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)、化脓性汗腺炎和红斑狼疮的风险显着 非裔美国人 (AA) 人群的比例较高，而非西班牙裔白人 (WNH) 人群的比例更高 牛皮癣的风险。不同炎症皮肤易感性差异的机制 人们对疾病知之甚少。在我们的试点实验中，使用经过广泛验证的 RNA-seq 分析 我们发现健康 AA 皮肤与 WNH 皮肤的基因表达存在显着差异，富含 促炎症信号传导。此外，由人体制成的 3D 人体皮肤等效培养物 (HSE) 接种在胶原蛋白基质上的原代角质形成细胞似乎对 TNFa 的促炎作用，一种参与多种炎症性皮肤的原型细胞因子 疾病。这些结果表明 AA 皮肤/角质形成细胞中的内在促炎回路可能 导致 AA 人群中某些炎症性皮肤病的增加 包括AD。然而，引发这种“矛盾”的疾病前期转变的分子机制 针对特定炎症性皮肤病的炎症信号传导仍有待研究。 3D健康安全环境 由原代人类角质形成细胞和免疫细胞以及经过处理的皮肤外植体培养物制成 Th1/Th17 细胞因子（TNFa、IL17、IL22）或 Th2 细胞因子（IL4、IL13）已成功用于 诱导银屑病和 AD 典型的体外形态和分子变化。因此，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 朝向促 AD 或促银屑病信号传导，我们可以描绘初始的重要阶段 在此过程中使用 AA 和 WNH 3D HSE 以及经过 Th1/Th17 处理的皮肤外植体培养物 (TNFa+IL17+IL22) 或 Th2 (IL3+IL14) 细胞因子。我们建议使用体外皮肤模型：3D HSE 制作 来自 AA 和 WNH 成人皮肤细胞（来自同一供体的角质形成细胞、成纤维细胞和 T 细胞）和 西北 SBDRC 的 STEM 核心提供 AA 和 WNH 人类皮肤的外植体培养物 细胞因子治疗。我们建议使用综合方法，包括 RNA-seq、Q-PCR、proximity 延伸免疫 PCR 测定（Olink 蛋白质组学），通过 Vectra 免疫染色证实 多光谱成像仪可识别 3D HSE/皮肤外植体基因/蛋白质表达变化的开始 在使用 AD 或银屑病相关细胞因子治疗期间。我们将把这些变化与已经发生的变化进行比较 识别/发表了 AD 和牛皮癣患者非病变和病变皮肤的分子特征。 我们还将确定表皮屏障的变化，这是牛皮癣和 AD 的一个组成部分 病理生理学。我们期望所获得的结果将揭示是否以及可能如何 健康皮肤的分子表型定义了向特定皮肤疾病的转变，并将奠定基础 为不同的少数群体制定个性化的预防方法，