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

摘要-项目2 我们研究计划的广泛，长期目标是推进病毒复制策略的知识， 与治疗干预相关的病毒-宿主细胞相互作用。我们研究的病毒是阳性的- 链RNA病毒，包括登革热病毒（DENV）、丙型肝炎病毒（HCV）和寨卡病毒（ZIKV）。到 作为对动物模型的补充，显然需要更好的体外模型来研究感染性疾病， 对人体神经系统等难以接近的器官造成很大影响。在当前的HTMID CRC提案中， 其总体目标是利用人类诱导多能干细胞开发基于类器官的平台 项目2的重点是ZIKV，ZIKV对人类健康构成了一个主要的新兴威胁，这是由于大量的 最近爆发的数量及其与小头畸形（一种神经发育性出生缺陷）的关系。 了解ZIKV如何与宿主相互作用以引起疾病对于ZIKV的发展至关重要。 有效的抗病毒药物和预防性疫苗，这两者都是迫切需要的。 我们最近发表的工作表明，ZIKV感染人类神经干细胞， 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，我们提出利用3D脑类器官模型来研究ZIKV发病机制 在人类神经发育（具体目标1和2）。我们还将使用类器官模型进行比较， 将ZIKV与西尼罗河病毒（WNV）进行对比，后者是另一种具有生物医学重要性的人类嗜神经病毒（特异性 目标3）。最后，我们将验证类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（具体目标4）。 随着项目的顺利完成，我们期望：积极识别细胞类型及其 ZIKV感染率在大脑发育的不同阶段;解开ZIKV感染的机制 感染导致神经元死亡和发育缺陷;揭示ZIKV之间的任何定量差异 和WNV在3D模型中的感染效率、细胞向性和病理学;并确认 在一些实施方案中，提供了用于在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脑类器官模型在ZIKV基础和转化研究中的应用， 直接和立即的影响，使命，以开发有效的治疗ZIKV感染及其 相关的神经系统疾病。

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.