Mechanistic analysis of Toxoplasma gondii sexual development in tissue culture and mouse models

组织培养和小鼠模型中弓形虫性发育的机制分析

• 批准号: 10533513
• 负责人: Nicole Marie Davis
• 金额: $ 6.81万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10533513
• 关键词: Ablation; Affect; Agreement; Biological Models; Biology; Brain; Cause of Death; Cell Culture Techniques; Cell Differentiation process; Cells; Data; Development; Disease; Environment; Epigenetic Process; Ethics; Eye; Family Felidae; Fatty Acids; Felis catus; Gene Proteins; Genes; Genetic Transcription; Goals; Growth; HDAC3 gene; Histone Deacetylase; Histone Deacetylase Inhibitor; Histones; Human; Immunocompromised Host; In Vitro; Individual; Integration Host Factors; Intestines; Knowledge; Linoleic Acids; Lipids; Literature; Logistics; Mammals; Measures; Mediating; Modeling; Modification; Molecular; Mus; Oocysts; Parasites; Polyunsaturated Fatty Acids; Prevalence; Production; Research; Role; Seroprevalences; Sex Differentiation; Sexual Development; Signal Pathway; Small Interfering RNA; Testing; Therapeutic; Toxoplasma gondii; Toxoplasmosis; Tumor stage; United States; Work; asexual; automated image analysis; cell type; companion animal; experimental study; foodborne illness; improved; in vivo; intestinal epithelium; knock-down; mouse model; overexpression; prevent; research study; response; sex; tissue culture; transmission process; vaccine development

Project Summary Toxoplasma gondii is the causative agent of toxoplasmosis, a leading cause of death due to foodborne illness that causes serious disease in immunocompromised individuals. The parasite develops asexually and sexually, and asexual parasite development is well-studied. However, the parasite completes its sexual stage only in the cat intestinal epithelium, presenting a significant ethical and logistical barrier to sexual stage research. To remove the need for companion animal research and study the unknown biology of sexual stage T. gondii, our lab developed murine and tissue culture models that support T. gondii sexual development. The polyunsaturated fatty acid linoleic acid is a critical factor for T. gondii to complete sexual development that is uniquely elevated in the feline intestine. The mechanisms by which linoleic acid acts on host or parasite remain unknown. The goal of this proposal is to determine the mechanism of linoleic acid in promoting T. gondii sexual development in cell culture and in vivo. Preliminary data from our group and others suggests that linoleic acid acts on both host and parasite to promote a permissive environment for T. gondii sexual development. I hypothesize that intracellular accumulation of linoleic acid activates T. gondii lipid signaling pathways to promote sexual development. I further hypothesize that sexual development relies on activation of similar signaling pathways in the host. In Aim 1, I will determine the mechanism of linoleic acid action on the parasite by measuring parasite transcriptional responses to linoleic acid treatment. I will also test our new mouse model of linoleic acid accumulation for its ability to promote efficient T. gondii sexual development in vivo. In Aim 2, I will use automated image analysis to determine the importance of host cell type in T. gondii sexual development. siRNA-mediated ablation of linoleic acid-responsive host factors will identify host genes that influence sexual development. Successful completion of these Aims will better define how linoleic acid enables growth of T. gondii sexual stages. The long-term implication of this work is a shareable model of T. gondii sexual development that will help reduce the burden of toxoplasmosis.

项目摘要 弓形虫Gondii是弓形虫病的致病药物，这是由于食源性疾病引起的主要死亡原因 这会导致免疫功能低下的个体严重疾病。寄生虫自然发展， 性行为和无性寄生虫的发展是充分研究的。但是，寄生虫完成了性阶段 仅在猫肠上皮，呈现出重大的道德和后勤障碍 研究。消除对伴侣动物研究的需求并研究性阶段的未知生物学 T. Gondii，我们的实验室开发了支持T. gondii性发展的鼠和组织培养模型。这 多不饱和脂肪酸亚油酸是T. gondii完成性发育的关键因素 猫肠中的独特升高。亚油酸对宿主或寄生虫作用的机制保留 未知。该提案的目的是确定亚油酸在促进gondii性方面的机制 细胞培养和体内的发展。我们小组和其他人的初步数据表明亚油酸 对宿主和寄生虫都作用，以促进gondii性发展的宽松环境。我 假设亚油酸的细胞内积累激活了T. gondii脂质信号传导途径 促进性发展。我进一步假设性发展依赖于类似的激活 主机中的信号通路。在AIM 1中，我将确定亚油酸在寄生虫上的机理 通过测量对亚油酸处理的寄生虫转录反应。我还将测试我们的新鼠标模型 亚油酸积累的促进体内有效T. gondii性发育的能力。在AIM 2中，我 将使用自动图像分析来确定T. gondii性宿主细胞类型的重要性 发展。 siRNA介导的亚油酸响应性宿主因子的消融将确定宿主基因 影响性发展。成功完成这些目标将更好地定义亚油酸如何启用 T. gondii性阶段的增长。这项工作的长期影响是T. gondii的可共享模型 性发育将有助于减轻弓形虫病的负担。