Linking Nicotinic Activation with Skin Innate Immunity and Atopic Dermatitis

将烟碱激活与皮肤先天免疫和特应性皮炎联系起来

• 批准号: 8836391
• 负责人: Katherine Amanda Radek
• 金额: $ 33.64万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8836391
• 关键词: Acetylcholine; Address; Advanced Development; Agonist; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotics; Area; Atopic Dermatitis; Bacteria; Bacterial Infections; Biochemical; Biological Models; Cells; Characteristics; Chromogranin A; Clinical; Cutaneous; Data; Defect; Development; Disease; Disease Progression; Economic Burden; Environment; Epidermis; Epithelial; Failure; Focal Infection; Functional disorder; Goals; Health; Homeostasis; Host Defense; Human; Immune; Immune response; Immunosuppression; In Vitro; Individual; Infection; Inflammation; Inflammatory; Injury; Knowledge; Link; Measures; Mediating; Microbe; Mission; Modality; Molecular; Multi-Drug Resistance; Mus; Natural Immunity; Neurosecretory Systems; Nicotinic Receptors; Pathogenesis; Pathway interactions; Patients; Play; Predisposition; Process; Production; Proteins; Proteomics; Public Health; Regimen; Regulation; Relative (related person); Research; Resistance; Role; Signal Transduction; Skin; Staphylococcus aureus; Stress; Symptoms; System; Techniques; Testing; Therapeutic Intervention; Tissues; Work; antimicrobial; antimicrobial peptide; base; biological adaptation to stress; cathelicidin; cholinergic; chromogranin A (344-364); clinically relevant; effective therapy; improved; in vivo; innovation; keratinocyte; microbial; microorganism; neuroimmunology; novel; relating to nervous system; response; skin disorder; social

DESCRIPTION (provided by applicant): An important breach exists in our understanding of how the negative regulation of antimicrobial peptides (AMPs) leads to the clinical symptoms associated with skin disease. The persistent lack of knowledge in this area of research signifies an important challenge to develop improved therapies for cutaneous infection and disease. The long-term goal is to identify how acetylcholine nicotinic receptor (nAChR) activation suppresses AMP expression and activity, and relate these changes in AMP regulation to the causal treatment of human skin diseases associated with AMP dysregulation. The objective of the proposed research is to identify the nAChR subtype(s) involved in AMP suppression in keratinocytes, evaluate the AMP profile altered by nAChR activation, and link this mechanism to the pathogenesis of Atopic Dermatitis (AD). Our hypothesis is that excess nAChR activation in keratinocytes impairs the normal processing and function of cutaneous AMPs, which contributes to the pathogenesis of AD. Our hypothesis was formulated based on preliminary data establishing that nAChR activation significantly reduced AMP activity and the resistance to cutaneous infection. Patients with AD develop a compromised skin barrier, which includes decreased AMP expression thought to precipitate inflammation and infection. Stress likely exacerbates the clinical manifestations of AD by increasing epidermal ACh and, consequently, nAChR activation to compromise normal AMP regulation and microbial susceptibility. Our rationale for these studies is that most research has focused on the positive regulation of AMPs to increase their expression and activity, yet identifying those molecules that inhibit or diminish AMP activity is critical to develop better clinical remedies to ameliorate the symptoms associated with AD and other inflammatory skin diseases. Driven by compelling preliminary data, our hypothesis will be evaluated by addressing three Specific Aims: 1) Identify which nAChR subtypes are involved in AMP suppression and identify the AMP profile in primary normal human epidermal keratinocytes (NHEKs) in vitro; 2) Determine the role of nAChR activation in the AMP response to infection in vivo; 3) Link mechanisms of nAChR signaling to the pathogenesis of AD. For Aim 1, NHEKs will be stimulated with nAChR agonists and antagonists to assess known AMPs and identify new antimicrobial molecules using established biochemical techniques. Aim 2 will use mice with altered states of nAChR activation to analyze the AMP response to infection using established proteomic approaches. Aim 3 will demonstrate that nAChR activation participates in the suppression of the AMP response to infection in skin from AD patients using molecular approaches. Our approach is innovative because it utilizes novel proteomic and molecular techniques to further define how nAChR activation influences the epidermal AMP response to infection. The proposed studies are significant because they are anticipated to identify the major nAChR signaling mechanism that negatively regulates AMP activity in skin, and identify alternative pathways for disease progression.

描述（由申请人提供）：我们对抗菌肽（AMP）负调控如何导致与皮肤病相关的临床症状的理解中存在着重要的违规行为。在这一研究领域，缺乏知识的持续缺乏，这是开发皮肤感染和疾病疗法的重要挑战。长期的目标是确定乙酰胆碱烟碱受体（NACHR）激活如何抑制AMP的表达和活性，并将AMP调节中的这些变化与与AMP失调相关的人类皮肤疾病的因果治疗相关。拟议的研究的目的是确定参与角质形成细胞中AMP抑制的NACHR亚型，评估随NACHR激活改变的AMP谱，并将此机制与特应性皮炎（AD）的发病机理联系起来。我们的假设是，角质形成细胞中的过量NACHR激活会损害皮肤AMP的正常处理和功能，这有助于AD的发病机理。我们的假设是根据初步数据提出的，该数据确定NACHR激活显着降低了AMP活性和对皮肤感染的抗性。 AD患者出现了受损的皮肤屏障，其中包括降低的AMP表达被认为会沉淀炎症和感染。压力可能通过增加表皮ACH，从而加剧AD的临床表现，从而损害正常AMP调节和微生物敏感性，从而加剧AD的临床表现。我们对这些研究的理由是，大多数研究都集中在AMP的积极调节上，以增加其表达和活性，但识别抑制或减少的分子 AMP活性对于开发更好的临床补救措施至关重要，以减轻与AD和其他炎症性皮肤疾病相关的症状。在引人注目的初步数据的驱动下，我们的假设将通过解决三个特定目的来评估：1）确定哪些NACHR亚型参与了AMP抑制，并在体外识别原发性人类表皮角质形成细胞（NHEKS）中的AMP谱。 2）确定NACHR激活在AMP对体内感染反应中的作用； 3）NACHR信号的机理与AD的发病机理。对于AIM 1，NACHR激动剂和拮抗剂将刺激NHEKS，以评估已知的AMP并使用已建立的生化技术鉴定新的抗菌分子。 AIM 2将使用改变NACHR激活状态的小鼠使用已建立的蛋白质组学方法分析AMP对感染的反应。 AIM 3将表明，使用分子方法，NACHR激活参与AD患者对皮肤感染的AMP反应。我们的方法具有创新性，因为它利用新型的蛋白质组学和分子技术来进一步定义NACHR激活如何影响表皮AMP对感染的反应。拟议的研究之所以重要，是因为预计它们可以识别负调节皮肤中AMP活性的主要NACHR信号传导机制，并确定疾病进展的替代途径。