Modeling Prenatal COVID-19 Exposure Using Human Cortical Organoids and Mouse Models

项目摘要/摘要 神经精神障碍的患病率正在增加，约占全球负担的28%。 非传染性疾病。这些疾病的一个主要原因是胎儿大脑发育异常，以及 越来越多的证据表明，产前暴露是一个驱动因素。一种常见的曝光，它具有 近年来发病率呈指数级增长的是病毒感染，部分原因是 新型冠状病毒病2019年(新冠肺炎)大流行。高达三分之一的新冠肺炎患者已经表现出 神经系统并发症，在脑组织活检中发现了来自SARS-CoV-2病原体的RNA 从致命病例中脱身。此外，早产、流产和其他缺陷的发生率一直在上升 在母体感染后被发现。SARS-CoV-2的神经亲和性及其与神经炎症的关系 对于成年和发育中的大脑，人们仍然知之甚少。因此，有必要深入了解 导致这些并发症的机制(S)。我的中心假设是SARS-CoV-2感染和 神经炎症引发神经发育改变，导致神经结构和神经回路中断 在发育中的皮质中起作用。由于无法接触到新冠肺炎患者的脑组织，我将在 这些问题使用了从人类胚胎干细胞分化而来的3D皮质器官。为了澄清 产前新冠肺炎暴露对胎儿脑细胞发育的影响 目的如下：(1)确定神经炎影响SARS-CoV-2感染的机制。 皮质类器官和(2)检查产前新冠肺炎感染的神经学后果 老鼠。我的初步数据显示，与神经细胞相比，胶质细胞类型更容易患上 SARS-CoV-2感染，病毒复制和细胞死亡不发生在这些受感染的细胞中。目标1将 将使用免疫染色和功能分析来评估病毒感染水平、病毒复制、 细胞完整性和神经元网络。在目标2中，我希望询问在这个体外模型中看到的效果 用SARS-CoV-2感染孕鼠建立完整的宫内系统。获得这笔奖学金将使我能够 对潜在神经精神障碍的特定细胞和分子病因学有了新的见解 从产前新冠肺炎接触中获得发展，以及建立一个可获得和伦理上可接受的 方法阐明人类胎儿大脑发育异常的潜在机制 为未来的治疗和干预做出贡献。

Project Summary/Abstract The prevalence of neuropsychiatric disorders is increasing, constituting approximately 28% of the global burden of non-communicable disease. A major cause of these disorders is that of aberrant fetal brain development, and there is growing evidence implicating prenatal exposures as a driving factor. A common exposure that has undergone an exponential increase in incidence during recent years is that of viral infections, due in part to the novel coronavirus disease 2019 (COVID-19) pandemic. Up to 1/3 of COVID-19 patients have manifested neurological complications, and RNA from the etiologic agent SARS-CoV-2 has been found in brain biopsies from fatal cases. Furthermore, increasing rates of pre-term birth, miscarriages, and other defects have been seen following a maternal infection. The neurotropism of SARS-CoV-2 and the implications of neuroinflammation on both the adult and developing brain are still poorly understood. Therefore, it is imperative to gain insight into the mechanism(s) responsible for these complications. My central hypothesis is that SARS-CoV-2 infection and neuroinflammation trigger neurodevelopmental changes leading to a disruption of neural structure and circuit function in the developing cortex. Due to the inaccessibility of brain tissue from COVID-19 patients, I will address these questions using 3D cortical organoids differentiated from human embryonic stem cells. To elucidate the cellular effects of prenatal COVID-19 exposure on the development of the fetal brain, I will investigate the following aims: (1) determine the mechanism by which neuroinflammation influences SARS-CoV-2 infection in cortical organoids and (2) examine the neurological consequences of prenatal COVID-19 infection in humanized mice. My preliminary data revealed that compared to neuronal cells, glial cell types were more susceptible to SARS-CoV-2 infection, and that viral replication and cell death does not occur in these infected cells. Aim 1 will be to use immunostaining and functional assays to evaluate alterations in viral infection levels, viral replication, cellular integrity, and neuronal networks. In Aim 2, I hope to interrogate the effects seen in this in vitro model in an intact in utero system by infecting pregnant mice with SARS-CoV-2. Receiving this fellowship will allow me to gain novel insight into the specific cellular and molecular etiology of potential neuropsychiatric disorder development from prenatal COVID-19 exposure, as well as establish an accessible and ethically acceptable method to elucidate the underlying mechanisms of abnormal human fetal brain development, potentially contributing to future therapies and interventions.