Role of healthy skin molecular phenotype in the switch to specific skin diseases

健康皮肤分子表型在向特定皮肤病转变中的作用

• 批准号: 10709874
• 负责人: Irina Budunova
• 金额: $ 18.21万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-23 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10709874
• 关键词: 3-Dimensional; Adult; African American; African American population; Atopic Dermatitis; Automobile Driving; Biological Assay; Biopsy; Cells; Coculture Techniques; Collagen; Dermatologic; Dermatology; Development; Disease; Disparity; Fibroblasts; Functional disorder; Gene Expression; Gene set enrichment analysis; Genes; Goals; Hidradenitis Suppurativa; Host Defense; Human; IL14 gene; IL17 gene; IL3 Gene; IL4 gene; IgE Receptors; Immune; Immunoglobulins; In Vitro; Incidence; Inflammation; Inflammatory; Interferons; Interleukin-13; Investigation; Lupus Erythematosus; Maps; Minority Groups; Minority Health Research; Modeling; Molecular; Molecular Profiling; Morphology; Not Hispanic or Latino; Pathway interactions; Patients; Phase; Population; Predisposition; Prevention; Prevention approach; Process; Proteomics; Psoriasis; Publishing; Risk; Role; Signal Induction; Signal Transduction; Skin; Skin Physiology; Skin Pigmentation; T-Lymphocyte; Testing; Thick; Underrepresented Minority; Up-Regulation; Western Blotting; cohesion; cytokine; defense response; experimental study; healthy volunteer; imager; individualized prevention; interleukin-22; keratinization; keratinocyte; keratinocyte differentiation; liquid crystal polymer; molecular phenotype; protein biomarkers; protein expression; prototype; receptor; response; skin barrier; skin color; skin disorder; transcriptome; transcriptome sequencing

The risk of atopic dermatitis (AD), hidradenitis suppurativa, and lupus erythematosus is significantly higher in African American (AA) population, while White Non-Hispanic (WNH) subjects have a greater risk of psoriasis. The mechanisms that underlie disparity in susceptibility to different inflammatory skin diseases are poorly understood. In our pilot experiments, using extensively validated RNA-seq analysis we revealed striking differences in gene expression in healthy AA versus WNH skin, enriched for proinflammatory signaling. In addition, 3D human skin equivalent cultures (HSE) made from human primary keratinocytes seeded on collagen matrix, appeared to have a much more robust response to the pro-inflammatory effects of TNFa, a prototype cytokine involved in multiple inflammatory skin diseases. These results suggested that intrinsic pro-inflammatory circuits in AA skin/keratinocytes may contribute to the increased development of certain inflammatory skin diseases in the AA population including AD. However, the molecular mechanisms that trigger the shift of this “ambivalent” pre-disease inflammatory signaling towards specific inflammatory skin diseases remain to be investigated. 3D HSE made from primary human keratinocytes and immune cells as well as skin explant cultures treated with Th1/Th17 cytokines (TNFa, IL17, IL22) or Th2 cytokines (IL4, IL13) have been successfully used to induce in vitro morphological and molecular changes typical for psoriasis and AD. Thus, we hypothesized that different molecular phenotypes of AA and WNH healthy skin define molecular switch towards either pro-AD or pro-psoriasis signaling and that we could delineate the initial significant stages in this process using AA and WNH 3D HSE and skin explant cultures treated with Th1/Th17 (TNFa+IL17+IL22) or Th2 (IL3+IL14) cytokines. We propose to use in vitro skin models: 3D HSE made from AA and WNH adult skin cells (keratinocytes, fibroblasts and T cells from the same donor) and explant cultures of AA and WNH human skin provided by STEM Core at Northwestern SBDRC for cytokine treatment. We propose to use comprehensive approach including RNA-seq, Q-PCR, proximity extension immuno-PCR assay (Olink proteomics), confirmed by immunostaining with Vectra Multispectral Imager to identify onset of changes in 3D HSEs/skin explants gene/protein expression during the treatment with AD- or psoriasis-related cytokines. We will compare these changes to already identified/published molecular signatures of non-lesional and lesional skin in AD and psoriasis patients. We will also determine changes in epidermal barrier that are an integral part of psoriasis and AD pathophysiology. We expect that the obtained results will reveal whether and potentially how the molecular phenotype of healthy skin defines the switch to specific skin diseases and will set the stage for the development of personalized prevention approaches for different minority populations,

特应性皮炎（AD），Hidradenitis purativa和狼疮红斑的风险显着 非洲裔美国人（AA）人口较高，而白人非西班牙裔（WNH）受试者的受试者更大 牛皮癣的风险。对不同炎症性皮肤敏感的差异构成的机制 疾病知之甚少。在我们的试点实验中，使用经过广泛验证的RNA-seq分析 我们揭示了健康AA与WNH皮肤中基因表达的差异，富含 促炎信号传导。此外，由人类制成的3D人类皮肤等效培养（HSE） 胶原蛋白基质的原代角质形成细胞似乎对 TNFA的促炎作用，一种与多种炎症皮肤有关的原型细胞因子 疾病。这些结果表明，AA皮肤/角质形成细胞中的内在促炎电路可能 有助于增加AA种群中某些炎症性皮肤疾病的发展 包括广告。但是，触发这种“矛盾”前疾病的转移的分子机制 针对特定炎症性皮肤疾病的炎症信号传导仍有待研究。 3D HSE 由原代人角质形成细胞和免疫细胞以及用皮肤外植体培养 TH1/TH17细胞因子（TNFA，IL17，IL22）或TH2细胞因子（IL4，IL13）已成功地用于 诱导牛皮癣和AD典型的体外形态和分子变化。那，我们 假设AA和WNH健康皮肤的不同分子表型定义了分子开关 朝着亲帕或亲孢子症信号传导，我们可以描述最初的重要阶段 在此过程中，使用th1/th17处理的AA和WNH 3D HSE和皮肤外植体培养物 （TNFA+IL17+IL22）或TH2（IL3+IL14）细胞因子。我们建议使用体外皮肤模型：制造3D HSE 来自AA和WNH成年皮细胞（角质形成细胞，成纤维细胞和来自同一供体的T细胞）和 西北SBDRC提供的茎核提供的AA和WNH人皮肤的外植体培养 细胞因子治疗。我们建议使用包括RNA-Seq，Q-PCR，接近性在内的全面方法 扩展免疫PCR分析（Olink蛋白质组学），通过用Vectra进行免疫染色证实 多光谱成像仪以识别3D HSE/皮肤外植体基因/蛋白质表达的变化发作 在用AD或牛皮癣相关的细胞因子治疗期间。我们将将这些更改与已经 在AD和牛皮癣患者中鉴定出/发表的非疗法皮肤和病变皮肤的分子特征。 我们还将确定表皮屏障的变化是牛皮癣和AD的组成部分 病理生理学。我们希望获得的结果将揭示是否以及可能是如何 健康皮肤的分子表型定义了转向特定的皮肤疾病，并将设置舞台 为了开发针对不同少数民族的个性化预防方法，