Mechanism of Infectivity Acquisition in African Trypanosomes

非洲锥虫感染性获得机制

• 批准号: 10356095
• 负责人: CHRISTIAN TSCHUDI
• 金额: $ 45.9万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-03-01 至 2024-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10356095
• 关键词: Africa South of the Sahara; African; African Trypanosomiasis; Animals; Biology; Blood; Blood Circulation; Cattle; Cells; Cellular Morphology; Characteristics; Communicable Diseases; Complement; Complex; Development; Disease; Domestic Pig; E2F transcription factors; Economic Development; Event; Exposure to; Funding; Gene Expression; Genes; Goals; High-Throughput DNA Sequencing; Human; Human Biology; In Vitro; Infection; Infectious Agent; Insect Vectors; Intervention; Investigation; Life Cycle Stages; Link; Livestock; Membrane Glycoproteins; Messenger RNA; Metabolism; Molecular; Parasites; Pathogenesis; Pathway interactions; Pharmaceutical Preparations; Phenotype; Play; Point Mutation; Process; Protozoa; Public Health; RNA; RNA Interference; RNA-Binding Proteins; Regulation; Research; Resistance; Role; Salivary Glands; Signal Transduction; Signaling Protein; Specificity; Surface; System; Therapeutic; Time; Transcript; Translations; Trypanosoma; Trypanosoma brucei brucei; Tsetse Flies; Vaccines; Variant; Work; base; combat; feeding; human pathogen; inducible gene expression; innovation; insight; metacyclogenesis; mutant; mutation screening; nagana; news; overexpression; programs; protein function; side effect; transcriptome sequencing; vector; wasting

Project Summary Unraveling the biology of human pathogens is fundamental toward understanding mechanisms of pathogenesis and identifying genes essential for survival in the host. This application focuses on the protozoan parasite Trypanosoma brucei, which causes devastating diseases in humans and animals in sub-Saharan Africa. There are no vaccines, and therapeutic drugs have serious side effects and decreasing efficacy. Thus, there is a pressing need for research to better understand the biology of these human pathogens and the mechanisms they use to survive within their hosts. T. brucei undergoes a complex life cycle between the mammalian host and the blood-feeding tsetse fly vector, which among others involves changes in cell morphology, surface coat composition, metabolism, signaling pathways and gene expression. Consequently, these parasites have evolved adaptations to allow for their survival in both the gut and salivary glands of the tsetse fly, as well as in the bloodstream of their mammalian host. By overexpressing a single RNA-binding protein (RBP6) in non-infectious trypanosomes, we recapitulated in vitro the events leading to acquisition of infectivity in the insect vector, including the expression of metacyclic variant surface glycoproteins (mVSGs). The overall goal of the proposed work here is to identify the mechanism by which RBP6 activates developmental progression to infectious metacyclics and to characterize the RNA binding protein network that regulates the developmental program leading to epimastigotes and infectious metacyclics. To accomplish these goals, we will build on our major findings in the previous funding period, namely the identification of initial targets of RBP6 function, the depiction of the molecular characteristics of metacyclics, and the identification of a large compendium of molecules linked to the developmental program leading to infective metacyclic parasites. We will use innovative approaches involving a screen for RBP6 function with deep mutational scanning by combining metacyclic selection and high-throughput DNA sequencing and analyze gene expression at the single cell level with RNA-Seq. Taken together our research plan provides unique opportunities to illuminate the developmental program leading from non-infective procyclics to infectious metacyclics, a crucial process in the T. brucei life cycle.

项目摘要 揭开人类病原体生物学的面纱是了解其发病机制的基础 致病机制和鉴定寄主生存所必需的基因。此应用程序侧重于 布鲁氏锥虫原生动物寄生虫，可导致人类和动物的毁灭性疾病 撒哈拉以南非洲。没有疫苗，治疗药物有严重的副作用和 药效下降。因此，迫切需要进行研究，以更好地了解这些疾病的生物学 人类病原体及其在宿主体内生存的机制。布氏毛滴虫经历了一个 哺乳动物宿主和食血采采蝇媒介之间的复杂生活史，其中 其他涉及细胞形态、表层成分、新陈代谢、信号通路的变化 和基因表达。因此，这些寄生虫进化出了适应能力，以允许它们生存 采采蝇的肠道和唾液腺以及它们的哺乳动物的血液中 主持人。通过在非传染性锥虫中过表达单一RNA结合蛋白(RBP6)，我们 在体外概述了导致在昆虫媒介中获得感染性的事件，包括 亚环变异体表面糖蛋白(MVSGs)的表达。拟议工作的总体目标 以下是确定RBP6激活发育进展为感染性疾病的机制 并表征调节发育的RNA结合蛋白网络。 导致上鞭毛和侵染性后循环的程序。为了实现这些目标，我们将在 我们在上一个供资期间的主要发现，即确定成果预算编制方案6的初步目标 功能，刻画了元环的分子特征，并鉴定了一个大的 与导致感染性后循环寄生虫的发育程序相关联的分子概要。 我们将使用创新的方法，包括RBP6功能的筛查和深度突变扫描 通过结合亚环选择和高通量DNA测序，分析基因表达 单细胞水平用RNA-Seq.总而言之，我们的研究计划提供了独特的机会 阐述了从非感染性前周期到感染性后周期的发展过程 布氏毛滴虫生活史中的关键过程。