Exploration of the oropharyngeal resistome as a reservoir of antimicrobial resistance in Neisseria gonorrhoeae

项目摘要/摘要 淋病是全球最常见的性传播感染之一，在COVID之前- 19大流行，美国第二常见的可报告感染。在过去的十年里，有 美国男男性接触者(MSM)淋病发病率显著上升，增幅达375% 观察时间为2010-2018年。同时，淋病奈瑟菌的耐药率 (AMR-NG)在MSM中继续显著增加。尽管AMR的患病率显著增加- NG，AMR-NG的驱动和储备者，尤其是MSM中常见的口咽部淋病 以及对所有头孢曲松治疗失败的报告病例的责任尚未完全阐明。这个 口咽部是AMR-NG的储存库，因为它拥有大量的微生物区系和AMR基因库。这个 这项建议的总体目标是全面描述男性口咽部抵抗组 性传播感染和艾滋病毒控制和预防的优先人群。在目标1中，我们将使用文化遗传学来识别NG和 共生菌，确定它们的AMR图谱，并使用表型AMR数据进行严格的质量控制 我们的口咽阻力组图谱(目标2)。在目标2中，我们将使用与文化无关的不偏不倚 元基因组测序和综合生物信息学分析确定口咽阻力组 MSM和MSW。在目标3中，我们将对150株共生奈瑟氏菌进行测序。与获得的NG分离株配对 分离时来自同一患者(目标1)，以证明它们共享相同的AMR遗传标记。 我们将明确多重耐药(MDR)共生奈瑟氏菌AMR的遗传机制， 并研究了从10个不同种类的共生体体外转移AMR的相对频率 奈瑟氏菌对完全敏感和基因多样化的NG受体菌株。我们预计结果来自于 这些研究将提供急需的有关AMR基因和遗传决定因素的数据 促进NG对建议用于淋病治疗的抗生素的耐药性的演变，包括 头孢曲松，最后一种治疗淋病的高效选择，为改进铺平了道路 主动识别和缓解新出现的AMR威胁。我们的数据将立即有助于 监测受淋病和艾滋病毒流行不成比例影响的男男性接触者的AMR威胁，并指导 努力开发用于对抗耐多药-NG的新型抗菌剂。 公共卫生意义。这项研究将确定人类口咽部AMR的大小和多样性， 一个被认为在AMR-NG的出现中发挥核心作用的解剖部位，并将为 哪些细菌和在什么情况下NG可以获得AMR。这一知识将提供重要的 对AMR-NG如何发展的见解，在制定遏制威胁的战略方面至关重要的信息 用于AMR-NG的研究和新型抗菌剂的开发。

Project Summary/Abstract Gonorrhea is one of the most common sexually transmitted infections (STIs) globally, and prior to the COVID- 19 pandemic, the second most common reportable infection in the U.S. In the past decade, there have been significant increases in gonorrhea among men who have sex with men (MSM) in the US, with a 375% increase observed from 2010–2018. At the same time, the percentages of antimicrobial-resistant Neisseria gonorrhoeae (AMR-NG) continues to increase markedly in MSM. Despite the significant increase in the prevalence of AMR- NG, the drivers and reservoirs of AMR-NG especially for oropharyngeal gonorrhea commonly diagnosed in MSM and responsible for all the reported cases of failed ceftriaxone treatment are yet to be fully elucidated. The oropharynx is reservoir for AMR-NG because it harbors a large microbiota and repertoire of AMR genes. The overall goal of this proposal is to describe comprehensively the oropharyngeal resistome of men who are a priority population for STI and HIV control and prevention. In Aim 1, we will use culturomics to identify NG and commensal bacteria, determine their AMR profiles, and use the phenotypic AMR data for stringent quality control of our oropharyngeal resistome profiling (Aim 2). In Aim 2, we will use an unbiased culture-independent metagenomic sequencing and comprehensive bioinformatic analysis to determine the oropharyngeal resistome of MSM and MSW. In Aim 3, we will sequence 150 commensal Neisseria spp. paired with NG isolates obtained from the same patient when isolated (Aim 1) to demonstrate that they share the same genetic markers of AMR. We will define the genetic mechanisms of AMR among multidrug-resistant (MDR) commensal Neisseria spp., and investigate the relative frequency of in vitro transfer of AMR from 10 different species of commensal Neisseria to fully susceptible and genetically diverse recipient strains of NG. We expect that the results from these studies will provide urgently needed data on the repertoire of AMR genes and genetic determinants facilitating the evolution of NG resistance to antibiotics recommended for gonorrhea treatment including ceftriaxone, the last remaining highly effective treatment option for gonorrhea, and paving the way for improved proactive identification and mitigation of emerging AMR threats. Our data will immediately contribute to the surveillance of AMR threats in MSM disproportionately impacted by gonorrhea and HIV epidemics, and guide efforts to develop novel antimicrobials for combatting MDR-NG. Public Health Significance. This study will define the magnitude and diversity AMR in the human oropharynx, an anatomic site thought to play a central role in the emergence of AMR-NG, and will provide new insights into which bacteria and under what circumstances NG may acquire AMR. This knowledge will provide important insights into how AMR-NG develops, critically important information in developing strategies to contain the threat of AMR-NG and for development of novel antimicrobials.