The Role of Bacterial Toxins in Human Skin Disease

细菌毒素在人类皮肤病中的作用

• 批准号: 9751642
• 负责人: Elena Goleva
• 金额: $ 33.11万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-07-17 至 2021-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9751642
• 关键词: 3-Dimensional; Adult; Affect; Allergens; Ankyrin Repeat; Ankyrins; Applications Grants; Asthma; Atopic Dermatitis; Bacterial Toxins; Binding; Binding Proteins; Calcium Binding; Cells; Cellular biology; Child; Complement; Cysteine; Data; Defect; Development; Diagnosis; Differentiation Antigens; Disease; Down-Regulation; Environment; Epidermis; Event; Exposure to; Expression Profiling; Food Hypersensitivity; Gene Expression; Gene Silencing; Gene Targeting; General Population; Genes; Genetic Transcription; Genus staphylococcus; Goals; Host Defense; Human; Hypersensitivity; Immune response; Infection; Inflammation; Interferon Type II; Interleukin-13; Interleukin-4; Laboratories; Lipids; Modality; Molecular; Monitor; Morbidity - disease rate; Notch Signaling Pathway; Occupational; Pathway interactions; Patients; Penetration; Process; Production; Program Sustainability; Proteins; Psoriasis; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; STC1 gene; STC2 gene; Severities; Signal Pathway; Signal Transduction; Skin; Skin colonization; Small Interfering RNA; Spinal; Staphylococcal Enterotoxin B; Staphylococcus aureus; Stratum Basale; Stratum Granulosum; System; Techniques; Therapeutic; Toxin; Undifferentiated; Up-Regulation; Western Blotting; antimicrobial; antimicrobial peptide; beta catenin; biomarker development; chronic inflammatory skin; cytokine; differential expression; disability; in vivo; inflammatory milieu; inhibitor/antagonist; interleukin-22; keratinization; keratinocyte; keratinocyte differentiation; knock-down; laser capture microdissection; lipoteichoic acid; notch protein; novel; novel strategies; novel therapeutics; overexpression; programs; promoter; protein expression; protein function; public health relevance; release of sequestered calcium ion into cytoplasm; respiratory; skin barrier; skin disorder; transcription factor; transcriptome; transcriptome sequencing; uptake

 DESCRIPTION (provided by applicant): Atopic dermatitis (AD) is the most common chronic inflammatory skin disease in the general population affecting 17% of children and nearly 3% of adults in the U.S. It is associated with significant morbidity and occupational disability. Recent studies have highlighted the importance of cytokine environment in the pathobiology of AD and Staphylococcus aureus colonization/infection contributing to the severity/exacerbation of this common disease. The skin of AD patients has significantly reduced epidermal barrier protein expression and low levels of antimicrobial peptide production, which result in abnormalities in the skin barrier function and antimicrobial host defense. In the majority of AD, these changes are caused by inhibition of epidermal keratinocyte differentiation. However, the molecular events that result in lack of keratinocyte differentiation in AD skin are poorly understood. The overall goal of this competing renewal of R01 grant application (5 R01 AR41256-25) will be to delineate the initial molecular and cellular events by which IL-4/IL-13 and IL-22, cytokines associated with AD skin environment, and staphylococcal toxins inhibit keratinocyte differentiation in AD skin leading to epidermal barrier abnormalities. Our new preliminary data indicates that both IL-4/IL-13 and IL-22 through different signaling pathways interfere with early differentiation events in keratinocytes by affecting Ca2+ signaling/mobilization, activating Wnt/beta-catenin pathway, while inhibiting Notch developmental pathway. We present evidence that staphylococcal lipoteichoic acid, LTA, selectively activates the expression of Notch- Regulated Ankyrin Repeat Protein, NRARP, that interferes with Notch pathway and inhibits the expression of early keratinocyte differentiation markers. Finally, we demonstrate that AD skin cytokine environment and staphylococcal products synergize and complement each other in the inhibition of keratinocyte differentiation. The specific aims of this proposal will be: 1) To establish the molecular mechanisms by which the immune response in AD skin alters differentiation of skin keratinocytes by examining gene and protein expression profiling in keratinocytes differentiated in the IL-4/IL-13 vs IL-22 vs IFNg environment; analyzing the role of the Ca2+- binding proteins, SMOC1 and STC2, regulated by IL-4/IL-13 in Ca2+ mobilization and signaling in keratinocytes; evaluating how interference with Ca2+ binding proteins expression and function may control early stages of keratinocyte differentiation and affect Wnt5a/beta-catenin signaling pathway in these cells. 2) To investigate the effects of staphylococcal toxins on keratinocyte differentiation In this aim, we will examine LTA interference with Notch signaling pathway, which controls keratinocyte differentiation. We will assess the role of LTA-induced NRARP in this process. 3) To investigate the synergistic effects of the AD immune response and staphylococcal LTA, on the keratinocyte differentiation program in vivo. We will assess keratinocyte differentiation program in the skin of AD patients in relationship to skin cytokine environment and colonization with S. aureus, examine transcriptional profile of different layers of keratinocytes in the skin by laser capture microdissection and RNAseq, evaluate keratinocyte differentiation in 3D organotypic skin cultures utilizing primary keratinocytes of AD patients and examine how differential expression of Ca2+ binding proteins SMOC1, STC2 and Notch regulating NRARP in these cultures influences keratinocyte differentiation. These studies will likely identify novel selective therapeutic approaches that can alter the keratinocyte differentiation program in AD skin at its earliest stages, providing the opportunity to restore and develop new approaches to enhance epidermal barrier function in AD.

