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： 识别多种耐药的细菌感染，特别是金黄色葡萄球菌引起的感染。 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感染 疾病表现，通常是致命的使命，这是我们最好的，最重要的疗法。器官脓肿是 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层。 脓肿的形成本身遵循不同的发育阶段，受宿主和细菌的积极支持，并且 最终为金黄色葡萄球菌创造了有利的利基市场。虽然过去的研究产生了 脓肿结构，我们缺乏有关脓肿的分子组成的信息，尤其是在上下文 不同的脓肿阶段。对脓肿形成过程中分子事件的这种有限的知识是 特别令人震惊的是，它阻碍了有意义的反峰星策略的目标设计。 我们的初步数据表明，脓肿的形成的特征是宿主广泛搬迁 在称为营养免疫的过程中，过渡金属靠近脓肿。因此，体内 成像表明，脓肿内的细菌因锌和铁而饿死。由于可用的金属水平可以 作为入侵病原体的生物标志物，我们假设这种波动的元素分布策划 与脓肿形成相关的细菌活性。沿着这些线，我们表明锌饥饿素数 金黄色葡萄球菌，随后与不同的免疫细胞种群接触。除了这些发现之外， 年代和金属搬迁涉及的因素，金黄色葡萄球菌对这些刺激的检测以及相应 细菌反应是完全出乎意料的。因此，我们计划在此提案中解决这些问题。 当前对葡萄球菌组织发展有意义研究的设计障碍 脓肿是极大程度的脓肿异质性，可能是由于不同的发育阶段的结果 同一器官中的个体病变。为了说明组织脓肿的非同步性质，我们有 确定了一组潜在的蛋白质标记，用于不同的脓肿阶段。这些蛋白质将作为 分子时钟，因此我们可以通过发育过程遵循单个脓肿的进展。 基于这些标记，我们将创建体内记者并表征开发的分子清单 脓肿，专注于元素和蛋白质成分的变化。在这里，我们将关联各种 体内成像方式，包括3D生物发光成像，MRI和成像质谱法，并具有 通过微液体提取表面分析的晚期蛋白质组学。一旦我们确定了脓肿 微环境在脓肿形成的不同阶段发生变化，我们将执行转录组分析 细菌亚群来评估环境刺激如何影响葡萄球菌病理生理学和在 转弯，脓肿的发展。合并，提出的实验将检查宿主病原体的事件 界面并为新颖和有针对性的治疗策略铺平道路，以对抗葡萄球菌感染。