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、长期目标是促进对病毒复制策略和 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(丙型肝炎病毒)和寨卡病毒(ZIKV)。至 作为动物模型的补充，显然需要更好的体外模型来研究传染病 影响到很大程度上无法接触到的器官，如人类神经系统。在当前的HTMID CRC提案中 其首要目标是利用人类诱导的多能干细胞开发一种基于有机化合物的平台 (IPSC)，项目2重点关注ZIKV，由于大量的 最近暴发的数量及其与小头畸形的关系，小头畸形是一种神经发育的出生缺陷。 了解ZIKV是如何与宿主相互作用导致疾病的对于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。 我们最近发表的研究表明，ZIKV感染人类神经干细胞，而且 进行了高通量筛选，以确定可以抑制ZIKV感染的小分子化合物。 在这里，我们建议利用3D脑器官模型来研究ZIKV的致病机制 在人类神经发育方面(具体目标1和2)。我们还将使用有机模型来比较和 将ZIKV与西尼罗河病毒(WNV)进行对比，西尼罗河病毒是另一种具有生物医学意义的人类嗜神经性病毒(特异性 目标3)。最后，我们将验证有机物模型作为药物测试工具的实用性 已证实具有抗病毒或抗细胞凋亡功能的化合物(特定目标4)。 随着该项目的成功完成，我们希望：积极确定细胞的类型和它们的 ZIKV在脑发育不同阶段的感染率；揭示ZIKV感染的机制 感染导致神经元死亡和发育缺陷；揭示ZIKV之间的任何数量差异 和西尼罗河病毒在3D模型中的感染效率、细胞趋向性和病理；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 三维脑器官模型在ZIKV的基础和翻译研究中的应用，并提供了 直接和直接影响到开发有效治疗寨卡病毒感染和其 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.