Investigating phage therapy for the treatment of urinary tract infections

研究噬菌体疗法治疗尿路感染

基本信息

批准号：
10677257
负责人：
Jacob Jeffrey Zulk
金额：
$ 4.77万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-09 至 2025-08-08
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10677257
关键词：
Affect Animal Model Antibacterial Response Antibiotic Resistance Antibiotic Therapy Antibiotics Antimicrobial Resistance Attenuated Automobile Driving Bacteria Bacterial Antibiotic Resistance Bacterial Infections Bacteriology Bacteriophages Binding Bladder Campylobacter coli Cells Clinic Clinical Collection Communities Complement Data Defect Development Diabetes Mellitus Diagnosis Disease model Elderly Environment Enzyme-Linked Immunosorbent Assay Epithelium Escherichia coli Fellowship Flow Cytometry Goals Growth Health Care Costs Human Immune Immune response Impairment In Vitro Infection Intracellular Space Knowledge Label Lipopolysaccharides Mammalian Cell Medical Medicine Membrane Membrane Proteins Mentorship Methods Microscopy Modeling Mus Mutation Nutrient Nutrient availability Organoids Pathogenesis Patients Phage Receptors Predisposition Proteins Public Health Publishing Recurrence Research Research Personnel Resistance Route Safety Techniques Testing Therapeutic Training Translating United States Urinary tract Urinary tract infection Urine Uropathogenic E. coli Virus Western Blotting Woman Work antimicrobial antimicrobial peptide bacterial fitness career career development college cost cytokine driving force environmental stressor experience extracellular fitness host microbiome human imaging in vivo in vivo Model infectious disease treatment innovation live cell imaging mouse model mutant novel pathogen pathogenic bacteria receptor recurrent infection skills standard of care superresolution microscopy tool uptake urinary

项目摘要

Urinary tract infections (UTIs) are responsible for a large public health burden with over half of women experiencing a UTI during their lifetime. These infections, primarily in women, older adults, and those with underlying conditions such as diabetes, are responsible for over $3.5 billion in health care costs annually in the USA alone. The majority of UTIs are caused by uropathogenic E. coli (UPEC) and are readily treated with antibiotics. However, global rise of antibiotic resistance amongst bacterial pathogens threatens the utility of the current standard of care. Furthermore, up to 30% of UPEC UTIs will have recurrence within 6 months of treatment due to intracellular UPEC reservoirs that are hidden from antibiotic therapy. Because of these reasons, alternatives to antibiotics are desperately needed. Bacteriophages (phage), viruses that infect bacteria, have long been hypothesized as a treatment for bacterial infection, predating the use of antibiotics by over a quarter of a century. Unfortunately, like antibiotics, bacteria can become resistant to phage. Preliminary work has identified several phage-resistant UPEC harboring mutations in lipopolysaccharide (LPS). These bacteria, although capable of evading phage, are poorer at growing in urine and at colonizing the bladder than their parental strains. The exact reason(s) for these fitness defects, however, are unknown. This work also aims to assess the utility of phage for treating intracellular UPEC reservoirs, which are untouched by antibiotic therapy, and which lead to UTI recurrence. Published and preliminary data suggests phage can interact and be internalized by bladder cells, although no group has tested the ability of phage to reduce intracellular UPEC burdens. Taken together, this preliminary data supports the hypothesis that phage therapy, through driving phage resistance that leaves bacteria less fit, and through phage interactions with the bladder, will be effective at treating both intracellular and extracellular UTIs. This hypothesis will be investigated by the following specific aims: 1) Identify mechanisms underlying decreased growth in urine and poor colonization of the murine bladder in phage resistant (LPS mutant) UPEC, 2) Characterize phage binding and internalization by the uroepithelium, reduction of intracellular reservoirs, and stimulation of bladder immune responses. In investigating these aims, several innovative tools such as primary bladder cell-derived organoids, super-resolution microscopy, and in vivo mouse models of disease and phage therapy will be used. This proposal will provide the candidate with training in UTI animal models, advanced microscopy techniques, protein isolation, and bladder organoid models. This training, along with excellent mentorship and career development goals, will provide the candidate with the necessary skills for an independent research career studying novel translational treatments for infectious disease. This training will take place at Baylor College of Medicine under the mentorship of leaders in the fields of antimicrobial resistance, phage therapy, and host-pathogen interactions. Taken together, this research will provide the information necessary to translate UTI phage therapy from the bench into the clinic.

尿路感染 (UTI) 造成巨大的公共健康负担，超过一半的女性患有尿路感染 (UTI) 一生中经历过尿路感染。这些感染主要发生在女性、老年人和患有糖尿病等潜在疾病每年造成超过 35 亿美元的医疗保健费用仅美国。大多数尿路感染是由尿路致病性大肠杆菌 (UPEC) 引起的，并且很容易用抗生素。然而，全球细菌病原体抗生素耐药性的上升威胁着抗生素的使用。目前的护理标准。此外，高达 30% 的 UPEC 尿路感染会在治疗后 6 个月内复发由于细胞内 UPEC 储库在抗生素治疗中是隐藏的。由于这些原因，迫切需要抗生素的替代品。噬菌体（噬菌体）是感染细菌的病毒，具有长期以来被假设为治疗细菌感染的方法，比抗生素的使用早了四分之一以上一个世纪。不幸的是，就像抗生素一样，细菌也会对噬菌体产生抗药性。前期工作有鉴定出几种具有脂多糖（LPS）突变的噬菌体抗性 UPEC。这些细菌，尽管能够逃避噬菌体，但它们在尿液中生长和在膀胱中定植的能力比它们的要差亲本菌株。然而，这些健康缺陷的确切原因尚不清楚。这项工作还旨在评估噬菌体治疗细胞内 UPEC 储库的效用，该储库未受到抗生素治疗的影响，并导致尿路感染复发。已发表的初步数据表明噬菌体可以相互作用并被被膀胱细胞内化，尽管没有研究小组测试过噬菌体减少细胞内 UPEC 的能力负担。总而言之，这些初步数据支持了这样的假设：噬菌体疗法通过驱动噬菌体抗性使细菌不太适合，并且通过噬菌体与膀胱的相互作用，将是有效的治疗细胞内和细胞外尿路感染。该假设将通过以下具体研究进行研究目标：1) 确定尿液生长减少和小鼠膀胱定植不良的机制在噬菌体抗性（LPS 突变体）UPEC 中，2) 表征噬菌体结合和尿上皮的内化，减少细胞内储库，刺激膀胱免疫反应。在研究这些目标时，几种创新工具，例如原代膀胱细胞衍生的类器官、超分辨率显微镜和体内将使用疾病小鼠模型和噬菌体疗法。该提案将为候选人提供培训 UTI 动物模型、先进显微镜技术、蛋白质分离和膀胱类器官模型。这培训以及出色的指导和职业发展目标将为候选人提供研究传染病新型转化疗法的独立研究生涯的必要技能疾病。该培训将在贝勒医学院在该领域领导者的指导下进行抗菌素耐药性、噬菌体治疗和宿主-病原体相互作用。总的来说，这项研究将提供将尿路感染噬菌体疗法从实验室转化为临床所需的信息。