Rapid Phenotyping of the ZIKV Genome

ZIKV 基因组的快速表型分析

• 批准号: 9263229
• 负责人: RICHARD YUQI ZHAO
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-07 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9263229
• 关键词: Adult; Alpha Cell; Americas; Animal Model; Biological Models; Biology; Cell Cycle Regulation; Cell Death; Cell Survival; Cells; Cellular biology; Centrosome; Chromosomes; Clinical; Cloning; Congenital Abnormality; Consensus; Development; Disease; Disease Outbreaks; Drug Targeting; Eukaryota; Fibroblasts; Fission Yeast; Flavivirus; Future; Gene Mutation; Genes; Genome; Goals; Guillain-Barré Syndrome; HIV-1; Human; Individual; Infection; Intrinsic factor; Link; Mammalian Cell; Measures; Microcephaly; Mitochondria; Mitotic Cell Cycle; Modeling; Neurologic Dysfunctions; Neurons; Newborn Infant; Outcome; Pathogenicity; Peptide Hydrolases; Pharmacotherapy; Phenotype; Preclinical Drug Evaluation; Property; Research; Shotguns; Site; Skin; System; Testing; Time; Translations; Variant; Violence; Viral; Viral Genome; Viral Physiology; Viral Proteins; Virulence Factors; Virus; Virus Diseases; Virus Replication; Yeast Model System; Yeasts; Zika Virus; base; cell growth; cell type; clinically relevant; design; drug discovery; drug testing; experience; experimental study; genome-wide; global health; multidisciplinary; nerve stem cell; relating to nervous system; success; therapeutic target; virology; virus development

1. Project Summary/Abstract The recent Zika virus (ZIKV) outbreaks has surprised the world because of its rapid spread through the America and its association with birth defects such as microcephaly in the newborns and other neurologic dysfunctions in adults such as the Guillain-Barré syndrome (GBS). However, we are currently challenged with the fact that we know very little about the ZIKV functionality, nor do we know why has ZIKV suddenly become so pathogenic. Attempts to rapidly assess the ZIKV functionality is further challenged by a wide range of host cells that ZIKV infects. Here, we propose a unique and combined research approach to meet these challenges. Specifically, we plan to use fission yeast as a surrogate system for the rapid functional analysis of the ZIKV genome followed by immediate translations in human neural cells and a ZIKV infection model. Fission yeast is a simple and single cell eukaryote that has been used extensively as a model organism to study human cell biology and virology. It is a very well-tested model to study highly conserved cellular activities such as those described ZIKV cytopathic effects on cellular growth, cell cycle regulation, chromosomal biology, and cell death. Thus, study of these ZIKV effects in fission yeast is not only clinically relevant to the ZIKV diseases but we can also expedite the functional characterization of the ZIKV genome. We are uniquely qualified for the proposed study because we have extensive experiences in functional analyses of viral genomes using fission yeast and in the associated mammalian studies on viral infections. In fact, we were the first to characterize the HIV-1 genome and to develop model systems in fission yeast to study HIV-1 viral protein R (Vpr) and proteases (PRs). Our study is further strengthen by the participation of a multidisciplinary team whose expertise covering the entire spectrum of the proposed study. Therefore, we should have a high likelihood of success in the proposed study. We hypothesize that the ZIKV diseases such as microcephaly or GBS are caused by viral pathogenicity factors that are intrinsic to all ZIKVs. Alternatively, new ZIKV variants have emerged as the results of gene mutations that have led to the adaptation of high pathogenicity. To test this hypothesis, we will focus on two Specific Aims (SAs). The SA1 is to quickly phenotype the ZIKV genome for the identification of pathogenicity factor(s), which will be further compared against genetically distinctive ZIKV viral variants to answer the question of whether the observed ZIKV pathogenicity is an intrinsic viral property or the newly acquired viral function. The SA2 is to develop anti-ZIKV drug testing and screening systems against the viral pathogenicity factor and the ZIKV NS2B/NS3 protease, the latter has already been used as a therapeutic target for other flaviviruses. The successful completion of the proposed experiments will 1) provide a genome-wide description of the ZIKV phenotypes, 2) identify viral pathogenic factors that are linked to individual ZIKV proteins, and 3) develop fission yeast cell-based systems for anti-ZIKV drug testing and screenings.

1。项目摘要/摘要 最近的寨卡病毒（Zikv）爆发使世界感到惊讶，因为它在 美国及其与新生儿小头畸形等出生缺陷的关联和其他神经系统 成年人（例如Guillain-Barré综合征（GBS））的功能障碍。但是，我们目前受到挑战 我们对ZIKV功能知之甚少，这一事实也不知道为什么Zikv突然变得 如此致病。迅速评估ZIKV功能的尝试进一步挑战了广泛的主机 zikv感染的细胞。在这里，我们提出了一种独特而合并的研究方法来应对这些挑战。 具体而言，我们计划将裂变酵母作为替代系统的快速功能分析 基因组，然后在人神经细胞和ZIKV感染模型中立即翻译。 裂变酵母是一种简单而单细胞的真核生物，已广泛用作模型生物 研究人类细胞生物学和病毒学。这是研究高度保守的细胞活性的经过良好测试的模型 例如描述的ZIKV细胞病毒对细胞生长，细胞周期调节，染色体生物学的影响， 和细胞死亡。这是对裂变酵母中这些ZIKV效应的研究不仅与ZIKV疾病有关 但是我们也可以加快ZIKV基因组的功能表征。 我们拥有拟议研究的独特资格，因为我们在功能方面有丰富的经验 使用裂变酵母和有关病毒感染的哺乳动物研究对病毒基因组进行分析。在 事实，我们是第一个表征HIV-1基因组并在裂变酵母中开发模型系统以研究的人 HIV-1病毒蛋白R（VPR）和蛋白酶（PRS）。通过参与，我们的研究进一步加强了 多学科团队的专业知识涵盖了拟议研究的整个范围。因此，我们应该 在拟议的研究中，成功​​的可能性很大。 我们假设ZIKV疾病（例如小头畸形或GBS）是由病毒致病性引起的 所有ZIKV固有的因素。另外，新的ZIKV变体已成为基因的结果 导致高致病性适应的突变。为了检验这一假设，我们将重点关注两个 具体目的（SAS）。 SA1是快速表型ZIKV基因组以识别致病性 因子，将进一步比较与遗传上独特的ZIKV病毒变异，以回答这个问题 观察到的ZIKV致病性是固有的病毒特性还是新获得的病毒功能。这 SA2是针对病毒致病因素和 ZIKV NS2B/NS3蛋白酶，后者已经用作其他黄病毒的治疗靶标。 提出的实验的成功完成将1）提供全基因组的描述 ZIKV表型，2）确定与单个ZIKV蛋白相关的病毒致病因素，3） 用于抗ZIKV药物测试和筛查的裂变酵母细胞系统。