Models of rhinovirus-C respiratory infection and asthma

丙型鼻病毒呼吸道感染和哮喘模型

• 批准号: 10682418
• 负责人: Marc B. Hershenson
• 金额: $ 42.21万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-22 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10682418
• 关键词: 4 year old; Adrenal Cortex Hormones; Air; Airway Disease; Alleles; Allergic; Animal Model; Asthma; Bronchiolitis; Cadherins; Cell surface; Cells; Child; Clone Cells; Cytoplasm; Data; Development; Dexamethasone; Diagnosis; Disease; Epithelial Cells; Epithelium; Family; Family member; Flow Cytometry; Gene Expression; Genotype; Hela Cells; Hospitalization; Human; IL2RA gene; IL7R gene; In Vitro; Individual; Infant; Infection; Inflammasome; Intercellular adhesion molecule 1; Interleukin-13; Life; Link; Liquid substance; Low-Density Lipoproteins; Lung; Lymphoid Cell; Macrophage; Measures; Metaplasia; Modeling; Mucins; Mucous body substance; Mus; Nasopharynx; Nose; Pathogenesis; Phenotype; Phosphotransferases; Physicians; Pilot Projects; Predisposition; Production; Prospective Studies; Pyroglyphidae; Refractory; Research Personnel; Resistance; Respiratory Tract Infections; Rhinovirus; Rhinovirus infection; Role; Sampling; Signal Pathway; Steroids; TSLP gene; Testing; Therapeutic Intervention; Universities; Variant; Viral; Virus; Virus Diseases; Wisconsin; airway epithelium; airway hyperresponsiveness; airway inflammation; aspirate; asthma exacerbation; asthmatic; cellular transduction; cytokine; eosinophil; eosinophilic inflammation; experimental study; high risk; human subject; in vivo; insight; neutralizing antibody; overexpression; receptor; response; virtual

PROJECT SUMMARY Accumulating evidence indicates that infections with a newly-discovered species of rhinovirus, RV-C, are asso- ciated with severe respiratory tract infections and asthma exacerbations often requiring hospitalization. In addi- tion, recent data suggest a possible role for early-life RV-C infections in asthma development. Despite increasing recognition of RV-C as a cause of asthmatic disease, virtually nothing is known about the pathogenesis of RV-C infections. To accomplish this, we infected mice with RV-C15 (the RV-C15 infec- tious clone and HeLa-E8 cells overexpressing a variant of the human RV-C receptor, cadherin related family member 3, were obtained from James Gern, University of Wisconsin). Our pilot studies show that RV-C15- infected mice show increased type 2 cytokine and mucin gene expression, BAL eosinophils and lineage-nega- tive, CD25+, CD127+ type 2 innate lymphoid cells (ILC2s) compared to RV-A1B-infected mice. In addition, pi- lot studies from children with natural RV-C infections show increased type 2 cytokine production. In this application, we will test the general hypothesis that, after RV-C infection, airway innate cytokine ex- pression drives ILC2 expansion and development of eosinophilic inflammation and mucous metaplasia. To test this hypothesis, we propose the following Specific Aims: Specific Aim 1. Determine the contribution of epithelial-derived innate cytokines to RV-C15-induced eosinophilic airway inflammation and hyperresponsiveness (AHR). We hypothesize that: 1) compared to RV-A, RV-C infection of mature mice induces greater lung expression of innate cytokines (IL-25, IL-33, TSLP); 2) IL-25 is produced by doublecortin-like kinase (DCLK)-1-positive airway tuft cells; 3) innate cytokines are re- quired for eosinophilic inflammation; 4) RV-C engagement of CDHR3 activates distinct signaling pathways leading to innate cytokine expression. Specific Aim 2. Determine the contribution of lung ILC2s and macrophages to RV-C-induced airway inflammation and AHR. We hypothesize that: 1) RV-C infection of mature mice induces innate cytokine-de- pendent expansion of ILC2s; 2) ILC2s promote eosinophilic inflammation, macrophage polarization and AHR; 3) house dust mite (HDM) and RV-C have additive effects on eosinophilic inflammation and AHR; 4) ILC2s convey corticosteroid resistance; and 5) nasal aspirates from human subjects infected with RV-C show in- creased expression of type 2 cytokines and ILC2s compared to samples from RV-A-infected subjects. Specific Aim 3. Determine the effects of early-life RV-C infection on the established asthma pheno- type. We have found that RV-A1B infection of six day-old mice, but not mature mice, induces long-lasting mu- cous metaplasia and AHR which is dependent on IL-13-producing ILC2s. We hypothesize that: 1) RV-C infec- tion of 6 day-old mice induces greater and more long-lasting mucous metaplasia than RV-A; 2) early-life RV-C infection increases the number of IL-25-producing airway tuft cells.

