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：
9125878
负责人：
HERBERT M LACHMAN
金额：
$ 41.75万
依托单位：
ALBERT EINSTEIN COLLEGE OF MEDICINE, INC
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 2018-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9125878
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 Family Study Foundations Gene Expression Genes Genetic Genetic study Genotype Grant Health Human In Vitro Individual Laboratories Libraries Mental Health Methods Modeling Molecular Molecular Genetics Monozygotic twins Morphology Mus NRG1 gene NRG3 gene National Institute of Mental Health Neuronal Differentiation Neurons Oligodendroglia Parents Patients Penetrance Phase Play Prader-Willi Syndrome Reading Regulation Role SNP array Schizophrenia Skin Spliced Genes Telencephalon Testing Therapeutic Topoisomerase Inhibitors Topotecan autism spectrum disorder base cell type chromatin immunoprecipitation developmental disease fetal genome-wide histone modification imprint induced pluripotent stem cell interest nerve stem cell neurogenesis neuropsychiatric disorder novel therapeutics 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）：随机和印迹单相表达的基因会影响细胞分化和发育。印迹基因以原始的方式表达，而在随机单相表达中，母体或父亲等位基因在细胞中具有活性。经典地，众所周知，烙印在神经精神疾病的发展中起着关键作用。此外，在精神分裂症（SZ），自闭症谱系障碍（ASD）和双相情感障碍（BD）的一部分中也发现了父母的效应。随机单位型表达和脑表达基因的印迹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，例如单卵双胞胎的不一致和降低的渗透率。已经出现了两个实验工具，这些工具提供了评估单相（也称为等位基因偏见）基因在神经元分化和神经精神疾病中的作用的手段。诱导多能干细胞（IPSC）技术和整个转录组测序（RNA-Seq）。为了鉴定单相表达的基因，我们使用范围内的人类SNP阵列6.0进行了源自对照IPSC系的神经元和基因分型DNA的神经元的初步RNA-Seq分析。鉴定了杂合子SNP，并分析了它们之间的RNA-seq读数。我们发现了801个基因中等位基因偏向表达的证据。此外，还发现了SZ和ASD候选基因的统计学意义富集，其中包括A2BP1（RBFOX1），ERBB4，NLGN4X，NRG1，NRG1，NRG3，NRXN1和NLGN1。 A2BP1特别有趣，因为作为神经元基因剪接破坏其表达的调节剂具有影响许多下游靶标。在当前的建议中，我们将探讨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的，还是通过表观遗传过程引起的，导致烙印或随机单相表达。通过进行全基因组DNA甲基化和染色质免疫沉淀研究，将探索分化人类神经元中等位基因偏见的表观基础偏置表达。完成这些研究后，我们将能够将大量SZ，ASD和BD候选基因分组为一个共同的功能伞：通过等位基因偏见的表达调节，这一发现将为基于表观遗传的治疗策略提供基础。