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

基本信息

批准号：
10565912
负责人：
JAMES E CASSAT
金额：
$ 74.07万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
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：
2020-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10565912
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 Macrophage 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 Technology Testing Therapeutic Therapeutic Intervention Time Tissue Sample Tissue imaging Tissues Translational Research Vaccines Work antimicrobial body system cell type comorbidity experimental study human pathogen imaging platform in vivo innovation laser capture microdissection mass spectrometric imaging microbial microbial host molecular imaging mouse model neutrophil novel novel therapeutics pathogen preservation response therapeutic candidate tissue processing transcriptome sequencing translational medicine

项目摘要

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.

项目概要/摘要金黄色葡萄球菌能够感染几乎所有脊椎动物器官系统，引发形成称为脓肿的特征性组织病变。了解金黄色葡萄球菌如何在脓肿中生存是对于新疗法的开发至关重要，因为这些病变代表了感染期间的病原体生态位。在此应用中，我们将利用强大的基于质谱的成像平台来识别宿主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学如何驱动由于脓肿中的分子异质性，我们将发现定制抗葡萄球菌疗法的新靶点。从历史上看，研究导致感染的细菌和宿主因素在技术上一直具有挑战性。脓肿生理学有两个主要原因。首先，寻求保留脓肿结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。这适用于技术例如免疫组织化学和荧光原位杂交，可以定义空间分布组织中的分析物，但依赖于预先存在的目标知识。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 RNA 测序或蛋白质组学等方法可以识别新的、与疾病相关的分析物，但需要破坏性组织处理，消除有关微生物和宿主空间方向的信息分子。为了克服这些技术限制，我们创建了一个基于质谱的成像平台鉴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（IMS）不需要探针或检测试剂，该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病机制的微生物和宿主因素。当应用于传播 S 模型时。金黄色葡萄球菌感染，这个基于 IMS 的平台能够对葡萄球菌进行三维分子成像宿主界面并推动了标记脓肿内病原体生态位的细菌蛋白的发现。尽管个别脓肿通常具有相似的组织学外观，但我们基于 IMS 的分析揭示了金黄色葡萄球菌病变之间显着的分子异质性。我们假设脓肿显示对组织生态位、抗菌免疫反应和共病宿主的分子异质性状况。为了验证这一假设，我们将 IMS 平台与激光捕获显微切割相结合，以实现葡萄球菌-宿主界面的空间分辨蛋白质组学。拟议的目标将定义分子金黄色葡萄球菌感染组织的脓肿结构，并确定先天免疫效应细胞和宿主合并症导致细菌生理学和脓肿分子结构的异质性。总共，这些实验将破译宿主生物学如何在侵入性感染过程中驱动分子异质性。