Molecular Biology of RV-C and its Asthma-related Receptor, CDHR3

RV-C 及其哮喘相关受体 CDHR3 的分子生物学

• 批准号: 10091396
• 负责人: ANN C. PALMENBERG
• 金额: $ 53.35万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10091396
• 关键词: Alleles; Allergic; Antiviral Agents; Antiviral Therapy; Asthma; Back; Binding; Binding Sites; Biochemical; Biochemistry; Biological Assay; Biology; C cadherin; Cadherins; Capsid; Cell Culture Techniques; Cell Differentiation process; Cell surface; Cells; Characteristics; Child; Childhood Asthma; Cilia; Clinic; Clinical; Common Cold; Complex; Cryoelectron Microscopy; Data; Development; Dimerization; Disease; Environmental Risk Factor; Etiology; Face; Family member; Frequencies; Funding; Genealogy; Genes; Genetic; Genetic Predisposition to Disease; Genotype; Growth; High Prevalence; Hospitalization; Human; Individual; Infection; Intervention; Investigation; Link; Location; Maps; Medical History; Modernization; Molecular; Molecular Biology; Mutation; Peptide Hydrolases; Predisposition; Preventive; Process; Proteins; Reagent; Recombinants; Resolution; Respiratory Tract Infections; Rhinovirus; Rhinovirus infection; Role; Severities; Severity of illness; Shapes; Site; Structural Biochemistry; Structure; Surface; Techniques; Technology; Time; Tissues; Translating; Variant; Viral; Virus; Virus Diseases; Virus Receptors; Wheezing; Work; asthma exacerbation; asthmatic; cell type; early childhood; experience; genetic element; glycosylation; immunogenicity; microbiome; microbiota; novel; particle; programs; prophylactic; protein expression; receptor; receptor binding; receptor expression

PROJECT SUMMARY/ABSTRACT Asthma is a highly complex disease because individual etiologies result from composite elements of genetic susceptibility, personal medical histories and a myriad of environmental factors. This Program will collect and evaluate data relevant to each component. Project II contributes experience and technologies to describe the molecular mechanisms by which virus infections, specifically by human rhinoviruses (RV), contribute and shape episodes of asthma. Canonically linked to the common cold, RV infections contribute 50-85% of asthma exacerbations and are the most frequently isolated viruses during the most severe respiratory infections and hospitalizations among children. More so than RV-A&B isolates, viruses in the RV-C species are particularly linked to illnesses in early childhood, and their infections are strongly linked to subsequent development of asthma. The RV-C require cadherin-related family member 3 (CDHR3), an unusual airway-specific protein, as their cell-entry receptor. The “A” allele of this gene (Tyr529 protein variant) is among the strongest known genetic correlates for a type of childhood asthma marked by severe episodic wheezing. Conversely, Cys529, encoded by the “G” allele, and of much higher prevalence in modern human lineages, is not an asthma correlate. Our fundamental hypothesis is that the CDHR3 Cys529 variant fails at the biochemical level to display this protein properly or extensively on cell surfaces, making their carriers more refractive to RV-C infections and virus-induced asthma-exacerbations. The three Aims of the Project will examine the structure, biochemistry and function of CDHR3 in recombinant, cell culture and native tissue formats. The expected information includes (a) a cryoEM structure determination of the receptor:RV-C complex, (b) cryoEM structure determinations of RV-C complexed with neutralizing and non-neutralizing antibodies, (c) biochemical descriptions of CDHR3 requirements for differential cell surface display (Cys/Tyr529), (d) biochemical descriptions of CDHR3 glycosylation sites, dimerization sites and virus interaction sites. We will also examine primary tissue, and tissue-derived differentiated cell cultures (e.g. ALI), to validate and localize the sites of CDHR3 display and determine how the genetics-determined allele (“A” vs “G”) confers susceptibility to RV-C infections. Our techniques and assays will also document for the first time, if and how common environmental factors, such as an individual's microbiome content, may influence the extent and severity of RV infections by altering RV receptor expression or a cell's infection susceptibility in its native primary context.

项目总结/摘要 哮喘是一种高度复杂的疾病，因为个体病因是由遗传因素的复合因素引起的。 易感性，个人病史和无数的环境因素。该计划将收集和 评估与每个组件相关的数据。项目二提供经验和技术， 病毒感染，特别是人鼻病毒（RV）感染的分子机制， 形成哮喘发作。RV感染通常与普通感冒有关，导致50 - 85%的哮喘 是最严重呼吸道感染期间最常分离到的病毒， 儿童住院。与RV-A和B分离株相比，RV-C种中的病毒特别多。 与幼儿期疾病有关，其感染与随后的疾病发展密切相关。 哮喘RV-C需要钙粘蛋白相关家族成员3（CDHR 3），这是一种不寻常的气道特异性蛋白， 它们的细胞进入受体该基因的"A"等位基因（Tyr529蛋白变体）是已知的最强等位基因之一。 一种以严重间歇性喘息为特征的儿童哮喘的遗传相关性。相反，Cys529， 由“G”等位基因编码的，在现代人类谱系中流行率高得多的，不是哮喘 相互关联。我们的基本假设是，CDHR 3 Cys529变体在生物化学水平上不能显示出 这种蛋白质适当或广泛地存在于细胞表面，使它们的载体对RV-C感染更敏感 和病毒引起的哮喘恶化。该项目的三个目标将审查结构， 在重组、细胞培养和天然组织形式中的CDHR 3的生物化学和功能。预期 信息包括（a）受体：RV-C复合物的cryoEM结构测定，（B）cryoEM结构 测定与中和和非中和抗体复合的RV-C，（c）生物化学 差异细胞表面展示（Cys/Tyr529）的CDHR 3要求的描述，（d）生物化学 CDHR 3糖基化位点、二聚化位点和病毒相互作用位点的描述。我们亦会研究 原代组织和组织来源的分化细胞培养物（例如ALI），以验证和定位 CDHR 3显示并确定遗传学决定的等位基因（"A" vs "G"）如何赋予RV-C易感性 感染.我们的技术和分析也将首次记录，如果和如何共同的环境 因素，如个体的微生物组含量，可能会影响RV感染的程度和严重程度， 改变RV受体表达或细胞在其天然原代环境中的感染易感性。