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.

修改后的项目摘要/摘要部分 SETD1A 中罕见的功能丧失 (LoF) 突变与精神分裂症 (SZ) 和其他严重的神经发育障碍密切相关，精神分裂症是一种影响 1% 人口的衰弱性精神障碍。 SETD1A 编码组蛋白甲基转移酶复合物的一个组成部分，在赖氨酸 4 (H3K4) 处产生单甲基化、二甲基化和三甲基化组蛋白 H3。 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，在主要精神疾病的常见基于变异的全基因组关联研究（GWAS）中，组蛋白甲基化也被认为是最丰富的基因途径之一。此外，最近的 SETD1A 杂合敲除小鼠模型表现出工作记忆缺陷，并显示出与神经发育障碍相关的转录变化重叠，但似乎独立于 H3K4me3 机制。因此，目前尚不清楚 SETD1A 是否以及如何引起人脑中与 SZ 相关的分子和细胞变化。我们的中心假设是，人类诱导多能干细胞 (hiPSC) 衍生的神经元细胞和皮质类器官概括了人脑中 SETD1A LoF 的关键 SZ 相关表观遗传、分子和细胞特性。使用 CRISPR/Cas9 基因编辑，我们生成了携带 SETD1A 杂合 LoF 突变（位于不同遗传背景的外显子 4 和外显子 16）的同基因 hiPSC 系。初步结果表明，突变系在皮质类器官发育方面存在缺陷，在早期发育阶段神经元过早分化。此外，携带 SETD1A LoF 突变的 hiPSC 神经元的形态学、电生理学和转录组学分析显示突触神经传递有缺陷。有趣的是，在来自突变系的 3D 皮质类器官和 2D 培养物中显示差异表达的基因在 SZ 和其他神经精神疾病/性状的常见 GWAS 风险变异中富集，表明 SETD1A LoF 和主要精神疾病的常见 GWAS 风险变异可能共享收敛途径。利用我们团队在 hiPSC 模型和神经发生、突触生理学和功能基因组学方面各自的专业知识，我们建议描述人类神经系统中 SETD1A 中 SZ 相关 LoF 突变相关缺陷的分子和细胞机制。我们将鉴定皮质类器官中与 SETD1A LoF 相关的细胞类型特异性和发育阶段特异性细胞和分子表型，然后研究人类神经元中 SETD1A LoF 突变的突触表型以及相关的转录组变化。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。