Defining the molecular inventory of staphylococcal tissue abscesses and its effects on the host-pathogen interface

定义葡萄球菌组织脓肿的分子库存及其对宿主-病原体界面的影响

• 批准号: 10533374
• 负责人: Andy Weiss
• 金额: --
• 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: 2020-12-01 至 2022-12-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10533374
• 关键词: 3-Dimensional; Abscess; Address; Affect; Architecture; Bacteria; Bacterial Endocarditis; Bacterial Infections; Biological Markers; Cells; Chronology; Cues; Data; Detection; Development; Developmental Process; Disease; Elements; Environment; Equipment and supply inventories; Event; Experimental Designs; Functional disorder; Genus staphylococcus; Heterogeneity; Hospitalization; Image; Immune; Immune system; In Vitro; Individual; Infection; Integration Host Factors; Invaded; Investigation; Iron; Knowledge; Lesion; Liquid substance; Magnetic Resonance Imaging; Metals; Micronutrients; Modeling; Molecular; Multi-Drug Resistance; Multimodal Imaging; Mus; Nature; Nutritional Immunity; Organ; Phase; Phenotype; Population; Process; Proteins; Proteome; Proteomics; Reporter; Resolution; Role; Sepsis; Shapes; Site; Staphylococcal Infections; Staphylococcus aureus; Starvation; Stimulus; Surface; System; Systemic infection; Technology; Testing; Time; Tissues; Transition Elements; United States; Zinc; combat; design; experimental study; imaging modality; imaging platform; in vivo; in vivo imaging; mass spectrometric imaging; molecular clock; molecular marker; molecular shape; mutant; novel; pathogen; prevent; response; spatiotemporal; targeted treatment; transcriptome; transcriptomics; treatment strategy

Summary: Multidrug-resistant bacterial infections, particularly those caused by Staphylococcus aureus, are recognized as one of the greatest threats of the 21st century. Bloodstream infections are the most severe staphylococcal disease manifestation and are often fatal despite our best and most current therapies. Organ abscesses are primary contributors to S. aureus systemic infections and serve as initial reservoirs for the invading pathogen. Abscess formation itself follows distinct developmental stages, is actively facilitated by host and bacterium, and ultimately creates an advantageous niche for S. aureus. While past studies have generated an overview of abscess architecture, we lack information on the molecular composition of abscesses, particularly in the context of different abscess stages. This limited knowledge of the molecular events during abscess formation is especially alarming, for it hinders meaningful attempts at targeted design of anti-staphylococcal strategies. Our preliminary data show that abscess formation is characterized by the host’s extensive relocation of transition metals in proximity to the abscess in a process known as nutritional immunity. Consequently, in vivo imaging reveals that bacteria within the abscess are starved for zinc and iron. Since available metal levels can serve as biomarkers for invading pathogens, we hypothesize that fluctuating elemental distributions orchestrate bacterial activities associated with abscess formation. Along these lines, we showed that zinc starvation primes S. aureus for subsequent contact with different immune cell populations. Beyond these findings, however, the chronology and factors involved in metal relocation, detection of these stimuli by S. aureus, and corresponding bacterial responses are entirely unexplored. We thus plan to address these questions in this proposal. One current barrier to the design of meaningful investigations into the development of staphylococcal tissue abscesses is a significant degree of abscess heterogeneity, likely a result of different developmental stages of individual lesions in the same organ. To account for the non-synchronous nature of tissue abscesses, we have identified a group of potential proteinaceous markers for different abscess stages. These proteins will serve as molecular clocks so we can follow the progression of individual abscesses through the developmental process. Based on these markers, we will create in vivo reporters and characterize the molecular inventory of developing abscesses, focusing on changes in elemental and proteinaceous compositions. Here, we will correlate various in vivo imaging modalities, including 3D-bioluminiscent imaging, MRI, and imaging mass spectrometry, with advanced proteomics via micro Liquid Extraction Surface Analysis. Once we have established how the abscess microenvironment changes during different phases of abscess formation, we will perform transcriptome analysis of bacterial subpopulations to assess how environmental stimuli affect staphylococcal pathophysiology and, in turn, abscess development. Combined, the proposed experiments will examine the events at the host-pathogen interface and pave the way for novel and targeted treatment strategies to combat staphylococcal infections.

概括： 多重耐药细菌感染，特别是由金黄色葡萄球菌引起的感染，已得到公认 作为 21 世纪最大的威胁之一，血液感染是最严重的葡萄球菌。 尽管我们目前最好和最新的治疗方法是治疗器官脓肿，但这种疾病的表现往往是致命的。 金黄色葡萄球菌全身感染的主要贡献者，并作为入侵病原体的初始储存库。 脓肿形成本身遵循不同的发育阶段，受到宿主和细菌的积极促进，并且 最终为金黄色葡萄球菌创造了一个有利的生态位，而过去的研究已经对金黄色葡萄球菌进行了概述。 脓肿结构，我们缺乏有关脓肿分子组成的信息，特别是在这种情况下 对脓肿形成过程中分子事件的有限了解。 尤其令人担忧的是，它阻碍了针对抗葡萄球菌策略的针对性设计的有意义的尝试。 我们的初步数据表明，脓肿形成的特点是宿主的广泛迁移 过渡金属在体内的营养免疫测试过程中接近脓肿。 成像显示脓肿内的细菌缺乏锌和铁，因为可用的金属水平可以。 作为入侵病原体的生物标志物，我们与波动的元素分布合作协调 沿着这些思路，我们发现锌饥饿引发了细菌活动。 然而，除了这些发现之外，金黄色葡萄球菌随后与不同的免疫细胞群接触。 金属重新定位所涉及的年代学和因素、金黄色葡萄球菌对这些刺激的检测以及相应的 因此，我们计划在本提案中解决这些问题。 当前设计对葡萄球菌组织发育有意义的研究的一个障碍 脓肿具有显着程度的异质性，可能是脓肿不同发育阶段的结果 为了解释组织脓肿的非同步性质，我们有 确定了一组不同脓肿阶段的潜在蛋白质标记物。 分子钟，这样我们就可以跟踪各个脓肿在发育过程中的进展情况。 基于这些标记，我们将创建体内生产并表征开发中的分子库存 脓肿，重点关注元素和蛋白质成分的变化在这里，我们将关联各种。 体内成像模式，包括 3D 生物发光成像、MRI 和成像质谱分析 一旦我们确定了脓肿如何发生，就可以通过微液体提取表面分析进行先进的蛋白质组学。 脓肿形成的不同阶段微环境发生变化，我们将进行转录组分析 细菌亚群，以评估环境刺激如何影响葡萄球菌病理生理学，并且 结合起来，拟议的实验将检查宿主病原体的事件。 接口并为对抗葡萄球菌感染的新颖和有针对性的治疗策略铺平道路。