Identification of novel immune and barrier genes in atopic dermatitis by means of laser capture microdissection

Hitokazu Esaki, David A. Ewald, Benjamin Ungar, Mariya Rozenblit, Xiuzhong Zheng, Hui Xu, Yeriel D. Estrada, Xiangyu Peng, Hiroshi Mitsui, Thomas Litman, Mayte Suárez-Fariñas, James G. Krueger, Emma Guttman-Yassky

Research output: Contribution to journalArticle

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Abstract

Background The molecular signature of atopic dermatitis (AD) lesions is associated with TH2 and TH22 activation and epidermal alterations. However, the epidermal and dermal AD transcriptomes and their respective contributions to abnormalities in respective immune and barrier phenotypes are unknown.

Objective We sought to establish the genomic profile of the epidermal and dermal compartments of lesional and nonlesional AD skin compared with normal skin.

Methods Laser capture microdissection was performed to separate the epidermis and dermis of lesional and nonlesional skin from patients with AD and normal skin from healthy volunteers, followed by gene expression (microarrays and real-time PCR) and immunostaining studies.

Results Our study identified novel immune and barrier genes, including the IL-34 cytokine and claudins 4 and 8, and showed increased detection of key AD genes usually undetectable on arrays (ie, IL22, thymic stromal lymphopoietin [TSLP], CCL22, and CCL26). Overall, the combined epidermal and dermal transcriptomes enlarged the AD transcriptome, adding 674 upregulated and 405 downregulated differentially expressed genes between lesional and nonlesional skin to the AD transcriptome. We were also able to localize individual transcripts as primarily epidermal (defensin, beta 4A [DEFB4A]) or dermal (IL22, cytotoxic T-lymphocyte antigen 4 [CTLA4], and CCR7) and link their expressions to possible cellular sources.

Conclusions This is the first report that establishes robust epidermal and dermal genomic signatures of lesional and nonlesional AD skin and normal skin compared with whole tissues. These data establish the utility of laser capture microdissection to separate different compartments and cellular subsets in patients with AD, allowing localization of key barrier or immune molecules and enabling detection of gene products usually not detected on arrays.

Original languageEnglish
Pages (from-to)153-163
Number of pages11
JournalJournal of Allergy and Clinical Immunology
Volume135
Issue number1
DOIs
Publication statusPublished - Jan 1 2015

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Laser Capture Microdissection
Atopic Dermatitis
Skin
Genes
Transcriptome
Claudins
CTLA-4 Antigen
Defensins
Dermis
Epidermis
Real-Time Polymerase Chain Reaction
Healthy Volunteers

All Science Journal Classification (ASJC) codes

  • Immunology and Allergy
  • Immunology

Cite this

Identification of novel immune and barrier genes in atopic dermatitis by means of laser capture microdissection. / Esaki, Hitokazu; Ewald, David A.; Ungar, Benjamin; Rozenblit, Mariya; Zheng, Xiuzhong; Xu, Hui; Estrada, Yeriel D.; Peng, Xiangyu; Mitsui, Hiroshi; Litman, Thomas; Suárez-Fariñas, Mayte; Krueger, James G.; Guttman-Yassky, Emma.

In: Journal of Allergy and Clinical Immunology, Vol. 135, No. 1, 01.01.2015, p. 153-163.

