Disappearance of rifampin resistance in MRSA foreign body osteomyelitis

MRSA异物骨髓炎利福平耐药性消失

• 批准号: 9272818
• 负责人: Robin Patel
• 金额: $ 23.85万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9272818
• 关键词: Affect; Animal Model; Animals; Antibiotics; Bacteria; Base Pairing; Clinical; Clinical Treatment; Combined Modality Therapy; Data; Debridement; Disadvantaged; Ensure; Evolution; Experimental Models; Exposure to; Foreign Bodies; Frequencies; Future; Gene Targeting; Genes; Genomics; Genus staphylococcus; Implant; In Vitro; Infection; Intuition; Joint Prosthesis; Kinetics; Mediating; Medical Technology; Methicillin Resistance; Microbial Biofilms; Modeling; Monitor; Mutation; Operative Surgical Procedures; Orthopedics; Osteomyelitis; Outcome; Patients; Pharmaceutical Preparations; Play; Population; Rattus; Regimen; Resistance; Rifampicin resistance; Rifampin; Role; Sampling; Site; Staphylococcus aureus; Staphylococcus epidermidis; Testing; Time; Treatment Protocols; Treatment outcome; Withholding Treatment; aging population; antimicrobial; antimicrobial drug; bacterial resistance; base; experimental study; fitness; genome sequencing; in vivo; joint infection; methicillin resistant Staphylococcus aureus; microbial; microorganism; novel; pathogen; retention rate; time use; whole genome

Project Summary Due to an aging population and advances in medical technology, prosthetic joint placement is increasing in frequency. Prosthetic joint infection (PJI) affects up to 2% of prosthetic joints. Staphylococci are the most common causes of PJI, with Staphylococcus aureus being the most common pathogen overall and Staphylococcus epidermidis being the second most common staphylococcal species involved. Treatment of PJI is problematic due to the limited activity of most antimicrobial agents against microbial biofilms. Rifampin, however, has anti-biofilm activity, and is a cornerstone drug in the management of staphylococcal PJI, when managed with débridement and implant retention (DAIR). Unfortunately, resistance to rifampin, mediated by single base pair mutations in the target gene, is readily selected and would be expected to compromise management of staphylococcal PJI using a DAIR strategy. Our preliminary data surprisingly suggest, however, that selected rifampin resistance may not necessarily negatively impact future treatment using rifampin-based therapy. Specifically, we have shown that, as expected, rifampin resistance is selected following rifampin monotherapy in a rat model of methicillin-resistant S. aureus foreign body osteomyelitis (FBO). But, surprisingly, rifampin resistance “disappears” two weeks following cessation of treatment, with only rifampin susceptible S. aureus being detectable. We have shown that rifampin resistance does not confer decreased fitness when bacteria are grown alone in vitro or in vivo; however, rifampin-resistant bacteria are out-competed by rifampin-susceptible bacteria when grown together in vitro and in vivo. And, when treating FBO with two rounds of rifampin monotherapy, resistance is observed at a lower frequency following the second than the first round of rifampin exposure. These observations suggest that rifampin may be a usable agent in biofilm- associated infections in which rifampin resistance has been previously selected. The central hypothesis of our proposed studies is therefore that in experimental FBO, treatment with rifampin can be successful even if rifampin resistance has been previously selected. Our objectives are to develop an experimental FBO model that allows sampling of bacteria in real time from the same animal and to use the model to define the kinetics of selection and disappearance of rifampin resistance. Whereas in traditional osteomyelitis models, microorganisms are analyzed at single time-points (i.e., when animals are sacrificed), the proposed model will allow analysis of bacteria from the same animal over time. We will perform whole genome sequencing on bacteria recovered at multiple time-points during and following one and two rounds of rifampin treatment. This will allow definition of rifampin resistance- and fitness-associated mutations that emerge during and following initial rifampin treatment, and subsequent re-exposure to rifampin. This approach will pinpoint mutations that may occur in rifampin-susceptible bacteria allowing their persistence or conferring upon them a selective advantage over their rifampin-resistant counterparts, even when challenged with rifampin. Finally, we will compare outcomes of FBO treated with either rifampin monotherapy followed by rifampin combination therapy or rifampin combination therapy alone. This will allow us to determine whether rifampin-containing regimens may be an option in the treatment of PJI in which rifampin resistance has been previously selected. Results of the proposed studies will define the kinetics of in vivo selection of rifampin resistance and identify mutations that play a role in out-competition of rifampin-resistant bacteria by those that are rifampin-susceptible. Additionally, our studies will inform clinical treatment options in the scenario of selected rifampin resistance and lay the groundwork for future experiments defining optimal therapy following selection of rifampin resistance.

