Evaluating teratogenic risk of anti-seizure medications in single-rosette brain organoids

项目摘要 神经管缺陷(NTD)是妊娠期间常见的神经系统畸形。 NTDS是由遗传变异或母体环境暴露引起的，它们往往会导致严重的 身体和/或认知障碍。药物有时会导致人类胚胎中的NTDS，尽管 人类特异性毒性在啮齿动物模型中的筛选。许多抗癫痫药物(ASM)，使用的药物 在癫痫的治疗中，增加怀孕期间NTDS的风险，但通常不会停止治疗 因为癫痫发作对母亲和胎儿有危险。因此，重要的是将 研究这些化合物的神经致畸风险，选择孕期最合适的ASM。拥有一个 人类特有的早期神经发育模型应该会增加这种筛查的预测性。试图 利用人脑有机体技术达到这一目的是有限的，因为结构的异质性和内部的 有机体的可变性。可复制自组织单玫瑰球体(SOSRS)的最新研究进展 从人类诱导的多能干细胞中分离出来，使我们能够用已知的神经致畸原和 观察与NTDS一致的明显结构变化。目标1中的目标是比较结构 所有20种常用ASM在不同浓度下在SOSRS中的结果 每个人都有浓度依赖的风险。在目标2中，我们将进行SOSRs的比较转录 用5种机制不同的神经致畸物质治疗。所有5个人共享的转录更改将是 被认为是一种神经致畸生物标记物，随后将由靶向NGS应用于来自 用每种常用的ASM治疗SOSR，以提供进一步的风险比较。在《目标3》中，我们将 采用SOSRS程序生成腰椎脊髓SOSRS，以比较 丙戊酸在吻部(皮质SOSRS)和尾部(腰部SOSRS)神经管模型之间的作用。我们的研究 也可能阐明ASM导致NTDS的不明机制。平台和生物标志物 由这项研究开发的还可以在人类特有的情况下筛查新药物的NTD风险 在未来的系统中。

Project Abstract Neural tube defects (NTDs) are common malformations of the nervous system that occur during pregnancy. NTDs are caused by genetic variants or maternal environmental exposures, and they often lead to severe physical and/or cognitive disabilities. Pharmaceuticals can sometimes lead to NTDs in human embryos despite screening in rodent models due to human-specific toxicity. Many anti-seizure medications (ASMs), drugs used in the management of epilepsy increase the risk of NTDs during pregnancy but are typically not discontinued because of danger to the mother and fetus from seizures. Therefore, it is important to compare the neuroteratogenic risk of these compounds to choose the most appropriate ASM during pregnancy. Having a human-specific model of early neurodevelopment should increase this screening predictiveness. Attempts to utilize human brain organoid technology to this end are limited due to structural heterogeneity and intra- organoid variability. Our recent development of reproducible self-organizing single rosette spheroids (SOSRS) from human induced pluripotent stem cells has allowed us to treat SOSRS with known neuroteratogens and observe distinct structural changes consistent with NTDs. The goal in Aim 1 is to compare the structural consequences of all 20 commonly used ASMs in SOSRS at multiple concentrations to determine a concentration dependent risk for each. In Aim 2, we will perform comparative transcriptomics of SOSRS treated with 5 mechanistically distinct neuroteratogens. Transcriptomic changes shared by all 5 will be considered a neuroteratogenic biomarker that will then be applied by targeted NGS to RNA samples from SOSRS treated with each of the commonly used ASMs to provide further risk comparison. In Aim 3, we will adapt the SOSRS protocol to generate lumbar spinal cord SOSRS in order to compare teratogenicity of valproic acid between rostral (cortical SOSRS) and caudal (lumbar SOSRS) neural tube models. Our study may also shed light on the undefined mechanisms by which ASMs cause NTDs. The platform and biomarkers developed by this study could also allow for screening novel pharmaceuticals for NTD risk in a human-specific system in the future.