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：
10015331
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 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10015331
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 Epithelium 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 epithelium 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.

项目摘要我们发现了甲酸（MA）途径脂质代谢物在介导中的作用哮喘疾病模型中的2型炎症。这一发现是新颖的，以前从工作中出现在我的K08奖中完成，并确定MA途径是过敏性注射不可或缺的在哮喘中。我们的初步数据表明，超过了被称为类分子的池 Farnesylylophophate（FPP）和黄烷基果仁酰PPP（GGPP）显着增强了白介素13诱导的STAT6 人类气道上皮细胞的激活和eotaxin -2和-3的产生，体外，关键分子途径哮喘中的Th2/2型注射。此外，用HMG-COA还原酶抑制FPP和GGPP合成抑制剂（“汀类药物”），将类化合物上游合成的酶抑制IL-13诱导的eotaxin 生产和细胞外分泌。 eotaxins是嗜酸性粒细胞募集到的主要趋化因子气道组织，我们的结果表明，高水平的类吸气介导并促进2型嗜酸性粒细胞气道炎症。此外，我们使用哮喘小鼠模型的体内数据确认介导过敏性嗜酸性注射的MA途径。药物耗尽，因此下游代谢物FPP和GGPP显着抑制肺和气道嗜酸性粒细胞。而这些数据暗示它们在疾病发病机理中的作用，仅限于人类气道上皮细胞培养和哮喘的小鼠模型。通过此R03提案，我们希望确定类异丙因在人类疾病中的作用并最终确定超过气道粘膜类异型与持续气道程度相关的水平是否相关哮喘中的嗜酸性炎症。因此，为了解决知识的差距，我们假设这超出了气道上皮类异戊二烯的水平通过进一步诱导的体内增强2型嗜酸性注射上皮JAK/STAT6磷酸化。我们将通过两个具体目的解决这一假设：（1）确定是否是否高水平的外源性气道类异戊二烯增加体内嗜酸性气道注射。（2）验证使用专门的质谱法测量气道和肺类异丙的定量方法现有的动物和人体组织。实现这些目标将建立超过的原则证明类体内类体型在疾病发病机理中起作用，最重要的是，提供了必要的工具将此技术应用于未来的授予R01赠款申请中的人类哮喘患者。