Cellular models of fetal neurodevelopment in maternal SARS-CoV-2 infection

项目总结 母亲感染SARS-CoV-2对发育中胎儿的影响尚不清楚，但仍处于初步阶段 已经开始积累的数据表明，后代会受到神经发育的影响。有令人信服的证据 胎儿大脑特别容易受到母体免疫激活和炎性暴露的影响 关键的发展窗口。鉴于数以百万计的胎儿最终将接触冠状病毒的预测- 19、了解和模拟这种风险是一个紧迫的科学和公共卫生问题。虽然是长期的 临床结果不可能在十年或更长时间内知道，这是对不利神经发育风险进行建模的工具 迫切需要结果，了解风险机制，并筛选干预措施。 小胶质细胞是驻留在大脑中的免疫细胞，在正常的大脑发育中发挥着关键作用，并且 已知受到宫内环境的影响。调查人员已经开发并验证了方法 从包括脐带血在内的外周血中高效地产生人小胶质细胞样细胞。他们 先前证明，这些细胞概括了小胶质细胞的形态、转录组和功能。 从死后大脑中提取的。此外，这些模型确定了与精神分裂症相关的修剪功能障碍， 使用可扩展的突触体模型以及长期共培养。 在这里，研究人员建议使用大型病毒来表征母亲感染SARS-CoV-2的影响 他们创建的Biospecimen库包括匹配的母亲血液和新生儿脐带血，与 丰富的临床细节和电子健康记录。该银行包括800多名母亲的新生儿 SARS-CoV-2阳性(547人)，或SARS-CoV-2阴性并在大流行期间怀孕(265人)。 具体地说，研究人员将通过多种细胞和血清来表征母体的免疫激活 措施。然后，他们将使用倾斜的脐带创建个性化的新生儿小胶质细胞功能模型 血液单个核细胞。他们将比较在形态、转录和功能上的差异 从SARS-CoV-2暴露和未暴露的新生儿中诱导小胶质细胞，以验证SARS-CoV-2感染的假设。 CoV-2相关的母体免疫激活启动子宫内小胶质细胞的炎症表型， 导致神经发育失调。最后，他们将创建一个更多的电子健康记录队列 超过10,000次分娩以检查SARS-CoV-2后代神经发育诊断的风险 积极的母亲与消极的母亲相比，最终获得了长达5年的随访。 总之，这些实验将量化母体病毒感染和免疫的潜在影响 对发育中的胎儿大脑的激活，检查潜在的风险生物标记物，并开发一个模型系统 可用于识别和测试干预措施，以将此类风险降至最低。该项目将实验室与 环境暴露对母体免疫激活和发育的影响的专门知识 结果，患者衍生的体外细胞模型，以及纵向电子健康记录的分析。

PROJECT SUMMARY The impact of maternal SARS-CoV-2 infection on the developing fetus remains unknown, but preliminary data has begun to accumulate suggesting neurodevelopmental effects in offspring. There is compelling evidence that the fetal brain is particularly vulnerable to maternal immune activation and inflammatory exposures during key developmental windows. In light of the projection that millions of fetuses will ultimately be exposed to COVID- 19, understanding and modeling this risk is a pressing scientific and public health concern. While long-term clinical outcomes cannot be known for a decade or more, tools to model risk for adverse neurodevelopmental outcomes, understand mechanisms of risk, and to screen for interventions, are urgently needed. Microglia, the resident brain immune cells, play a critical role in normal brain development, and are known to be impacted by the intrauterine environment. The investigators have developed and validated methods to efficiently generate human microglia-like cells from peripheral blood, including umbilical cord blood. They demonstrated previously that these cells recapitulate morphology, transcriptome, and function of microglia derived from postmortem brain. Further, these models identify schizophrenia-associated pruning dysfunction, using a scalable synaptosome model as well as long-term co-culture. Here, the investigators propose to characterize effects of maternal SARS-CoV-2 infection using the large biospecimen bank they have created that includes matched maternal blood and neonatal cord blood, linked to abundant clinical detail and electronic health records. The bank includes more than 800 neonates of mothers who are SARS-CoV-2 positive (547), or SARS-CoV-2 negative and pregnant during the pandemic (265). Specifically, the investigators will characterize maternal immune activation via multiple cellular and serum measures. They will then create personalized neonatal models of microglial function using banked umbilical cord blood mononuclear cells. They will compare morphologic, transcriptomic, and functional differences between induced microglial cells from SARS-CoV-2-exposed and unexposed neonates, to test the hypothesis that SARS- CoV-2-related maternal immune activation primes microglial cells in utero toward an inflammatory phenotype, leading to dysregulated neurodevelopment. Finally, they will create an electronic health records cohort of more than 10,000 deliveries to examine risk for neurodevelopmental diagnoses among offspring of SARS-CoV-2 positive compared to negative mothers, ultimately capturing up to 5 years of follow-up. Together, these experiments will quantify the potential impact of maternal viral infection and immune activation on the developing fetal brain, examine a potential biomarker of risk, and develop a model system that may be used to identify and test interventions to minimize such risk. The project integrates laboratories with expertise in the impact of environmental exposures on maternal immune activation and developmental outcomes, patient-derived in vitro cellular modeling, and analysis of longitudinal electronic health records.