Identifying novel players in Toxoplasma autophagy during chronic infection”

识别慢性感染期间弓形虫自噬的新参与者 –

基本信息

批准号：
10223735
负责人：
Vernon Bruce Carruthers
金额：
$ 19.5万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-15 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10223735
关键词：
Acquired Immunodeficiency Syndrome Automobile Driving Autophagocytosis Biological Assay Birth Blindness Brain CRISPR/Cas technology Cells Cellular biology Chemicals Chronic Code Cyst Defect Disease Eating Eye Genes Genetic Screening Goals Health Heart Homeostasis Human Immune Immune system Individual Infant Infection Intervention Investigation Life Literature Lung Measures Mitotic Molecular Genetics Mus Mutagenesis Mutation Ocular Toxoplasmosis Organ Organ Transplantation Parasites Pathway interactions Phase Phenotype Populations at Risk Pregnancy Property Proteins Publishing Reporter Risk Role Seeds Single Nucleotide Polymorphism Site-Directed Mutagenesis Testing Time Tissues Toxoplasma Toxoplasma gondii Toxoplasmosis Vision Work acute infection base cancer therapy chronic infection defined contribution experience genome sequencing in vivo mouse model mutant novel operation protein degradation spatiotemporal transplantation therapy whole genome

项目摘要

The protozoan parasite Toxoplasma gondii renders nearly one third of the global human population at risk of experiencing disease due to reactivation of chronic infection. Current treatments fail to eliminate the slow replicating, persistent Toxoplasma bradyzoite cysts that seed reactivation and ensuing disease in the eyes, brain, heart or lungs. Individuals with ocular toxoplasmosis are particularly at risk of experiencing progressive loss of vision due to sequential periods of reactivation. Our long-term goal is to identify critical liabilities for disrupting Toxoplasma persistence, thereby precluding reactivated disease in at risk individuals. Toward this goal, we have recently demonstrated that the viability of bradyzoite cysts in culture and in infected mice relies on the parasite having a functional autophagy pathway. However, little is known about autophagy in Toxoplasma and exploiting parasite autophagy to impact infection requires identifying new and divergent components in the pathway. To meet this need, leveraging our recent genetic screen we will identify novel components of the autophagy pathway by developing for the first time in T. gondii a “prime editing” approach for pooled identification of phenotype driving single nucleotide variants, define the general roles of novel components in parasite autophagy, and measure their impact on acute and chronic infection. Completing the proposed studies is expected to identify at least 4 new and divergent components of T. gondii autophagy that can be prioritized for deeper analysis and open novel avenues of investigation in the next phase of this work.

原生动物寄生虫弓形虫使全球近三分之一的人口面临感染的风险由于慢性感染重新激活而导致的疾病，目前的治疗无法消除缓慢复制，持久性弓形虫缓殖子包囊会在眼睛、大脑、心脏或眼睛中重新激活并引发疾病患有眼部弓形体病的人特别容易因以下原因而出现进行性视力丧失的风险。我们的长期目标是确定破坏弓形虫的关键责任。为了实现这一目标，我们最近采取了一些措施。证明缓殖子包囊在培养物和受感染小鼠中的生存能力依赖于具有然而，人们对弓形虫的自噬和利用寄生虫知之甚少。自噬影响感染需要识别该途径中新的和不同的成分来满足这一需求。利用我们最近的基因筛选，我们将通过开发来识别自噬途径的新成分首次在弓形虫中采用“启动编辑”方法来汇总鉴定驱动单核苷酸的表型变体，定义新成分在寄生虫自噬中的一般作用，并测量它们对急性自噬的影响完成拟议的研究预计将确定至少 4 种新的和不同的感染。弓形虫自噬的组成部分可以优先进行更深入的分析并开辟新的途径这项工作的下一阶段的调查。