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Development of a novel mouse model with progesterone-independent susceptibility to genital HSV infection

开发一种对生殖器 HSV 感染具有独立孕酮敏感性的新型小鼠模型

基本信息

批准号：
10615599
负责人：
Takeshi Egawa
金额：
$ 7.8万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-05-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10615599
关键词：
Address Adverse event Affect Animal Model Animals Birth Breeding Cell surface Code Contraceptive Agents Dependence Depo Provera Development Diestrus Ensure Enterobacteria phage P1 Cre recombinase Environment Epithelial Cells Epithelium Estrogens Estrus Estrus Synchronization Female Future Genetic Transcription Goals Gonadal Steroid Hormones Herpes Simplex Infections Hormones Human Human Herpesvirus 2 Immune system Immunity Immunologic Surveillance Infection Injectable Keratin Link LoxP-flanked allele Medroxyprogesterone Medroxyprogesterone 17-Acetate Microbe Modeling Modification Molecular Mus Mutation Outcome PVRL1 Pathogenesis Phase Pre-Clinical Model Predisposition Progesterone Regulation Research Resistance Risk Role Sexually Transmitted Diseases Simplexvirus Site Surface Testing Time Tissues Transgenic Mice Vaccines Vagina Vertical Transmission Viral Viral Load result Viral Pathogenesis Virus Diseases Woman Women&apos s Health Work antiviral immunity co-infection commensal bacteria constitutive expression dysbiosis female sex hormone fungus genital herpes hormonal contraception microbial mouse model new therapeutic target novel pathogen posttranscriptional prevent promoter receptor reproductive tract sex sexually transmitted virus vaginal infection vector

项目摘要

Genital herpes, an incurable sexually transmitted infection (STI) caused by herpes simplex virus (HSV), disproportionately affects women and can cause sex-specific adverse events such as increased risk of acquiring other STIs as well as vertical transmission during birth. The mouse is commonly used as a preclinical model to study immunity and pathogenesis of vaginal HSV infection. However, one major drawback of the murine model is that unlike humans, mice are susceptible to infection only at certain stages of the estrus cycle. Thus, female mice are typically pre-treated with depot medroxyprogesterone acetate (DMPA, Depo-Provera), a progesterone-based hormonal contraceptive, to hold the mice in diestrus and ensure uniform susceptibility to HSV infection. However, the use of DMPA limits the study of host-pathogen interactions a single stage of the estrus cycle, despite abundant evidence that female sex hormones have a major impact on antiviral defense and immunosurveillance of the female reproductive tract. Furthermore, use of DMPA precludes co-inoculation studies with commensal bacteria or fungi, as these microbes often require an estrogen-high environment to robustly colonize the murine vagina. One major difference between the mouse and human vagina is that expression of the viral entry receptor nectin-1 on epithelial cells is hormone dependent in mice, but constitutively expressed in humans. To overcome this challenge, this proposal will develop a novel mouse model in which nectin-1 is constitutively expressed in the murine vaginal epithelium by addressing transcriptional and post-translational regulatory mechanisms. Nectin-1 expression will be placed under the control two different constitutive promoters, and two mutations will be introduced to prevent ectodomain cleavage. Epithelium-specific expression of nectin-1 will be achieved through the inclusion of a stop-flox cassette and breeding to mice in which Cre recombinase expression is controlled by the human keratin 14 promoter. Finally, the HSV susceptibility of these novel mice will be tested at different stages of the estrus cycle. The successful development of a novel, more translational mouse model of HSV infection would help achieve our long-term goal of exploring the role of sex hormones in modulating host-pathogen interactions at the vaginal barrier as well as investigating the molecular impact of vaginal microbial dysbiosis or STI co- infection on HSV pathogenesis. We expect future studies to reveal unknown aspects of protective immunity against genital herpes, including that induced by vaccines, and potentially lead to the discovery of novel targets for therapeutics aimed at reducing susceptibility against HSV infection.

生殖器疱疹是由单纯疱疹病毒（HSV）引起的一种无法治愈的性传播感染（STI），对妇女的影响尤为严重，并可能导致性别特异性不良事件，如感染其他性传播感染的风险增加以及分娩期间的垂直传播。小鼠通常被用作研究阴道HSV感染的免疫和发病机制的临床前模型。然而，小鼠模型的一个主要缺点是，与人类不同，小鼠仅在发情周期的某些阶段易受感染。因此，雌性小鼠通常使用一种基于黄体酮的激素避孕药醋酸甲羟孕酮（DMPA, Depo-Provera）进行预处理，以使小鼠处于妊娠期，并确保对HSV感染的均匀易感性。然而，尽管有大量证据表明雌性性激素对雌性生殖道的抗病毒防御和免疫监视有重要影响，但DMPA的使用将宿主-病原体相互作用的研究限制在发情周期的单个阶段。此外，使用DMPA排除了与共生细菌或真菌的共接种研究，因为这些微生物通常需要高雌激素的环境才能在小鼠阴道中稳稳地定植。小鼠和人类阴道的一个主要区别是，小鼠上皮细胞上的病毒进入受体nectin-1的表达是激素依赖性的，而人类则是组成性的。为了克服这一挑战，本研究将开发一种新的小鼠模型，其中通过解决转录和翻译后调控机制，连接蛋白-1在小鼠阴道上皮中组成性表达。将Nectin-1的表达置于两个不同的组成启动子的控制之下，并引入两个突变来防止外畴切割。连接蛋白-1的上皮特异性表达将通过包含一个停止絮体盒和繁殖到Cre重组酶表达受人类角蛋白14启动子控制的小鼠中来实现。最后，这些新型小鼠的HSV易感性将在发情周期的不同阶段进行测试。成功开发一种新的，更具可翻译性的HSV感染小鼠模型将有助于实现我们探索性激素在调节阴道屏障宿主-病原体相互作用中的作用，以及研究阴道微生物生态失调或性传播感染对HSV发病机制的分子影响的长期目标。我们期望未来的研究能够揭示针对生殖器疱疹的保护性免疫的未知方面，包括由疫苗诱导的免疫，并可能导致发现旨在降低对HSV感染易感性的治疗新靶点。