Toxoplasma effector-mediated modulation of innate immune pathways in non-murine macrophages

弓形虫效应介导的非鼠巨噬细胞先天免疫途径的调节

• 批准号: 10827548
• 负责人: Yifan Wang
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10827548
• 关键词: Acquired Immunodeficiency Syndrome; Acute; Affect; Animals; Binding; Biochemical; Breeding; CRISPR screen; Cells; Cessation of life; Cytoplasmic Granules; Data; Development; Disease; Disease Outcome; Drug Targeting; Fetus; Future; Gene Expression Profile; Genes; Genetic Polymorphism; Genetic Screening; Genetic Transcription; Goals; Growth; Human; Immune system; Immunocompromised Host; Immunotherapy; In Vitro; Individual; Infection; Inflammasome; Innate Immune Response; Interferon Type II; Knowledge; Laboratory mice; Leadership; Libraries; Ligase; Macrophage; Mediating; Mentors; Modeling; Molecular; Mus; Names; Organelles; Outcome; Parasites; Parasitic infection; Pathogenesis; Patients; Phase; Physiology; Play; Predisposition; Proliferating; Proteins; Proteomics; Rat Strains; Rattus; Rattus norvegicus; Research Personnel; Resistance; Role; Technical Expertise; Testing; Toxoplasma; Toxoplasma gondii; Toxoplasmosis; Training; Ubiquitination; adaptive immune response; cell type; chronic infection; cytokine; fitness; follow-up; genome-wide; in vivo; innate immune pathways; insight; new therapeutic target; novel; novel therapeutics; obligate intracellular parasite; pathogen; persistent symptom; receptor; response; rhoptry; secretory protein; single-cell RNA sequencing; skills; ubiquitin-protein ligase

PROJECT SUMMARY Toxoplasma gondii is an obligate intracellular parasite that causes severe disease in immunocompromised individuals (e.g., AIDS patients) and fetuses. Its abilities to proliferate inside all nucleated cells and establish an infection in almost all warm-blooded animals make the parasite an ideal model to study the mechanisms involving in host-pathogen interaction. The outcome of Toxoplasma infection varies between host species. For example, mice generally succumb to acute Toxoplasma infection while most non-murine hosts, such as rats and humans, usually do not display symptoms but the chronic infection is established. As one of the cell types determining the infection outcome, macrophages play an essential role in the early innate immune response against Toxoplasma and coordinate the adaptive immune response. To successfully establish infection, Toxoplasma co-opts host macrophages via parasite effectors secreted from its unique secretory organelles (e.g., rhoptry and dense granule), named ROPs and GRAs, respectively. The role of Toxoplasma effectors has been predominantly studied in murine macrophages, however, the effectors contributing to the infection in rat and human macrophages are mostly unknown. My previous studies, together with preliminary data of this project, identified several Toxoplasma effectors that specifically modulate the innate immune response in rat but not murine macrophages. My central hypothesis is that Toxoplasma secretes a different set of parasite effectors involved in the modulation of various innate immune responses to establish an infection in macrophages from non-murine hosts. The goals during the K99 mentored phase are: 1) to understand the mechanism of Toxoplasma effector-mediated activation of the NLRP1 inflammasome in rat macrophages; 2) to mechanistically determine the role of Toxoplasma effectors that are required for parasite proliferation in naïve rat macrophages. In the independent R00 phase, I will apply the training from the mentored phase to study the interaction between Toxoplasma effectors and the innate immune response in human macrophages. Specifically, I will identify fitness-conferring Toxoplasma secreted effectors and mainly determine parasite effector-regulated host transcriptional responses in human macrophages. To accomplish these goals, valuable training from a highly complementary mentor team with scientific and mentoring skills will guide my path to an independent researcher. Furthermore, I will develop my leadership skills and strengthen my technical skills through the proposed training. Collectively, the completion of this project will enhance our understanding of the molecular mechanisms controlling host susceptibility to Toxoplasma infection. Also, knowing the parasite effectors important for host modulation will provide better drug targets against toxoplasmosis.

项目摘要 Toxoplasma gondii是一种强制性细胞内寄生虫，在免疫功能低下会引起严重疾病 个体（例如，艾滋病患者）和胎儿。它的能力在所有核细胞内扩散并建立 几乎所有温血动物的感染使寄生虫成为研究机制的理想模型 涉及宿主病原体相互作用。弓形虫感染的结局在宿主物种之间有所不同。为了 例如，小鼠通常屈服于急性弓形虫感染，而大多数非乳腺宿主（例如大鼠） 人类通常不显示符号，但建立了慢性感染。作为细胞类型之一 确定感染结果，巨噬细胞在早期先天免疫反应中起着至关重要的作用 反对弓形虫并协调适应性免疫反应。为了成功建立感染， 弓形虫通过其独特的分泌细胞器分泌的寄生虫效应托管巨噬细胞 （例如，Rhoptry和茂密的颗粒），分别命名为Rops和Gras。弓形虫效应的作用具有 然而，主要是在鼠巨噬细胞中研究的，这种影响导致了大鼠感染 人类巨噬细胞大多未知。我以前的研究，以及此的初步数据 项目，确定了几个毒素效应器 但不是鼠巨噬细胞。我的中心假设是弓形虫分泌不同的寄生虫 调节各种先天免疫反应以建立感染的影响 来自非乳腺宿主的巨噬细胞。 K99修正阶段的目标是：1）了解 大鼠巨噬细胞中NLRP1炎症体的毒素效应介导的激活的机理； 2）到 机械学上确定寄生虫增殖所必需的弓形虫生效的作用 大鼠巨噬细胞。在独立的R00阶段，我将应用修改阶段的培训来研究 人类巨噬细胞中的弓形虫效应与先天免疫反应之间的相互作用。 具体而言，我将确定育种弓形虫分泌的效果，主要确定寄生虫 人类巨噬细胞中效应子调节的宿主转录反应。为了实现这些目标，价值 具有科学和心理技能的高度完成者团队的培训将指导我的道路 独立研究员。此外，我将发展自己的领导能力并增强我的技术技能 通过拟议的培训。总的来说，该项目的完成将增强我们对 控制宿主对弓形虫感染的易感性的分子机制。另外，知道寄生虫 对宿主调节重要的效应子将提供更好的药物靶标针对弓形虫病。