 描述（由申请人提供）：特应性皮炎（AD）是普通人群中最常见的慢性炎症性皮肤病，影响美国17%的儿童和近3%的成人。最近的研究已经强调了细胞因子环境在AD的病理生物学中的重要性，并且金黄色葡萄球菌定殖/感染有助于这种常见疾病的严重性/恶化。AD患者的皮肤具有显著降低的表皮屏障蛋白表达和低水平的抗菌肽产生，这导致皮肤屏障功能和抗菌宿主防御的异常。在大多数AD中，这些变化是由表皮角质形成细胞分化抑制引起的。然而，导致AD皮肤中角质形成细胞分化缺乏的分子事件知之甚少。R 01授权申请（5 R 01 AR 41256 -25）的竞争性更新的总体目标是描述IL-4/IL-13和IL-22、与AD皮肤环境相关的细胞因子和葡萄球菌毒素抑制AD皮肤中角质形成细胞分化导致表皮屏障异常的初始分子和细胞事件。我们的新的初步数据表明，IL-4/IL-13和IL-22通过不同的信号通路，通过影响Ca 2+信号/动员，激活Wnt/β-catenin通路，同时抑制Notch发育通路，干扰角质形成细胞的早期分化事件。我们提供了葡萄球菌脂磷壁酸（LTA）选择性激活Notch调节的锚蛋白重复序列蛋白（NRARP）的表达的证据，NRARP干扰Notch途径并抑制早期角质形成细胞分化标志物的表达。最后，我们证明了AD皮肤细胞因子环境和葡萄球菌产物在抑制角质形成细胞分化中相互协同和互补。1）通过检测在IL-4/IL-13 vs IL-22 vs IFNg环境中分化的角质形成细胞中的基因和蛋白质表达谱，建立AD皮肤中的免疫应答改变皮肤角质形成细胞分化的分子机制;分析由IL-4/IL-13调节的Ca 2+结合蛋白SMOC 1和STC 2在角质形成细胞中Ca 2+动员和信号传导中的作用;评估干扰Ca 2+结合蛋白的表达和功能如何控制角质形成细胞分化的早期阶段并影响这些细胞中的Wnt 5a/β-连环蛋白信号通路。2)为了研究葡萄球菌毒素对角质形成细胞分化的影响，我们将研究LTA对控制角质形成细胞分化的Notch信号通路的干扰。我们将评估LTA诱导的NRARP在这一过程中的作用。3)研究AD免疫应答和葡萄球菌LTA对体内角质形成细胞分化程序的协同作用。我们将评估AD患者皮肤中角质形成细胞分化程序与皮肤细胞因子环境和S。金黄色葡萄球菌，检查皮肤中角质形成细胞的不同层的转录谱， 激光捕获显微切割和RNAseq，在利用AD患者的原代角质形成细胞的3D器官型皮肤培养物中评估角质形成细胞分化，并检查这些培养物中调节NRARP的Ca 2+结合蛋白SMOC 1、STC 2和Notch的差异表达如何影响角质形成细胞分化。这些研究可能会确定新的选择性治疗方法，可以在AD皮肤的最早阶段改变角质形成细胞分化程序，为恢复和开发增强AD表皮屏障功能的新方法提供机会。