Harnessing polymicrobial interactions in the catheterized urinary tract to identify novel inhibitors of Proteus mirabilis urease activity

利用插管尿路中的多种微生物相互作用来鉴定奇异变形杆菌尿素酶活性的新型抑制剂

• 批准号: 10678389
• 负责人: Lauren Beryl Guterman
• 金额: $ 3.73万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10678389
• 关键词: Acids; Active Sites; Ammonia; Animals; Antibiotics; Bacteremia; Bacteria; Bathing; Biological Availability; Biomass; Bladder; Carbon Dioxide; Catheterization; Catheters; Cells; Cessation of life; Clinical; Clinical Trials; Communicable Diseases; Cytoplasm; Data; Development; Dose; Drainage procedure; Enterococcus faecalis; Enzymes; Extracellular Space; Foundations; Future; Glass; Goals; Histamine; Hospitalization; Human; Imidazole; In Vitro; Incidence; Incubated; Individual; Indwelling Catheter; Infection; Innate Immune Response; Internal Medicine; Investigation; Ions; Klebsiella aerogenes; Knowledge; Laboratories; Length of Stay; Leu-Gly; Measures; Mentors; Messenger RNA; Microbial Biofilms; Modeling; Morbidity - disease rate; Morganella morganii; National Research Service Awards; Nickel; Organism; Pain; Pathogenesis; Patient-Focused Outcomes; Patients; Physicians; Physiological; Preceptorship; Precipitation; Process; Production; Proteins; Proteus mirabilis; Providencia; Publications; Research; Risk; Science; Scientist; Sepsis; Severities; Severity of illness; Staphylococcus aureus; System; Temperature; Testing; Therapeutic; Time; Tissues; Toxic effect; Training; United States Food and Drug Administration; Urea; Urease; Urinary Calculi; Urinary Catheterization; Urinary tract; Urinary tract infection; Urine; Virulence Factors; Water; Work; antimicrobial; catheter associated UTI; drug discovery; efficacy evaluation; efficacy testing; enhancing factor; experimental study; improved; in vitro Model; in vitro activity; inhibitor; insight; instrumentation; interest; medical schools; metabolomics; metalloenzyme; microbial; mortality; mouse model; new therapeutic target; novel; novel therapeutics; pathogen; phenylpyruvate; pre-clinical assessment; prevent; side effect; translational potential; urinary

Project Summary Catheter-associated urinary tract infection (CAUTI) is the leading cause of secondary nosocomial bloodstream infections. Urease-producing organisms such as Proteus mirabilis are common causes of CAUTI and associated with numerous complications. Urease hydrolyzes urea to ammonia, which raises the urine pH and causes precipitation of ions into crystals, ultimately leading to catheter encrustation and blockage and urinary stone formation and increasing the risk of bacteremia, sepsis and death. The only urease inhibitor approved by the Food and Drug Administration (FDA), acetohydroxamic acid (AHA), shows efficacy in preventing urinary stone formation but has severe side effects that limit its clinical use. Therefore, alternative strategies are needed for targeting bacterial urease activity to prevent catheter encrustation and stone formation. In patients with long-term indwelling catheters, CAUTI is often polymicrobial. As demonstrated in our prior publications and preliminary data, we found that common constituents of polymicrobial CAUTI secrete molecules that modulate P. mirabilis urease activity and infection severity. We therefore performed untargeted global metabolomics analysis on cell-free supernatants of urease-modulating bacterial strains and identified 36 candidate urease dampening metabolites. We validated the activity of four compounds: histamine (CAS 51-45- 6), leucylglycine (CAS 686-50-0), phenylpyruvate (CAS 156-06-9) and imidazole lactate (CAS 14403-45-3). Our preliminary data suggests that leucylglycine and imidazole lactate dampen urease activity, at least in part, by acting directly on the urease active site. In contrast, histamine and phenylpyruvate appear to indirectly inhibit urease activity through an uncharacterized mechanism. The goal of this NRSA F30 proposal is to determine the mechanism of action of urease modulation and conduct pre-clinical assessment of the therapeutic potential of these dampening compounds. Aim 1 will determine the mechanism of indirect urease modulation for histamine and phenylpyruvate and explore whether compounds of different mechanisms of action have synergistic effects on urease activity when administered in combination against a panel of urease- producing pathogens. Aim 2 will assess the translational potential of dampening compounds for preventing Foley catheter encrustation, crystalline biofilm formation, and blockage. The experiments outlined in this proposal will provide a framework for harnessing polymicrobial interactions for drug discovery and the development of novel therapeutics to prevent CAUTI associated morbidity and mortality. This work will take place at the Jacobs School of Medicine and Biomedical Sciences in the laboratory of Dr. Chelsie Armbruster, who is an expert in CAUTI and microbial pathogenesis research. The training plan is tailored for my development as a physician-scientist in the field of infectious disease, and includes mentoring by successful physician-scientists and clinical preceptorships in infectious disease and internal medicine.

项目摘要 导管相关性尿路感染（CAUTE-associated urinary tract infection，CAUTE-TI）是引起继发性院内血流的主要原因 感染.产尿素酶的微生物如奇异变形杆菌（Proteusmirabilis）是引起尿路感染的常见原因， 与许多并发症有关。尿素酶将尿素水解为氨，这会提高尿液pH值， 导致离子沉淀成晶体，最终导致导管结壳和堵塞， 结石形成和增加菌血症、败血症和死亡的风险。唯一的尿素酶抑制剂批准由 美国食品和药物管理局（FDA），乙酰异羟肟酸（AHA），显示出预防尿 但具有严重的副作用，限制了其临床应用。因此，替代策略是 所需的靶向细菌尿素酶活性，以防止导管结壳和结石形成。患者 对于长期留置导管而言，ESTTI通常是多微生物的。正如我们以前的出版物所展示的那样， 和初步的数据，我们发现，共同组成的多微生物分泌的分子， 调节奇异变形杆菌尿素酶活性和感染严重程度。因此，我们在全球范围内 对尿素酶调节细菌菌株的无细胞上清液进行代谢组学分析，并鉴定了36种 候选脲酶抑制代谢物。我们验证了四种化合物的活性：组胺（CAS 51-45- 2000）。 6）、亮氨酰甘氨酸（CAS 686-50-0）、苯丙酮酸盐（CAS 156-06-9）和咪唑乳酸盐（CAS 14403-45-3）。 我们的初步数据表明亮氨酰甘氨酸和咪唑乳酸盐至少部分抑制了脲酶活性， 通过直接作用于脲酶活性部位。相比之下，组胺和苯丙酮酸盐似乎间接 通过未表征的机制抑制脲酶活性。NRSA F30提案的目标是 确定尿素酶调节的作用机制，并进行临床前评估， 这些阻尼化合物的治疗潜力。目的1将确定间接脲酶的作用机理 调节组胺和苯丙酮酸，并探讨是否化合物的不同机制， 当与一组尿素酶联合给药时， 产生病原体。目标2将评估阻尼化合物的转化潜力， Foley导管结壳、结晶生物膜形成和堵塞。本文中概述的实验 该提案将为利用多微生物相互作用进行药物发现提供一个框架， 开发新的治疗方法，以预防与TBI相关的发病和死亡。这项工作将需要 放置在雅各布斯医学和生物医学科学学院的Chelsie Armbruster博士实验室， 他是一位抗真菌感染和微生物致病机理研究方面的专家。培训计划是为我量身定制的。 发展作为一个医生，科学家在传染病领域，并包括指导成功的 传染病和内科的医生科学家和临床导师。