Biofilms and Bacteriophages in Chronic Wound Infections

慢性伤口感染中的生物膜和噬菌体

基本信息

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
项目类别：
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

项目摘要

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.

项目概要铜绿假单胞菌 (Pa) 是一种主要的人类病原体，其毒力取决于其形成细菌的能力。生物膜——由多糖和细菌组成的粘稠层，使 Pa 能够在伤口上定居并逃避抗生素。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。这种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有助于抗生素耐受性：结合和隔离抗生素。这种效应取决于抗生素的电荷特性；积极地带电荷的抗生素，如妥布霉素，被有效地隔离在带负电荷的晶体束内噬菌体，而带中性电荷的抗生素，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。总之，这些数据表明 Pa 细菌和 Pf 噬菌体可能以有助于促进生物膜感染的致病性。与此一致的是，我们的初步数据表明，在 Pa 的小鼠模型中生物膜伤口感染，Pf噬菌体的存在有助于慢性感染。这些对 Pf 噬菌体致病贡献的新见解有可能开辟一条新的前沿对抗 Pa 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 噬菌体水平与临床结果（包括抗生素耐药性）之间的关系。鉴于这些对生物膜发病机制的贡献，靶向 Pf 噬菌体的治疗可能是有益的。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组织并促进抗生素的产生体外渗透率。这表明通过对糖尿病患者进行免疫接种来预防 Pa 伤口感染是可能的。个人对抗Pf噬菌体。然而，首先有必要证明这些治疗在体内的影响。在目标 2 中，我们将测试针对 Pf 噬菌体的抗体是否能在小鼠模型中提高抗生素功效。 Pa生物膜感染。总之，这些目标代表了一种大胆且全新的方法来治疗 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针对 Pf 噬菌体的新疗法。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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