Evaluating Human Pluripotent Stem Cell-Derived Neural Rosette Arrays as a Neural Tube Defect Risk Screening Platform

评估人类多能干细胞衍生的神经花环阵列作为神经管缺陷风险筛查平台

基本信息

批准号：
10369044
负责人：
Randolph S Ashton
金额：
$ 19.44万
依托单位：
UNIVERSITY OF WISCONSIN-MADISON
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2023-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10369044
关键词：
Affect Anencephaly and spina bifida X linked Biological Assay Biomedical Engineering Birth Cell Line Cells Chemical Exposure Chemicals Clinical Complex Congenital Abnormality Consensus Cryopreserved Cell DNA Sequence Alteration Defect Derivation procedure Development Diet Dietary Supplementation Drug Screening Embryo Environmental Risk Factor Epidemiology Etiology Evaluation FDA approved Failure Fibrinogen Folic Acid Future Genes Genetic Genetic Carriers Genetic Enhancement Genetic Models Genetic Predisposition to Disease Genotype Government Grain Human In Vitro Incontinence Investigation Lead Libraries Location Methodology Modeling Mutation Neural Tube Closure Neural Tube Defects Neural tube Neuraxis Neuroepithelial Cells Neuroepithelial Tissue Pathology Patients Perinatal mortality demographics Phenotype Postdoctoral Fellow Pregnant Women Process Research Risk Rodent Sales Screening procedure Sensitivity and Specificity Signal Pathway Spinal Dysraphism Standardization Structure Testing Tissues To specify Validation Work analog carrier testing cell bank clinical translation clinically relevant experimental study fortification genome wide association study high reward high risk human pluripotent stem cell in vivo induced pluripotent stem cell insight motor impairment mutant new technology novel precision medicine prevent relating to nervous system screening small molecule libraries stem cell derived tissues tool

项目摘要

Project Summary Neural tube defects (NTDs) are the second leading cause of congenital malformations affecting ~3000 births per year in the US alone and with significantly higher rates elsewhere. NTDs occur within the first month post- conception and are caused by failed closure of the neural tube, which is the developmental anlage of the entire central nervous system (CNS). The most prevalent NTDs are anencephaly and spina bifida. These defects are embryonic lethal or result in lifelong motor impairment with possible incontinence, respectively. While folic acid dietary supplementation has been clinically proven to reduce NTD risk in expecting mothers, the persistence of NTD occurrences despite proper diet suggest that multifactorial environmental and genetic factors are the primary etiological origins of remaining NTD cases. Furthermore, while rodent studies provide significant insights into NTD etiology and pathology, they do not provide the requisite throughput to efficiently screen environmental factors, i.e. chemical exposures, for associated NTD risks. Moreover, incongruence between rodent genetic models and clinical observations from human epidemiological and genome-wide association studies (GWAS) has limited development of precision medicine approaches for predicting and minimizing a patients’ NTD risk. Here, we propose to evaluate whether Neural Rosette Arrays (NRAs) can serve as a quantitative, high-throughput, human pluripotent stem cell (hPSC)-derived screening platform for assessing a chemical’s or genetic mutation’s NTD risk. While NRAs are not an exact recapitulation of in vivo primary neurulation, the bioengineered assay is the first to spatially and temporally standardize neural tube analog formation, i.e. singular neural rosette tissues, in a microarray format. Rosette tissues within NRAs possess the requisite cell phenotypes, tissue cytoarchitecture, and are derived using morphogenetic signaling pathways endogenous to the in vivo neural tube formation process. Thus, we hypothesize that the NRA platform will be an effective and efficient screening platform to detect NTD risk. Aim 1 test this hypothesis using a small chemical library screen to evaluate the NRA assay’s sensitivity and specificity. Aim 2 test this hypothesis using screens of hPSC lines genetically modified with NTD-associated genetic mutations and compared to the wild- type, isogenic control. If this high risk/high reward proposal is successful, then future research efforts would use this novel technology for systematic investigation of NTD multifactorial etiologies and clinical translation via forward screens of EPA/FDA-approved compound libraries and reverse screens using NTD patient induced pluripotent stem cell lines. In this manner, the NRA platform could have a transformative effect on NTD research, regulatory efforts to prevent commercial sale of NTD-causing agents, and future development of precision medicine approaches to minimize NTD risk in genetically predisposed patients.

项目概要神经管缺陷 (NTD) 是影响约 3000 名新生儿的先天性畸形的第二大原因仅在美国，NTD 的发病率在其他地区就明显更高。受孕是由于神经管闭合失败引起的，神经管是整个身体的发育原基。最常见的 NTD 是无脑畸形和脊柱裂。叶酸分别会导致胚胎致命或导致终生运动障碍并可能导致失禁。临床证明膳食补充剂可以降低准妈妈罹患 NTD 的风险，尽管饮食适当，NTD 的发生表明多因素环境和遗传因素是此外，啮齿动物研究提供了重要的证据。虽然它们无法深入了解 NTD 病因学和病理学，但无法提供有效筛查所需的通量相关 NTD 风险的环境因素，即化学品暴露。啮齿动物遗传模型和人类流行病学和全基因组关联的临床观察研究（GWAS）限制了用于预测和最小化疾病的精准医学方法的发展在这里，我们建议评估神经玫瑰阵列（NRA）是否可以作为一种治疗方法。定量、高通量、人类多能干细胞 (hPSC) 衍生筛选平台，用于评估化学物质或基因突变的 NTD 风险，而 NRA 并不是体内原发性的精确再现。神经溶解术，生物工程测定是第一个在空间和时间上标准化神经管模拟的方法 NRA 内的微阵列形式的形成，即单一神经花结组织，具有以下特征。必要的细胞表型、组织细胞结构，并使用形态发生信号通路衍生因此，我们追求 NRA 平台将是体内神经管形成过程的内源性。目标 1 使用一个小样本来测试这一假设。目标 2 使用化学库筛选来评估 NRA 的敏感性和特异性来测试该假设。筛选经过 NTD 相关基因突变进行基因改造的 hPSC 系，并与野生型进行比较如果这种高风险/高回报的提议成功，那么未来的研究工作将是。使用这项新技术对 NTD 多因素病因进行系统研究并进行临床转化对 EPA/FDA 批准的化合物库进行正向筛选，并使用 NTD 患者诱导的反向筛选通过这种方式，NRA 平台可以对 NTD 产生变革性影响。研究、防止 NTD 致病剂商业销售的监管工作以及未来的发展精准医疗方法可最大限度地降低遗传易感患者的 NTD 风险。