项目概要 越来越多的证据表明，新发现的鼻病毒 RV-C 的感染与 患有严重呼吸道感染和哮喘恶化通常需要住院治疗。另外， 最近的数据表明，早期 RV-C 感染在哮喘发展中可能发挥作用。 尽管越来越多的人认识到 RV-C 是哮喘病的一个原因，但实际上对 RV-C 却一无所知。 RV-C 感染的发病机制。为了实现这一目标，我们用 RV-C15 感染小鼠（RV-C15 infec- 过表达人类 RV-C 受体、钙粘蛋白相关家族变体的 Tious 克隆和 HeLa-E8 细胞 成员 3，获自威斯康星大学 James Gern）。我们的试点研究表明 RV-C15- 受感染的小鼠表现出 2 型细胞因子和粘蛋白基因表达增加、BAL 嗜酸性粒细胞和谱系阴性增加 与 RV-A1B 感染的小鼠相比，CD25+、CD127+ 2 型先天淋巴细胞 (ILC2) 明显增加。此外，pi- 对自然感染 RV-C 的儿童进行的大量研究表明，2 型细胞因子的产生增加。 在此应用中，我们将测试一般假设，即 RV-C 感染后，气道先天细胞因子 ex- 压力驱动 ILC2 扩张以及嗜酸性炎症和粘膜化生的发展。测试 根据这一假设，我们提出以下具体目标： 具体目标 1. 确定上皮源性先天细胞因子对 RV-C15 诱导的贡献 嗜酸性粒细胞性气道炎症和高反应性（AHR）。我们假设：1）相比 成熟小鼠的 RV-A、RV-C 感染诱导先天细胞因子（IL-25、IL-33、TSLP）的肺部表达增加； 2) IL-25由双皮质素样激酶(DCLK)-1阳性气道簇细胞产生； 3）先天细胞因子被重新 嗜酸性粒细胞性炎症需要； 4) CDHR3 的 RV-C 参与激活不同的信号通路 导致先天细胞因子的表达。 具体目标 2. 确定肺 ILC2 和巨噬细胞对 RV-C 诱导的气道的贡献 炎症和 AHR。我们假设：1) 成熟小鼠的 RV-C 感染会诱导先天性细胞因子去- ILC2 的悬而未决的扩展； 2）ILC2促进嗜酸性炎症、巨噬细胞极化和AHR； 3）屋尘螨（HDM）和RV-C对嗜酸性炎症和AHR有相加作用； 4) ILC2 传达皮质类固醇抵抗； 5) 感染 RV-C 的人类受试者的鼻吸出物显示- 与 RV-A 感染受试者的样本相比，2 型细胞因子和 ILC2 的表达增加。 具体目标 3. 确定生命早期 RV-C 感染对已确定的哮喘表型的影响 类型。我们发现，六日龄小鼠（而非成熟小鼠）的 RV-A1B 感染会诱导持久的 mu- 库斯化生和 AHR 依赖于产生 IL-13 的 ILC2。我们假设：1) RV-C 感染- 与 RV-A 相比，6 日龄小鼠的化生诱导更大且更持久的粘液化生； 2) 早期RV-C 感染会增加产生 IL-25 的气道簇细胞的数量。