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：
8544624
负责人：
Stephen B Liggett
金额：
$ 34.68万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
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/8544624
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）：人类鼻病毒（HRV）感染至少导致50％的哮喘加重。气道上皮细胞（AEC）感染唤起了炎症，生长和支气管疗法因子的释放，以及其他自分泌/旁分泌介质导致广泛的AEC和气道平滑肌（ASM）“促进”病理学。 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菌株的差异会影响哮喘的恶化表型，我们最近完成了所有99个参考HRV-A和-b血清型的测序为了将其他HRV整合到系统发育树中。这将由三个目标完成。在AIM 1中，我们将使用大量平行测序方法确定来自200个现代临床分离株的HRV的完整基因组序列。在AIM 2中，该数据将集成到我们基于结构的参考基因组支架中，以定义菌株中相似且不同的基因组区域，从而提供了一种严格的机制来为体外功能研究选择HRV。在AIM 3中，将使用高通量方法在细胞培养模型中确定HRV的GPCR信号表型（SUB AIM 1）。并且，这些信号表型将使用具有内部和外部验证的贝叶斯技术与HRV基因组特征相关（SUB AIM 2）。这样的研究将为那些不引起AEC和ASM表型性状的HRV提供基因组基础，从而确立了病毒诱导的哮喘加剧的某些机制。这些发现可能会提供诊断和预后的信息以及药理策略，以管理哮喘恶化的最常见原因。公共卫生相关性：人类鼻病毒（HRV）感染导致约50％的哮喘发作（和COPD加剧）。但是，由于尚不清楚的原因，HRV感染的哮喘攻击的临床特征的性质和严重程度有很大的可变性。但是，现在很明显，有许多不同的HRV菌株，该建议将定义哪些HRV，以及它们的哪些部分，在HRV感染期间施加了导致哮喘发作的病理和症状的气道受体功能的变化。