Global Transcriptome Analysis of Mucosal Gonoccal Infection

 描述(申请人提供)：淋球菌是性传播感染(STI)淋病的病原体，这是一种全球范围内的高发病率疾病，估计每年有1.06亿例。淋病奈瑟菌感染男性和女性生殖道，并经常可以逃避宿主的免疫机制，直到抗生素干预。淋球菌抗药性菌株的惊人增加，这种疾病在女性中往往没有症状，以及缺乏针对关键毒力决定因素的疫苗，这说明迫切需要在人类自然疾病期间研究这种病原体，以指导治疗感染的新疗法。这里提出的研究旨在确定淋球菌“体内表达因子”(IVEF)，长期目标是将它们开发为新的治疗靶点。我们的研究集中在南京国家性病预防控制中心的一组受试者，中国，因为该地区的疾病水平很高，并且有报告说淋球菌菌株增加了耐药性。利用南京地区人群的尿路和阴道灌洗标本，结合RNA深度测序，我们首次发现了在自然感染过程中表达的基因，这些基因在体外培养的淋球菌株中没有检测到。在男性和女性自然黏膜感染过程中检测到的高水平表达的常见淋球菌基因组中，有一组以前未描述的淋球菌噬菌体和假设基因，以及编码抗菌素耐药决定因素的基因。我们假设淋球菌在体内表达以前没有在体外鉴定过的基因。尽管女性和男性在自然感染过程中表达的淋球菌转录组不同，但可以识别共同的基因，并有可能代表新的治疗靶点。我们提出以下目标来解决这一假说：目的1.确定淋球菌在体内感染男性和女性生殖道时表达的转录组，并鉴定淋球菌IVEF。目的2.明确IVEF亚型在淋球菌致病中的作用。在这项申请中提出的研究是第一次检查淋球菌在自然粘膜感染期间的全球转录组，并识别和表征在体内表达的尚未被假设存在的新基因。

 DESCRIPTION (provided by applicant): Neisseria gonorrhoeae is the causative agent of the sexually transmitted infection (STI) gonorrhea, a high morbidity disease worldwide with an estimated 106 million cases annually. N. gonorrhoeae infects the male and female genital tract and can often evade host immune mechanisms to persist until antibiotic intervention. The alarming increase in antibiotic resistant strains of N. gonorrhoeae, the often-asymptomatic nature of this disease in women, and the lack of a vaccine directed at crucial virulence determinants, speaks to the urgent need to study this pathogen during natural disease in humans to guide new therapies to treat infection. The studies proposed here are designed to identify gonococcal "in vivo expressed factors" (IVEFs) with the long- term goal to develop these as new therapeutic targets. Our studies have focused on a cohort of subjects attending the National Center for STD Control (NCSTD) in Nanjing, China due to the high level of disease in this region and reports of gonococcal strains with increased resistance. Using urethral and vaginal lavage samples obtained from the Nanjing cohort, together with RNA deep sequencing we were the first to identify genes expressed during natural infection, which were not detected in the corresponding gonococcal strains cultured under in vitro conditions. Within the group of common gonococcal genes detected with high-level expression during natural mucosal infection in men and women, was a group of previously uncharacterized gonococcal phage and hypothetical genes as well as genes encoding antimicrobial resistant determinants. We hypothesize that the gonococcus expresses genes in vivo that have not previously been identified in vitro. Despite differences in the gonococcal transcriptome expressed during natural infection in female and male subjects, common genes can be identified and have the potential to represent new therapeutic targets. We propose the following Aims to address this hypothesis: Aim 1. To define the N. gonorrhoeae transcriptome expressed during in vivo infection of the male and female genital tract and identify gonococcal IVEFs. Aim 2. To define the role of subsets of IVEF in N. gonorrhoeae pathogenesis. The studies proposed in this application are the first to examine the global transcriptome of N. gonorrhoeae during natural mucosal infection and to identify and characterize novel genes expressed in vivo that have not been yet hypothesized to exist.