Multiscale analysis of MyosinA-based motility in Toxoplasma gondii

弓形虫基于肌球蛋白 A 的运动的多尺度分析

• 批准号: 10064612
• 负责人: GARY E WARD
• 金额: $ 53.97万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-12-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10064612
• 关键词: 3-Dimensional; Acquired Immunodeficiency Syndrome; Acute; Adverse drug effect; Affect; Animal Model; Animals; Behavior; Biochemical; Biological; Biology; Biomechanics; Biophysics; Brain; Cell Shape; Cells; Characteristics; Chemicals; Chronic; Clinical; Complex; Data; Development; Disease; Disease Progression; Drug Targeting; Eukaryotic Cell; Evaluation; Extracellular Matrix; Goals; HIV; Human; Impairment; Individual; Infection; Invaded; Life; Life Cycle Stages; Methods; Molecular Genetics; Motor; Mus; Myosin ATPase; Parasites; Pathogenesis; Patients; Pharmaceutical Preparations; Pharmacology; Play; Process; Proteins; Risk; Role; Structure; Suggestion; System; Technology; Testing; Therapeutic; Three-dimensional analysis; Time; Tissues; Toxoplasma gondii; Toxoplasmosis; Virulence; Work; antiretroviral therapy; appendage; base; biophysical analysis; biophysical properties; cell motility; drug development; druggable target; high throughput screening; improved; in vivo; inhibitor/antagonist; innovation; insight; migration; mouse model; opportunistic pathogen; prevent; protein function; side effect; small molecule; small molecule inhibitor; three-dimensional modeling; toxoplasmic encephalitis

Toxoplasmic encephalitis (TE) is a life-threatening infection of the brain in AIDS patients caused by the opportunistic pathogen, Toxoplasma gondii. Drugs are available to treat acute T. gondii infection in these patients, and to suppress its re-emergence in those who are chronically infected, but for many patients the adverse effects of the drugs are severe, resulting in their discontinuation. Thus, there is a need to develop new, better-tolerated drugs to treat AIDS-related TE. This, in turn, requires a better understanding of the biology of T. gondii and the mechanisms underlying its virulence, so that critical points of vulnerability in the parasite’s life cycle can be identified and targeted. The life cycle stage of T. gondii responsible for disease pathogenesis, the tachyzoite, is highly motile. Tachyzoite motility is required for host cell invasion, migration across biological barriers, and dissemination through host tissues. T. gondii MyosinA (TgMyoA) is an unconventional myosin motor protein that plays a central role in parasite motility, and tachyzoites lacking TgMyoA are completely avirulent. The overarching goals of this project are to advance our mechanistic understanding of tachyzoite motility and to test whether small molecules targeting the motility machinery can ameliorate disease in an animal model of infection. The Specific Aims are to: (1) Determine how altering specific aspects of TgMyoA motor function affects parasite motility, by characterizing how recently identified small-molecule inhibitors of the TgMyoA motor affect its biomechanical activity and connecting these changes in motor function to effects on parasite 3D motility; and (2) Determine how inhibiting TgMyoA impacts parasite dissemination and disease progression, in vivo, to better understand the role of TgMyoA and parasite motility during infection and to provide the first direct evaluation of the TgMyoA motor as a drug target for preventing or treating toxoplasmosis. Recent technological advances have created an unprecedented opportunity to manipulate and study parasite motility in a truly integrated way. This project will capitalize on that opportunity across the full range of scales – from the biochemical and biophysical properties of the TgMyoA motor, to the characteristics of parasite motility within a model 3D extracellular matrix, to the ability of parasites to disseminate and cause disease in infected animals. The results will therefore significantly enhance our mechanistic understanding of how T. gondii moves within its hosts to cause disease. Because TgMyoA is both essential for virulence and distinctly different from human myosins it is also a potential target for drug development; by directly testing the druggability of TgMyoA in an animal model of infection, this work will contribute to ongoing efforts to develop new and improved chemotherapeutics for managing T. gondii infections in AIDS patients.

弓形体脑炎（TE）是艾滋病患者的一种危及生命的脑部感染， 条件致病菌弓形虫目前已有治疗急性T细胞白血病的药物。弓形虫感染 病人，并抑制其重新出现在那些谁是慢性感染，但对许多病人来说， 药物的副作用很严重，导致停药。因此，需要开发新的、 耐受性更好的药物来治疗艾滋病相关TE。这反过来又需要更好地理解T. 弓形虫及其毒力机制，因此，在寄生虫的生命中， 周期可以确定和针对性。 对T.负责疾病发病机制的弓形虫速殖子是高度能动的。 速殖子的运动性是宿主细胞侵入、跨越生物屏障和传播所必需的 通过宿主组织t.刚地肌球蛋白A（TgMyoA）是一种非传统的肌球蛋白马达蛋白， 在寄生虫运动中的作用，缺乏TgMyoA的速殖子是完全无毒的。本项目的总体目标是 该项目旨在推进我们对速殖子运动机制的理解，并测试小分子 靶向运动机制可以改善感染动物模型中的疾病。具体目标是 目的：（1）确定改变TgMyoA运动功能的特定方面如何影响寄生虫运动， 表征最近发现的TgMyoA运动的小分子抑制剂如何影响其生物力学 活动，并将运动功能的这些变化与对寄生虫3D运动的影响联系起来;以及（2）确定如何 抑制TgMyoA在体内影响寄生虫传播和疾病进展，以更好地了解其作用 感染期间TgMyoA和寄生虫运动的影响，并提供TgMyoA运动的第一个直接评估 作为预防或治疗弓形虫病的药物靶标。 最近的技术进步为操纵和研究寄生虫创造了前所未有的机会 以一种真正整合的方式。该项目将在整个范围内利用这一机会- 从TgMyoA运动的生物化学和生物物理特性，到寄生虫运动的特征， 在模型3D细胞外基质中，寄生虫传播和在感染者中引起疾病的能力 动物因此，这些结果将大大提高我们对T。弓形虫运动 在宿主体内引起疾病因为TgMyoA是毒力所必需的，并且与 人肌球蛋白也是药物开发的潜在靶点;通过直接测试TgMyoA的可药用性， 在感染的动物模型中，这项工作将有助于开发新的和改进的 用于控制T.艾滋病患者的弓形虫感染。