Genetic Determinants of Aspergillus host-pathogen interactions

曲霉菌宿主-病原体相互作用的遗传决定因素

基本信息

批准号：
10724816
负责人：
Jarrod R. Fortwendel
金额：
$ 23.1万
依托单位：
UNIVERSITY OF TENNESSEE HEALTH SCI CTR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-23 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10724816
关键词：
Address Allergic Bronchopulmonary Aspergillosis Anabolism Aspergillosis Aspergillus Aspergillus fumigatus Asthma Automobile Driving Caspase Cell Line ChIP-seq Chronic Clinical Coculture Techniques Complex Cystic Fibrosis Development Fungal Components Future Gene Deletion Gene Expression Profile Gene Expression Profiling Genetic Determinism Genetic Epistasis Genetic Transcription Goals Human Immune response Immunity Immunocompetent Immunocompromised Host In Vitro Individual Infection Inflammasome Inflammation Inflammatory Response Interleukin-1 beta Libraries Macrophage Masks Measures Mediating Molecular Morbidity - disease rate Organism Pathogen detection Pathogenicity Pathway interactions Patients Pattern Peripheral Blood Mononuclear Cell Phagocytosis Process Protein Kinase Proteins Regulation Research Research Design Role Screening Result Severities Signal Transduction Stimulus Stress Response Signaling Syndrome Tissue-Specific Gene Expression Virulence Work biological adaptation to stress chronic infection diagnostic criteria fungus gene complementation immunological status immunopathology improved outcome in vivo molecular assembly/self assembly monocyte mortality mouse model mutant new therapeutic target novel novel therapeutic intervention pathogen pathogenic fungus protein-histidine kinase respiratory response sensor histidine kinase therapeutic target tool transcription factor transcriptome sequencing

项目摘要

The long-term goal of our work is to reduce the morbidity and mortality associated with Aspergillus infections by improving outcomes of host-pathogen interactions. Aspergillus fumigatus is the major airborne fungal pathogen and is responsible for a range of clinical syndromes, the severity of which is dependent on host immune status. Invasive pulmonary aspergillosis and disseminated infections can occur in severely immunocompromised hosts and are often associated with mortality rates of up to 90%. In immune competent hosts, A. fumigatus colonization can lead to chronic pulmonary aspergillosis (CPA), a clinical manifestation with recently defined diagnostic criteria that impacts upwards of 1.6 million individuals per year. Many individuals with CPA have underlying respiratory conditions, such as asthma or cystic fibrosis, that are further exacerbated by the presence of the fungus. Although multiple Pathogen-Associated Molecular Patterns (PAMPs) driving host recognition and response are known, uncovering novel fungal molecular pathways that could serve as therapeutic targets to enhance or mask PAMP display and/or biosynthesis could prove useful therapeutic targets to modulate host response for improved outcomes. To address this, we utilized activation of the inflammasome, a multiprotein intracellular complex that detects pathogenic organisms to initiate inflammatory responses, as a tool to uncover Aspergillus mutants with altered abilities to activate host response. We have completed a preliminary screen to measure IL-1β secretion from a macrophage differentiated human monocyte cell line employing an A. fumigatus protein kinase disruption library recently generated in our lab. Of 118 protein kinase disruption mutants screened, we identified seven A. fumigatus protein kinases that either significantly increased (n=2) or decreased (n=5) inflammasome-dependent IL-1β secretion upon disruption. Strikingly, both of the protein kinase disruption mutants that induced increased IL-1β release encode phospho-relay sensor histidine kinases (phkA and fhk3), a sub-class of protein kinases that directly sense environmental and intracellular changes and subsequently activate stress response signaling. The molecular pathways controlled by PhkA and Fhk3 are unknown, as both HKs are largely uncharacterized. Although a major consequence of HK signaling is activation of stress responses pathways that upregulate transcription factor activity to respond to environmental stimuli, the downstream transcriptional effectors for all A. fumigatus HKs remain undescribed. Our work will delineate A. fumigatus histidine kinase (HK)-dependent mechanisms of host response (Aim 1) and identify A. fumigatus transcriptional regulators of inflammasome activation (Aim 2). Through completion of the proposed Aims, we will delineate fungal molecular pathways mediating Aspergillus-induced host response. This work is essential for future development of novel therapeutic approaches targeted towards activating protection against invasive infections or mitigating harmful host-response during chronic infections.

我们工作的长期目标是减少与曲霉感染相关的发病率和死亡率改善宿主 - 病原体相互作用的结果。曲曲霉富马图斯是主要的空中真菌病原体并负责一系列临床综合征，其严重程度取决于宿主免疫状态。在严重免疫功能低下的宿主中可能发生侵入性肺曲霉病和传播感染并且通常与高达90％的死亡率有关。在免疫能力的宿主中，烟曲霉定植可以导致慢性肺曲霉菌病（CPA），这是一种临床表现，具有最近定义的诊断每年影响160万个人的标准。许多患有CPA的人的基础呼吸条件，例如哮喘或囊性纤维化，由于存在而进一步加剧真菌。尽管多个病原体相关的分子模式（PAMP）驱动宿主识别和响应是已知的，发现了新型的真菌分子途径，这些途径可以用作治疗靶标的增强或掩盖pamp显示器和/或生物合成可以证明有用的治疗靶标可调节宿主改善结果的响应。为了解决这个问题，我们利用了炎性蛋白的激活细胞内复合物检测致病生物以启动炎症反应，作为揭露的工具曲霉突变体具有改变能力激活宿主反应的能力。我们已经完成了一个初步屏幕测量巨噬细胞分化的人类单核细胞系的IL-1β分泌，该细胞系采用烟曲霉的蛋白激酶破坏库最近在我们的实验室中生成。筛选的118个蛋白激酶破坏突变体的我们确定了七个明显增加（n = 2）或减少（n = 5）的五张烟曲霉蛋白激酶炎症体依赖性IL-1β分泌发生时。令人惊讶的是，两种蛋白激酶破坏诱导IL-1β释放增加的突变体编码磷酸 - 雷（Phka和FHK3）的磷酸 - 雷（Phka和FHK3），蛋白激酶的子类，直接感知环境和细胞内变化，然后激活应力响应信号。由PHKA和FHK3控制的分子途径是未知的，因为 HK在很大程度上没有表征。尽管香港信号传导的主要结果是激活应力反应上调转录因子活性以响应环境刺激的途径，下游所有A. fumigatus hks的转录效应仍未描述。我们的工作将描绘A. Fumigatus 组氨酸激酶（HK）依赖于宿主反应的机制（AIM 1）并识别烟曲霉转录炎性体激活的调节剂（AIM 2）。通过完成拟议的目标，我们将描述真菌分子途径介导曲霉诱导的宿主反应。这项工作对于未来至关重要开发针对激活侵入性感染保护的新型热方法或减轻慢性感染期间有害宿主反应。