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

基本信息

批准号：
10530647
负责人：
GARY E WARD
金额：
$ 57.42万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10530647
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 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 Play Process Proteins Risk Role Structure Suggestion System Technology Testing Therapeutic Three-dimensional analysis Time Tissues Toxoplasma gondii Toxoplasmosis Virulence Work antiretroviral therapy appendage biophysical analysis biophysical properties cell motility drug development druggable target high throughput screening improved in vivo inhibitor innovation insight migration mouse model opportunistic pathogen pharmacologic prevent protein function side effect small molecule small molecule inhibitor technology development three-dimensional modeling toxoplasmic encephalitis

项目摘要

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.

弓形虫性脑炎（TE）是由艾滋病患者的威胁生命的大脑感染机会性病原体，弓形虫弓形虫。药物可用于治疗急性T. gondii感染患者，并抑制其在长期感染的患者中的重新出现，但对于许多患者药物的不良影响很严重，导致其中断。那是有必要开发新的更耐受耐受性的药物来治疗与艾滋病相关的TE。反过来，这需要更好地了解T的生物学。 Gondii和其病毒的基础机制，因此寄生虫一生中脆弱性的关键点可以识别和针对周期。造成疾病发病机理的T. gondii的生命周期阶段是高度运动。宿主细胞侵袭，跨生物屏障和传播需要tachyzoite运动性通过宿主组织。 T. gondii肌酸（tgmyoa）是一种非常规的肌球蛋白运动蛋白，可发挥中心在寄生虫运动中的作用和缺乏tgmyoa的速二鼠是完全活泼的。总体目标项目是为了提高我们对tachyzoite运动性的机械理解，并测试是否小分子靶向运动机制可以在动物感染模型中改善疾病。具体目的是至：（1）确定TGMYOA运动功能的特定方面如何影响寄生虫运动。表征最近鉴定出TGMYOA电动机的小分子抑制剂如何影响其生物力学活动并将这些变化的运动功能与对寄生虫3D运动的影响相关联；（2）确定如何抑制tgmyoa会影响寄生虫的传播和疾病进展，以更好地理解该作用感染过程中TGMYOA和寄生虫运动性的首次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学习寄生虫创造了前所未有的机会以一种真正整合的方式运动。该项目将在整个规模上利用这一机会 - 从TGMYOA电动机的生化和生物物理特性到寄生虫运动的特征在3D模型的细胞外基质中，寄生虫传播并引起感染的疾病的能力动物。因此，结果将显着增强我们对T. gondii如何移动的机械理解在其宿主内引起疾病。因为tgmyoa既是病毒必不可少的，又与人肌球蛋白也是药物开发的潜在目标。通过直接测试TGMYOA的可药用性在感染动物模型中，这项工作将有助于发展新的和改进的努力用于管理艾滋病患者中gondii感染的化学治疗药。