Bacteriophage to treat multidrug‐resistant UTI in Persons with Spinal Cord Injury

噬菌体治疗脊髓损伤患者的多重耐药性尿路感染

• 批准号: 10623140
• 负责人: BARBARA Wells TRAUTNER
• 金额: --
• 依托单位: MICHAEL E DEBAKEY VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10623140
• 关键词: Activities of Daily Living; Aftercare; Animals; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacteremia; Bacteria; Bacterial Antibiotic Resistance; Bacteriophages; Behavioral; Biological; Bladder; Bladder Control; Blood Circulation; Catheter Management; Catheters; Cause of Death; Charge; Clinical; Clinical Management; Clostridium difficile; Collection; Dangerousness; Directed Molecular Evolution; Dose; Drainage procedure; Environment; Epidemiology; Escherichia coli; Escherichia coli drug resistance; Evolution; Health; Healthcare; Hospitals; Human; Image; Impairment; In Vitro; Indwelling Catheter; Infection; Infective cystitis; Laboratories; Length of Stay; Libraries; Life; Lytic; Measures; Medical Device; Metagenomics; Methods; Microbial Biofilms; Microbiology; Modeling; Morbidity - disease rate; Multi-Drug Resistance; Multiple Bacterial Drug Resistance; Mus; Nature; Neurogenic Bladder; Organism; Outcome; Patients; Personal Satisfaction; Persons; Phenotype; Population; Procedures; Property; Quality of life; Recommendation; Recurrence; Reporting; Research; Resistance; Resources; Risk; Role; Sepsis; Severity of illness; Site; Source; Spinal cord injury; Spinal cord injury patients; Sterilization; Structure; Surface; Testing; Time; Tissues; United Nations; United States National Institutes of Health; Urinary tract; Urinary tract infection; Urine; Veterans; Virus; Work; bacterial resistance; catheter associated UTI; clinical outcome measures; combat; current pandemic; efficacy evaluation; efficacy testing; experience; experimental study; gut microbiome; health assessment; improved; in vivo; in vivo evaluation; indexing; infection risk; microbiome; military veteran; mortality; mouse model; novel; novel therapeutics; pandemic disease; pathogen; prevent; prospective; resistant strain; self-renewal; therapy resistant; urinary; urologic; virology

Our overall purpose is to take aim at two urgent clinical problems for persons with spinal cord injury (SCI). The first problem is ongoing morbidity from urinary tract infections (UTI) and mortality from bacteremia and sepsis arising from pathogens in the urinary tract. Urinary tract infections and associated complications are a common threat to the wellbeing and even survival of persons with spinal cord injury (SCI). The second clinical dilemma is the disproportionate harm persons with SCI are experiencing from multidrug -resistant bacteria, particularly Escherichia coli, the most common urinary pathogen. We plan to develop bacteriophage, viruses that kill specific strains of bacteria, as a treatment for multidrug-resistant E. coli UTI and sepsis in SCI. Phage have numerous properties that suggest they will be effective in this role, including lack of cross resistance with antibiotics, ability to attack biofilms, evolvability in vitro, and being a self-renewing agent at the site of infection. Phage therapy for resistant urinary organisms has potential applications in the SCI population beyond UTI treatment and also for the Veteran population in general, such as for bladder sterilization prior to urologic procedures. Phage therapy aligns with VHA antibiotic stewardship efforts and may alleviate the burden of Clostridium difficile experienced by Veterans. The work we propose here with the most common urinary pathogen in SCI, E. coli, would subsequently be applied to the other SCI urinary pathogens to create broadly effective phage cocktails. Project Objectives: We will develop E. coli-specific phages that are efficacious against a broad range of SCI-specific E. coli isolates from our VA hospital in a mouse model of catheter-associated urinary tract infection (CAUTI) and/or sepsis. Aim 1 is to determine the efficacy of phage in killing antibiotic-resistant E. coli in murine infection models of CAUTI (1A), bacteremia (1B), and in combination with antibiotics to which the infecting E. coli pathogen is resistant (1C). Aim 2 is to develop and test a bank of phages effective against contemporary E. coli isolates from Veterans with SCI. In Aim 2A, we will actively collect E. coli clinical isolates from Veterans with SCI, creating an SCI-specific bacterial strain bank. In Aim 2B, we will collect and experimentally generate phages with broad host range against E. coli, creating a bank of sequenced phage that cover over 80% of the SCI E. coli strains. In Aim 2C we will create phage cocktails from our phage bank, and test the efficacy of the cocktails in the mouse model of CAUTI induced by SCI E. coli. Aim 3 is to examine the emergence of phage-resistant E. coli during treatment of CAUTI and determine how to overcome phage resistance. Aim 3A: Determine if E. coli persistence after phage treatment results from bacterial resistance to phage. Aim 3Bb: Determine if a “recharge” dose of phage several days into therapy prevents recurrence of E. coli. Methods: We propose to study bacteriophage in vitro and in vivo using our established models of mouse CAUTI and bacteremia for all three aims. In these experiments we will track microbiological outcomes and clinical outcomes. The microbiological outcomes include quantitative counts of bacteria and phage, [microbiome diversity before and after treatment with phage and/or antibiotics, and images of urinary catheter-associated biofilms.] Our clinical outcomes are intended to recapitulate the human experience, particularly the ability to perform activities of daily living. We have two measures of clinical outcomes: the disease severity index (assesses health) and the behavioral score (assesses whether animals still perform routine activities). The experiments in Aim 1 will use bacteria (E. coli JJ2528) and phage (HP3) from our existing collections, so that the work can begin immediately. At the end of this proposal, we will have a bacterial strain bank specific to SCI, and we also will have developed, characterized, and tested in vivo a library of phage that are effective at lysing over 80% of these SCI isolates. Our team brings together complementary expertise in clinical infections, epidemiology, SCI clinical management, SCI research, microbiology, virology, [metagenomics, biofilm imaging,] and animal studies to accomplish the work proposed.

