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

评估单莲座脑类器官中抗癫痫药物的致畸风险

• 批准号: 10491352
• 负责人: Andrew M Tidball
• 金额: $ 9.75万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10491352
• 关键词: Aminopterin; Anencephaly; Animals; Apical; Apoptosis; Apoptotic; Area; Biological Assay; Biological Markers; Brain; CASP3 gene; Cell Line; Confocal Microscopy; Congenital Abnormality; Congenital neurologic anomalies; Development; Dose; Drug Prescriptions; Drug usage; Embryo; Environmental Exposure; Epilepsy; Fetus; Folic Acid; Future; GDF11 gene; Gene Expression; Genes; Goals; Heterogeneity; Homeobox Genes; Human; Image; Label; Lead; Left; Light; Live Birth; Lumbar spinal cord structure; Measures; Messenger RNA; Modeling; Mothers; Neural Tube Defects; Neural tube; Neuraxis; Organoids; Outcome; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacologic Substance; Phenotype; Pluripotent Stem Cells; Predisposition; Pregnancy; Pregnant Women; Process; Protocols documentation; Quantitative Reverse Transcriptase PCR; RNA; Radial; Recurrence; Reproducibility; Rho-associated kinase; Risk; Rodent Model; Sampling; Seizures; Spinal Cord; Spinal Dysraphism; Structural Models; Structure; Surface; System; TNP470; Techniques; Technology; Teratogens; Testing; Thalidomide; Tight Junctions; Toxic effect; Transcript; Tretinoin; Tubulin; Valproic Acid; Withdrawal; Woman; base; cognitive disability; comparative; constriction; epidemiology study; experimental study; genetic variant; imaging platform; induced pluripotent stem cell; insight; kinase inhibitor; mRNA sequencing; nestin protein; neurodevelopment; neuroteratogen; novel; physically handicapped; planar cell polarity; potential biomarker; response; screening; transcriptome sequencing; transcriptomics

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.

项目摘要 神经管缺陷（NTD）是妊娠期间发生的神经系统常见畸形。 NTD是由遗传变异或母体环境暴露引起的，它们通常会导致严重的 身体和/或认知残疾。药物有时会导致人类胚胎中的NTD， 由于人类特异性毒性，在啮齿动物模型中进行筛选。许多抗癫痫药物（ASMs），使用的药物 在癫痫的管理中，增加了妊娠期间NTD的风险，但通常不会中断 因为癫痫发作会对母亲和胎儿造成危险。因此，比较 这些化合物的神经致畸风险，以选择最合适的ASM在怀孕期间。具有 早期神经发育人类特异性模型应该增加这种筛选的预测性。试图 利用人脑类器官技术达到这一目的是有限的，由于结构异质性和内部， 类器官变异性我们最近开发的可再生自组织单玫瑰花结球体（SOSRS） 从人类诱导多能干细胞中分离出来的细胞，使我们能够用已知的神经致畸剂治疗SOSRS， 观察到与NTD一致的明显结构变化。目标1中的目标是比较结构 在多种浓度下，对SOSRS中所有20种常用ASM的结果进行分析，以确定 每个人的风险集中度。在目标2中，我们将进行SOSRS的比较转录组学 用5种机械性不同的神经畸形剂治疗。所有5个基因共享的转录组学变化将是 被认为是一种神经致畸生物标志物，然后将通过靶向NGS应用于来自 使用每种常用ASM治疗SOSRS，以提供进一步的风险比较。在目标3中，我们 调整SOSRS方案以生成腰髓SOSRS，以比较 在吻侧（皮质SOSRS）和尾侧（腰椎SOSRS）神经管模型之间使用丙戊酸。我们的研究 也可能揭示ASM导致NTD的未定义机制。平台和生物标志物 这项研究开发的药物也可以允许在人类特异性 系统在未来。