Identifying enhancers in schizophrenia and bipolar disorder-associated noncoding eQTLs during cortical synaptic organization

识别皮质突触组织过程中精神分裂症和双相情感障碍相关非编码 eQTL 的增强子

• 批准号: 10585909
• 负责人: Tracy Leigh Warren
• 金额: $ 4.01万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-04-01 至 2024-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10585909
• 关键词: Adolescence; Adolescent; Adult; Alleles; Basic Science; Biological Assay; Biology; Bipolar Disorder; Brain; Brain imaging; Brain region; Calcium Channel; Code; Collection; Complex; DNA; Dependovirus; Development; Developmental Process; Diagnostic; Disease; Enhancers; Etiology; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Enhancer Element; Genetic Transcription; Genomics; Goals; Image; Individual; Injections; Introns; Label; Libraries; Link; Literature; Mental disorders; Meta-Analysis; Modeling; Molecular Target; Mus; Mutation; Nature; Neurons; Physiological; Plasmids; Process; Production; Protocols documentation; Psychotic Disorders; Quantitative Trait Loci; RNA; Regulatory Pathway; Reporter; Reporter Genes; Research; Risk; Role; Schizophrenia; Specificity; Synapses; Testing; Time; Transcriptional Activation; Transcriptional Regulation; Untranslated RNA; Variant; Vertebral column; Viral Packaging; Virus; Work; cell type; disorder risk; excitatory neuron; experimental study; genetic variant; genome wide association study; genomic variation; improved; in vitro Model; in vivo; inhibitory neuron; postnatal; postsynaptic neurons; primary outcome; protein aminoacid sequence; psychiatric genomics; psychotic; risk variant; schizophrenia risk; synaptogenesis; therapeutic development; voltage

PROJECT SUMMARY Genome-wide association studies of psychiatric disease have repeatedly linked risk for schizophrenia and bipolar disorder to noncoding regions that are in or near genes relevant in synaptic organization, including an interval in the third intron of the gene CACNA1C, which encodes the pore-forming subunit of a pan-neuronal postsynaptic voltage-gated calcium channel. As these variants are located in noncoding DNA, they do not alter the subsequent amino acid sequence of the protein. Rather, many of the risk variants are expected to be expression quantitative trait loci (eQTLs) of synaptic genes, and it is most likely that the risk-containing sequences are enhancers that regulate transcription of a targeted synaptic gene. This adds to a wide body of research that have identified schizophrenia and bipolar disorder as “disorders of the synapse.” However, though there is strong evidence linking the risk eQTLs and synaptic organization, the nature of this relationship is unclear. In this project, I aim to test the hypotheses that (1) non-coding psychotic disorder risk eQTLs of synaptic genes are enhancers during juvenile or adolescent cortical synaptic development and (2) risk- associated sequence variation in high-priority enhancers in CACNA1C disrupts the cell-type specific ability of the sequence to act as an enhancer during cortical synaptogenesis. I will test the first hypothesis by conducting a massively parallel reporter assay (MPRA) of candidate genomic enhancers that contain eQTLs that contribute transdiagnostic risk for bipolar disorder and schizophrenia in vivo during cortical synaptogenesis in mice. I will conduct this MPRA at two time points representing juvenile (P7) and adolescent (P56) synaptic developmental processes. These experiments will allow me to determine the functional role of top psychotic disorder-associated risk regions across development during mammalian cortical synaptogenesis. I will also test for allelic effects, which may allow me to identify eQTLs whose risk allele has a strong effect on transcriptional regulation. I will test the second hypothesis by conducting a reduced complexity MPRA of identified enhancers in CACNA1C in vivo during mouse cortical synaptogenesis, including sequences containing the risk allele, the reference allele, and a deletion centered at the risk-associated SNP. I will also test for cell-type or regional specificity of expression of CACNA1C enhancers. By testing fewer sequences in my second aim, my sensitivity to detect allelic differences in activity will be strongly improved and I anticipate being able to determine any potential differences in activity related to risk-associated sequence variation. The results of these experiments will clarify the link between risk-related noncoding genomic variation and mechanisms of disease etiology in mammalian cortical synaptogenesis. Future directions from this research include determining how expression changes associated with risk variants impact synaptic biology and the study of transdiagnostic genomic or molecular targets identified in this research for future therapeutic development.

项目摘要 全基因组疾病的基因组关联研究反复将精神分裂症的风险与 对突触组织中相关基因或附近的非编码区域的躁郁症，包括 基因cacna1c的第三个内含子的间隔，该内含子编码泛神经元的孔形成亚基 突触后电压门控钙通道。由于这些变体位于非编码DNA中，它们不会改变 随后的蛋白质氨基酸序列。相反，预计许多风险变体将是 突触基因的表达定量性状位置（EQTL），很可能是含风险的 序列是调节靶向突触基因转录的增强子。这增加了宽阔的身体 将精神分裂症和躁郁症确定为“突触的疾病”的研究。然而， 尽管有有力的证据与风险EQTL和突触组织联系起来，但这种关系的性质 不清楚。在这个项目中，我旨在检验（1）非编码精神病风险eqtls的假设 突触基因是少年或青少年皮质突触发育期间增强子，（2）风险 - CACNA1C中高优先级增强子的相关序列变化破坏了细胞类型的特异性能力 在皮质突触发生过程中充当增强子的序列。我将通过进行第一个假设来检验 候选基因组增强子的大规模平行记者测定法（MPRA），其中包含EQTL 在皮质突触发生过程中，在体内造成了躁郁症和精神分裂症的转诊风险 老鼠。我将在代表少年（P7）和青少年（p56）突触的两个时间点进行此MPRA 发展过程。这些实验将使我能够确定顶级精神病的功能作用 哺乳动物皮质突触发生期间，跨发育过程中与疾病相关的风险区域。我也会测试 对于等位基因效应，这可能使我能够识别其风险等位基因对转录有很强影响的EQTL 规定。我将通过降低已识别增强剂的复杂性MPRA来检验第二个假设 在小鼠皮质突触发生过程中的cacna1c中，包括含有风险等位基因的序列， 参考等位基因，并以风险相关的SNP为中心。我还将测试细胞类型或区域 CACNA1C增强子表达的特异性。通过测试我的第二个目标中的较少序列，我的敏感性 要检测活动差异的活动差异将得到很大的改善，我预计能够确定任何 与风险相关序列变化有关的活动的潜在差异。这些实验的结果 将阐明与风险相关的非编码基因组变异与疾病病因机制之间的联系 哺乳动物皮质突触发生。这项研究的未来方向包括确定如何表达 与风险变异相关的变化会影响突触生物学以及转诊基因组或 在这项研究中确定的未来治疗性发育的分子靶标。