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

基本信息

批准号：
10511569
负责人：
Irina Budunova
金额：
$ 21.77万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
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/10511569
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 Fibroblasts Functional disorder Gene Expression Gene Proteins 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 Molecular Molecular Profiling Morphology Not Hispanic or Latino Pathway interactions Patients Phase Pigments Population Predisposition Prevention Prevention approach Process Proteomics Psoriasis Publishing Risk Role Signal Transduction Skin Skin Physiology T-Lymphocyte TNF gene Testing Thick Underrepresented Minority Up-Regulation Western Blotting cohesion cytokine defense response experimental study healthy volunteer imager individualized prevention keratinocyte keratinocyte differentiation liquid crystal polymer molecular phenotype protein biomarkers protein expression prototype receptor response skin barrier skin color skin disorder skin lesion three-dimensional modeling transcriptome transcriptome sequencing

项目摘要

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) 人群的比例更高不同炎症皮肤易感性差异的机制。在我们的试点实验中，我们使用经过普遍验证的RNA-seq分析来了解疾病。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初级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然而，引发这种“矛盾”疾病前期转变的分子机制。 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。 AA和WNH健康肌肤定义分子开关促 AD 或促银屑病信号传导，我们可以描绘出最初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我们建议使用综合方法，包括RNA-seq、Q-PCR、邻近法。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我们将在使用 AD 或牛皮癣相关细胞因子治疗期间将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 的一个组成部分我们期望所获得的结果将揭示是否以及如何可能。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，