我们的总体目标是针对脊髓损伤（SCI）患者的两个紧迫的临床问题。 第一个问题是尿路感染（UTI）的持续发病率以及菌血症和败血症的死亡率 由尿路病原体引起的感染和相关并发症是常见的。 威胁脊髓损伤（SCI）患者的健康甚至生存第二个临床困境。 SCI 患者正遭受多重耐药细菌带来的不成比例的伤害，尤其是 大肠杆菌是最常见的泌尿道病原体，我们计划开发噬菌体，即杀死特定细菌的病毒。 作为治疗多重耐药大肠杆菌 UTI 和 SCI 脓毒症的噬菌体菌株有很多。 表明它们将有效发挥这一作用的特性，包括缺乏与抗生素的交叉耐药性、能力 攻击生物膜，体外进化，并作为感染部位的自我更新剂。 耐药性泌尿微生物在 SCI 人群中除 UTI 治疗外还有潜在应用 一般退伍军人群体，例如在泌尿外科手术之前进行膀胱绝育。 与 VHA 抗生素管理工作保持一致，可能减轻艰难梭菌的负担 我们在此提出的针对 SCI 中最常见泌尿道病原体大肠杆菌的研究将 随后应用于其他 SCI 尿路病原体，以创建广泛有效的噬菌体混合物项目。 目标：我们将开发能够有效对抗多种 SCI 特异性大肠杆菌的大肠杆菌特异性噬菌体。 在导管相关尿路感染 (CAUTI) 小鼠模型中从我们 VA 医院分离出的大肠杆菌和/或 目的 1 是确定噬菌体在小鼠感染模型中杀死抗生素耐药性大肠杆菌的功效。 CAUTI (1A)、菌血症 (1B) 以及与感染大肠杆菌病原体所用的抗生素联合使用 抗性 (1C) 目标 2 是开发和测试一组对当代大肠杆菌分离株有效的噬菌体。 在目标 2A 中，我们将积极收集患有 SCI 的退伍军人的大肠杆菌临床分离株，创建 在 Aim 2B 中，我们将收集并通过实验生成具有广泛用途的噬菌体。 大肠杆菌的宿主范围，创建了一个测序噬菌体库，覆盖了 80% 以上的 SCI 大肠杆菌菌株。 目标 2C，我们将从我们的噬菌体库中创建噬菌体鸡尾酒，并在小鼠中测试鸡尾酒的功效 SCI 大肠杆菌诱导的 CAUTI 模型目的 3 是检查噬菌体抗性大肠杆菌的出现。 治疗 CAUTI 并确定如何克服噬菌体耐药性 目标 3A：确定大肠杆菌是否持续存在。 噬菌体处理后由于细菌对噬菌体产生耐药性而导致的结果 目标 3Bb：确定是否需要“补充”剂量。 治疗几天后使用噬菌体可预防大肠杆菌复发。 方法：我们建议研究噬菌体。 使用我们建立的小鼠 CAUTI 和菌血症模型进行体外和体内研究以实现这三个目标。 我们将跟踪微生物结果和临床结果的实验​​，微生物结果包括。 细菌和噬菌体的定量计数，[用噬菌体和/或处理之前和之后的微生物组多样性 抗生素，以及导尿管相关生物膜的图像。]我们的临床结果旨在 概括人类的经验，特别是进行日常生活活动的能力，我们有两个。 临床结果的衡量标准：疾病严重程度指数（评估健康状况）和行为评分（评估健康状况） 动物是否仍进行日常活动）目标 1 中的实验将使用细菌（大肠杆菌 JJ2528）和 噬菌体（HP3）来自我们现有的收藏，以便工作可以在本提案结束时立即开始， 我们将拥有一个针对 SCI 的菌株库，我们还将开发、表征和测试 我们的团队汇集了一个可有效裂解 80% 以上 SCI 分离株的体内噬菌体文库。 临床感染、流行病学、SCI 临床管理、SCI 研究、 微生物学、病毒学、[宏基因组学、生物膜成像]和动物研究来完成拟议的工作。