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.

项目摘要/摘要 阴道单细胞寄生虫Trichomonas造成最普遍，非病毒，性 - 全世界传播感染，每年约有1/4亿人患有毛刺病。 trichomoniasis是美国最常见的寄生虫感染，估计每年约500万 案例。然而，这种寄生虫是广泛理解的。因此，阴道链球菌是由中心分类的 疾病控制和预防是2014年美国被忽视的感染。除了是常见原因外 阴道，毛诺病与不良炎症后遗症有关，这可能有助于妊娠 并发症和新生儿死亡率，HIV的传播以及泌尿生殖器癌的转移增加。这 感染的发病率，人们对阴道链球菌与长期健康后果相关的越来越多的认识 阴道托氏杆菌的抗药性临床分离株的数量增加了增长的需求 新的化学治疗和疫苗设计策略。对涉及的过程的更好理解 感染性和发病机理（例如这里提出的）必须实现这些目标。一些 多年前 中介宿主：寄生虫相互作用。我们已经表明，寄生虫蛋白通过EV转移到宿主细胞， 反过来，这既可以调节寄生虫遵守宿主细胞和宿主细胞反应。宿主：病原体跨 - 电动汽车介导的谈话可能导致宿主的寄生虫殖民和细胞因子的下调 将免疫细胞引入感染部位。该提案重点介绍了所使用的分子和细胞机制 对于电动汽车的宿主细胞内在化：寄生虫与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检查电动汽车对体内寄生虫生存的影响 模型。为此，我们建议：表征EV配体的生化特性（TV 4-Alpha- 葡萄糖转移酶结合宿主细胞硫酸肝素蛋白聚糖并驱动宿主细胞的EV内在化 （目标1）;分离并鉴定宿主细胞上的EV受体，并测试EV是否需要受体 内在化（目标2）并确定与宿主细胞内在化有关的EV蛋白并测试EV是否影响 在体内早期感染的寄生虫生存使用宿主细胞中不同的阴道菌株在体内感染。 依从性（目标3）。我们将发现促进寄生虫感染和的生化和细胞机制 将揭示我们的体外发现是否支持EV在宿主细胞定植中的作用。 可能会发现新型机制，因为关于任何相互作用或吸收的机械数据很少 寄生虫衍生的EV与宿主细胞有报道。除了扩大我们的宿主知识：病原体 相互作用，这些研究将增加我们对电动汽车如何介导细胞的总体知识：细胞通信和 将有助于更好地理解电动汽车在传染病中的作用。这些发现也可能 实现新的治疗方法的未来发展。