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

基本信息

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
项目类别：
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

项目摘要

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）的负调节如何导致与皮肤病相关的临床症状的理解存在重大突破。该研究领域知识的持续缺乏意味着开发改进的皮肤感染和疾病疗法面临着重要挑战。长期目标是确定乙酰胆碱烟碱受体 (nAChR) 激活如何抑制 AMP 表达和活性，并将 AMP 调节的这些变化与与 AMP 失调相关的人类皮肤病的因果治疗联系起来。本研究的目的是确定参与角质形成细胞 AMP 抑制的 nAChR 亚型，评估 nAChR 激活改变的 AMP 谱，并将这种机制与特应性皮炎 (AD) 的发病机制联系起来。我们的假设是，角质形成细胞中 nAChR 的过度激活会损害皮肤 AMP 的正常处理和功能，从而导致 AD 的发病机制。我们的假设是根据初步数据制定的，该数据证实 nAChR 激活显着降低 AMP 活性和对皮肤感染的抵抗力。 AD 患者的皮肤屏障受损，其中 AMP 表达减少，被认为会加速炎症和感染。压力可能会通过增加表皮 ACh 以及 nAChR 的激活从而损害正常的 AMP 调节和微生物敏感性而加剧 AD 的临床表现。我们进行这些研究的基本原理是，大多数研究都集中在 AMP 的正向调节上，以增加其表达和活性，同时识别那些抑制或减弱的分子 AMP 活性对于开发更好的临床疗法以改善与 AD 和其他炎症性皮肤病相关的症状至关重要。在令人信服的初步数据的驱动下，我们的假设将通过解决三个具体目标进行评估：1）确定哪些 nAChR 亚型参与 AMP 抑制，并在体外确定原代正常人表皮角质形成细胞（NHEK）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信号传导与AD发病机制的联系机制。对于目标 1，将用 nAChR 激动剂和拮抗剂刺激 NHEK，以评估已知的 AMP 并使用现有的生化技术识别新的抗菌分子。目标 2 将使用 nAChR 激活状态改变的小鼠，利用已建立的蛋白质组学方法分析 AMP 对感染的反应。目标 3 将使用分子方法证明 nAChR 激活参与抑制 AD 患者皮肤感染的 AMP 反应。我们的方法是创新的，因为它利用新颖的蛋白质组学和分子技术来进一步确定 nAChR 激活如何影响表皮 AMP 对感染的反应。拟议的研究意义重大，因为它们预计将确定负调节皮肤中 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。