Rhinovirus C Infection in Normal and Asthmatic Human Airway Epithelium at Single Cell Resolution

总结 作为大多数反复呼吸道病毒感染的病原体和最常见的触发因素， 在肺部疾病恶化的情况下，鼻病毒是对公共健康的主要威胁和显著的经济负担。 没有疫苗或直接激活的抗病毒疗法可用于预防鼻病毒感染或减轻 疾病相对于其他毒株，鼻病毒C种（RV-C）与更严重的疾病相关， 尤其是哮喘患者。此外，在基因编码的单核苷酸多态性（SNP）， RV-C受体CDHR 3最近被确定为严重哮喘急性发作的危险因素。 然而，作为一种最近发现的病原体，RV-C生物学的许多方面，如宿主的细节， CDHR 3基因变异对感染动力学的影响仍然不确定。因此 本申请的总体目标是（i）定义正常和非正常宿主对RV-C感染的应答， 哮喘人气道上皮（HAE）和（ii）确定CDHR 3 rs6967330 SNP开启 生理环境中的病毒-宿主动力学。我们的中心假设是，先天免疫反应， 与正常HAE相比，RV-C在哮喘HAE中将改变，并且CDHR 3 rs6967330[A]风险等位基因 通过改变纤毛细胞的病毒感染性来改变这种反应。 我们将通过以下方式来验证这些假设： 两个具体目标：目标1将在单细胞水平上定义正常和哮喘HAE中对RV-C的宿主应答 目的2将确定CDHR 3哮喘风险等位基因对RV-C感染和宿主的贡献。 反应我们的研究计划建立在Scull和Rosenberg之间正在进行的合作之上 实验室，在概念和技术上都是创新的，它整合了HAE培养模型， 单细胞RNA-Seq，以前所未有的速度探索生理相关的宿主对RV-C的反应 分辨率此外，我们提出了新的基因编辑实验，将直接确定如何CDHR 3 遗传变异 影响 等基因背景下原发性HAE中的RV-C感染。这项工作意义重大， 我们的研究结果将对理解特定气道上皮细胞类型在气道炎症中的作用具有广泛的意义。 抗病毒反应，特别是解决基础疾病状态和宿主基因型如何影响 对RV-C的易感性。

SUMMARY As the causative agents in a majority of recurrent respiratory virus infections and the most common trigger of lung disease exacerbations, rhinoviruses are a major threat to public health and a significant economic burden. There are no vaccines or direct-activating antiviral therapies available to prevent rhinovirus infection or mitigate disease. Relative to other strains, rhinoviruses species C (RV-C) are associated with more severe disease, especially in individuals with asthma. In addition, a single nucleotide polymorphism (SNP) in the gene encoding CDHR3, a receptor for RV-C, was recently identified as a risk factor for severe asthma exacerbations. However, as a recently identified pathogen, many aspects of RV-C biology, such as details of the host response and the effects of CDHR3 genetic variation on infection dynamics remain undefined. Thus, the overall goals of this application are to (i) define the host response to RV-C infection in both normal and asthmatic human airway epithelium (HAE) and (ii) determine the impact of the CDHR3 rs6967330 SNP on virus–host dynamics in physiological context. Our central hypotheses are that the innate immune response to RV-C will be altered in asthmatic HAE compared to normal HAE, and that the CDHR3 rs6967330[A] risk allele modifies this response by altering viral infectivity of ciliated cells. We will test these hypotheses by pursuing two Specific Aims: Aim 1 will define the host response to RV-C in normal and asthmatic HAE at single cell resolution; Aim 2 will determine the contribution of the CDHR3 asthma risk allele to RV-C infection and host response. Our research plan builds upon an ongoing collaboration between the Scull and Rosenberg laboratories and is both conceptually and technically innovative in its integration of HAE culture models and single cell RNA-Seq to explore the physiologically-relevant host response to RV-C at unprecedented resolution. Furthermore, we propose novel gene editing experiments that will directly determine how CDHR3 genetic variation affects RV-C infection in primary HAE on an isogenic background. This work is significant as our results will have broad implications for understanding the roles of specific airway epithelial cell types in the antiviral response and specifically address how underlying disease status and host genotype impact susceptibility to RV-C.