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贡迪使几乎全球人口的三分之一处于风险由于慢性感染的重生而导致疾病。当前的治疗方法无法消除缓慢的复制，持续的弓形虫刺激性囊肿，使种子重新激活并确保眼睛，大脑，心脏或肺。眼科毒素病的患者尤其面临着由于重新激活的顺序时期。我们的长期目标是确定破坏毒品的关键责任持久性，从而排除在风险个体中重新激活疾病。达到这个目标，我们最近有了证明了培养物和感染小鼠中的胸肌囊肿的生存能力取决于具有A的寄生虫功能自噬途径。但是，关于弓形虫中的自噬和利用寄生虫知之甚少自噬要影响感染，需要识别途径中的新成分和不同的成分。为了满足这种需求，利用我们最近的遗传屏幕，我们将通过开发为第一次在T. gondii中是汇总表型驱动单核苷酸的“主要编辑”方法变体，定义新成分在寄生虫自噬中的一般作用，并测量其对急性的影响和慢性感染。预计完成拟议的研究将至少确定4个新的和不同的分歧可以优先进行更深入的分析和开放的新型途径，将gondii自噬的成分优先考虑在这项工作的下一阶段进行调查。