权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Human Chromosome 14 Analysis in Neuronal Cells

神经元细胞中的人类 14 号染色体分析

基本信息

批准号：
9360000
负责人：
Stewart A Anderson
金额：
$ 25.2万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
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：
2016-09-30 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9360000
关键词：
3-Dimensional Affect Animal Model Architecture Binding Brain Brain Pathology Cell Cycle Cell Line Cell Nucleus Cell Proliferation Cell Survival Cell model Cells Cerebral cortex Cerebrum Chromatin Chromatin Loop Chromatin Structure Chromosome Arm Chromosome Positioning Chromosome Structures Chromosome Territory Chromosome abnormality Chromosomes Chromosomes, Human, Pair 14 Clinical Corpus striatum structure DNA Development Developmental Delay Disorders Developmental Gene Differentiation and Growth Disease Drug resistance Electrophysiology (science)Embryo Epigenetic Process Epilepsy Event Eye Eye Abnormalities FOXG1B gene Family Family member Feeling Fluorescent in Situ Hybridization Forebrain Development Foundations Future Gene Expression Gene Expression Regulation Generations Genes Genetic Materials Goals Growth Human Human Chromosomes Immune system Impairment In Vitro Intellectual functioning disability Interneurons Interphase Investigation Laboratories Lead Learning Life Maintenance Maps Measures Memory Microcephaly Mitotic Modeling Molecular Abnormality Molecular Cytogenetics Morphology Movement Disorders Mus Muscle hypotonia Neuraxis Neurological Models Neuronal Differentiation Neurons Nuclear Organoids Patients Pattern Phenotype Pigmentation physiologic function Population Positioning Attribute Prosencephalon Publishing Reagent Recurrence Reproducibility Research Personnel Resources Respiratory Tract Infections Retinal Ring Chromosomes Role Sampling Seizures Stress Structure Study models Syndrome System Systems Analysis Telencephalon Testing Tissues Work biobank body system brain abnormalities cell growth chromatin modification cis acting element clinical phenotype early onset epigenetic regulation hippocampal pyramidal neuron histone modification impaired brain development induced pluripotent stem cell innovation mammalian genome nerve stem cell neural circuit neurodevelopment neuron development neuronal patterning neuropathology precursor cell progenitor psychologic telomere tool

项目摘要

Project Summary/Abstract Human ring chromosomes are abnormal structures formed by intrachromosomal fusions, creating a circular chromosome. Ring chromosome syndrome is a debilitating disorder resulting in severe drug-resistant epilepsy, intellectually disabilities (IQ <70), and developmental delay. Ring chromosome patients and families deal with life-long challenges of prolonged seizures, learning and memory issues, severe psychological disruption, and a feeling of powerlessness, resulting in a great deal of stress and strain. Clinically this disorder affects multiple organ systems including the brain (microcephaly, early onset epilepsy, and intellectual disabilities), the eye, the immune system, and growth. Our laboratory and others, have focused on mapping and characterizing the molecular and cytogenetics of ring chromosome structures; however, to date no published studies have focused on investigating the cellular neuropathology due to lack of appropriate models. To investigate this disorder, we have created a large biobank of cell lines from ring chromosome 14 (r(14)) patients and family members. From these cell lines, we have produced the only model available by generating patient-specific induced pluripotent stem cells, which provide the foundation for studying this devastating disorder that lacks alternative models. In our preliminary work, we find that by differentiating ring chromosome iPSCs toward a forebrain fate we can generate populations of neuronal precursor cells (NPCs) that can be further differentiated into post-mitotic neurons. Neuronal precursor cells harboring the ring chromosome 14 have reduced cellular growth and decreased expression of key telencephalon neuronal markers. Interestingly, a number of genes located on chromosome 14 are involved in neuronal differentiation or maintenance, leading us to postulate that the ring structure modifies expression of this key gene responsible for in vitro cortical cell generation. We have also begun to generate an in vitro cerebral cortex organoid model, which will be important for studying the early embryonic events perturbed in a R(14) developmental model. Mammalian genomes are organized in distinct architectures which allow for the orchestration of proper gene and epigenetic regulation. Our proposed studies will identify ring chromosome 14 orientation in the nucleus, epigenetic marks on chromosome 14, and establish a 3-dimensional (3D) cerebral cortex developmental model for studying phenotypes of Ring chromosome 14 syndrome. We hypothesize that the altered higher order chromosome structure resulting from ring formation disrupts normal chromatin compaction, looping, localization within the nucleus and ultimately gene regulation. The proposed work will provide important tools for studying R(14) syndrome, and yield a comprehensive view of the role a ring chromosome has on DNA positioning, chromatin modifications, and in vitro neuronal development. In addition, this innovative model should provide a strong foundation for future work fully characterizing 3D organoid development in R(14) and provide essential reagents (neuronal tissues) to study the neural circuits and electrophysiological patterns in R(14) patient samples.

项目概要/摘要人类环染色体是由染色体内融合形成的异常结构，形成环状染色体。环染色体综合征是一种使人衰弱的疾病，导致严重的耐药性癫痫，智力障碍（智商<70）和发育迟缓。环染色体患者及家属应对长期癫痫发作、学习和记忆问题、严重的心理障碍以及无力感，导致巨大的压力和压力。临床上这种疾病影响多种器官系统，包括大脑（小头畸形、早发性癫痫和智力障碍）、眼睛、免疫系统和生长。我们的实验室和其他实验室专注于绘制和表征环染色体结构的分子和细胞遗传学；然而，迄今为止还没有发表的研究由于缺乏合适的模型，专注于研究细胞神经病理学。为了调查此事疾病，我们创建了一个来自环染色体 14 (r(14)) 患者和家人的大型细胞系生物库成员。从这些细胞系中，我们通过生成患者特异性而产生了唯一可用的模型诱导多能干细胞，为研究这种缺乏的破坏性疾病提供了基础替代模型。在我们的初步工作中，我们发现通过将环染色体 iPSC 分化为前脑命运我们可以产生可以进一步分化的神经元前体细胞（NPC）群体进入有丝分裂后神经元。含有环 14 号染色体的神经元前体细胞减少了细胞关键端脑神经元标志物的生长和表达减少。有趣的是，许多基因位于 14 号染色体上的基因参与神经元分化或维持，使我们推测环状结构改变了负责体外皮质细胞生成的关键基因的表达。我们有还开始生成体外大脑皮层类器官模型，这对于研究早期大脑皮层类器官具有重要意义 R(14) 发育模型中胚胎事件受到干扰。哺乳动物基因组以不同的方式组织允许编排适当的基因和表观遗传调控的架构。我们提出的研究将识别环 14 号染色体在细胞核中的方向、14 号染色体上的表观遗传标记，并建立用于研究 14 号环染色体表型的 3 维 (3D) 大脑皮层发育模型综合症。我们假设环导致了高阶染色体结构的改变形成会破坏细胞核内正常的染色质压缩、循环和定位最终是基因调控。拟议的工作将为研究 R(14) 综合征提供重要工具，并且全面了解环状染色体对 DNA 定位、染色质修饰、和体外神经元发育。此外，这种创新模式应该为未来的工作充分表征 R(14) 中 3D 类器官的开发，并提供必要的试剂（神经元组织）来研究 R(14) 患者样本中的神经回路和电生理模式。