Monoallelic expression in neurons derived from induced pluripotent stem cells

诱导多能干细胞衍生的神经元中的单等位基因表达

基本信息

批准号：
8720821
负责人：
HERBERT M LACHMAN
金额：
$ 41.42万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-08-15 至 2018-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8720821
关键词：
3-Dimensional Alleles Allelic Imbalance Angelman Syndrome Animal Model Astrocytes Autistic Disorder Bipolar Disorder Blood Cells Brain Brain region Candidate Disease Gene Cell Line Cells ChIP-seq DNA DNA Methylation Development Disease Disease model Embryo Environment Epidemiology Epigenetic Process ErbB4 gene Family Foundations Gene Expression Genes Genetic Genotype Grant Human In Vitro Individual Laboratories Libraries Mental Health Methods Modeling Molecular Molecular Genetics Monozygotic Twinning Monozygotic twins Morphology Mus NRG1 gene NRG3 gene National Institute of Mental Health Neuronal Differentiation Neurons Oligodendroglia Parents Patients Penetrance Pharmaceutical Preparations Phase Play Prader-Willi Syndrome Reading Regulation Relative (related person)Role Schizophrenia Skin Spliced Genes Telencephalon Testing Therapeutic Topoisomerase Inhibitors Topotecan autism spectrum disorder base cell type chromatin immunoprecipitation developmental disease family genetics fetal genome-wide histone modification imprint induced pluripotent stem cell interest nerve stem cell neurogenesis neuropsychiatry public health relevance stem cell technology tool transcriptome sequencing treatment strategy

项目摘要

DESCRIPTION (provided by applicant): Stochastic and imprinted monoallelically expressed genes influence cellular differentiation and development. Imprinted genes are expressed in a parent-of-origin manner, whereas in stochastic monoallelic expression either the maternal or paternal allele is active in a cell. Classically, imprinting is known to play a key role in the development of the neuropsychiatric disorders Prader-Willi Syndrome and Angelman Syndrome. In addition, parent-of-origin effects have also been found in a subset of families with schizophrenia (SZ), autism spectrum disorders (ASD) and bipolar disorder (BD). Both stochastic monoallelic expression and imprinting of brain-expressed genes could help explain some interesting epidemiological features of neuropsychiatric disorders, such as discordance in monozygotic twins and reduced penetrance. Two experimental tools have emerged that provide the means to evaluate the role of monoallelic (also known as allele-biased) gene expression in neuronal differentiation and neuropsychiatric disorders; induced pluripotent stem cell (iPSC) technology, and whole transcriptome sequencing (RNA-Seq). To identify monoallelically expressed genes, we carried out a preliminary RNA-Seq analysis of neurons derived from a control iPSC line and genotyped DNA using the Affymetrix Genome-Wide Human SNP Array 6.0. Heterozygous SNPs were identified and RNA-Seq reads across them were analyzed. We found evidence for allele-biased expression in 801 genes. In addition, a statistically significant enrichment for SZ and ASD candidate genes was found, which included A2BP1 (RBFOX1), ERBB4, NLGN4X, NRG1, NRG3, NRXN1, and NLGN1. A2BP1 is particularly interesting because as a regulator of neuronal gene splicing disrupting its expression has the capacity to influence numerous downstream targets. In this current proposal, we will explore the mechanism of allele-biased expression and determine whether the phenomenon is caused by cis-acting genetic factors, or by an epigenetic process leading to either imprinting or stochastic monoallelic expression. The epigenetic basis underlying allele-biased expression in differentiating human neurons will be explored by carrying out genome-wide DNA methylation and chromatin immunoprecipitation studies. Upon completion of these studies we will be able to group a large number of SZ, ASD and BD candidate genes into a common functional umbrella: regulation by allele-biased expression, a finding that will provide the foundation for epigenetic-based treatment strategies.

描述（由申请人提供）：随机和印记单等位基因表达的基因影响细胞分化和发育。印记基因以亲本方式表达，而在随机单等位基因表达中，母本或父本等位基因在细胞中活跃。传统上，印记在神经精神疾病普瑞德-威利综合症和天使综合症的发展中发挥着关键作用。此外，在一部分患有精神分裂症（SZ）、自闭症谱系障碍（ASD）和双相情感障碍（BD）的家庭中也发现了父母的影响。随机单等位基因表达和大脑表达基因的印记都可以帮助解释神经精神疾病的一些有趣的流行病学特征，例如同卵双胞胎的不一致和外显率降低。已经出现的两种实验工具提供了评估单等位基因（也称为等位基因偏向）基因表达在神经元分化和神经精神疾病中的作用的方法；诱导多能干细胞（iPSC）技术和全转录组测序（RNA-Seq）。为了鉴定单等位基因表达的基因，我们使用 Affymetrix 全基因组人类 SNP 阵列 6.0 对来自对照 iPSC 系的神经元和基因分型 DNA 进行了初步 RNA 测序分析。鉴定了杂合 SNP，并分析了它们的 RNA-Seq 读数。我们在 801 个基因中发现了等位基因偏向表达的证据。此外，还发现了 SZ 和 ASD 候选基因的统计显着富集，其中包括 A2BP1 (RBFOX1)、ERBB4、NLGN4X、NRG1、NRG3、NRXN1 和 NLGN1。 A2BP1 特别有趣，因为作为神经元基因剪接的调节因子，破坏其表达有能力影响许多下游靶标。在当前的提案中，我们将探索等位基因偏向表达的机制，并确定该现象是由顺式作用遗传因素引起，还是由导致印记或随机单等位基因表达的表观遗传过程引起。通过进行全基因组 DNA 甲基化和染色质免疫沉淀研究，将探索人类神经元分化中等位基因偏向表达的表观遗传基础。完成这些研究后，我们将能够将大量 SZ、ASD 和 BD 候选基因分组到一个共同的功能伞中：通过等位基因偏向表达进行调节，这一发现将为基于表观遗传学的治疗策略提供基础。