Lung HRV: G-Protein Coupled Signaling Interactions in Asthma

肺 HRV：哮喘中 G 蛋白耦合信号传导相互作用

基本信息

批准号：
8010837
负责人：
Stephen B Liggett
金额：
$ 38.9万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
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：
2010-01-01 至 2013-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8010837
关键词：
Agonist Amino Acid Sequence Asthma Base Sequence Blood Circulation Bronchial Hyperreactivity Cell Culture Techniques Cell Line Cells Chronic Obstructive Airway Disease Clinical Common Cold Coupled Data Diagnostic Epithelial Epithelial Cells Event Family Picornaviridae G Protein-Coupled Receptor Signaling G-Protein-Coupled Receptors GTP-Binding Proteins Genetic Drift Genetic Recombination Genome Genomics Genotype Goals Growth Heterogeneity Human Impairment In Vitro Infection Inflammatory Ions Irrigation Liquid substance Lung Mediator of activation protein Methods Modeling Morbidity - disease rate Muscle Contraction Muscle function Muscle relaxation phase Mutation Nature Pathology Patients Peptide Sequence Determination Phenotype Phylogenetic Analysis Phylogeny Physiological RNA Relaxation Resistance Rhinovirus Sampling Sequence Alignment Serotyping Severities Signal Transduction Smooth Muscle Structure Symptoms Techniques Trees Validation Variant Viral asthmatic airway autocrine base cell type genome sequencing genome-wide analysis in vivo methacholine novel paracrine patient population prognostic protein structure public health relevance receptor coupling receptor function repository respiratory respiratory smooth muscle response scaffold trait

项目摘要

DESCRIPTION (provided by applicant): Human rhinovirus (HRV) infection causes at least 50% of asthma exacerbations. Airway epithelial cell (AEC) infection evokes the release of inflammatory, growth and bronchospastic factors, and other autocrine/paracrine mediators leading to generalized AEC and airway smooth muscle (ASM) "pro-exacerbation" pathology. This includes altered AEC lining fluid/ion content, ASM hyperreactivity, and ASM resistance to relaxation by (-agonists, which are controlled by G-protein coupled receptors (GPCRs) on these cells. The heterogeneity of these asthmatic responses is, in part, thought to be dependent on the HRV strain (serotype). There are over 100 HRV strains, yet little is known about how genomic differences in strains impact asthma exacerbation phenotypes. We have very recently completed sequencing the genomes of all 99 reference HRV-A and -B serotypes from a banked historical repository. This revealed previously unknown aspects of HRV RNA and protein structure, phylogenetic relationships, recombination, and extensive diversity among the canonical serotypes. It also provided structure-based sequence alignments which are a scaffold for integration of additional HRVs into the phylogenetic tree. The broad long-term objectives of this revised proposal are to ascertain the genomic features of modern HRV strains that contribute to specific asthmatic airway GPCR phenotypes and their heterogeneity. This will be accomplished by three aims. In Aim 1, we will determine the complete genome sequences of HRVs from 200 modern clinical isolates using massively parallel sequencing methods. In Aim 2, this data will be integrated into our structure-based reference genomic scaffold so as to define genomic regions that are similar and dissimilar amongst the strains, providing a rigorous mechanism to select the HRVs for in vitro functional studies. In Aim 3, GPCR signaling phenotypes of HRVs in the context of AEC and ASM will be ascertained in cell culture models using high-throughput methods (Sub Aim 1). And, these signaling phenotypes will be correlated to HRV genome features using Bayesian techniques with internal and external validations (Sub Aim 2). Such studies will provide a genomic basis for those HRVs that do, and do not, evoke AEC and ASM phenotypic traits, and thus establish some of the mechanisms of heterogeneity of viral induced asthma exacerbations. These findings may provide diagnostic and prognostic information, and pharmacologic strategies, for managing the most common cause of asthma exacerbations. PUBLIC HEALTH RELEVANCE: Human rhinovirus (HRV) infection causes about 50% of asthma attacks (and COPD exacerbations). There is, though, substantial variability in the nature and severity of the clinical features of asthma attacks from HRV infections, for reasons that are not known. However, it is now clear that there are many distinct HRV strains, and this proposal will define which HRVs, and which parts of their genomes, impose changes in airway receptor function that contribute to the pathology and symptoms of asthmatic attacks during HRV infection.

描述（由申请人提供）：至少 50% 的哮喘恶化是由人鼻病毒 (HRV) 感染引起的。气道上皮细胞 (AEC) 感染引起炎症、生长和支气管痉挛因子以及其他自分泌/旁分泌介质的释放，导致全身 AEC 和气道平滑肌 (ASM)“促恶化”病理。这包括 AEC 衬里液体/离子含量的改变、ASM 过度反应性以及 ASM 对 (β-激动剂) 松弛的抵抗，这些激动剂由这些细胞上的 G 蛋白偶联受体 (GPCR) 控制。这些哮喘反应的异质性部分是，被认为取决于 HRV 毒株（血清型）有超过 100 种 HRV 毒株，但我们对毒株的基因组差异如何影响哮喘恶化表型知之甚少。最近完成了对历史存储库中所有 99 种参考 HRV-A 和 -B 血清型的基因组测序，这揭示了以前未知的 HRV RNA 和蛋白质结构、系统发育关系、重组以及典型血清型之间的广泛多样性。基于结构的序列比对，这是将其他 HRV 整合到系统发育树中的支架。该修订提案的广泛长期目标是确定现代 HRV 的基因组特征。导致特定哮喘气道 GPCR 表型及其异质性的菌株。这将通过三个目标来实现。在目标 1 中，我们将使用大规模并行测序方法确定 200 个现代临床分离株的 HRV 的完整基因组序列。在目标 2 中，这些数据将被整合到我们基于结构的参考基因组支架中，以便定义菌株之间相似和不相似的基因组区域，为选择 HRV 进行体外功能研究提供严格的机制。在目标 3 中，将使用高通量方法在细胞培养模型中确定 AEC 和 ASM 背景下 HRV 的 GPCR 信号表型（子目标 1）。并且，这些信号表型将使用贝叶斯技术以及内部和外部验证与 HRV 基因组特征相关联（子目标 2）。此类研究将为那些引起或不引起 AEC 和 ASM 表型特征的 HRV 提供基因组基础，从而建立病毒诱导哮喘恶化的一些异质性机制。这些发现可能提供诊断和预后信息以及药理学策略，用于管理哮喘恶化的最常见原因。公共卫生相关性：约 50% 的哮喘发作（和 COPD 恶化）由人鼻病毒 (HRV) 感染引起。然而，HRV 感染引起的哮喘发作临床特征的性质和严重程度存在很大差异，原因尚不清楚。然而，现在已经清楚存在许多不同的 HRV 毒株，该提案将定义哪些 HRV 及其基因组的哪些部分会引起气道受体功能的变化，从而导致 HRV 感染期间哮喘发作的病理和症状。