Development of an influenza A virus and Aspergillus fumigatus coinfection model

甲型流感病毒和烟曲霉共感染模型的建立

基本信息

批准号：
10089411
负责人：
JOSHUA J OBAR
金额：
$ 20.5万
依托单位：
DARTMOUTH COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2023-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10089411
关键词：
Address Alveolar Macrophages Animal Model Animals Antifungal Agents Aspergillus Aspergillus fumigatus Automobile Driving Bacterial Pneumonia C57BL/6 Mouse Cells Clinical Data Data Set Development Ensure Experimental Models Face Future Genetic Transcription Goals Growth Heterogeneity Immunocompetent Impairment Infection Influenza A virus Laboratories Leukocytes Lung Modeling Molecular Monitor Morbidity - disease rate Mus Mycoses Nose Patients Phagocytosis Population Predisposition Prevalence Reporter Reporting Reproducibility Reproduction spores Research Risk Secondary to Serotyping Staphylococcus aureus Streptococcus pneumoniae System Techniques Testing Tissues Virulence Virus Diseases Work co-infection experience experimental study in vivo influenza virus strain innovation insight macrophage mortality mouse model novel prevent pulmonary function secondary infection single-cell RNA sequencing transcriptome sequencing

项目摘要

ABSTRACT Influenza A virus (IAV) infected patients admitted to the ICU face severe secondary infection complications. There is increasingly strong evidence that secondary fungal infections with Aspergillus fumigatus (Af) present a significant risk for ICU patients with severe IAV infection. Overall, approximately 20% of severe IAV patients in the ICU may develop secondary Af infection. Currently, there is a critical gap in understanding how and why these fungal secondary infections develop in IAV patients. This R21 application aims to fill this gap by generating a robust small animal IAV-Af coinfection model that can be used to dissect the molecular mechanism(s) by which IAV infection increases susceptibility to Af challenge. Our initial pilot experiments demonstrate that IAV infection four days prior to Af challenge results in significant fungal growth and tissue invasion, which is absent in naïve mice challenged with Af. In SA1 of this R21 proposal we will further optimize and develop this IAV-Af coinfection model to ensure it is highly reproducible and define the importance of IAV serotype and Af clinical strain heterogeneity in susceptibility to coinfection. In SA2, we will test the hypothesis that IAV infection impairs pulmonary leukocyte anti-fungal activity using two innovative techniques: 1) Fluorescent Aspergillus reporter (FLARE) strains to directly quantify anti-fungal activity of leukocyte populations at a cell single level and 2) Single cell RNA sequencing (scRNA-Seq) to monitor transcriptional changes in pulmonary leukocyte populations in an unbiased manner. These data will provide the first insights into mechanisms driving Af susceptibility following IAV infection. Overall, this research fills a critical gap by providing the field with a unique and highly needed IAV-Af coinfection model and provides a unique and robust dataset for the development of future focused, mechanistic R01 applications.

抽象的流感病毒（IAV）感染了ICU的患者面临严重的继发感染并发症。有越来越有力的证据表明，曲霉的次生真菌感染 Fumigatus（AF）对患有严重IAV感染的ICU患者带来了重大风险。全面的， ICU中约有20％的严重IAV患者可能会出现继发性自动AF感染。现在，了解这些真菌二次感染如何以及为什么发展在IAV患者中。该R21应用程序旨在通过产生坚固的小动物IAV-AF来填补这一空白可用于剖析IAV感染的分子机制的共感染模型增加对AF挑战的敏感性。我们最初的试点实验表明IAV感染四个在AF挑战之前的几天导致大量真菌生长和组织侵袭，这在天真的小鼠受到AF的挑战。在此R21提案的SA1中，我们将进一步优化并开发 IAV-AF共感染模型，以确保其高度可重复并定义IAV血清型的重要性和AF临床应变异质性在对共感染的敏感性中。在SA2中，我们将检验假设 IAV感染使用两种创新技术损害了肺白细胞抗真菌活性：1）荧光曲霉报告基因（火炬）菌株直接量化白细胞的抗真菌活性单个单个水平的种群和2）单细胞RNA测序（SCRNA-SEQ）监测肺白细胞种群的转录变化以公正的方式变化。这些数据将提供对IAV感染后驱动AF敏感性的机制的第一个见解。总体而言，这研究通过为该领域提供独特且急需的IAV-AF共感染来填补关键空白模型并提供了一个独特而强大的数据集，以开发未来的注重，机械 R01应用程序。