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，其毒力取决于其形成 生物膜--粘糊糊的多糖和细菌层，使Pa能够在伤口上定植并逃避抗生素。 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，由Pa产生的Pf噬菌体（Pf噬菌体）自发地组装宿主的松散网络， 和存在于Pa感染部位的微生物聚合物形成致密、高度稳定的液晶。该结晶 结构增强了生物膜的粘附性、粘度和抗干燥性。 我们最近还报道了Pf噬菌体和这种晶体结构通过以下方式促进抗生素耐受性： 结合和隔离抗生素。这种作用取决于抗生素的电荷性质;积极的 带电荷的抗生素，如妥布霉素，被有效地隔离在带负电荷的晶体束中， 而中性电荷的抗生素如环丙沙星则不是。这些数据表明Pf的存在 噬菌体可能是决定哪些抗生素对Pa有效的重要因素。 总而言之，这些数据表明Pa细菌和Pf噬菌体可能以某种方式合作，从而有助于 生物膜感染的致病性。与此一致，我们的初步数据表明，在Pa的小鼠模型中， 在生物膜伤口感染中，Pf噬菌体的存在有助于慢性感染。 这些对Pf噬菌体致病作用的新见解有可能开辟一个新的前沿， 对抗帕的生物膜然而，首先重要的是确定它们与人类慢性伤口的相关性 感染.在目标1中，我们将评估人类伤口中的Pa生物膜是否具有晶体结构，并定义 Pf噬菌体水平和临床结果之间的关系，包括抗生素耐药性。 考虑到这些对生物膜致病的贡献，靶向治疗Pf噬菌体可能是有益的。到 为此，我们开发了针对Pf噬菌体外壳蛋白的单克隆抗体和疫苗， CoaB.我们的初步实验表明，抗体可以干扰生物膜的组织，并促进抗生素的产生。 体外培养。这表明，通过免疫糖尿病患者， 抗Pf噬菌体。然而，首先有必要证明这些治疗在体内的影响。 在目的2中，我们将测试针对Pf噬菌体的抗体是否在感染性肺炎的小鼠模型中促进抗生素功效。 Pa生物膜感染。 总之，这些目标代表了一种大胆的、全新的方法来治疗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。 伤口感染的病理生理学和靶向Pf噬菌体的新疗法。

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