Global transcriptome analysis of mucosal gonococcal infection

描述（由申请方提供）：淋病奈瑟菌是性传播疾病（STD）淋病的病原体，淋病是一种高发病率疾病，全球每年估计有6200万例（WHO）。虽然男性尿道感染通常有症状，但近80%的女性感染了N。淋病是无症状的。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），这是不孕症和异位妊娠的常见原因。通过与NIAID资助的U19赠款（AI 084048）合作的现有临床核心，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，以解决对N.淋病感染NCSTD诊所目前从报告有淋球菌症状的男性获得尿道标本。这些人往往同时感染一种或多种性传播感染。NCSTD还从可能感染淋球菌或淋球菌/衣原体菌株的女性中获得匹配的宫颈阴道灌洗标本，这些菌株是从其匹配的男性伴侣处感染的。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转变，以及不同的男性和女性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测序分析来确定N.在体内感染期间和在与宿主细胞的体外孵育期间，我们的分析揭示了一个独特的签名淋球菌应激反应，代谢和呼吸系统的基因在体内感染过程中表达。基于这些结果，我们假设N。淋病在体内感染过程中表达一组独特的基因，其受男性和女性生殖道感染过程中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个假设：目标1。比较N.淋病在女性生殖道的体内感染到与宫颈上皮细胞一起孵育以及在培养基中体外生长的过程中。目标二。比较N.淋病在体内感染男性生殖道至与尿道上皮细胞一起孵育以及在培养基中体外生长的过程中。目标3：比较N.在体内感染男性和女性生殖道以及与男性尿道上皮细胞和子宫颈内上皮细胞孵育期间，

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.