eur Acad Dermatol Venereol, 26, pp.1067~1073 (2012)
- Tost J, "Analysis and quantification of multiple methylation variable positions in CpG islands by Pyrosequencing", Biotechniques, 35, pp.152~156 (2003)
- Rakyan VK, "Epigenome-wide association studies for common human diseases", Nat Rev Genet, 12, pp.529~541 (2011)
- Rakyan VK, "DNA methylation profiling of the human major histocompatibility complex : a pilot study for the human epigenome project", PLoS Biol, 2, pp.e405~ (2004)
- Huang Q, "Methylation variable position profiles of hMLH1 promoter CpG islands in human sporadic colorectal carcinoma", Diagn Mol Pathol, 21, pp.24~33 (2012)