项目概要 由于人口老龄化和医疗技术的进步，假体关节植入术在世界范围内不断增加 频率。假体关节感染 (PJI) 影响高达 2% 的假体关节。葡萄球菌是最多的 PJI 的常见原因，金黄色葡萄球菌是最常见的病原体 表皮葡萄球菌是第二常见的葡萄球菌种类。治疗 由于大多数抗菌剂对微生物生物膜的活性有限，PJI 存在问题。利福平， 然而，具有抗生物膜活性，并且是治疗葡萄球菌 PJI 的基础药物，当 通过清创和种植体保留（DAIR）进行管理。不幸的是，对利福平的耐药性是由 目标基因中的单碱基对突变很容易被选择，并且预计会受到损害 使用 DAIR 策略管理葡萄球菌 PJI。然而，我们的初步数据令人惊讶地表明， 选定的利福平耐药性不一定会对未来使用基于利福平的治疗产生负面影响 治疗。具体来说，我们已经表明，正如预期的那样，利福平耐药性是在利福平之后选择的 耐甲氧西林金黄色葡萄球菌异物骨髓炎（FBO）大鼠模型的单一疗法。但， 令人惊讶的是，停止治疗两周后，利福平耐药性“消失”，只剩下利福平 可检测到易感金黄色葡萄球菌。我们已经证明利福平耐药性不会降低 细菌在体外或体内单独生长时的适应性；然而，利福平耐药细菌在竞争中被击败 当体外和体内一起生长时，由利福平敏感细菌产生。并且，当用两个治疗 FBO 时 在利福平单药治疗的几轮中，第二轮后观察到耐药性的频率低于第一轮 一轮利福平暴露。这些观察结果表明利福平可能是生物膜中的有用药物。 先前已选择利福平耐药的相关感染。 因此，我们提出的研究的中心假设是，在实验性 FBO 中，利福平治疗 即使先前已选择利福平耐药，也可以成功。我们的目标是开发一个 实验性 FBO 模型，允许从同一动物实时采样细菌并使用 模型来定义利福平耐药性的选择和消失的动力学。而在传统 骨髓炎模型，在单个时间点（即处死动物时）分析微生物， 所提出的模型将允许随着时间的推移对同一动物的细菌进行分析。我们将全程表演 对在一轮和两轮期间和之后的多个时间点恢复的细菌进行基因组测序 利福平治疗。这将允许定义利福平耐药性和适应性相关突变， 在初始利福平治疗期间和之后以及随后再次暴露于利福平期间出现。这种做法 将查明利福平敏感细菌中可能发生的突变，使其持续存在或赋予 即使面临挑战，他们也比利福平耐药的同行具有选择性优势 利福平。最后，我们将比较使用利福平单药治疗和随后的 FBO 治疗的结果 利福平联合治疗或单独利福平联合治疗。这将使我们能够确定是否 含利福平的方案可能是治疗 PJI 的一种选择，其中利福平耐药已被证实。 之前选择的。拟议研究的结果将定义利福平体内选择的动力学 耐药性并识别在利福平耐药细菌与利福平耐药细菌的竞争中发挥作用的突变 对利福平敏感。此外，我们的研究将为以下情况下的临床治疗选择提供信息： 选择利福平耐药性，并为未来确定最佳治疗方案的实验奠定基础 利福平耐药性的选择。

项目成果

Global spread of three multidrug-resistant lineages of Staphylococcus epidermidis.

• DOI: 10.1038/s41564-018-0230-7
• 发表时间: 2018-10
• 期刊: Nature microbiology
• 影响因子: 28.3
• 作者: Lee JYH;Monk IR;Gonçalves da Silva A;Seemann T;Chua KYL;Kearns A;Hill R;Woodford N;Bartels MD;Strommenger B;Laurent F;Dodémont M;Deplano A;Patel R;Larsen AR;Korman TM;Stinear TP;Howden BP
• 通讯作者: Howden BP

A novel rat model of foreign body osteomyelitis for evaluation of antimicrobial efficacy.

用于评估抗菌功效的新型异物骨髓炎大鼠模型。

• DOI: 10.20454/jeaas.2019.1555
• 发表时间: 2019
• 期刊: Journal of experimental and applied animal sciences
• 作者: Brinkman,CassandraL;Schmidt-Malan,SuzannahM;Karau,MelissaJ;Patel,Robin
• 通讯作者: Patel,Robin

Rifampicin resistance in Staphylococcus epidermidis: molecular characterisation and fitness cost of rpoB mutations.

• DOI: 10.1016/j.ijantimicag.2017.12.019
• 发表时间: 2017-12
• 期刊: International journal of antimicrobial agents
• 影响因子: 10.8
• 作者: Y. Wi;K. Greenwood-Quaintance;C. Brinkman;Jean Y. H. Lee;B. Howden;Robin Patel

Evaluation of Oritavancin Combinations with Rifampin, Gentamicin, or Linezolid against Prosthetic Joint Infection-Associated Methicillin-Resistant Staphylococcus aureus Biofilms by Time-Kill Assays.

通过时间灭活测定评估奥利万星与利福平、庆大霉素或利奈唑胺的组合对假体关节感染相关的耐甲氧西林金黄色葡萄球菌生物膜的影响。

• DOI: 10.1128/aac.00943-18
• 发表时间: 2018
• 期刊: Antimicrobial agents and chemotherapy
• 影响因子: 4.9
• 作者: Yan,Qun;Karau,MelissaJ;Raval,YashS;Patel,Robin
• 通讯作者: Patel,Robin