Isoprenoid Biology in Asthma: Proof-of-Principle and Method Development

哮喘中的类异戊二烯生物学：原理验证和方法开发

基本信息

批准号：
9810509
负责人：
Amir A. Zeki
金额：
$ 7.85万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-10 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9810509
关键词：
3-hydroxy-3-methylglutaryl-coenzyme A Acute Address Airway Resistance Alcohols Allergic Applications Grants Asthma Award Biology Blood Bronchoalveolar Lavage CCL24 gene CCL26 gene CD8B1 gene Cell Count Cell Culture Techniques Cell membrane Cells Cholesterol Chronic Coenzyme A Collagen Data Detection Development Diphosphates Disease Disease model Enzymes Eosinophilia Eotaxin Epithelial Epithelial Cells Exposure to Extrinsic asthma Family Farnesol Fluorescence Fluorescence-Activated Cell Sorting Future Gene Silencing Goals Goblet Cells Grant Guanosine Triphosphate Phosphohydrolases Harvest High Pressure Liquid Chromatography House Dust Mite Allergens Human Hydroxymethylglutaryl-CoA Reductase Inhibitors Hyperplasia Immune In Vitro Inflammation Inhalation Inhalators Interleukin-13 Knowledge Lead Lipids Lung Mass Spectrum Analysis Measures Mediating Methods Molecular Monomeric GTP-Binding Proteins Mucous Membrane Mus Natural Killer Cells Oxidoreductase Pathogenesis Pathway interactions Pharmacology Phosphorylation Plasma Cells Play Population Post-Translational Protein Processing Production Proteins Recurrence Regulatory T-Lymphocyte Research Support Respiratory physiology Role STAT6 gene Signal Transduction Small inducible cytokine A24 Squalene Stains Sterols Structure of parenchyma of lung Suggestion Tail Techniques Technology Testing Tissues Western Blotting Work airway hyperresponsiveness airway inflammation airway remodeling animal tissue asthma model asthmatic biobank chemokine cytokine empowered eosinophil eosinophilic inflammation experimental study extracellular farnesyl pyrophosphate geranylgeraniol human disease human tissue in vivo inhibitor/antagonist isopentenyl pyrophosphate isoprenoid isoprenylation method development mevalonate mouse model novel persistent symptom prevent recruit response rho targeted treatment tool translation to humans

项目摘要

PROJECT SUMMARY We have discovered a role for the mevalonate (MA) pathway lipid metabolites known as isoprenoids in mediating type 2 inflammation in asthma disease models. This finding is novel and emerges from work previously accomplished in my K08 award, and establishes the MA pathway as integral to allergic eosinophilic inflammation in asthma. Our preliminary data show that excess pools of the isoprenoid molecules known as farnesylpyrophosphate (FPP) and geranylgeranyl-PP (GGPP) markedly augment interleukin-13-induced STAT6 activation and eotaxin-2 and -3 production in human airway epithelial cells in vitro, key molecular pathways in Th2/type 2 inflammation in asthma. Further, inhibition of FPP and GGPP synthesis with HMG-CoA reductase inhibitors (‘statins’), the enzyme that synthesizes MA upstream of the isoprenoids, inhibits IL-13-induced eotaxin production and extracellular secretion. The eotaxins being a major chemokine for eosinophil recruitment into airway tissues, our results suggest that high levels of airway isoprenoids mediate and promote type 2 eosinophilic airway inflammation. Furthermore, our in vivo data using mouse models of asthma, confirm the critical role of the MA pathway in mediating allergic eosinophilic inflammation. Depleting MA pharmacologically, and therefore the downstream metabolites FPP and GGPP, significantly inhibits lung and airway eosinophilia. While these data are suggestive of their role in disease pathogenesis, they are limited to human airway epithelial cell culture and mouse models of asthma. Via this R03 proposal, we wish to establish the role of isoprenoids in human disease and ultimately determine if excess levels of airway mucosal isoprenoids correlate with degree of persistent airway eosinophilic inflammation in asthma. Therefore to address this gap in knowledge, we hypothesize that excess levels of airway epithelial isoprenoids enhance type 2 eosinophilic inflammation in vivo by further inducing epithelial JAK/STAT6 phosphorylation. We will address this hypothesis via two specific aims: (1) To determine if high levels of exogenous airway isoprenoids augment eosinophilic airway inflammation in vivo. (2) To validate a quantitative method of measuring airway and lung isoprenoids using specialized mass spectrometry in pre- existing animal and human tissues. Accomplishing these goals will establish the proof-of-principle that excess isoprenoids in vivo play a role in disease pathogenesis, and most importantly, provide the tools necessary to apply this technology to human asthmatics in future grant R01 grant applications.

项目概要我们发现甲羟戊酸 (MA) 途径脂质代谢物（称为类异戊二烯）在介导哮喘疾病模型中的 2 型炎症这一发现是新颖的，是从之前的工作中得出的。在我的 K08 奖中取得了成就，并将 MA 途径确立为过敏性嗜酸性粒细胞炎症的组成部分我们的初步数据表明，过量的类异戊二烯分子被称为哮喘。法呢基焦磷酸 (FPP) 和香叶基香叶基-PP (GGPP) 显着增强白细胞介素 13 诱导的 STAT6 体外人气道上皮细胞的激活和eotaxin-2和-3的产生，关键分子途径哮喘中的 Th2/2 型炎症此外，用 HMG-CoA 还原酶抑制 FPP 和 GGPP 合成。抑制剂（“他汀类药物”）是类异戊二烯上游合成 MA 的酶，可抑制 IL-13 诱导的嗜酸细胞趋化因子嗜酸细胞趋化因子是嗜酸性粒细胞募集的主要趋化因子。在气道组织中，我们的结果表明，高水平的气道类异戊二烯介导和促进 2 型嗜酸性粒细胞此外，我们使用哮喘小鼠模型的体内数据证实了气道炎症的关键作用。 MA 途径介导过敏性嗜酸性粒细胞炎症，因此在药理学上消耗 MA。下游代谢物 FPP 和 GGPP 显着抑制肺和气道嗜酸性粒细胞增多。提示它们在疾病发病机制中的作用，它们仅限于人类气道上皮细胞培养和通过这个 R03 提案，我们希望确定类异戊二烯在人类疾病中的作用。并最终确定气道粘膜类异戊二烯的过量水平是否与持续性气道程度相关因此，为了解决这一知识空白，我们捕获了过量的嗜酸性粒细胞炎症。气道上皮类异戊二烯水平通过进一步诱导增强体内 2 型嗜酸性粒细胞炎症我们将通过两个具体目标来解决这一假设：(1) 确定是否存在上皮 JAK/STAT6 磷酸化。 (2) 验证使用专门的质谱法测量气道和肺类异戊二烯的定量方法现有的动物和人体组织的实现将建立过量的原理证明。体内类异戊二烯在疾病发病机制中发挥作用，最重要的是，提供必要的工具在未来的 R01 拨款申请中将该技术应用于人类哮喘患者。