Nonhuman Primate Model to Assess Fetal Zika Virus Infection Complications

摘要 寨卡病毒（ZIKV）感染最近被认为与寨卡病毒感染的发病率增加有关。 然而，胎儿中ZIKV的存在对神经发育的实际影响， 包括小头畸形症，我们对此知之甚少虽然持续的流行病可能会加强怀孕 结果与ZIKV的相关性，并可能揭示母体感染的动态， 妊娠结局，直接询问驱动胎儿结局的因素在人类中将是不可能的 临床环境。非人灵长类动物提供了一个绝佳的机会， 胎儿感染ZIKV的病理生理过程。我们的总体目标是建立胎儿ZIKV感染 通过羊水分娩模型研究其对胎儿生长和CNS发育的影响。通过直接感染 胎儿与病毒，我们将绕过潜在的瓶颈，经胎盘传播，这可能是一个 这是研究胎儿结局的限速步骤，因为在大多数情况下，胎盘会阻止， 促进病原体向胎儿的传播。因此，将病毒直接引入羊水将 是一种更有效的方法，用于直接定义胎儿室中的病理事件。我们提出了两 明确目标，实现我们的目标。 具体目标1。为了确定猕猴胎儿是否对ZIKV感染敏感，我们将绕过 潜在的胎盘屏障，并直接覆盖羊水，监测胎儿的健康和发育 病毒学免疫学超声波和核磁共振分析 具体目标2。为了确定胎儿感染的个体发生和组织嗜性，我们将依次收集胎儿 宫内感染后的组织，以绘制病毒嗜性和胎儿发病机制，重点是全面的 胎儿组织病理学 该模型的开发和验证将建立一种快速筛查胎儿感染对胎儿发育的影响的方法。 中枢神经系统以及整个胎儿发育。非人灵长类动物的直接验证 研究胎儿神经病理学、生长受限和可能的胎儿死亡的模型将是一个非常有价值的方法。 在建立非人类灵长类动物平台方面向前迈进了一步，以获得对严重的 神经病理学在巴西爆发。可预测的胎儿结局将允许选择性滴定 通过组织病理学判断的剂量相关的神经影响，使模型也将揭示更微妙的 神经行为损害比小头畸形，可能是一个极端的结果。

ABSTRACT Infection with Zika virus (ZIKV) has been very recently implicated in an increase in the incidence of microcephaly in Brazil, however the actual impact of the presence of ZIKV in the fetus on neural development, including microcephaly, is poorly understood. While ongoing epidemiology will likely strengthen pregnancy outcome associations with ZIKV and possibly reveal the dynamics of maternal infection that influence pregnancy outcome, direct interrogation of factors that drive fetal outcomes will not be possible in human clinical settings. The nonhuman primate offers an outstanding opportunity to gain insight into these pathophysiological processes in fetal infection with ZIKV. Our overall goal is to establish a fetal ZIKV infection model by amniotic fluid delivery to study its effects on fetal growth and CNS development. By directly infecting the fetus with the virus, we will bypass the potential bottleneck of transplacental transmission, which may be a rate-limiting step in studying fetal outcomes, since in most circumstances the placenta prevents, and does not facilitate, transmission of pathogens to the fetus. Thus, direct introduction of the virus into the amniotic fluid will be a more efficient approach for defining pathological events directly in the fetal compartment. We propose two Specific Aims to address our goal. Specific Aim 1. To determine whether the macaque fetus is sensitive to ZIKV infection, we will bypass the potential placental barrier and directly inoculate the amniotic fluid, monitoring fetal well-being and development with virological, immunological, ultrasound and MRI analyses. Specific Aim 2. To define the ontogeny and tissue tropism of fetal infection, we will sequentially collect fetal tissues following intrauterine infection to map viral tropism and fetal pathogenesis, focusing on comprehensive fetal histopathology. The development and validation of this model will establish a rapid screen for the impact of fetal infection on the central nervous system as well as overall fetal development. Direct validation of the nonhuman primate model to study fetal neuropathology, growth restriction, and possibly fetal demise will be an incredibly valuable step forward in building the nonhuman primate platform to obtain a mechanistic understanding of the severe neuropathology seen in the outbreak in Brazil. A predictable fetal outcome will allow selective titration of the dose-related neurological impact as judged by histopathology, so that the model will also reveal more subtle neurobehavioral insults than microcephaly, likely to be an extreme outcome.