The affect of hormones and oxygen-limitation on gonococcal pathophysiology

描述(申请人提供)：临床数据表明女性淋球菌感染有激素成分；然而，类固醇激素和氧气限制对淋球菌感染的影响是一个未被充分研究的重要研究领域。我们的长期目标是阐明淋球菌和宫颈上皮之间发生的动态相互作用，这是导致女性无症状疾病的原因，最终目标是改善女性的健康。这项应用的目的是提供淋球菌如何对类固醇激素以及在体内发生的氧气限制(缺氧)的反应的详细了解，在淋球菌感染期间，宫颈微环境中会发生这种情况。我们的中心假设是，在体内，淋球菌性宫颈炎症是由氧气限制以及激素诱导的淋球菌和宫颈上皮的变化所控制的。无症状的宫颈炎症是导致妇女患慢性淋球菌性疾病后遗症的主要因素，这转化为大量相关的健康成本。因此，拟议研究的理由是，在月经周期的背景下更好地了解淋球菌的发病机制，对于未来制定抗击淋病的治疗战略和为改善妇女健康提供框架是至关重要的。因此，拟议的研究直接适用于NIH任务中与基础知识的发展有关的部分，这将减少人类疾病的负担。在强大的初步数据的指导下，将寻求两个特定的目标来验证这一假设：1)确定在宫颈感染期间淋球菌对类固醇激素和氧气限制的反应的表达谱的变化；以及2)在原代人类宫颈上皮细胞模型中确定类固醇激素和氧气限制对补体-淋球菌相互作用的影响。在目标1中，我们将确定可能导致体内淋球菌疾病的淋病成分。这些分子的一个子集将被进一步分析，以确定它们对促进宫颈疾病的潜在贡献。补体的产生对类固醇激素有反应，并在介导宫颈感染中起着关键作用。因此，目标2包括进一步确定补体-淋球菌在体内可能遇到的条件下的相互作用的分析。我们还将鉴定一个假定的淋球菌补体结合蛋白NGO0033。各种细胞、分子和生化技术将被用来完成我们特定目标的目标。这些研究首次通过使用原代人类上皮细胞模型来研究类固醇激素和低氧的组合、可变的生理水平如何潜在地调节细菌的发病，因此是创新的。这项拟议的研究意义重大，因为它有望在粘膜上皮感染的背景下产生关于淋球菌如何响应外源压力(即激素、补体和/或氧气限制)的非常有意义的数据。与公共卫生相关：妇女更容易由于宫颈感染淋病奈瑟菌而产生慢性后果，通常是严重的后果，淋病是导致淋病的细菌。本应用的重点是确定类固醇激素和氧气限制对淋球菌疾病的影响。拟议的研究是细菌发病机制的一个重要且未被充分研究的领域，适用于人类健康。

DESCRIPTION (provided by applicant): Clinical data indicate that there is a hormonal component to gonococcal infection in women; however, the effect of steroid hormones and oxygen limitation to gonococcal infection is an under-investigated, important, area of study. Our long-term goal is to elucidate the dynamic interactions, occurring between the gonococcus and the cervical epithelium, which contribute to asymptomatic disease in women, with the ultimate goal of improved women's health. The intent of this application is to provide a detailed understanding of how gonococci respond to steroid hormones, as well as to oxygen limitation (hypoxia), as would occur, in vivo, within the cervical microenvironment during gonococcal infection. It is our central hypothesis that, in vivo, gonococcal cervicitis is governed by oxygen limitation as well as by hormone-induced changes to the gonococcus and to the cervical epithelium. Asymptomatic cervicitis is the primary factor contributing to the propensity of women to develop chronic gonococcal disease sequelae, which translates into substantial associated health cost. Therefore, the rationale for the proposed studies is that a greater understanding of gonococcal pathogenesis in the context of the menses cycle is imperative to the future development of therapeutic strategies to combat gonococcal disease and to provide the framework for improved women's health. Thus, the proposed research is directly applicable to that part of the NIH's mission that pertains to the development of fundamental knowledge that will reduce the burden of human illness. Guided by strong preliminary data, two specific aims will be pursued to test this hypothesis: 1) Define changes occurring during cervical infection in the N. gonorrhoeae expression profile in response to steroid hormones and oxygen limitation; and 2) Define the effect of steroid hormones and oxygen limitation on the complement-gonococcus interaction in a primary human cervical epithelial cell model. Within Aim 1, we will identify gonococal constituents that are likely to contribute to gonococcal disease in vivo. A subset of these molecules will be analyzed further to define their potential contribution to promoting cervical disease. Complement production is responsive to steroid hormones and plays a critical role in mediating cervical infection. Thereby, Aim 2 comprises analyses to further define the complement-gonococcus interaction under conditions likely to be encountered in vivo. We will also characterize a putative gonococcal complement binding protein, NGO0033. A variety of cellular, molecular, and biochemical techniques will be used to complete the objectives of our Specific Aims. These investigations are the first to examine how combined, variable, physiological levels of steroid hormones and hypoxia potentially modulate bacterial pathogenesis by using a primary, human, epithelial cell model, and, thus, are innovative. The proposed research is significant because it is expected to generate exceedingly meaningful data regarding how gonococci respond to exogenous stresses (i. e. hormones, complement, and/or oxygen limitation) in the context of mucosal epithelial infection. PUBLIC HEALTH RELEVANCE: Women are more prone to develop chronic, often severe, consequences as a result of cervical infection with Neisseria gonorrhoeae, the bacterium that causes gonorrhea. The focus of this application is to define the effect of steroid hormones and oxygen limitation to gonococcal disease. The proposed studies are an important and under-investigated area of bacterial pathogenesis and are applicable to human health.