Synergistic killing of bacterial pathogens by histones

项目总结/摘要 皮肤、深部软组织和伤口的细菌感染的有效治疗仍然是一个挑战。 这是医疗保健环境中尚未解决的重大挑战，特别是在慢性糖尿病患者中。葡萄球菌 金黄色葡萄球菌和铜绿假单胞菌是最常见的细菌，分离自慢性，非- 愈合伤口抗生素耐药性已经出现在这些特定的细菌，导致这些感染， 变得越来越难以治疗，并产生多重耐药菌株，包括耐甲氧西林菌株 金黄色葡萄球菌（MRSA）。 拟议工作的目标是开发下一代抗菌剂， 是受到了对哺乳动物抗菌防御途径更好的机械理解的启发。我们集中我们 注意中性粒细胞胞外陷阱（NETs）的抗微生物活性，它使用组蛋白杀死或 抑制微生物增殖。组蛋白的抗微生物机制还不清楚。的 Siryaporn和Gross实验室最近报告说，组蛋白与一种额外的成分配对， NET-抗菌肽（AMP）LL-37（抗菌肽）-产生有效的抗菌协同作用。LL-37 在细菌膜上形成孔，使组蛋白能够进入细菌并干扰基因。 表情这对细菌有不可逆的杀菌（杀死）作用。这里提出的工作将利用 通过鉴定人组蛋白和膜/细胞壁靶向抗菌剂的组合， （MTA），其产生有效的抗微生物活性和协同作用。该项目的总体目标是更好地 了解组蛋白和MTA之间的抗菌协同作用机制，并利用它建立 一类用于治疗皮肤感染和伤口的新疗法。 我们将通过鉴定人组蛋白与LL-37和其他蛋白的组合来实现这一目标。 产生最大抗菌活性和协同作用的MTA。我们将针对S测试这些。aureus、黑腹拟单胞菌P. 铜绿假单胞菌和体外皮肤细菌群落（目的1）。我们会尝试增加抗生素 通过工程改造影响NET中组蛋白功能的因素，特别是化学修饰， 通过瓜氨酸和将组蛋白与DNA纤维连接（Aim 2）。为了验证我们的方法，我们将测试 目的1和2中鉴定的组蛋白和MTA的组合在标准化小鼠皮肤感染模型中 (Aim 3）。为了进一步解决治疗糖尿病患者皮肤感染和伤口的挑战， 我们将在糖尿病小鼠模型中进行测试。 这项工作的结果将提供一个抗菌协同作用的机制的理解和发展 一项对抗抗生素耐药性上升的战略。这项研究的结果可能会创造一个新的类 用于治疗糖尿病和非糖尿病患者的皮肤感染和伤口的抗微生物治疗剂。 这将代表抗菌治疗方法的游戏规则改变者。

Project Summary/Abstract The effective treatment of bacterial infections of skin, deep soft tissues and wounds continues to be a major unmet challenge in healthcare settings, especially among patients with chronic diabetes. Staphylococcus aureus and Pseudomonas aeruginosa are the most common bacteria that are isolated from chronic, non- healing wounds. Antibiotic resistance has arisen in these particular bacteria, causing these infections to become increasingly difficult to treat and giving rise to multi-drug resistant strains, including Methicillin-resistant Staphylococcus aureus (MRSA). The goal of the proposed work is to develop the next generation of antimicrobials for which the design is inspired by a better mechanistic understanding of mammalian antimicrobial defense pathways. We focus our attention on the antimicrobial activities of neutrophil extracellular traps (NETs), which use histones to kill or suppress microbial proliferation. The antimicrobial mechanism of histones has not been understood. The Siryaporn and Gross labs recently reported that the pairing of histones with an additional component found in NETs – the antimicrobial peptide (AMP) LL-37 (cathelicidin) – produces potent antimicrobial synergy. LL-37 forms pores in the bacterial membrane, which enable histones to enter the bacterium and interfere with gene expression. This has an irreversible bactericidal (killing) effect on bacteria. The work proposed here will exploit this discovery by identifying combinations of human histones and membrane-/cell wall-targeting antimicrobials (MTAs) that produce potent antimicrobial activity and synergy. The overall objective of the project is to better understand the mechanism of antimicrobial synergy between histones and MTAs, and to harness it to establish a class of new therapeutics for the treatment of skin infections and wounds. We will accomplish this objective by identifying combinations of human histones with LL-37 and other MTAs that produce the greatest antimicrobial activities and synergies. We will test these against S. aureus, P. aeruginosa, and communities of skin bacteria in vitro (Aim 1). We will attempt to augment the antimicrobial activity by engineering in factors that impact histone function in NETs, specifically chemical modification through citrullination and tethering histones to DNA fibers (Aim 2). To validate our approach, we will test the combinations of histones and MTAs identified in Aims 1 and 2 in a standardized mouse skin infection model (Aim 3). To additionally address the unmet challenge of treating skin infection and wounds in diabetes patients, we will perform the tests in a diabetic mouse model. The results of this work will provide a mechanistic understanding of antimicrobial synergy and develop a strategy to combat the rise of antibiotic resistance. The results of the study could create a new class of antimicrobial therapeutics for the treatment of skin infections and wounds in diabetic and non-diabetic patients. This would represent a game-changer in the approach to antimicrobial treatments.