Global transcriptome analysis of mucosal gonococcal infection

描述（由申请人提供）：淋病奈瑟菌是性传播疾病（STD）淋病的病原体，是一种全球高发病率疾病，估计每年有6200万例（世卫组织）。虽然男性尿道感染通常有症状，但近80%感染淋病奈瑟菌的女性是无症状的。这是特别危险的，因为长期未经治疗的淋球菌感染可导致盆腔炎（PID），这是不孕和异位妊娠的常见原因。通过现有的临床核心与NIAID资助的U19基金（AI084048）合作，我们有独特的机会与中国南京国家性病控制中心（NCSTD）的合作者合作，解决淋病奈撒菌感染症状反应的差异。NCSTD诊所目前从报告有淋球菌症状的男性获取尿道标本。这些男性往往同时感染一种或多种性传播感染。国家性病中心还从可能感染了淋球菌或淋球菌/衣原体菌株的女性那里获得匹配的宫颈阴道灌洗标本，这些女性可能从匹配的男性伴侣那里感染了淋球菌或淋球菌/衣原体菌株。在这项研究中，我们建议使用RNA深度测序来定义生殖器粘膜对淋病的反应，它们在淋球菌感染期间如何变化，以及男性和女性的不同反应如何导致两者的不同症状结果。在新的研究中，我们利用RNA序列分析来确定淋病奈瑟菌在体内感染和与宿主细胞体外孵育期间的全局转录组。我们的分析揭示了体内感染期间淋球菌应激反应、代谢和呼吸基因表达的独特特征。基于这些结果，我们假设淋病奈瑟菌在体内感染过程中表达了一组独特的基因，这些基因受男性和女性生殖道感染期间存在的特定宿主环境的影响。此外，宿主性别特异性差异导致不同的淋球菌转录组。我们提出以下目标来解决这一假设：目标1。比较淋病奈瑟菌在女性生殖道体内感染、宫颈上皮细胞孵育和体外培养基生长过程中的转录组学特征。目标2。比较淋病奈瑟菌在男性生殖道体内感染、尿道上皮细胞孵育和培养基中体外生长期间的转录组谱。目标3。比较淋病奈瑟菌在男性和女性生殖道体内感染以及与男性尿道上皮细胞和宫颈上皮细胞孵育期间的转录组学特征。

DESCRIPTION (provided by applicant): Neisseria gonorrhoeae is the causative agent of the sexually transmitted disease (STD) gonorrhea, a high morbidity disease worldwide with an estimated 62 million cases annually (WHO). Although infection of the male urethra is usually symptomatic, nearly 80% of females infected with N. gonorrhoeae are asymptomatic. This is particularly dangerous as prolonged untreated gonococcal infections can lead to pelvic inflammatory disease (PID), a common cause of infertility and ectopic pregnancy. Through an existing Clinical Core in partnership with a NIAID funded U19 grant (AI084048), we have the unique opportunity to work with our collaborators at the National Center for STD Control (NCSTD) in Nanjing, China to address the disparity in symptomatic responses to N. gonorrhoeae infection. The NCSTD clinic currently obtains urethral specimens from males who report with gonococcal symptoms. These men are often co-infected with one or more STIs. The NCSTD also obtains matched cervico-vaginal lavage specimens from females who may be infected with gonococcal or gonococcal/chlamydial strains contracted from their matched male partners. We propose in this study to use RNA deep sequencing to define genital mucosal responses to gonorrhea, how they shift during gonococcal infection, and how differential male and female responses lead to different symptomatic outcomes in both. In new studies we have utilized RNA seq analysis to define the global transcriptome of N. gonorrhoeae during in vivo infection and during in vitro incubation with host cells. Our analysis revealed a unique signature of gonococcal stress response, metabolism, and respiratory genes expressed during in vivo infection. Based on these results we hypothesize N. gonorrhoeae expresses a unique set of genes during in vivo infection that is influenced by the specific host environment present during infection of the male and female genital tract. Furthermore, host gender-specific differences result in distinct gonococcal transcriptomes. We propose the following Aims to address this hypothesis: Aim 1. To compare the transcriptomic profile of N. gonorrhoeae during in vivo infection of the female genital tract to incubation with endocervical epithelial cells and growth i vitro in media. Aim 2. To compare the transcriptomic profile of N. gonorrhoeae during in vivo infection of the male genital tract to incubation with urethral epithelial cells and growth in vitr in media. Aim 3. To compare the transcriptomic profile of N. gonorrhoeae during in vivo infection of both male and female genital tract and incubation with male urethral epithelial cells and endocervical epithelial cells.