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

• 批准号: 8493771
• 负责人: Sally E Wenzel
• 金额: $ 34.15万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 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/8493771
• 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; public health relevance; raf Kinases; receptor; response

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.

描述（由适用提供）：尤其是人类哮喘和严重哮喘的病理生物学，对TH2感染的作用和重要性的考虑，仍然对人类哮喘和严重哮喘的病理学仍然知之甚少。我们的初步数据表明，15脂氧合酶（LO）1，其产物15羟基羟基二烯酸（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TH2炎症和Damning€2 partennoreceptor（Ar）的较高（Ar）的升级和扩增的含量（BP）1可能是核心。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结合使用的作用，在人体气道上皮细胞中在体外和EX VIVO中的IL-13/IL-4RA信号扩增中发挥了作用，以及该过程是否进一步影响2受体信号。在AIM 1中，我们将确定长期暴露于IL-13（以及15 LO1的上调）是否导致上皮细胞，其中MEK-ERK途径长期激活，而PI3K途径下调，以及IL-4RA多态性是否会影响这些过程。我们将解决15个LO1/15 Hete-PE在此过程中发挥作用的作用，以及这种上调对下游基因表达的影响，无论是在急性（低15 LO1）状态以及慢性（高15 LO1）状态下。我们假设活跃的15 LO1/15 Hete-PE途径的存在将大大增强IL-13诱导的基因表达。基因阵列将在急性和慢性刺激下以及在存在和不存在敲低的情况下进行评估，以及药物抑制，以确定15 LO1/15 HETE-PE途径的总体影响。在AIM 2中，我们将解决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。该目标将仅集中在15 LO1/15 Hete-PE与PEBP1的相互作用上，进一步定义了相互作用的亚细胞位置，物理相互作用的机制以及PEBP1的光谱物的作用和机制。在AIM 3中，我们将确定15 Hete-PE与PEBP1的相互作用是否竞争抑制了GRK2与PEBP1的结合，从而导致人类哮喘患者在体外和体内的脱敏增强。我们认为，从哮喘和正常受试者中评估原代人细胞中的这些过程将揭开一种新的令人兴奋的途径，该途径似乎严格调节PERK，“ Th2”型型反应，也许是哮喘上皮细胞中的2AR途径。我们认为，对这些互动的更好理解将为新的和新颖的治疗方法提供许多机会。