Research output: Contribution to journalArticle

Esaki, H, Ewald, DA, Ungar, B, Rozenblit, M, Zheng, X, Xu, H, Estrada, YD, Peng, X, Mitsui, H, Litman, T, Suárez-Fariñas, M, Krueger, JG & Guttman-Yassky, E 2015, 'Identification of novel immune and barrier genes in atopic dermatitis by means of laser capture microdissection', Journal of Allergy and Clinical Immunology, vol. 135, no. 1, pp. 153-163. https://doi.org/10.1016/j.jaci.2014.10.037
Esaki, Hitokazu ; Ewald, David A. ; Ungar, Benjamin ; Rozenblit, Mariya ; Zheng, Xiuzhong ; Xu, Hui ; Estrada, Yeriel D. ; Peng, Xiangyu ; Mitsui, Hiroshi ; Litman, Thomas ; Suárez-Fariñas, Mayte ; Krueger, James G. ; Guttman-Yassky, Emma. / Identification of novel immune and barrier genes in atopic dermatitis by means of laser capture microdissection. In: Journal of Allergy and Clinical Immunology. 2015 ; Vol. 135, No. 1. pp. 153-163.
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abstract = "Background The molecular signature of atopic dermatitis (AD) lesions is associated with TH2 and TH22 activation and epidermal alterations. However, the epidermal and dermal AD transcriptomes and their respective contributions to abnormalities in respective immune and barrier phenotypes are unknown.Objective We sought to establish the genomic profile of the epidermal and dermal compartments of lesional and nonlesional AD skin compared with normal skin.Methods Laser capture microdissection was performed to separate the epidermis and dermis of lesional and nonlesional skin from patients with AD and normal skin from healthy volunteers, followed by gene expression (microarrays and real-time PCR) and immunostaining studies.Results Our study identified novel immune and barrier genes, including the IL-34 cytokine and claudins 4 and 8, and showed increased detection of key AD genes usually undetectable on arrays (ie, IL22, thymic stromal lymphopoietin [TSLP], CCL22, and CCL26). Overall, the combined epidermal and dermal transcriptomes enlarged the AD transcriptome, adding 674 upregulated and 405 downregulated differentially expressed genes between lesional and nonlesional skin to the AD transcriptome. We were also able to localize individual transcripts as primarily epidermal (defensin, beta 4A [DEFB4A]) or dermal (IL22, cytotoxic T-lymphocyte antigen 4 [CTLA4], and CCR7) and link their expressions to possible cellular sources.Conclusions This is the first report that establishes robust epidermal and dermal genomic signatures of lesional and nonlesional AD skin and normal skin compared with whole tissues. These data establish the utility of laser capture microdissection to separate different compartments and cellular subsets in patients with AD, allowing localization of key barrier or immune molecules and enabling detection of gene products usually not detected on arrays.",
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AU - Esaki, Hitokazu

AU - Ewald, David A.

AU - Ungar, Benjamin

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AU - Zheng, Xiuzhong

AU - Xu, Hui

AU - Estrada, Yeriel D.

AU - Peng, Xiangyu

AU - Mitsui, Hiroshi

AU - Litman, Thomas

AU - Suárez-Fariñas, Mayte

AU - Krueger, James G.

AU - Guttman-Yassky, Emma

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N2 - Background The molecular signature of atopic dermatitis (AD) lesions is associated with TH2 and TH22 activation and epidermal alterations. However, the epidermal and dermal AD transcriptomes and their respective contributions to abnormalities in respective immune and barrier phenotypes are unknown.Objective We sought to establish the genomic profile of the epidermal and dermal compartments of lesional and nonlesional AD skin compared with normal skin.Methods Laser capture microdissection was performed to separate the epidermis and dermis of lesional and nonlesional skin from patients with AD and normal skin from healthy volunteers, followed by gene expression (microarrays and real-time PCR) and immunostaining studies.Results Our study identified novel immune and barrier genes, including the IL-34 cytokine and claudins 4 and 8, and showed increased detection of key AD genes usually undetectable on arrays (ie, IL22, thymic stromal lymphopoietin [TSLP], CCL22, and CCL26). Overall, the combined epidermal and dermal transcriptomes enlarged the AD transcriptome, adding 674 upregulated and 405 downregulated differentially expressed genes between lesional and nonlesional skin to the AD transcriptome. We were also able to localize individual transcripts as primarily epidermal (defensin, beta 4A [DEFB4A]) or dermal (IL22, cytotoxic T-lymphocyte antigen 4 [CTLA4], and CCR7) and link their expressions to possible cellular sources.Conclusions This is the first report that establishes robust epidermal and dermal genomic signatures of lesional and nonlesional AD skin and normal skin compared with whole tissues. These data establish the utility of laser capture microdissection to separate different compartments and cellular subsets in patients with AD, allowing localization of key barrier or immune molecules and enabling detection of gene products usually not detected on arrays.

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