Identification of key players mediating internalization of Trichomonas vaginalis extracellular vesicles by host cells and parasite adherence and survival in vivo

宿主细胞介导阴道毛滴虫细胞外囊泡内化的关键参与者的鉴定以及寄生虫在体内的粘附和存活

• 批准号: 10177862
• 负责人: Patricia Jean Johnson
• 金额: $ 65.6万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-02 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10177862
• 关键词: Adherence; Affect; American; Annual Reports; Antigen Presentation; Binding; Biochemical; Case Study; Cell Communication; Cell physiology; Cells; Centers for Disease Control and Prevention (U.S.); Cervicitis; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Communication; Contracts; Data; Development; Disease; Down-Regulation; Drug resistance; Future; Glycoside Hydrolases; Goals; HIV; Health; Helminths; Heparan Sulfate Proteoglycan; Heparitin Sulfate; Homeostasis; Human; Immune; Immune response; In Vitro; Incidence; Infection; Inflammatory; Knock-out; Knowledge; Lead; Learning; Leishmania; Ligands; Malignant neoplasm of cervix uteri; Malignant neoplasm of prostate; Mammalian Cell; Mass Spectrum Analysis; Mediating; Membrane; Molecular; Neonatal Mortality; Neoplasm Metastasis; Outcome; Parasites; Parasitic infection; Pathogenesis; Pelvic Inflammatory Disease; Pharmaceutical Preparations; Plasmodium; Play; Pregnancy Complications; Process; Property; Proteins; Proteomics; RNA; Reporting; Research; Risk; Role; Severities; Sexually Transmitted Diseases; Site; Testing; Therapeutic; Transferase; Trichomonas Infections; Trichomonas vaginalis; Trypanosoma; Urethritis; Urogenital Cancer; Vaccine Design; Vaginitis; Vesicle; adverse pregnancy outcome; cytokine; design; exosome; extracellular; extracellular vesicles; host colonization; human pathogen; immunoregulation; in vivo; insight; male; mouse model; neglect; novel; novel therapeutic intervention; pathogen; receptor; response; transmission process; uptake; urogenital tract; vaccine candidate

Project Summary/Abstract The unicellular parasite Trichomonas vaginalis is responsible for the most prevalent, non-viral, sexually- transmitted infection worldwide, with approximately ¼ billion people contracting trichomoniasis annually. Trichomoniasis is the most common parasitic infection in the US, with an annual incidence estimated at ~5 million cases. Nevertheless, this parasite is vastly understudied. As such, T. vaginalis was classified by the Center of Disease Control and Prevention as a neglected infection in the US in 2014. In addition to being a common cause of vaginitis, trichomoniasis is associated with adverse inflammatory sequelae, that can contribute to pregnancy complications and neonatal mortality, the spread of HIV, and increased metastasis of urogenital cancers. The incidence of infection, the growing recognition that T. vaginalis is associated with long-term health consequences and an increase in the number of drug resistant clinical isolates of T. vaginalis underscore the need to develop new chemotherapeutic and vaccine design strategies. A much better understanding of processes involved in infectivity and pathogenesis, such as those proposed here, will be imperative to achieve these goals. Several years ago we discovered that T. vaginalis secretes small, membrane-bound extracellular vesicles (EVs), that mediate host:parasite interactions. We have shown that parasite proteins are transferred to host cells via EVs, which in turn, modulates both parasite adherence to host cells and the host cell responses. Host:pathogen cross- talk mediated by EVs likely contributes to parasite colonization of the host and down-regulation of cytokines that elicit immune cells to the site of infection. This proposal focuses on the molecular and cellular mechanisms used for host cell internalization of EVs: a process required for EV-mediated communication between the parasite and host. We will also examine the effect of EVs on the survival of parasites in vivo using a newly-developed mouse model. To this end, we propose to: characterize biochemical properties of an EV ligand (Tv 4-alpha- glucanotransferase) that binds host cell heparan sulfate proteoglycans and drives EV internalization by host cells (Aim 1); isolate and identify EV receptor(s) on the host cell and test whether the receptor(s) are required for EV internalization (Aim 2) and to identify EV proteins involved in host cell internalization and test whether EVs affect parasite survival during early stage infection in vivo using isogenic T. vaginalis strains that differ in host cell adherence (Aim 3). We will uncover biochemical and cellular mechanisms that promote parasite infection and will reveal whether our in vitro findings supporting a role for EVs in host cell colonization are confirmed in vivo. Novel mechanisms are likely to be found, as very little mechanistic data on the interaction or uptake of any parasite-derived EV with host cells have been reported. In addition to expanding our knowledge of host:pathogen interactions, these studies will increase our overall knowledge of how EVs mediate cell:cell communication and will contribute to a better understanding of the role of EVs in infectious diseases. These findings could also enable future development of new therapeutic approaches.

项目总结/摘要 单细胞寄生虫阴道毛滴虫是最普遍的，非病毒性的，性- 在世界范围内传播感染，每年约有1/4亿人感染滴虫病。 滴虫病是美国最常见的寄生虫感染，每年的发病率估计约为500万 例然而，这种寄生虫的研究还远远不够。因此，T。迷走神经病被分类的中心 2014年，疾病控制和预防在美国被忽视。除了作为一个共同的事业 阴道炎，滴虫病是与不良炎症后遗症，可有助于怀孕 并发症和新生儿死亡率，艾滋病毒的传播，泌尿生殖系统癌症的转移增加。的 感染的发病率，越来越多的人认识到，T。迷走神经病与长期健康后果有关 耐药临床分离株的数量增加。流浪汉强调需要发展 新的化疗和疫苗设计策略。更好地理解涉及的过程 传染性和发病机制，如这里提出的那些，将是实现这些目标的必要条件。几 几年前我们发现T.迷走神经分泌小的、膜结合的细胞外囊泡（EV）， 介导宿主：寄生虫相互作用。我们已经证明寄生虫蛋白质通过EV转移到宿主细胞， 其又调节寄生虫对宿主细胞的粘附和宿主细胞的反应。宿主：病原体交叉- 由EV介导的谈话可能有助于宿主的寄生虫定殖和细胞因子的下调， 将免疫细胞引到感染部位。这项建议的重点是分子和细胞的机制， 对于EV的宿主细胞内化：寄生虫和宿主细胞之间EV介导的通信所需的过程。 主持人我们还将使用新开发的小鼠研究EV对体内寄生虫存活的影响 模型为此，我们建议：表征EV配体（Tv 4-α- 葡聚糖转移酶），其结合宿主细胞硫酸乙酰肝素蛋白聚糖并驱动宿主细胞的EV内化 (Aim 1）;分离和鉴定宿主细胞上的EV受体，并测试该受体是否为EV所需 目的2），并鉴定参与宿主细胞内化的EV蛋白，并测试EV是否影响 使用同基因T.宿主细胞不同的迷走神经菌株 坚持（目标3）。我们将揭示促进寄生虫感染的生化和细胞机制， 将揭示我们的体外研究结果是否支持EV在宿主细胞定植中的作用在体内得到证实。 可能会发现新的机制，因为很少有关于任何药物相互作用或摄取的机制数据。 已经报道了寄生虫来源的EV与宿主细胞。除了扩大我们对宿主的了解：病原体 这些研究将增加我们对EV如何介导细胞：细胞通信的整体知识， 将有助于更好地了解电动汽车在传染病中的作用。这些发现也可能 使未来开发新的治疗方法成为可能。