Fullard John F, Giambartolomei Claudia, Hauberg Mads E, Xu Ke, Voloudakis Georgios, Shao Zhiping, Bare Christopher, Dudley Joel T, Mattheisen Manuel, Robakis Nikolaos K, Haroutunian Vahram, Roussos Panos
Department of Psychiatry, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
Department of Biomedicine.
Hum Mol Genet. 2017 May 15;26(10):1942-1951. doi: 10.1093/hmg/ddx103.
Open chromatin provides access to DNA-binding proteins for the correct spatiotemporal regulation of gene expression. Mapping chromatin accessibility has been widely used to identify the location of cis regulatory elements (CREs) including promoters and enhancers. CREs show tissue- and cell-type specificity and disease-associated variants are often enriched for CREs in the tissues and cells that pertain to a given disease. To better understand the role of CREs in neuropsychiatric disorders we applied the Assay for Transposase Accessible Chromatin followed by sequencing (ATAC-seq) to neuronal and non-neuronal nuclei isolated from frozen postmortem human brain by fluorescence-activated nuclear sorting (FANS). Most of the identified open chromatin regions (OCRs) are differentially accessible between neurons and non-neurons, and show enrichment with known cell type markers, promoters and enhancers. Relative to those of non-neurons, neuronal OCRs are more evolutionarily conserved and are enriched in distal regulatory elements. Transcription factor (TF) footprinting analysis identifies differences in the regulome between neuronal and non-neuronal cells and ascribes putative functional roles to a number of non-coding schizophrenia (SCZ) risk variants. Among the identified variants is a Single Nucleotide Polymorphism (SNP) proximal to the gene encoding SNX19. In vitro experiments reveal that this SNP leads to an increase in transcriptional activity. As elevated expression of SNX19 has been associated with SCZ, our data provide evidence that the identified SNP contributes to disease. These results represent the first analysis of OCRs and TF-binding sites in distinct populations of postmortem human brain cells and further our understanding of the regulome and the impact of neuropsychiatric disease-associated genetic risk variants.
开放染色质为DNA结合蛋白提供了对基因表达进行正确时空调控的通道。绘制染色质可及性图谱已被广泛用于识别顺式调控元件(CRE)的位置,包括启动子和增强子。CRE具有组织和细胞类型特异性,与疾病相关的变异通常在与特定疾病相关的组织和细胞中的CRE中富集。为了更好地理解CRE在神经精神疾病中的作用,我们应用转座酶可及染色质测序分析(ATAC-seq),通过荧光激活核分选(FANS)从冷冻的死后人类大脑中分离出神经元和非神经元细胞核。大多数已识别的开放染色质区域(OCR)在神经元和非神经元之间具有不同的可及性,并显示出与已知细胞类型标记、启动子和增强子的富集。相对于非神经元,神经元OCR在进化上更保守,并且在远端调控元件中富集。转录因子(TF)足迹分析确定了神经元和非神经元细胞之间调控组的差异,并将许多非编码精神分裂症(SCZ)风险变异赋予了假定的功能作用。在已识别的变异中,有一个位于编码SNX19基因附近的单核苷酸多态性(SNP)。体外实验表明,该SNP导致转录活性增加。由于SNX19的表达升高与SCZ有关,我们的数据提供了证据表明所识别的SNP促成了疾病。这些结果代表了对死后人类脑细胞不同群体中的OCR和TF结合位点的首次分析,并进一步加深了我们对调控组以及神经精神疾病相关遗传风险变异影响的理解。
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