Epitranscriptomics in the AIDS-opportunistic pathogen Toxoplasma gondii

艾滋病机会致病菌弓形虫的表观转录组学

• 批准号: 9889878
• 负责人: William J Sullivan
• 金额: $ 23.28万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-08 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889878
• 关键词: 4-thiouracil; 5' Untranslated Regions; AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Address; Adenosine; Affect; Antiparasitic Agents; Area; Automobile Driving; Brain; Cells; Cellular Stress; Chronic; Chronic Disease; Clinical; Communities; Complex; Congenital Abnormality; Cyst; Data Set; Development; Epigenetic Process; Eukaryotic Initiation Factor-2; Felis catus; Follow-Up Studies; Food Contamination; Future; Gene Expression; Gene Expression Regulation; Genetic Transcription; Heat-Shock Response; Homologous Gene; Human; Immune response; Immunity; Immunocompromised Host; Immunoprecipitation; In Vitro; Individual; Infection; Investigation; Label; Life; Link; Location; Lytic Phase; Messenger RNA; Methylation; Modification; Molecular; Multienzyme Complexes; Muscle; Myocardium; Nuclear; Nucleotides; Opportunistic Infections; Parasites; Pathogenesis; Patients; Pharmaceutical Preparations; Phosphorylation; Physiologic pulse; Population; Positioning Attribute; Post-Transcriptional Regulation; Process; Proliferating; Proteins; RNA; RNA Processing; RNA Splicing; Reagent; Recurrence; Refractory; Regulation; Research; Resolution; Role; Site; Skeletal Muscle; Stress; Therapeutic Intervention; Tissues; Toxoplasma; Toxoplasma gondii; Toxoplasmosis; Transcript; Translations; Water; clinically relevant; crosslink; epitranscriptome; epitranscriptomics; insight; knock-down; pathogen; polysome profiling; protein complex; response; transcription factor; transcriptome; transcriptome sequencing

Toxoplasma gondii is an intracellular parasite that has infected up to one-third of the population. The pathogen can cause congenital birth defects and life-threatening opportunistic infection in HIV/AIDS. The replicative stage (tachyzoite) develops into a latent stage (bradyzoite) that resides inside brain, heart, and skeletal muscle tissues, and is impervious to immunity and currently approved antiparasitic drugs. Tissue cysts give rise to recurrent reactivation of infection in immunocompromised patients, creating chronic disease in HIV/AIDS patients. Despite its clinical importance, we know very little about the molecular details orchestrating the conversion between tachyzoites and bradyzoites. Studies have revealed that the development of bradyzoites, which can be triggered in vitro by cellular stress, is a complex process subject to regulation at transcriptional and post-transcriptional levels. Recently, it has been shown in other species that mRNA modifications, specifically post-transcriptional methylation of adenosines at position 6 (m6A), influence gene expression by altering RNA processing or translation. Changes in m6A marks have been associated with modulating cellular stress, development, and differentiation. The discovery of m6A represents a new layer of gene regulation called epitranscriptomics that has not been investigated in protozoan parasites. In preliminary studies, we establish that Toxoplasma RNA is subject to m6A, and we have begun characterizing the enzyme complex that delivers this RNA modification. The study of m6A in Toxoplasma in the context of HIV/AIDS is significant as this modification has been linked to stress-induced changes in cells, making it highly likely that m6A participates in bradyzoite development. In this new R21, we will address our hypothesis that m6A mRNA modifications are required for tachyzoite differentiation into bradyzoites by identifying changes in the m6A epitranscriptome during stage conversion (Aim 1) and determining the “writer” protein complex that delivers m6A onto Toxoplasma mRNA (Aim 2). Our proposed studies will be the first analysis of m6A on mRNA in parasites, and will generate valuable datasets and reagents needed to interrogate this vital new area of investigation relevant to the development of the tissue cysts, which give rise to chronic toxoplasmosis in HIV/AIDS patients.

弓形虫Gondii是一种细胞内寄生虫，已感染多达三分之一的人口。病原体 可能导致先天性的先天出生缺陷和艾滋病毒/艾滋病生命的机会感染。复制 阶段（tachyzoite）发展成一个潜在的阶段（Bradyzoite），该阶段位于大脑，心脏和骨骼肌肉内 组织，并且对免疫学不透水，目前批准了抗寄生虫药物。组织囊肿会产生 免疫功能低下的患者的感染复发，在艾滋病毒/艾滋病中造成慢性疾病 患者。尽管它的临床重要性，但我们对策划的分子细节知之甚少 速氮和曲二核之间的conversion依。研究表明，布拉迪二核的发展， 可以通过细胞应力在体外触发的，这是一个复杂的过程，受转录时的调节。 和转录后级别。最近，在其他物种中已显示mRNA修饰， 特定于位置6（M6A）的腺苷的转录后甲基化影响基因表达 改变RNA处理或翻译。 M6A标记的变化与调节细胞有关 压力，发展和分化。 M6A的发现代表了基因调节的新层 在原生动物寄生虫中尚未研究的称为表交流组学。在初步研究中，我们 确定弓形虫RNA受M6a的约束，我们已经开始表征酶复合物 提供此RNA修饰。在艾滋病毒/艾滋病背景下，对弓形虫中M6a的研究非常重要，因为 这种修饰已与压力诱导的细胞变化有关，使M6A很有可能 参加Bradyzoite开发。在这个新的R21中，我们将解决M6A mRNA的假设 通过识别M6A的变化，需要修改将速氮化物分化为bradyzoite 舞台转换期间的表面参考组（AIM 1）并确定提供的“作家”蛋白质复合物 M6A上弓形虫mRNA（AIM 2）。我们提出的研究将是对m6a对mRNA的首次分析 寄生虫，并将生成需要询问这个至关重要的新领域所需的宝贵数据集和试剂 与组织囊肿的发展有关的研究，这些囊肿引起了慢性弓形虫病在 艾滋病毒/艾滋病患者。