Pathological Mechanisms of Human Cerebellar Malformations

人类小脑畸形的病理机制

基本信息

批准号：
10467630
负责人：
Kathleen Joyce Millen
金额：
$ 35.36万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-01 至 2022-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10467630
关键词：
Address Affect Affective Anatomy Animal Model Atlases Attention deficit hyperactivity disorder Automobile Driving Behavioral Biological Models Biology Birth Blood Vessels Cell Cycle Cell Proliferation Cells Cellular Assay Cerebellar Diseases Cerebellar malformation Cerebellar vermis structure Cerebellum Chick Clinical Cognition Cognitive Complement Conceptions Congenital Abnormality Counseling Dandy-Walker Syndrome Data Data Set Development Diagnosis Diagnostic Family Fetal Development Gene Expression Genes Goals Gold Head Heterogeneity Histologic Human Image Immunohistochemistry In Situ In Situ Hybridization In Vitro Intellectual functioning disability Knowledge Lateral Light Linguistics Lip structure Macaca mulatta Medial Meninges Mesenchyme Methods Microscopy Modeling Molecular Morbidity - disease rate Morphology Motor Mus Neurodevelopmental Disorder Neurons Nuclear Outcome Output Pathogenesis Pathologic Play Posterior Fossa Pregnancy Process Property RNA Sequences Regulation Reporting Research Role Sampling Schizophrenia Source Specific qualifier value Structure Ventricular autism spectrum disorder base brain malformation cell type data standards design developmental disease experimental study human fetal cerebellar tissue human model humanized mouse imaging study improved in vivo Model insight medulloblastoma mortality motor control mouse model neurogenesis nonhuman primate novel prenatal progenitor programs sensory integration single cell analysis single-cell RNA sequencing social deficits stem cells subventricular zone transcriptome sequencing transcriptomics vascular bed

项目摘要

Abstract of the funded parent grant: Numerous cerebellar malformations have been described in humans. Most cause cognitive, in addition to motor and sensory integration deficits. Surprisingly little is understood regarding the developmental basis of these malformations, particularly since little human specific data is available for normal or abnormal fetal cerebellar development. This proposal seeks to advance knowledge of human cerebellar development and malformations using human fetal samples and mouse models. The human-specific data will directly test the validity of our working mouse-derived hypotheses regarding the causes these disorders and strengthen the foundation of normal developmental data which will inform our ongoing genetic analyses of human cerebellar malformations. We will conduct the first in-depth analysis of normal human fetal cerebellar development from 4-23 Gestational Weeks, covering major developmental events. We will then examine the pathology of human fetal Dandy-Walker malformation the most common human cerebellar malformation, affecting ~1/3000 live births. Mouse models will be generated in conjunction with these experiments to assess the mechanisms of the developmental pathology. Finally, we will generate the first transcriptome data for normal human fetal cerebellar neurons. These cell-type specific data are critically missing from current publicly available brain resources. Our human fetal cerebellar neuron data will be compared to transcriptome data from existing datasets of endogenous mouse developing cerebellar neurons as well as mES and hPSC-derived cerebellar neurons to development to assess their validity as model systems. Further, the data will also be integrated with exome data from human cerebellar malformation patients to facilitate gene discovery for these important and understudied birth defects. Abstract of Requested Supplement: This application is being submitted for PA-19-056 in accordance with NOT-OD-19-071. The purpose of this research supplement is to define the cellular, molecular and morphological cerebellar developmental trajectories in human Down Syndrome samples. Developmental profiles will be generated through a combination of single cell sequencing, histological and immunohistochemical analyses and complemented with cell culture assays defining the mitogenic properties of cerebellar granule progenitors. Data from Down Syndrome samples will then be directly compared to profiles from normal and Dandy-Walker malformation developmental cerebellar samples available in the lab and generated under the parent R01. Cerebellar hypoplasia is one of the most consistent phenotypes in Down Syndrome patients that is a significant contributor to neurological phenotypes in these patients. Yet, very little is understood about the developmental disruption of cerebellar development that underlies the congenital hypoplasia. We will produce a multi-modal description of human cerebellar development in Down Syndrome, comparable to data we are already generating to define normal cerebellar development. An understanding how and when Down Syndrome cerebellar developmental trajectories differ from normal and other cerebellar malformations will elucidate the cellular and circuit underpinnings of pediatric and adult Down Syndrome neurological phenotypes. The studies are of high impact with considerable translational potential to identify new therapeutic approaches for neurological deficits in Down Syndrome. They will also generate baseline data human data to the developmental stage-, cell type-, and molecular-specificity of model systems (hiPSCs, organoids, animal models). These experiments specifically address Component 1 and Component 2 of the INCLUDE Project research objectives.

资助父母赠款的摘要：人类已经描述了许多小脑畸形。除运动和感觉整合缺陷外，大多数引起认知。令人惊讶的是，关于这些畸形的发育基础知之甚少，特别是因为很少有人类特定数据可用于正常或异常的胎儿小脑发育。该建议旨在使用人类胎儿样本和小鼠模型来提高人类小脑发育和畸形的知识。人类特异性数据将直接测试我们工作的小鼠衍生的假设的有效性，这些假设在这些原因方面，并增强正常发育数据的基础，这将为我们对人类小脑畸形的持续遗传分析提供信息。从4-23周开始，我们将对正常人类小脑发育进行首次深入分析，涵盖重大的发育事件。然后，我们将检查人类胎儿dandy-walker畸形的病理学最常见的人类小脑畸形，影响〜1/3000的活产。小鼠模型将与这些实验结合生成，以评估发育病理的机制。最后，我们将生成正常人胎儿小脑神经元的第一个转录组数据。这些细胞类型的特定数据在当前可公开的大脑资源中严重缺少。我们的人类小脑神经元数据将与来自内源性小鼠的现有数据集的转录组数据进行比较，开发小脑神经元，MES和HPSC衍生的小脑神经元，以评估其作为模型系统的有效性。此外，数据还将与人类小脑畸形患者的外显子数据集成在一起，以促进这些重要且研究不足的先天缺陷的基因发现。请求补充的摘要：根据NOT-OD-19-071，正在为PA-19-056提交此申请。该研究补充的目的是定义人唐氏综合症样本中的细胞，分子和形态的小脑发育轨迹。发育曲线将通过单细胞测序，组织学和免疫组织化学分析的结合，并与定义小脑颗粒祖细胞有丝分裂特性的细胞培养试验相互补充。然后，来自唐氏综合症样本的数据将直接与实验室中可用的正常和dandy-Walker畸形的小脑样品进行比较，并在父级R01下生成。小脑发育不全是唐氏综合症患者中最一致的表型之一，这是这些患者神经系统型的重要贡献。然而，关于基于先天性下降症的小脑发育的发展中断，几乎没有理解。我们将对唐氏综合症的人类小脑发育产生多模式描述，与我们已经生成的数据相媲美，以定义正常的小脑发育。了解唐氏综合症小脑发育轨迹的方式和何时与正常和其他小脑畸形不同，将阐明小儿和成人唐氏综合征神经系统型的细胞和电路基础。这些研究具有很高的影响，具有巨大的转化潜力，可以鉴定唐氏综合症神经缺陷的新治疗方法。他们还将生成基线数据人类数据，以使模型系统的发育阶段，细胞类型和分子特异性（HIPSC，类器官，动物模型）生成基线数据。这些实验专门针对包括项目研究目标的组件1和组件2。