Molecular mechanisms linking viral replication and neuropathogenesis

连接病毒复制和神经发病机制的分子机制

基本信息

批准号：
10660340
负责人：
Priya Shirish Shah
金额：
$ 39.1万
依托单位：
UNIVERSITY OF CALIFORNIA AT DAVIS
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-02-08 至 2028-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10660340
关键词：
Alleles Amino Acids Biochemical Biological Assay Brain Cell division Chemicals Collaborations Congenital Abnormality Cytomegalovirus Development Disease Drosophila genus Etiology Fetal Tissues Future Genes Genetic Determinism Genetic Predisposition to Disease Goals Head Hereditary Disease Human In Vitro Individual Infection Inherited Integration Host Factors Intellectual functioning disability Link Mediating Methods Microcephaly Mitosis Modeling Molecular Mus Mutagenesis Mutation Neuropathogenesis Nonstructural Protein Pathogenesis Pathogenicity Pathway interactions Phenotype Phosphorylation Physiological Process Proteins Proteomics Publishing Replication-Associated Process Reporter Resolution Role Rubella virus Scaffolding Protein Site System Testing Transgenic Organisms Variant Viral Viral Pathogenesis Virus Virus Replication Work ZIKV infection Zika Virus brain size developmental disease drug development experimental study human pathogen in vivo insight knock-down mouse model mutant mutation screening nerve stem cell nervous system disorder recruit therapeutic target virus host interaction

项目摘要

PROJECT SUMMARY/ABSTRACT Viruses that infect the developing brain, including Zika virus (ZIKV), cytomegalovirus, and rubella virus cause major birth defects. Microcephaly is one such birth defect, in which head and brain size are severely reduced, and is often accompanied by intellectual disability. This virally-inflicted neurological disease, or viral neuropathogenesis, can be caused by multiple mechanisms. One recently identified way ZIKV non-structural protein 4A (NS4A) causes microcephaly is by disrupting the human ANKLE2 protein. Interestingly, individuals with mutations in the gene encoding ANKLE2 suffer from microcephaly. ANKLE2 is conserved from worms to humans, and is essential for coordinating cell division during brain development. ANKLE2 derives this function in cell division and development by mediating protein interactions. NS4A physically interacts with ANKLE2 and disrupts brain development in an ANKLE2-dependent manner in a fruit fly model of brain development. ANKLE2 also promotes ZIKV replication. Taken together, these studies show that in the process of coopting a host protein for replication, ZIKV dysregulates an important developmental pathway. Thus, the NS4A-ANKLE2 protein interaction represents an important model to study viral neuropathogenesis and how it is connected to viral replication and hereditary disorders at the molecular level. The long-term goal of this work is to decipher how virus-host protein interactions impact virus replication and pathogenesis, as these discoveries will fuel therapeutic target identification and drug development. The objective of this proposal is to dissect the mechanisms by which the protein interaction between ZIKV NS4A and human ANKLE2 promote ZIKV replication and inhibit brain development. To accomplish this objective, we will test the central hypothesis that ANKLE2 promotes viral replication through its interaction with NS4A and by recruiting other host factors involved in ZIKV replication to sites of replication, and this disrupts physiological ANKLE2 protein interactions required for brain development. The following specific aims will test this hypothesis: Aim 1: Dissect the impact of the NS4A-ANKLE2 protein interaction in ZIKV replication and pathogenesis. Aim 2: Unravel the molecular function of ANKLE2 in ZIKV replication and pathogenesis. When completed, this work will delineate how a single virus-host protein interaction rewires a developmental pathway to facilitate virus replication and inflict neurological disease at the molecular level. This will reveal detailed biochemical insight into a virus-host protein interaction with amino acid-level resolution, new host factors that play a role in ZIKV replication, and previously unknown proteins key to brain development and disrupted in other hereditary developmental disorders. In the long term, the methods established here could be employed to uncover the molecular mechanisms behind other diseases with viral and hereditary etiologies.

项目概要/摘要感染发育中大脑的病毒，包括寨卡病毒 (ZIKV)、巨细胞病毒和风疹病毒，会导致重大出生缺陷。小头畸形就是这样一种出生缺陷，其中头部和大脑的尺寸严重缩小，并且常伴有智力障碍。这种由病毒引起的神经系统疾病，或病毒性神经病理机制，可由多种机制引起。最近发现的一种 ZIKV 非结构性方法蛋白 4A (NS4A) 通过破坏人类 ANKLE2 蛋白而导致小头畸形。有趣的是，个人编码 ANKLE2 的基因发生突变，患有小头畸形。 ANKLE2 在蠕虫中是保守的人类，对于大脑发育过程中协调细胞分裂至关重要。 ANKLE2 导出该函数通过介导蛋白质相互作用来参与细胞分裂和发育。 NS4A 与 ANKLE2 发生物理相互作用，在果蝇大脑发育模型中，以 ANKLE2 依赖性方式破坏大脑发育。脚踝2 还促进 ZIKV 复制。总而言之，这些研究表明，在选择宿主蛋白的过程中对于复制而言，ZIKV 失调了一条重要的发育途径。因此，NS4A-ANKLE2 蛋白相互作用是研究病毒神经发病机制及其与病毒的关系的重要模型分子水平上的复制和遗传性疾病。这项工作的长期目标是破译病毒-宿主蛋白相互作用如何影响病毒复制和发病机制，因为这些发现将促进治疗靶点的识别和药物的开发。这该提案的目的是剖析 ZIKV NS4A 之间的蛋白质相互作用的机制人类ANKLE2促进ZIKV复制并抑制大脑发育。为了实现这一目标，我们将检验 ANKLE2 通过与 NS4A 相互作用促进病毒复制的中心假设通过将参与 ZIKV 复制的其他宿主因子招募到复制位点，这会破坏生理学大脑发育所需的 ANKLE2 蛋白质相互作用。以下具体目标将检验这一假设：目标 1：剖析 NS4A-ANKLE2 蛋白相互作用对 ZIKV 复制和发病机制的影响。目标 2：揭示 ANKLE2 在 ZIKV 复制和发病机制中的分子功能。完成后，这项工作将描述单个病毒-宿主蛋白相互作用如何重新连接发育促进病毒复制并在分子水平上造成神经系统疾病的途径。这将揭示通过氨基酸水平分辨率、新宿主因素对病毒-宿主蛋白相互作用进行详细的生化观察在 ZIKV 复制中发挥作用，以及以前未知的对大脑发育至关重要并在其他遗传性发育障碍。从长远来看，这里建立的方法可以用来揭示其他具有病毒和遗传病因的疾病背后的分子机制。