A comprehensive investigation of Pseudomonas quorum sensing regulatory relationships and the consequences on quorum sensing inhibitors in complex communities

复杂群落中假单胞菌群体感应调控关系及其对群体感应抑制剂影响的全面研究

• 批准号: 10716869
• 负责人: LAURENCE G RAHME
• 金额: $ 75.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10716869
• 关键词: 16S ribosomal RNA sequencing; Acute; Address; Antibiotic Resistance; Antibiotics; Architecture; Bacteria; Bacterial DNA; Bacterial Infections; Cells; Chronic; Clinical; Collection; Communication; Communities; Complex; Disease Progression; ESKAPE pathogens; Environment; Feces; Frequencies; Gene Mutation; Goals; Healthcare; Homeostasis; Human; Immune; Immunofluorescence Immunologic; Immunologics; In Vitro; Infection; Inflammation; Inflammatory Response; Intestinal permeability; Investigation; Lead; Lung; Metabolic; Microbe; Microbial Biofilms; Modeling; Molecular; Monitor; Multi-Drug Resistance; Mus; Mutation; Pathogenesis; Pathogenicity; Pathway interactions; Proliferating; Pseudomonas; Pseudomonas aeruginosa; Pseudomonas aeruginosa infection; Publishing; Regulation; Relapse; Resistance; Resistance development; Role; Severities; Signal Transduction; Stains; System; Systemic infection; Therapeutic; Time; Trachea; Transcriptional Regulation; Virulence; Virulence Factors; Virulent; Work; antagonist; antibiotic tolerance; antimicrobial; bacterial resistance; bactericide; clinically relevant; combat; cytokine; dysbiosis; fighting; gut microbiota; health care settings; host-associated microbial communities; in vivo; inhibitor; insight; microbial; microbial community; microbiota; microorganism interaction; multi-drug resistant pathogen; mutant; novel; pathogen; preservation; pressure; quorum sensing; small molecule; synergism; traditional therapy; transcriptome sequencing

Our overarching goal is to develop efficacious strategies to combat multidrug-resistant (MDR) pathogens, a growing healthcare threat. Quorum sensing (QS) inhibition is an attractive anti-microbial approach, as it represents an anti-virulence strategy fundamentally different from current traditional therapies. QS a highly conserved bacterial communication signaling regulatory mechanism controlling numerous virulence functions and inter- and intraspecies interactions in mono- and polymicrobial communities. The recalcitrant ESKAPE pathogen Pseudomonas aeruginosa (PA) exemplifies a highly problematic nosocomial pathogen with astounding complex interconnected QS systems controlling multiple virulence functions and regulating antagonism or synergism with other microbes. QS relevance in pathogenesis has been recognized; however, the regulatory interrelationships of these systems during infection and in polymicrobial settings have been studied primarily in vitro, limiting relevance to the human environment during infection and leaving answered fundamental questions that will aid in combatting MDR infections by utilizing the anti-virulence/anti-QS approach. We hypothesize that the interrelationships between the three QS systems are significant drivers of infection severity, interspecies interactions, and microbial community composition. In turn, we anticipate that our anti-QS lead compound will restrict infection exacerbations and promote microbial homeostasis. We propose to assess this hypothesis paradigmatically by leveraging an extensive collection of PA QS mutants and a novel anti-QS compound in dynamic and human-relevant host environments to identify specific infection-relevant host-microbe and microbe-microbe interactions and dissect the mechanisms underlying these interactions. Specifically, in the proposed three Aims, we will address three fundamental questions that will fill existing gaps: 1) What are the regulatory relationships and functions of the three major PA QS systems? 2) How do anti-QS therapies impact infection severity and QS regulatory interactions in PA mono- and polymicrobial settings? and 3) How do the QS systems and inhibitors impact the development of resistance and limit perturbations of host-associated microbial communities? Answering these questions will not only provide critical insights into long-standing questions about the anti-virulence approach and the role of QS mutations in PA virulence but will also uncover how the in vivo environment impacts the regulatory interrelationships of the three QS systems and QS-dependent inter- and intraspecies interactions. By tapping into bacterial QS pathways with novel potent and highly specific anti-QS compounds, we aim to treat infections while preserving the composition and protective function of the microbiota. Results from these dynamic settings may aid in the fight against antibiotic resistance and provide fundamental insights into the molecular mechanisms underlying the virulence of complex infections and microbial interactions in vivo.

我们的首要目标是制定有效的策略来对抗多重耐药（MDR）病原体， 日益严重的医疗保健威胁。群体感应（QS）抑制是一种有吸引力的抗微生物方法，因为它 代表了一种与当前传统疗法根本不同的抗毒力策略。 QS 高度 保守的细菌通讯信号调节机制控制着多种毒力功能 以及单一和多微生物群落中的种间和种内相互作用。顽固的埃斯卡普 病原体 铜绿假单胞菌 (PA) 是一种高度成问题的医院病原体，其危害性令人震惊 复杂的相互关联的 QS 系统控制多种毒力功能并调节拮抗作用或 与其他微生物有协同作用。 QS 与发病机制的相关性已得到认可；然而，监管 这些系统在感染期间和多种微生物环境中的相互关系主要在以下领域进行了研究： 体外，限制感染期间与人类环境的相关性并留下基本问题的答案 这将有助于通过利用抗毒力/抗 QS 方法来对抗 MDR 感染。 我们假设三个 QS 系统之间的相互关系是重要的驱动因素 感染严重程度、种间相互作用和微生物群落组成。反过来，我们预计我们的 抗 QS 先导化合物将限制感染恶化并促进微生物稳态。 我们建议通过利用广泛的 PA QS 集合来范例性地评估这一假设 突变体和新型抗 QS 化合物在动态和人类相关的宿主环境中识别 特定感染相关的宿主-微生物和微生物-微生物相互作用并剖析其机制 这些相互作用的基础。具体来说，在拟议的三个目标中，我们将解决三个基本目标 将填补现有空白的问题：1）三大PA的监管关系和职能是什么 QS系统？ 2) 抗 QS 疗法如何影响 PA 单克隆感染的严重程度和 QS 调节相互作用 和多种微生物环境？ 3) QS系统和抑制剂如何影响耐药性的发展 并限制宿主相关微生物群落的扰动？回答这些问题不仅 对有关抗毒力方法和 QS 作用的长期存在的问题提供重要见解 PA 毒力的突变，但也将揭示体内环境如何影响监管 三个 QS 系统的相互关系以及依赖于 QS 的种间和种内相互作用。 通过利用新型有效且高度特异性的抗 QS 化合物来利用细菌 QS 途径，我们的目标是 治疗感染，同时保留微生物群的组成和保护功能。这些结果 动态设置可能有助于对抗抗生素耐药性，并提供对抗生素耐药性的基本见解 复杂感染的毒力和体内微生物相互作用的分子机制。