Modeling Zika virus pathogenesis and potential intervention during neural development using a human brain organoid system

使用人脑类器官系统模拟寨卡病毒发病机制和神经发育过程中的潜在干预

• 批准号: 9913466
• 负责人: HENGLI TANG
• 金额: $ 35.83万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9913466
• 关键词: 3-Dimensional; Acute; Address; Adherent Culture; Affect; Animal Model; Antiviral Agents; Apoptotic; Asians; Astrocytes; Basic Science; Biological Assay; Brain; Caspase Inhibitor; Cell Communication; Cell Death; Cell Proliferation; Cells; Cellular Tropism; Cerebrum; Chemicals; Chronic; Clinical; Collaborations; Communicable Diseases; Complement; Congenital Abnormality; Country; Cyclin-Dependent Kinase Inhibitor; Data; Defect; Dengue Virus; Development; Disease; Disease Outbreaks; Engineering; Flavivirus; Florida; Goals; Health; Hepatitis C virus; Hour; Human; Infection; Inflammatory Response; International; Intervention; Knowledge; Lead; Luciferases; Measures; Mediating; Microcephaly; Mission; Modeling; Molecular; Molecular Profiling; Monitor; Nervous system structure; Neuraxis; Neuroglia; Neurons; Nuclear Pore Complex; Organ; Organoids; Outcome; Pathogenesis; Pathology; Pharmaceutical Preparations; Phenotype; Preventive vaccine; Proteins; Public Health; Publishing; Puerto Rico; RNA Viruses; Reporter; Reporting; Research; Severities; Standardization; Structure; System; Testing; Therapeutic Intervention; Tissue Model; Translational Research; Tropism; Universities; Viral; Viral Nonstructural Proteins; Viral Pathogenesis; Virion; Virus; Virus Diseases; Virus Replication; West Nile virus; Work; World Health Organization; ZIKV infection; Zika Virus; acute infection; base; cell killing; cell type; cytokine; drug testing; effective therapy; experimental study; fetal; high throughput screening; in utero; in vitro Model; induced pluripotent stem cell; infection rate; longitudinal analysis; member; mosquito-borne; nerve stem cell; nervous system disorder; neurodevelopment; neuron development; neuron loss; neurotropic virus; programs; public health emergency; response; single-cell RNA sequencing; small molecule; three-dimensional modeling; tool; virology

SUMMARY – Project 2 The broad, long-term goal of our research program is to advance knowledge of viral replication strategies and virus–host cell interactions that are relevant for therapeutic intervention. The viruses that we study are positive- strand RNA viruses including dengue virus (DENV), hepatitis C virus (HCV), and Zika virus (ZIKV). To complement animal models, there is a clear need for better in vitro models to study infectious diseases that affect largely inaccessible organs such as the human nervous system. In the current HTMID CRC proposal with the overarching goal to develop an organoid-based platform using human induced pluripotent stem cells (iPSCs), Project 2 focuses on ZIKV, which poses a major emerging threat to human health due to a large number of recent outbreaks and its association with microcephaly, a neurodevelopmental birth defect. Understanding how ZIKV interacts with the host to cause disease is of critical importance to the development of effective antiviral drugs and a prophylactic vaccine, both of which are urgently needed. We have recently published work showing that ZIKV infects human neural stem cells and also performed a high-throughput screening to identify small molecule compounds that can inhibit ZIKV infection. Here we propose to utilize a 3D cerebral organoid model to investigate the mechanism of ZIKV pathogenesis in human neural development (Specific Aims 1 & 2). We will also use the organoid model to compare and contrast ZIKV with West Nile virus (WNV), another human neurotropic virus of biomedical importance (Specific Aim 3). Finally, we will validate the utility of the organoid model as a drug testing tool using chemical compounds with confirmed antiviral or anti-apoptotic functions (Specific Aim 4). With the successful completion of the project, we expect to: positively identify the type of cells and their ZIKV infection rate during the different stages of brain development; unravel the mechanisms by which ZIKV infection causes neuronal death and developmental defects; reveal any quantitative differences between ZIKV and WNV in infection efficiencies, cellular tropism, and pathology in the 3D model; and confirm lead compounds for inhibiting ZIKV replication in both 2D culture and 3D tissue models. These results will validate the utility of the 3D brain organoid model for both basic and translational research of ZIKV and also provide direct and immediate impact on the mission to develop effective therapy to treat ZIKV infection and its associated neurological diseases.

摘要 - 项目2 我们的研究计划的广泛长期目标是提高有关病毒复制策略的知识和 与治疗干预有关的病毒 - 霍斯特细胞相互作用。我们研究的病毒是阳性的 链RNA病毒在内，包括登革热病毒（DENV），乙型肝炎病毒（HCV）和Zika病毒（ZIKV）。到 补充动物模型，显然需要更好地体外模型来研究传染病 影响很大程度上无法访问的器官，例如人类神经系统。在当前的HTMID CRC提案中 以人类诱导的多能干细胞开发基于器官的平台的总体目标 （IPSCS），项目2专注于ZIKV，该ZIKV构成了对人类健康的重大威胁 近期暴发及其与小头畸形的关联，这是一种神经发育的先天缺陷。 了解ZIKV如何与宿主互动以引起疾病对于发展至关重要 有效的抗病毒药物和预防性疫苗，迫切需要两者。 我们最近发表了工作，表明ZIKV感染了人类神经干细胞，也是 进行了高通量筛选，以鉴定可以抑制ZIKV感染的小分子化合物。 在这里，我们建议利用3D大脑器官模型研究ZIKV发病机理的机理 在人类神经发育中（特定目标1和2）。我们还将使用器官模型比较和 与西尼罗河病毒（WNV）对比ZIKV，这是另一种人类的生物医学重要性神经性病毒（特定的 目标3）。最后，我们将使用化学物质验证器官模型作为药物测试工具的实用性 具有确认的抗病毒或抗凋亡功能的化合物（特定目标4）。 随着项目的成功完成，我们期望：积极地识别细胞的类型及其它们的类型 大脑发育不同阶段的ZIKV感染率；解开zikv的机制 感染会导致神经元死亡和发育缺陷；揭示ZIKV之间的任何定量差异 3D模型中的感染效率，细胞向潮流和病理学中的WNV；并确认铅 在2D培养和3D组织模型中抑制ZIKV复制的化合物。这些结果将验证 3D脑器官模型的实用性，用于ZIKV的基本和翻译研究，还提供 直接和直接影响开发有效治疗ZIKV感染及其的疗法的使命 相关的神经疾病。