Global transcriptome analysis of mucosal gonococcal infection

描述(申请人提供)：淋球菌是性传播疾病(STD)淋病的病原体，这是一种全世界范围内的高发病率疾病，估计每年有6200万例(世卫组织)。虽然男性尿路感染通常是有症状的，但近80%感染淋病奈瑟菌的女性没有症状。这尤其危险，因为长期未经治疗的淋球菌感染可能导致盆腔炎(PID)，这是不孕不育和宫外孕的常见原因。通过现有的临床核心和NIAID资助的U19赠款(AI084048)，我们有独特的机会与我们位于南京的国家性病控制中心的合作者中国合作，解决淋球菌感染对症反应的差异。NCSTD诊所目前从报告有淋病症状的男性那里获得尿路样本。这些男性通常会同时感染一种或多种性传播感染。NCSTD还从可能感染从匹配的男性伴侣那里感染淋球菌或淋球菌/衣原体菌株的女性那里获得匹配的宫颈阴道灌洗液样本。在这项研究中，我们建议使用RNA深度测序来确定生殖器粘膜对淋病的反应，它们在淋球菌感染期间是如何转移的，以及男性和女性的不同反应如何导致两者不同的症状结果。在新的研究中，我们利用RNA序列分析来确定淋球菌在体内感染和体外与宿主细胞孵育期间的整体转录组。我们的分析揭示了淋球菌应激反应、代谢和呼吸道基因在体内感染期间表达的独特特征。根据这些结果，我们推测淋球菌在体内感染时表达一组独特的基因，该基因受感染男性和女性生殖道的特定宿主环境的影响。此外，宿主性别差异导致不同的淋球菌转录本。目的1.比较淋球菌体内感染女性生殖道与宫颈上皮细胞孵育和体外生长的转录谱。目的2.比较淋病奈瑟菌在体内感染男性生殖道、与尿路上皮细胞孵育和在培养基中生长的转录谱。目的3.比较淋病奈瑟菌在体内感染男性和女性生殖道以及与男性尿路上皮细胞和宫颈上皮细胞孵育过程中的转录图谱。

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.