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）是妊娠期间发生的常见神经系统畸形。 NTD 是由遗传变异或母亲环境暴露引起的，通常会导致严重的疾病 身体和/或认知障碍。尽管药物有时会导致人类胚胎出现 NTD 由于人类特异性毒性而在啮齿动物模型中进行筛查。许多抗癫痫药物 (ASM)、使用的药物 癫痫治疗会增加怀孕期间患 NTD 的风险，但通常不会停药 因为癫痫发作会对母亲和胎儿造成危险。因此，比较重要的是 这些化合物有神经致畸风险，因此在妊娠期间选择最合适的ASM。拥有一个 早期神经发育的人类特异性模型应该会提高这种筛查的预测性。尝试 由于结构异质性和内部结构，利用人脑类器官技术来实现这一目标受到限制。 类器官的变异性。我们最近开发了可重复的自组织单玫瑰花状球体（SOSRS） 来自人类诱导多能干细胞的研究使我们能够用已知的神经致畸剂治疗 SOSRS 观察与 NTD 一致的明显结构变化。目标 1 的目标是比较结构 SOSRS 中所有 20 种常用 ASM 在多个浓度下的结果，以确定 每个的浓度依赖性风险。在目标 2 中，我们将进行 SOSRS 的比较转录组学 用 5 种机制不同的神经致畸剂治疗。所有 5 人共享的转录组变化将是 被认为是一种神经致畸生物标志物，然后通过靶向 NGS 将其应用于来自以下来源的 RNA 样本： SOSRS 使用每种常用的 ASM 进行处理，以提供进一步的风险比较。在目标 3 中，我们将 采用 SOSRS 协议生成腰脊髓 SOSRS，以比较致畸性 丙戊酸在头侧（皮质 SOSRS）和尾侧（腰椎 SOSRS）神经管模型之间。我们的研究 还可能揭示 ASM 导致 NTD 的未定义机制。平台和生物标志物 这项研究开发的方法还可以用于筛选针对人类特定的 NTD 风险的新型药物 未来的系统。