Amplification of IL-4Ralpha signaling pathways in human airways through 15 LO1

通过 15 LO1 放大人类气道中的 IL-4Rα 信号通路

基本信息

批准号：
8098018
负责人：
Sally E Wenzel
金额：
$ 36.64万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8098018
关键词：
1-Phosphatidylinositol 3-Kinase ADRBK1 gene Acute Address Adrenergic Agents Agonist Allergic inflammation Applications Grants Arachidonate 15-Lipoxygenase Asthma Binding Binding Proteins Cells Chronic Code Data Development Disease Down-Regulation Epithelial Cells Epithelium Ethanolamines Exposure to GTP-Binding Proteins Gene Expression Genes Genetic Polymorphism Genotype Human Hydroxyeicosatetraenoic Acids IL4R gene In Vitro Inflammation Interleukin-13 Interleukin-4 Lead Lipoxygenase 1 Location MEKs MUC5AC gene Mediating Mitogen-Activated Protein Kinase Kinases Molecular Pathway interactions Pharmaceutical Preparations Phosphatidylethanolamine Phosphatidylethanolamine Binding Protein Phosphorylation Phosphotransferases Play Process Proteins Proto-Oncogene Proteins c-raf Receptor Signaling Role Severities Signal Pathway Signal Transduction Site Up-Regulation adrenergic asthmatic airway base cytokine desensitization drug development extracellular in vivo innovation novel novel therapeutic intervention phosphatidylethanolamine public health relevance raf Kinases receptor response

项目摘要

DESCRIPTION (provided by applicant): The pathobiology of human asthma and severe asthma in particular, remains poorly understood, with considerable controversy as to the role and importance of Th2 inflammation. Our preliminary data suggest that 15 lipoxygenase (LO)1, its product 15 hydroxyeicosatetraenoic acid (HETE) and their interactions with phosphatidyl-ethanolamine (PE) binding protein (BP)1 may be central to promoting and amplifying Th2 inflammation and dampening ¿2 adrenoreceptor (AR) responses in the absence of high levels of Th2. Based on our preliminary data, we propose 3 aims to better understand the role that 15 HETE-PE/15 LO1, in combination with PEBP1, plays in amplification of IL-13/IL-4Ra signaling in human airway epithelial cells in vitro and ex vivo, and whether the process further impacts ¿2 receptor signaling. In Aim 1, we will determine whether prolonged exposure to IL-13 (and upregulation of 15 LO1) leads to epithelial cells in which MEK-ERK pathways are chronically activated, while PI3K pathways are downregulated and whether IL-4Ra polymorphisms impact these processes. We will address the role that 15 LO1/15 HETE-PE plays in this process and the impact this upregulation has on downstream gene expression, both in an acute (low 15 LO1) state, as well as in a chronic (high 15 LO1) state. We hypothesize that the presence of an active 15 LO1/15 HETE-PE pathway will greatly enhance IL-13 induced gene expression. Gene arrays will be evaluated under both acute and chronic stimulation, and in the presence and absence of knockdown, as well as pharmacologic inhibition to determine the overall impact of the 15 LO1/15 HETE-PE pathway. In Aim 2, we will address the mechanisms by which the 15 LO1/15 HETE-PE pathway interacts with the pERK pathway to modulate gene expression. This aim will focus exclusively on the interactions of 15 LO1/15 HETE-PE with PEBP1, further defining the subcellular location of the interaction, the mechanism of the physical interaction and the role for and mechanism by which phosphorylation of PEBP1 occurs. In Aim 3, we will determine whether the interaction of 15 HETE-PE with PEBP1 competitively inhibits the binding of GRK2 to PEBP1, leading to enhanced desensitization of 2¿AR in vitro and in vivo in human asthmatics. We believe that evaluating these processes in primary human cells, from asthmatic and normal subjects, will unravel a new and exciting pathway which appears to tightly regulate pERK, "Th2"-type responses and perhaps the ¿2AR pathway in asthmatic epithelial cells. We believe a better understanding of these interactions will open up many opportunities for new and novel therapeutic approaches. PUBLIC HEALTH RELEVANCE: Asthma is an extremely common disease which remains poorly understood. Although Th2 cytokines (IL-4/-13) have been believed to be important, initial studies using agents which block this pathway have been disappointing. Despite that, there is evidence in the airways of asthmatic people that portions of this pathway are highly active. This grant application will build on new and highly novel interactions identified in human asthmatic airway cells between the 15 lipoxygenase pathway and phosphatidylethanolamine binding protein-1. These interactions could serve to amplify and sustain a "Th2"-type of inflammation in the absence of high levels of IL-14/-13, as well as alter responses to b2 agonist drugs, mainstays of asthma therapy. Finally, these interactions open up new opportunities for innovative molecular approaches to drug development.

