Assessing Klebsiella pneumoniae invasion of the intact gut microbiome

评估肺炎克雷伯菌对完整肠道微生物组的入侵

• 批准号: 10780120
• 负责人: Jay Vornhagen
• 金额: $ 24.88万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-05 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10780120
• 关键词: 16S ribosomal RNA sequencing; Address; Antibiotic Resistance; Antibiotics; Bacteremia; Bacteria; Bacterial Physiology; Biological Assay; Categories; Cells; Citrate (si)-Synthase; Clinic; Complement; Data; Development; Disease; Ecology; Extended-spectrum β-lactamase; Foundational Skills; Health; Hospitals; Human; In Vitro; Indigenous; Infection; Inflammation; Invaded; Investigation; Klebsiella pneumoniae; Knowledge; Libraries; Life; Mediating; Mentors; Metabolic; Michigan; Mission; Nosocomial pneumonia; Operon; Pathogenesis; Patients; Physiological; Pneumonia; Population; Process; Production; Public Health; Publications; Research; Resistance; Risk; Role; Stress; System; Tellurium; Testing; Therapeutic; Therapeutic Intervention; Training; United States National Institutes of Health; Universities; Urinary tract infection; Work; antibiotic resistant infections; antimicrobial peptide; biological adaptation to stress; carbapenemase; density; design; enhancing factor; fitness; gut colonization; gut microbes; gut microbiome; gut microbiota; host microbiota; infection risk; inflammatory milieu; innovation; member; microbial; microbial composition; microbial host; microbiome; microbiota; mouse model; novel; novel diagnostics; novel therapeutics; pathogen; prevent; resilience; response; stress resilience; stress tolerance; therapeutic development; therapeutic target; therapy design; transposon sequencing

Project summary/abstract The bacteria Klebsiella pneumoniae is an important cause of hospital-acquired and antibiotic-resistant infections, including pneumonia, bacteremia, and urinary tract infection. Infection risk sharply increases when K. pneumoniae is present in the gut, and K. pneumoniae must overcome many barriers to successfully colonize the gut, including the gut microbiome. While disruption of the gut microbiome through antibiotics can permit colonization, K. pneumoniae frequently colonizes the gut in the absence of antibiotics when an intact microbiome is present. There is a fundamental gap in our understanding of how K. pneumoniae invades the intact gut microbiome. Furthermore, the specific mechanisms underlying the ability of K. pneumoniae to compete with the endogenous gut microbiota remain unexplored. The objective of this proposal is to identify and characterize factors necessary for K. pneumoniae gut colonization in an intact gut microbiome and determine how these factors influence K. pneumoniae fitness in the gut. The central hypothesis of this proposal is that Kp employs microbiome dependent and specific factors to enhance their ability to directly compete with the host microbiota or survive microbiota-mediated modulation of host inflammation to invade the intact gut microbiome, which is critical for Kp gut colonization. I will test this hypothesis in three specific aims: 1) define the role of a previously identified gut fitness factor, the tellurium resistance (ter) operon during invasion of an intact gut microbiome; 2) determine the role of the ter operon in response to physiologically relevant stresses and; 3) systematically identify K. pneumoniae fitness factors during invasion of intact gut microbiomes. The work in this proposal is innovative, as it requires the development of novel transposon sequencing application, and it addresses a gap in understanding that can shift the current paradigm of K. pneumoniae gut colonization. Completion of this work will have sustained positive impact through the identification, characterization, and prioritization of K. pneumoniae factors necessary for invasion of the intact gut microbiome, which will help guide the development of therapeutics targeted at the disruption of K. pneumoniae gut colonization. Finally, the research in this proposal is complemented by a comprehensive training plan designed to develop a foundational skillset for the pursuit of research focused on mechanisms of bacterial pathogenesis directly informed by and applicable to the clinic. This project will be undertaken within the University of Michigan in conjunction with a distinguished team of co- mentors. This team of co-mentors will complement that candidate's expertise in host-pathogen interaction by providing training in microbial ecology, exploration of bacterial physiology, and comprehensive analyses of host- microbial systems necessary for the candidate's transition to independence. These state-of-the-art studies will lead to the identification and characterization of K. pneumoniae factors that permit the invasion of the intact gut microbiome and the corresponding role of the microbiota in this process to enable the design of therapies to reduce the impact of K. pneumoniae disease.

项目概要/摘要 肺炎克雷伯菌是医院获得性感染和抗生素耐药性感染的重要原因， 包括肺炎、菌血症和尿路感染。当 K. 感染风险急剧增加。 肺炎克雷伯菌存在于肠道中，肺炎克雷伯菌必须克服许多障碍才能成功定植于肠道。 肠道，包括肠道微生物组。虽然通过抗生素破坏肠道微生物组可以允许 在没有抗生素的情况下，当完整的微生物组存在时，肺炎克雷伯菌经常在肠道中定殖 存在。我们对肺炎克雷伯菌如何侵入完整肠道的理解存在根本性差距 微生物组。此外，肺炎克雷伯菌与肺炎克雷伯菌竞争的能力的具体机制 内源性肠道微生物群仍未被探索。该提案的目的是确定并描述 肺炎克雷伯菌在完整肠道微生物组中肠道定植所必需的因素，并确定这些因素如何 影响肺炎克雷伯菌在肠道内适应性的因素。该提案的中心假设是 Kp 采用 微生物组依赖性和特定因素可增强其与宿主微生物群直接竞争的能力 或者在微生物介导的宿主炎症调节中存活下来，侵入完整的肠道微生物组，这是 对于 Kp 肠道定植至关重要。我将从三个具体目标来检验这个假设：1）定义先前的角色 确定了肠道健康因子，即在完整肠道微生物组入侵期间​​的碲抗性（ter）操纵子； 2） 确定三元操纵子在响应生理相关应激中的作用； 3）系统地识别 肺炎克雷伯菌入侵完整肠道微生物群期间的适应性因素。本提案中的工作具有创新性， 因为它需要开发新型转座子测序应用程序，并且它解决了 了解这可以改变目前肺炎克雷伯菌肠道定植的模式。完成这项工作 将通过识别、描述和优先考虑 K 来产生持续的积极影响。 入侵完整肠道微生物组所必需的肺炎因子，这将有助于指导发展 针对破坏肺炎克雷伯菌肠道定植的治疗方法。最后，本提案的研究 还辅以一项全面的培训计划，旨在培养追求以下目标的基本技能： 研究重点是直接了解并适用于临床的细菌发病机制。这 该项目将在密歇根大学内与一个杰出的合作团队一起进行 导师。该联合导师团队将通过以下方式补充候选人在宿主与病原体相互作用方面的专业知识： 提供微生物生态学、细菌生理学探索以及宿主的综合分析方面的培训 候选人过渡到独立所必需的微生物系统。这些最先进的研究将 导致肺炎克雷伯菌因子的鉴定和表征，这些因子允许侵入完整的肠道 微生物组以及微生物组在此过程中的相应作用，使疗法的设计能够 减少肺炎克雷伯菌疾病的影响。