Biofilms and Bacteriophages in Chronic Wound Infections

慢性伤口感染中的生物膜和噬菌体

• 批准号: 9375747
• 负责人: Paul L Bollky
• 金额: $ 31.94万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-06-26 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9375747
• 关键词: Adhesiveness; Alginates; Amputation; Animal Model; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antibodies; Architecture; Bacteria; Bacteriophages; Binding; Capsid Proteins; Charge; Chronic; Ciprofloxacin; Clinical; DNA; Data; Desiccation; Diabetes Mellitus; Diabetic wound; Diffusion; Effectiveness; Entropy; Functional disorder; Goals; Human; Immune system; Immunize; In Vitro; Individual; Infection; Investigation; Light; Liquid substance; Microbial Biofilms; Monoclonal Antibodies; Outcome; Pathogenicity; Patients; Penetrance; Play; Polymers; Polysaccharides; Property; Pseudomonas Infections; Pseudomonas aeruginosa; Reporting; Resistance; Role; Severities; Site; Small Business Innovation Research Grant; Structure; Testing; Therapeutic; Tissues; Tobramycin; Treatment Failure; Ulcer; Vaccines; Virulence; Viscosity; Wound Infection; base; chronic wound; crystallinity; diabetic; experimental study; fight against; improved; in vivo; insight; liquid crystal; microbial; mouse model; new therapeutic target; novel; novel strategies; novel therapeutics; novel vaccines; pathogen; prevent; wound

Project Summary Pseudomonas aeruginosa (Pa) is a major human pathogen whose virulence is predicated upon its ability to form biofilms - slimy layers of polysaccharides and bacteria that allow Pa to colonize wounds and to evade antibiotics. My lab has recently uncovered novel roles for bacteriophages in the formation and function of Pa biofilms. We reported that Pf bacteriophages (Pf phages) produced by Pa spontaneously assemble the loose network of host and microbial polymers present at sites of Pa infection into a dense, highly stable, liquid crystal. This crystalline architecture enhances biofilm adhesiveness, viscosity, and resistance to desiccation. We also recently reported that Pf phage and this crystalline architecture contribute to antibiotic tolerance by binding and sequestering antibiotics. This effect is dependent on the charge properties of the antibiotic; positively charged antibiotics, like tobramycin, are efficiently sequestered within crystalline bundles of negatively charged phage while neutrally charged antibiotics, like ciprofloxacin, are not. These data suggest that presence of Pf phage may be an important factor in determining which antibiotics are effective against Pa. Together, these data suggest that Pa bacteria and Pf phage may partner in ways that contribute to the pathogenicity of biofilm infections. Consistent with this, our preliminary data suggest that, in a murine model of Pa biofilm wound infection, the presence of Pf phage contributes to chronic infection. These novel insights into the pathogenic contributions of Pf phage have the potential to open up a new front in the fight against Pa biofilms. However, first it is important to determine their relevance to human chronic wound infections. In Aim 1, we will assess whether Pa biofilms in human wounds have crystalline structure and define the relationship between Pf phage levels and clinical outcomes, including antibiotic resistance. Give these contributions to biofilm pathogensis, it may be beneficial to to target Pf phage therapeutically. To this end, we have developed monoclonal antibodies and vaccines directed against the Pf phage coat protein, CoaB. Our initial experiments suggest that antibodies can interfere with biofilm organization and promote antibiotic penetrance in vitro. This suggests that it may be possible to prevent Pa wound infections by immunizing diabetic individuals against Pf phage. However, first it is necessary to demonstrate the impact of these treatments in vivo. In Aim 2, we will test whether antibodies directed against Pf phage promote antibiotic efficacy in a mouse model of Pa biofilm infection. Together, these aims represent a bold and radically novel approach to Pa biofilm infections and chronic wound infections. If successful, the data generated from these studies will support investigations into the role of Pf phage in the pathophysiology of wound infections and novel therapies targeting Pf phage.

项目摘要 铜绿假单胞菌（PA）是一种主要的人类病原体，其毒力是基于形成能力的 生物膜 - 多糖和细菌的粘层，可允许PA定殖并逃避抗生素。 我的实验室最近发现了噬菌体在PA生物膜的形成和功能中的新作用。我们 报道PA产生的PF噬菌体（PF噬菌体）自发地组装了宿主的宽松网络 以及在PA感染部位存在于致密，高度稳定的液晶中的微生物聚合物。这个结晶 建筑提高了生物膜粘附性，粘度和对干燥的抗性。 我们最近还报道了PF噬菌体和这种结晶构建导致抗生素的耐受性 结合和隔离抗生素。该作用取决于抗生素的电荷特性。积极 带电抗生素（例如毒素）在带负电荷的结晶束中有效隔离 噬菌体虽然中性充电的抗生素（如环丙沙星）不是。这些数据表明存在PF 噬菌体可能是确定哪些抗生素有效抗PA的重要因素。 这些数据一起表明，PA细菌和PF噬菌体可能以有助于 生物膜感染的致病性。与此相一致，我们的初步数据表明，在PA的鼠模型中 生物膜伤口感染，PF噬菌体的存在导致慢性感染。 这些对PF噬菌体致病贡献的新颖见解有可能在 与PA生物膜的斗争。但是，首先确定它们与人类慢性伤口的相关性很重要 感染。在AIM 1中，我们将评估人伤口中的PA生物膜是否具有结晶结构并定义 PF噬菌体水平与临床结局之间的关系，包括抗生素耐药性。 对生物膜病原体提供这些贡献，以治疗靶向PF可能是有益的。到 这端，我们开发了针对PF噬菌体蛋白的单克隆抗体和疫苗 驾驶。我们最初的实验表明抗体可以干扰生物膜组织并促进抗生素 体外渗透。这表明可以通过免疫糖尿病来预防PA伤口感染 个人反对PF噬菌体。但是，首先有必要证明这些治疗方法在体内的影响。 在AIM 2中，我们将测试针对PF噬菌体的抗体是否在小鼠模型中促进抗生素功效 PA生物膜感染。 这些目的共同代表了一种大胆而根本新颖的方法，用于PA生物膜感染和慢性伤口 感染。如果成功，这些研究产生的数据将支持对PF噬菌体作用的研究 在靶向PF噬菌体的伤口感染和新型疗法的病理生理中。

项目成果

The Rationale for Using Bacteriophage to Treat and Prevent Periprosthetic Joint Infections.

• DOI: 10.3389/fmicb.2020.591021
• 发表时间: 2020
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Van Belleghem JD;Manasherob R;Miȩdzybrodzki R;Rogóż P;Górski A;Suh GA;Bollyky PL;Amanatullah DF
• 通讯作者: Amanatullah DF