HOST-VIRUS INTERACTIONS AT THE CELLULAR LEVEL

细胞水平上的宿主-病毒相互作用

• 批准号: 8470003
• 负责人: John Yin
• 金额: $ 26.44万
• 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 2018-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8470003
• 关键词: Address; Antiviral Agents; Area; Asthma; Basic Science; Biology; Biopsy; Bronchiolitis; Bronchitis; Cell Culture Techniques; Cell Density; Cells; Characteristics; Chronic lung disease; Clinical; Clinical Research; Color; Common Cold; Complex; Data; Development; Environment; Epithelial Cells; Gene Expression; Growth; Heterogeneity; Human; Image; Immune; Infection; Instruction; Integration Host Factors; Knowledge; Lead; Lower Respiratory Tract Infection; Measures; Methods; Molecular; Patients; Population; Predisposition; Production; Reporter; Resources; Respiratory Tract Infections; Rhinovirus; Sampling; Severity of illness; Signal Transduction; Symptoms; Technology; Testing; Time; Tissues; Upper respiratory tract; Viral; Virus; Virus Diseases; Virus-Cell Membrane Interaction; Wheezing; Work; cell behavior; experience; extracellular; improved; in vivo; insight; new technology; novel; novel strategies; particle; respiratory; response; standard measure; tool; treatment strategy; virus host interaction

PROJECT SUMMARY (See instructions): Infections by human rhinovirus (HRV) contribute to a diversity of human illnesses that include the common cold, bronchitis, exacerbation of asthma and other chronic lung diseases. Patient biopsies of HRV-infected tissues from the upper and lower airways reveal patches of robust infection dispersed among areas of healthy tissue. We hypothesize that differing tissue susceptibility to HRV infection reflects an underlying and intrinsic cell-level heterogeneity in responses to virus infection and antiviral immune signals. Testing this hypothesis poses a number of challenges: standard measures of cell behavior sampled from large cell populations provide an average, not single-cell measures; spreading infections are by definition complex and heterogeneous environments; and most mechanistic studies of viral growth and infection spread lack host factors that potentially promote virus spread. To address these challenges we will build on our significant experience measuring single-cell infections, as well as implementing and quantifying infection spread in controlled cell-culture environments. We propose to employ: (i) a dual-color fluorescent reporter method to provide simultaneous readouts of viral and cellular gene expression from HRV-infected cells, (ii) a quantitative imaging acquisition and analysis platform for high-throughput measures of dynamic single-cell behaviors, and (iii) a micro-scale cell-culture technology that offers unprecedented environmental control over cell density, localized virus inoculation, and flow conditions. Preliminary results indicate the feasibility of these technologies and significant opportunity for elucidating mechanisms of viral and cellular gene expression, virus growth, and infection spread. Our project aims to: (1) advance a data-driven integrated understanding of HRV single-cycle growth in cell cultures, (2) elucidate the impact of cell-virus interactions on HRV infection spread, and (3) isolate, characterize and utilize host factors that promote growth and spread of HRV in vivo. These aims will make extensive use of molecular and clinical tools in the other two projects, including airway epithelial cells and chimeric viruses. Results from this project will provide novel insights into how some tissues resist infection while others are susceptible, and this information will guide the development of improved treatment strategies for HRV-related illnesses.

项目摘要（参见说明）：人鼻病毒 (HRV) 感染会导致多种人类疾病，包括普通感冒、支气管炎、哮喘恶化和其他慢性肺部疾病。对患者上呼吸道和下呼吸道感染 HRV 的组织进行活组织检查，发现健康组织区域中散布着严重的感染斑块。我们假设不同的组织对 HRV 感染的易感性反映了对病毒感染和抗病毒免疫信号反应的潜在和内在的细胞水平异质性。测试这一假设提出了许多挑战：从大细胞群中采样的细胞行为的标准测量提供了平均而非单细胞测量；根据定义，传播感染是复杂且异构的环境；大多数关于病毒生长和感染传播的机制研究缺乏可能促进病毒传播的宿主因素。为了应对这些挑战，我们将利用测量单细胞感染以及在受控细胞培养环境中实施和量化感染传播的丰富经验。我们建议采用：（i）双色荧光报告方法，同时读取 HRV 感染细胞的病毒和细胞基因表达，（ii）定量成像采集和分析平台，用于动态单通道高通量测量细胞行为，以及（iii）微型细胞培养技术，可对细胞密度、局部病毒接种和流动条件提供前所未有的环境控制。初步结果表明这些技术的可行性以及阐明病毒和细胞基因表达、病毒生长和感染传播机制的重要机会。我们的项目旨在：(1) 推进对细胞培养物中 HRV 单周期生长的数据驱动综合理解，(2) 阐明细胞-病毒相互作用对 HRV 感染传播的影响，以及 (3) 分离、表征和利用促进 HRV 在体内生长和传播的宿主因素。这些目标将在其他两个项目中广泛使用分子和临床工具，包括气道上皮细胞和嵌合病毒。该项目的结果将为了解某些组织如何抵抗感染而另一些组织如何易感提供新的见解，并且这些信息将指导开发针对 HRV 相关疾病的改进治疗策略。