The affect of hormones and oxygen-limitation on gonococcal pathophysiology

描述（由申请人提供）：临床数据表明，女性淋球菌感染有激素成分；然而，类固醇激素和限氧对淋球菌感染的影响是一个未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，这有助于妇女无症状疾病，最终目标是改善妇女的健康。本应用的目的是详细了解淋球菌对类固醇激素的反应，以及在体内，淋球菌感染期间在宫颈微环境中发生的氧限制（缺氧）。我们的中心假设是，在体内，淋球菌性宫颈炎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。无症状宫颈炎是导致妇女倾向于发展慢性淋球菌病后遗症的主要因素，这转化为大量相关的卫生费用。因此，拟议研究的基本原理是，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发展对抗淋球菌疾病的治疗策略和为改善妇女健康提供框架是必不可少的。因此，拟议的研究直接适用于NIH的使命，即发展将减轻人类疾病负担的基础知识。在强有力的初步数据的指导下，将追求两个具体目标来验证这一假设：1)确定宫颈感染期间淋病奈瑟菌表达谱在类固醇激素和氧气限制下发生的变化；2)在原代人宫颈上皮细胞模型中确定类固醇激素和限氧对补体-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球菌成分。这些分子的一个子集将进一步分析，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，在介导宫颈感染中起关键作用。因此，目的2包括进一步定义体内可能遇到的条件下补体-淋球菌相互作用的分析。我们还将鉴定一种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的具体目标。这些研究是第一次通过使用原发性人上皮细胞模型来研究类固醇激素和缺氧的联合、可变、生理水平如何潜在地调节细菌的发病机制，因此是创新的。拟议的研究意义重大，因为它有望产生关于淋球菌如何在粘膜上皮感染背景下对外源应激（即激素，补体和/或氧气限制）作出反应的极有意义的数据。公共卫生相关性：由于宫颈感染淋病奈瑟菌（引起淋病的细菌），妇女更容易产生慢性、往往是严重的后果。本应用的重点是确定类固醇激素和限氧对淋球菌疾病的影响。提出的研究是细菌发病机制的一个重要和未被充分研究的领域，并适用于人类健康。

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.