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.

总结： 多重耐药细菌感染，特别是由金黄色葡萄球菌引起的感染， 是21世纪世纪最大的威胁之一血液感染是最严重的葡萄球菌感染 尽管我们有最好和最新的治疗方法，但这些疾病往往是致命的。器官移植是 主要贡献者S。金黄色葡萄球菌全身感染，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，由宿主和细菌积极促进， 最终为S创造了一个有利的利基。金黄色。虽然过去的研究已经产生了一个概述， 脓肿结构，我们缺乏脓肿分子组成的信息，特别是在上下文中， 不同的脓肿阶段。这种对脓肿形成过程中分子事件的有限认识， 尤其令人担忧的是，它阻碍了有针对性地设计抗葡萄球菌策略的有意义的尝试。 我们的初步资料显示，脓肿形成的特点是宿主的广泛迁移， 过渡金属在接近脓肿的过程中被称为营养免疫。因此，在体内 成像显示脓肿内的细菌缺乏锌和铁。由于可用的金属水平可以 作为入侵病原体的生物标志物，我们假设波动的元素分布协调了 与脓肿形成有关的细菌活动。沿着这些路线，我们发现锌饥饿引发 S.金黄色葡萄球菌用于随后与不同的免疫细胞群体接触。然而，除了这些发现， 年表和因素参与金属搬迁，检测这些刺激的S。金黄色葡萄球菌，以及相应的 细菌的反应是完全未知的。因此，我们计划在本提案中解决这些问题。 目前对葡萄球菌组织发育进行有意义研究的一个障碍是 脓肿的异质性程度很高，可能是由于不同的发展阶段， 同一器官的单个病变。为了解释组织扩张的非同步性质，我们有 鉴定了一组不同脓肿阶段的潜在蛋白质标记物。这些蛋白质将作为 分子钟，这样我们就可以通过发育过程跟踪个体的发育进程。 基于这些标记，我们将创建体内报告基因，并表征开发中的分子库存。 主要研究元素和蛋白质组成的变化。在这里，我们将各种 体内成像模式，包括3D生物发光成像、MRI和成像质谱， 先进的蛋白质组学通过微液萃取表面分析。一旦我们确定了脓肿是如何 在脓肿形成的不同阶段，我们将进行转录组分析 的细菌亚群，以评估环境刺激如何影响葡萄球菌的病理生理学， 转，脓肿发展。结合起来，拟议的实验将检查在宿主-病原体 接口和铺平道路的新的和有针对性的治疗策略，以打击葡萄球菌感染。