描述（由申请人提供）：人类哮喘尤其是严重哮喘的病理学仍然知之甚少，对于 Th2 炎症的作用和重要性存在相当大的争议。我们的初步数据表明 15 脂氧合酶 (LO)1（其产物 15）。羟基二十碳四烯酸 (HETE) 及其与磷脂酰乙醇胺 (PE) 结合蛋白 (BP)1 的相互作用可能是促进和放大 Th2 炎症和阻尼 ¿ 2 在缺乏高水平 Th2 的情况下肾上腺素受体 (AR) 反应根据我们的初步数据，我们提出 3 旨在更好地了解 15 HETE-PE/15 LO1 与 PEBP1 组合在 IL- 扩增中所起的作用。体外和离体人气道上皮细胞中的 13/IL-4Ra 信号传导，以及该过程是否进一步影响 ¿在目标 1 中，我们将确定长期暴露于 IL-13（以及 15 LO1 上调）是否会导致上皮细胞中 MEK-ERK 通路长期激活，而 PI3K 通路下调，以及 IL-4Ra 多态性是否存在。我们将讨论 15 LO1/15 HETE-PE 在此过程中发挥的作用以及这种上调对下游基因表达的影响，两者都在急性期。（低 15 LO1）状态以及慢性（高 15 LO1）状态我们发现，活性 15 LO1/15 HETE-PE 途径的存在将大大增强 IL-13 诱导的基因表达。在急性和慢性刺激、存在或不存在敲低以及药物抑制的情况下进行评估，以确定 15 LO1/15 HETE-PE 途径的总体影响。我们将探讨 15 LO1/15 HETE-PE 途径与 pERK 途径相互作用以调节基因表达的机制。这一目标将专门关注 15 LO1/15 HETE-PE 与 PEBP1 的相互作用，进一步研究 15 LO1/15 HETE-PE 的亚细胞定位。相互作用、物理相互作用的机制以及PEBP1磷酸化发生的作用和机制在目标3中，我们将确定15 HETE-PE是否与磷酸化发生相互作用。 PEBP1 竞争性抑制 GRK2 与 PEBP1 的结合，从而增强 2¿我们相信，在哮喘患者和正常受试者的原代人类细胞中评估这些过程，将揭示出一条新的、令人兴奋的途径，该途径似乎严格调节 pERK、“Th2”型反应，或许还有 pERK。 ¿我们相信，更好地了解这些相互作用将为开发新的治疗方法带来更多机会。公共健康相关性：哮喘是一种极其常见的疾病，人们对它知之甚少，尽管人们认为 Th2 细胞因子 (IL-4/-13) 很重要，但使用阻断该途径的药物进行的初步研究却令人失望。哮喘患者气道中的证据表明，该途径的某些部分高度活跃，这项拨款申请将建立在人类哮喘气道细胞中 15 脂氧合酶途径和 15 脂氧合酶途径之间发现的高度新颖的相互作用的基础上。磷脂酰乙醇胺结合蛋白-1。这些相互作用可以在缺乏高水平 IL-14/-13 的情况下放大和维持“Th2”型炎症，并改变对哮喘治疗主要药物 b2 激动剂的反应。最后，这些相互作用为药物开发的创新分子方法开辟了新的机会。