Defining the impact of host factors on the molecular architecture and bacterial physiology of Staphylococcus aureus abscesses

确定宿主因素对金黄色葡萄球菌脓肿分子结构和细菌生理学的影响

• 批准号: 10115595
• 负责人: JAMES E CASSAT
• 金额: $ 75.55万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10115595
• 关键词: 3-Dimensional; Abscess; Address; Animals; Anti-Bacterial Agents; Appearance; Architecture; Attention; Bacteria; Bacterial Infections; Bacterial Physiology; Bacterial Proteins; Binding; Biology; Cells; Characteristics; Complex; Cues; Data; Detection; Development; Diabetes Mellitus; Disease; Effector Cell; Evolution; Fluorescence Microscopy; Fluorescent in Situ Hybridization; Genus staphylococcus; Heterogeneity; Histologic; Histology; Hyperglycemia; Image; Imaging Device; Immune; Immune response; Immunohistochemistry; In Situ; Individual; Infection; Inflammatory; Innate Immune Response; Integration Host Factors; Knowledge; Lesion; Link; Maintenance; Mass Spectrum Analysis; Medical; Methods; Microbe; Modeling; Molecular; Murine Tissue Type; Myelosuppression; Organ; Osteomyelitis; Outcome; Pathogenesis; Pathology; Patients; Peptides; Physiology; Population; Predisposition; Proteomics; Publishing; Reagent; Role; Shapes; Space Perception; Spatial Distribution; Staphylococcal Infections; Staphylococcus aureus; Staphylococcus aureus infection; Techniques; Testing; Therapeutic; Therapeutic Intervention; Time; Tissue Sample; Tissue imaging; Tissues; Translational Research; Vaccines; Work; antimicrobial; base; body system; cell type; comorbidity; experimental study; human pathogen; imaging platform; in vivo; innovation; laser capture microdissection; macrophage; microbial; microbial host; molecular imaging; mouse model; neutrophil; novel; novel therapeutics; pathogen; preservation; response; therapeutic candidate; tissue processing; transcriptome sequencing; translational medicine

PROJECT SUMMARY / ABSTRACT Staphylococcus aureus is capable of infecting nearly every vertebrate organ system, triggering the formation of characteristic tissue lesions known as abscesses. Understanding how S. aureus survives within abscesses is critical to the development of new therapeutics, as these lesions represent the pathogen niche during infection. In this application, we will leverage a powerful mass spectrometry-based imaging platform to identify host and bacterial factors that contribute to staphylococcal disease. By defining how tissue niche and host biology drive molecular heterogeneity in abscesses, we will uncover new targets for tailored anti-staphylococcal therapeutics. Historically, it has been technically challenging to study the bacterial and host factors that contribute to abscess physiology for two primary reasons. First, approaches that seek to preserve abscess architecture within a tissue sample are inherently limited to the study of known microbial and host targets. This applies to techniques such as immunohistochemistry and fluorescence in situ hybridization, which can define the spatial distribution of analytes in a tissue but rely on pre-existing knowledge of targets. These approaches, by definition, cannot be used to discover unknown bacterial or host factors that contribute to disease. Conversely, discovery-based methods such as RNA sequencing or proteomics can identify novel, disease-associated analytes, but require destructive tissue processing that eliminates information regarding the spatial orientation of microbial and host molecules. To overcome these technical limitations, we created a mass spectrometry-based imaging platform to identify host and microbial analytes in abscessed tissue during invasive S. aureus infection. Because imaging mass spectrometry (IMS) does not require probes or detection reagents, this platform can define the localization and abundance of abscess-associated analytes in a spatially-defined manner, thereby enabling the discovery of microbial and host factors that contribute to disease pathogenesis. When applied to a model of disseminated S. aureus infection, this IMS-based platform enabled three-dimensional molecular imaging of the staphylococcal- host interface and powered the discovery of bacterial proteins that mark the pathogen niche within abscesses. Although individual abscesses typically have a similar histologic appearance, our IMS-based analysis revealed significant molecular heterogeneity between S. aureus lesions. We hypothesize that abscesses display molecular heterogeneity in response to tissue niche, antibacterial immune responses, and comorbid host conditions. To test this hypothesis, we will couple our IMS platform with laser capture microdissection to enable spatially-resolved proteomics of the staphylococcal-host interface. The proposed Aims will define the molecular architecture of abscesses across S. aureus infected tissues and determine how innate immune effector cells and host comorbidities drive heterogeneity in bacterial physiology and abscess molecular architecture in situ. In total, these experiments will decipher how host biology drives molecular heterogeneity during invasive infection.

项目摘要/摘要 金黄色葡萄球菌能够感染几乎所有脊椎动物的器官系统，触发形成 一种称为脓肿的特征性组织损伤。了解金黄色葡萄球菌是如何在脓肿中生存的 对新疗法的开发至关重要，因为这些损伤代表了感染期间的病原体生态位。 在这一应用中，我们将利用一个强大的基于质谱学的成像平台来识别主机和 导致葡萄球菌疾病的细菌因素。通过定义组织生态位和宿主生物如何推动 在脓肿中的分子异质性，我们将发现量身定制的抗葡萄球菌疗法的新靶点。 从历史上看，研究影响细菌和宿主的因素在技术上一直具有挑战性。 脓肿生理上有两个主要原因。首先，寻求在内部保留脓肿架构的方法 组织样本本质上仅限于对已知微生物和宿主目标的研究。这适用于技术 如免疫组织化学和荧光原位杂交，可以确定细胞的空间分布。 组织中的分析物，但依赖于对目标的预先存在的知识。根据定义，这些方法不能 用于发现导致疾病的未知细菌或宿主因素。相反，基于发现的 RNA测序或蛋白质组学等方法可以识别与疾病相关的新分析物，但需要 破坏性组织处理，消除有关微生物和宿主的空间取向的信息 分子。为了克服这些技术限制，我们创建了一个基于质谱学的成像平台 确定侵袭性金黄色葡萄球菌感染期间脓肿组织中的宿主和微生物分析物。因为成像 质谱学(IMS)不需要探针或检测试剂，这个平台可以定义定位 和丰富的脓肿相关分析物以空间定义的方式，从而能够发现 导致疾病发生的微生物和宿主因素。当应用于播散性S。 金黄色葡萄球菌感染，这个基于IMS的平台使三维分子成像成为可能- 宿主界面，并推动了细菌蛋白质的发现，这些蛋白质标记了脓肿内的病原体生态位。 尽管单个脓肿通常具有相似的组织学外观，但我们基于IMS的分析 显示金黄色葡萄球菌病变之间存在显著的分子异质性。我们假设脓肿表现为 组织生态位、抗菌免疫反应和共病宿主的分子异质性 条件。为了验证这一假设，我们将把我们的IMS平台与激光捕获显微解剖结合起来，以实现 葡萄球菌-宿主界面的空间分辨蛋白质组学。提议的目标将定义分子 金黄色葡萄球菌感染组织中脓肿的结构和确定先天免疫效应细胞和 宿主共生导致细菌生理和原位脓肿分子构型的异质性。总的来说， 这些实验将破译宿主生物如何在侵入性感染过程中驱动分子异质性。