Modeling genetic anencephaly with human brain organoids

用人脑类器官模拟遗传性无脑畸形

基本信息

批准号：
10605362
负责人：
Andrew M Tidball
金额：
$ 11.69万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-07 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10605362
关键词：
Amniocentesis Anencephaly Apical Area Biological Models Blood Blood specimen Brain CELSR1 gene CRISPR correction CRISPR/Cas technology Cell Line Cell Reprogramming Cells Cephalic Chemicals Chorionic Villi Sampling Chorionic villi Clinical Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Confocal Microscopy Congenital Abnormality Congenital neurologic anomalies DNA DNA Sequence Alteration Development Disease Engineering Enrollment Environmental Exposure Etiology Failure Family Fertility Fetal Death Fetus Fibroblasts Fluorescence Microscopy Folic Acid Frameshift Mutation Future Gene Mutation Genes Genetic Genetic Models Genome Genome Stability Goals Heterogeneity Human Human Genetics Image Informed Consent Investigation Label Lead Measurement Measures Methods Modeling Molecular Conformation Mononuclear Mutagenesis Mutation Neonatal Mortality Neural Tube Closure Neural Tube Defects Neural tube Organoids Parents Pathway interactions Patients Phase Phenotype Pregnancy Prophylactic treatment Proteins Publishing Quantitative Reverse Transcriptase PCR Recommendation Reproducibility Research Personnel Severities Surface Techniques Technology Teratogens Umbilical Cord Blood Validation Variant causal variant cell bank clinical practice constriction cranium de novo mutation exome exome sequencing expectation fetal fetal medicine folic acid supplementation gene correction genetic variant human model in utero induced pluripotent stem cell inhibitor knockout gene member mouse model neonate neurodevelopment neuroteratogen novel planar cell polarity pluripotency proband screening self organization

项目摘要

Abstract Neural tube defects (NTDs) are common malformations of the nervous system that occur during pregnancy. Anencephaly is one of the most dramatic and devastating NTDs characterized by failure of the brain and skull to close during neurulation. It is always fatal. Recent trio exome sequencing studies have identified de novo genetic mutations (DNMs) in anencephalic fetuses, and some of these mutations have been in genes associated with anencephaly in genetic mouse models. For these reasons, DNMs are beginning to be considered an important factor in the etiology of anencephaly and other NTDs. However, current model systems do not easily allow for directly validating that these variants lead to NTDs. Having a human-specific model of early neurodevelopment would allow for these types of conformational studies and further mechanistic investigations. 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 two known neuroteratogens and observe distinct structural changes consistent with NTDs. The goal in Aim 1 is to generate a model of anencephaly by knocking out genes known to lead to anencephaly (SHROOM3 and CELSR1) in SOSRS and characterize structural signatures indicative of NTDs. In Aim 2, we will generate fetal-specific models. To this end, we will enroll families with anencephalic fetuses to obtain blood samples, cord blood, and/or amniocentesis. Blood DNA will be used to perform trio exome sequencing to identify DNMs. Cells from the cord blood or amniocentesis from anencephalic fetuses with likely causative DNMs will be reprogrammed into iPSCs. The potentially causative mutations will also be corrected by CRISPR/Cas9 to generate isogenic controls. Finally, SOSRS from these models will be generated and measured for phenotypes identified in the proof-of-principle anencephalic models. Our study will likely shed light on the mechanisms of human anencephaly and allow for screening novel DNMs for NTD causality, thus, greatly expanding our understanding of human NTD genetics. Furthermore, our collaboration of basic researchers with clinicians specializing in genetics of maternal and fetal medicine will provide a framework for rapid modeling of clinical cases, potentially resulting in altered clinical practices including fertility recommendations and the utility of prophylactic treatments, such as folate supplementation.

抽象的神经管缺陷（NTD）是妊娠期间发生的常见神经系统畸形。无脑畸形是最严重和最具破坏性的 NTD 之一，其特征是大脑和头骨衰竭在神经调节期间关闭。这总是致命的。最近的三重外显子组测序研究已经确定了从头无脑胎儿的基因突变（DNM），其中一些突变发生在基因中与遗传小鼠模型中的无脑畸形有关。由于这些原因，DNM 开始被被认为是无脑畸形和其他 NTD 病因的重要因素。然而，目前的模型系统不容易直接验证这些变异是否会导致 NTD。具有人类特有的早期神经发育模型将允许进行这些类型的构象研究并进一步机制研究。为此目的利用人脑类器官技术的尝试受到限制，因为结构异质性和类器官内的变异性。我们最近开发的可重复自组织来自人类诱导多能干细胞的单莲座球体 (SOSRS) 使我们能够治疗 SOSRS 与两种已知的神经致畸剂进行实验，并观察到与 NTD 一致的明显结构变化。目标在目标 1 是通过敲除已知导致无脑畸形的基因来生成无脑畸形模型（SHROOM3 和 CELSR1）在 SOSRS 中并表征了指示 NTD 的结构特征。在目标 2 中，我们将生成胎儿特定的模型。为此，我们将招募有无脑胎儿的家庭来获取血液样本、脐带血和/或羊膜穿刺术。血液 DNA 将用于进行三重外显子组测序识别 DNM。来自脐带血或无脑胎儿羊膜穿刺术的细胞，可能是致病原因 DNM 将被重新编程为 iPSC。潜在的致病突变也将通过以下方式得到纠正： CRISPR/Cas9 用于生成等基因对照。最后，将生成来自这些模型的 SOSRS 并测量原理验证无脑模型中识别的表型。我们的研究可能会减少阐明人类无脑畸形的机制，并允许筛选新的 DNM 以发现 NTD 因果关系，因此，极大地扩展了我们对人类 NTD 遗传学的理解。此外，我们的基本合作研究人员与专门从事母婴医学遗传学的临床医生合作，将提供一个框架临床病例的快速建模，可能会改变包括生育在内的临床实践建议和预防性治疗的效用，例如补充叶酸。

项目成果

期刊论文数量（2）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A Shared Pathogenic Mechanism for Valproic Acid and SHROOM3 Knockout in a Brain Organoid Model of Neural Tube Defects.

神经管缺陷脑类器官模型中丙戊酸和 SHROOM3 敲除的共同致病机制。

DOI：
发表时间：
2023-04-11
期刊：
影响因子：
0
作者：
Takla, Taylor N;Luo, Jinghui;Sudyk, Roksolana;Huang, Joy;Walker, J Clayton;Vora, Neeta L;Sexton, Jonathan Z;Parent, Jack M;Tidball, Andrew M
通讯作者：
Tidball, Andrew M

A Shared Pathogenic Mechanism for Valproic Acid and SHROOM3 Knockout in a Brain Organoid Model of Neural Tube Defects.

神经管缺陷脑类器官模型中丙戊酸和 SHROOM3 敲除的共同致病机制。

DOI：
发表时间：
2023-06-23
期刊：
影响因子：
6
作者：
Takla, Taylor N;Luo, Jinghui;Sudyk, Roksolana;Huang, Joy;Walker, John Clayton;Vora, Neeta L;Sexton, Jonathan Z;Parent, Jack M;Tidball, Andrew M
通讯作者：
Tidball, Andrew M