Loss-of-Function Analyses of SETD1A in Human Neural Models

人类神经模型中 SETD1A 的功能丧失分析

基本信息

批准号：
10616753
负责人：
Guo-li Ming
金额：
$ 62.33万
依托单位：
RUTGERS BIOMEDICAL AND HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10616753
关键词：
3-Dimensional Affect Axon Brain CRISPR/Cas technology Cell Line Cells Complex Data Defect Development Electrophysiology (science)Engineering Enhancers Epigenetic Process Exhibits Exons Family Functional disorder Genes Genetic Genetic Transcription Genetic study Goals Heterozygote Histone H3 Human Human Genetics Image Induced pluripotent stem cell derived neurons Knock-out Loss of Heterozygosity Lysine Memory impairment Mental disorders Methylation Modeling Molecular Morphology Neurodevelopmental Disorder Neuronal Differentiation Neurons Organoids Pathway interactions Patients Phenotype Physiology Population Prevalence Property Resolution Role SET Domain Schizophrenia Short-Term Memory Societies Stress Structure Supporting Cell Synapses System Techniques Variant cell type chromatin immunoprecipitation differential expression epigenomics excitatory neuron experimental study functional genomics genome wide association study genome-wide histone methylation histone methyltransferase induced pluripotent stem cell inhibitory neuron loss of function loss of function mutation molecular phenotype mouse model mutant neural neural model neurodevelopment neurogenesis neuropsychiatric disorder neurotransmission novel therapeutics overexpression pharmacologic premature promoter public health relevance risk variant single nucleus RNA-sequencing socioeconomics stem cell model synaptic function therapeutically effective trait transcriptome transcriptome sequencing transcriptomics

项目摘要

Modified Project Summary/Abstract Section Rare loss-of-function (LoF) mutations in SETD1A are strongly associated with schizophrenia (SZ), a debilitating mental disorder affecting 1% of the population, and other severe neurodevelopmental disorders. SETD1A encodes a component of the histone methyltransferase complex producing mono-, di, and trimethylated histone H3 at Lysine 4 (H3K4). H3K4 trimethylation (H3K4me3) and H3K4me1 are epigenomic marks of active gene transcriptional promoters and enhancers, respectively. Interestingly, histone methylation has also been suggested as one of the most enriched gene pathways in common variant-based genome-wide associations studies (GWAS) of major psychiatric disorders. Furthermore, a recent mouse model with heterozygous knockout of SETD1A exhibited working memory deficits and showed transcriptional changes that overlap with those implicated in neurodevelopmental disorders, however, seemingly independent from a H3K4me3 mechanism. Therefore, it remains largely unclear whether and how SETD1A causes SZ-relevant molecular and cellular changes in a human brain. Our central hypothesis is that human induced pluripotent stem cell (hiPSC)-derived neuronal cells and cortical organoids recapitulate key SZ-relevant epigenetic, molecular and cellular properties of SETD1A LoF in the human brain. Using CRISPR/Cas9 gene editing, we have generated isogenic hiPSC lines carrying heterozygous LoF mutations (in exon 4 and exon 16, on different genetic backgrounds) of SETD1A. Preliminary results showed that mutant lines were defective in cortical organoid development with premature neuronal differentiation at early developmental stages. Furthermore, morphological, electrophysiological and transcriptomic analyses of hiPSC neurons carrying SETD1A LoF mutation showed defective synaptic neurotransmission. Interestingly, genes showing differential expression in both 3D cortical organoids and 2D cultures from mutant lines are enriched for common GWAS risk variants of SZ and other neuropsychiatric disorders/traits, suggesting possible convergent pathways shared by SETD1A LoF and common GWAS risk variants of major psychiatric disorders. Leveraging our respective expertise in hiPSC models and neurogenesis, synaptic physiology and functional genomics within our team, we propose to characterize the molecular and cellular mechanisms underlying the deficits associated with SZ-associated LoF mutations in SETD1A in human neural systems. We will identify the cell-type-specific and developmental stage-specific cellular and molecular phenotypes associated with SETD1A LoF in cortical organoids, and then investigate the synaptic phenotype(s) of SETD1A LoF mutations in human neurons and associated transcriptome changes. The proposed study will enable us to perform a well-controlled assessment of the impact of SETD1A LoF mutations on the molecular and cellular mechanisms underlying deficits in early neurodevelopment and synaptic properties.

修改的项目摘要/摘要部分 SETD1A中罕见的功能丧失（LOF）突变与精神分裂症（SZ）密切相关，这是一种影响人群1％的令人衰弱的精神障碍，以及其他严重的神经发育障碍。 SETD1A编码在赖氨酸4（H3K4）的组蛋白甲基转移酶复合酶复合酶复合酶复合物的成分（H3K4）。 H3K4三甲基化（H3K4ME3）和H3K4ME1分别是活性基因转录启动子和增强子的表观基因组标记。有趣的是，还建议组蛋白甲基化是主要精神疾病基于常见的全基因组相关研究（GWAS）中最丰富的基因途径之一。此外，最近具有杂合敲除setD1a的小鼠模型表现出工作记忆缺陷，并显示出转录变化，与与神经发育障碍有关的变化重叠，但是看似与H3K4ME3机制独立。因此，在很大程度上尚不清楚setD1a是否以及如何引起与SZ相关的分子和细胞变化。我们的中心假设是人类诱导的多能干细胞（HIPSC）衍生的神经元细胞和皮质器官概括了人脑中SETD1A LOF的关键SZ相关表观遗传，分子和细胞性质。使用CRISPR/CAS9基因编辑，我们已经生成了SETD1A的杂合LOF突变（在外显子4和外显子16中，在外显子4和外显子16中）产生的等源HIPSC系。初步结果表明，在早期发育阶段，突变线在皮质器官发育中有缺陷。此外，携带setD1A LOF突变的HIPSC神经元的形态，电生理和转录组分析显示突触神经传递缺陷。有趣的是，在3D皮质器官中显示出差异表达的基因和来自突变系的2D培养物富含SZ的常见GWAS风险变异和其他神经精神疾病/性状/性状，这表明SETD1A LOF和COMMON GWAS风险风险变体的主要精神疾病变体可能会收敛途径。利用我们团队内部的HIPSC模型以及神经发生，突触生理和功能基因组学的专业知识，我们建议表征人类神经系统中与SETD1A中与SZ相关LOF突变相关的缺陷的分子和细胞机制。我们将鉴定与皮质器官中与setD1a LOF相关的细胞类型特异性和发育阶段特异性的细胞和分子表型，然后研究人类神经元和相关转录组变化中SETD1A LOF突变的突触表型。拟议的研究将使我们能够对SETD1A LOF突变对早期神经发育和突触特性缺陷的分子和细胞机制的影响进行良好的控制评估。