Identifying novel players in Toxoplasma autophagy during chronic infection”

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 由于慢性感染重新激活而经历疾病的。目前的治疗方法无法消除缓慢的复制， 持续的弓形虫缓殖子包囊，可导致眼、脑、心脏或其他器官的重新激活和随后的疾病 肺部。患有眼弓形体病的人尤其有可能经历进行性视力丧失，原因是 重新激活的连续周期。我们的长期目标是找出破坏弓形虫的关键责任 持久性，从而在高危人群中排除重新激活的疾病。为了实现这一目标，我们最近 证明了在培养和受感染的小鼠中，慢殖子包囊的生存依赖于寄生虫有一个 功能自噬途径。然而，关于弓形虫中的自噬和利用寄生虫的情况知之甚少。 影响感染的自噬需要识别途径中新的和不同的成分。为了满足这一需求， 利用我们最新的基因筛查，我们将通过开发 弓形虫首次采用“主编辑”方法共同鉴定驱动单核苷酸的表型 变异体，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 和慢性感染。完成拟议的研究预计将确定至少4个新的和不同的 弓形虫自噬的成分，可优先用于更深入的分析和开辟新的途径 在这项工作的下一阶段进行调查。

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.