Perturbation of Host DNA Replication and Cell Cycle Progression by Zika Virus

寨卡病毒对宿主 DNA 复制和细胞周期进程的干扰

基本信息

批准号：
10189506
负责人：
HENGLI TANG
金额：
$ 37.89万
依托单位：
FLORIDA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10189506
关键词：
3-Dimensional ATM activation Adherent Culture Antiviral Agents Area Biology Brain Cell Communication Cell Cycle Cell Cycle Arrest Cell Cycle Progression Cells Chimera organism DNA DNA Damage DNA Double Strand Break DNA biosynthesis DNA replication fork Data Defect Dengue Dengue Virus Development Disease Down-Regulation Embryo Ensure Flavivirus Goals Growth Health Hepatitis C virus Human Hybrids In Vitro Infection Knowledge Lead Life Cycle Stages Mediating Medical Microcephaly Mission Molecular Molecular Biology Neuropathogenesis Nuclear Nucleotides Organoids Pathologic Pathway interactions Phenotype Preventive vaccine Process Public Health Publishing RNA Viruses Reporting Research Role S Phase S Phase Arrest Stress System Techniques Testing Validation Viral Proteins Virulent Virus Virus Replication Work ZIKA ZIKV infection Zika Virus coping disorder prevention effective therapy exhaustion experimental study human pathogen hydroxyurea mosquito-borne nerve stem cell nervous system disorder neural growth neurodevelopment neurogenesis new therapeutic target programs replication stress response stem cell biology vector-borne virology

项目摘要

The broad, long-term goal of our research program is to advance knowledge of virus–host cell interactions that are relevant for disease prevention. The viruses that we study are positive- strand RNA viruses such as dengue virus (DENV), hepatitis C virus (HCV), and Zika virus (ZIKV). The current proposal focuses on ZIKV, which has re-emerged worldwide and poses a major emerging threat to human health. Understanding how ZIKV interacts with the host cell is of critical importance to the development of antiviral drugs and a prophylactic vaccine, both of which are currently lacking. Building upon our recently published work on the infection and impact of ZIKV on human neural stem cells, we propose to unravel the mechanism by which ZIKV impedes the growth of human cortical neural progenitor cells (hNPCs) and leads to defects in cortex development. Our preliminary data indicate that part of the mechanism for cell cycle arrest is a ZIKV-induced DNA damage response (DDR) which blocks DNA replication and leads to S-phase arrest. Unexpectedly, the ATR/Chk1 DNA checkpoint pathway that normally functions to deal with DNA replication stress in S-phase was not activated, suggesting that ZIKV suppresses ATR/Chk1 activation during DNA replication. The ability to increase DNA replication stress while simultaneously inhibiting ATR responds constitute a potent “one-two punch” that exacerbates replication defects and ultimately leads to cell cycle arrest. We will carry out experiments to investigate the mechanisms by which ZIKV achieve these feats and identify the viral proteins responses. We will also cross-validate our results with brain organoids and infectious clones of Zika/Dengue chimeric viruses. We expect to clearly understand the mechanisms by which ZIKV perturbs the cell cycle to achieve its pathological effect on hNPC-mediated neural development in vitro. We also expect to reveal the virulent determinant of ZIKV critical for its effect on brain development. These results will not only fundamentally advance our understanding of this important human pathogen but also provide direct and immediate impact on the mission to develop effective therapy to treat ZIKV infection and its associated diseases.

我们的研究计划的广泛，长期目标是促进与预防疾病有关的病毒 - 霍斯特细胞相互作用的了解。我们研究的病毒是阳性链RNA病毒，例如登革热病毒（DENV），丙型肝炎病毒（HCV）和Zika病毒（ZIKV）。当前的提议着重于ZIKV，ZIKV已重新出现在全球范围内，并对人类健康构成了重大新兴威胁。了解ZIKV与宿主细胞的相互作用对于抗病毒药物的开发和预防性疫苗至关重要，目前缺乏两种预防性疫苗。在我们最近发表的有关ZIKV对人神经元干细胞的感染和影响的工作的基础上，我们建议揭示ZIKV阻碍人皮质神经祖细胞（HNPC）生长的机制，并导致皮质发育中的缺陷。我们的初步数据表明，细胞周期停滞机制的一部分是ZIKV诱导的DNA损伤响应（DDR），该反应阻断了DNA复制并导致S期停滞。出乎意料的是，未激活通常在S阶段处理DNA复制应力的ATR/CHK1 DNA检查点途径，这表明ZIKV抑制DNA复制过程中ATR/CHK1的激活。同时抑制ATR反应的同时增加DNA复制应力的能力构成了有效的“两次打孔”，从而加剧了复制缺陷并最终导致细胞周期停滞。我们将进行实验，以研究ZIKV实现这些功能并确定病毒蛋白反应的机制。我们还将通过Zika/Zika/登革热嵌合病毒的脑器官和感染性克隆进行跨验证我们的结果。我们期望清楚地了解ZIKV在细胞周期中实现其对HNPC介导的神经发育的病理效应的机制。我们还期望揭示ZIKV对其对大脑发育的影响至关重要的有力确定。这些结果不仅从根本上可以提高我们对这种重要人类病原体的理解，而且还可以直接而直接地影响开发有效治疗ZIKV感染及其相关疾病的疗法。