Functional Studies of Variants of the Bipolar Risk Gene CACNA1C in Human Neurons

人类神经元中双向情感障碍风险基因 CACNA1C 变体的功能研究

基本信息

批准号：
8550832
负责人：
Jen Qian Pan
金额：
$ 20.76万
依托单位：
BROAD INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-25 至 2016-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8550832
关键词：
12p13 Address Adoption Adverse effects Affect Alleles Animals Behavioral Biochemical Biological Models Bipolar Disorder Brain Brain Diseases Brain imaging Calcium Calcium Channel Cell Line Cell Survival Cell membrane Cell physiology Cells Chemicals Chromosomes, Human, Pair 12 Chronic Code Complex Data Disease Drug Targeting Ectopic Expression Engineering Exons Exploratory/Developmental Grant Family Fibroblasts Functional RNA Functional disorder Future Gene Expression Genes Genetic Genetic Polymorphism Genetic Risk Genetic Variation Genome Genomics Genotype Haplotypes Heart Hereditary Disease Human Hypertension Impaired cognition In Vitro Individual Introns L-Type Calcium Channels L-type calcium channel alpha(1C)Lead Life Link Lithium Major Depressive Disorder Manic Mental Depression Mental disorders Meta-Analysis Modeling Molecular Mood Disorders Mus Muscle Contraction Mutation Neurons Organ Patch-Clamp Techniques Patients Phenotype Play Population Predisposition Prevalence Process Recurrence Research Personnel Risk Role Sampling Schizophrenia Second Messenger Systems Signal Transduction Single Nucleotide Polymorphism Site Susceptibility Gene Synapses Techniques Technology Testing Time Timothy syndrome Transcription Coactivator Twin Multiple Birth United States Variant Wit Work base cell growth cohort density disorder risk gain of function genome wide association study genome-wide high risk human ASCL1 protein human disease insight neurotransmitter release novel novel therapeutics nuclease painful neuropathy psychogenetics risk variant second messenger severe mental illness tool transcription factor valproate voltage

12p13 Address Adoption Adverse effects Affect Alleles Animals Behavioral Biochemical Biological Models Bipolar Disorder Brain Brain Diseases Brain imaging Calcium Calcium Channel Cell Line Cell Survival Cell membrane Cell physiology Cells Chemicals Chromosomes, Human, Pair 12 Chronic Code Complex Data Disease Drug Targeting Ectopic Expression Engineering Exons Exploratory/Developmental Grant Family Fibroblasts Functional RNA Functional disorder Future Gene Expression Genes Genetic Genetic Polymorphism Genetic Risk Genetic Variation Genome Genomics Genotype Haplotypes Heart Hereditary Disease Human Hypertension Impaired cognition In Vitro Individual Introns L-Type Calcium Channels L-type calcium channel alpha(1C)Lead Life Link Lithium Major Depressive Disorder Manic Mental Depression Mental disorders Meta-Analysis Modeling Molecular Mood Disorders Mus Muscle Contraction Mutation Neurons Organ Patch-Clamp Techniques Patients Phenotype Play Population Predisposition Prevalence Process Recurrence Research Personnel Risk Role Sampling Schizophrenia Second Messenger Systems Signal Transduction Single Nucleotide Polymorphism Site Susceptibility Gene Synapses Techniques Technology Testing Time Timothy syndrome Transcription Coactivator Twin Multiple Birth United States Variant Wit Work base cell growth cohort density disorder risk gain of function genome wide association study genome-wide high risk human ASCL1 protein human disease insight neurotransmitter release novel novel therapeutics nuclease painful neuropathy psychogenetics risk variant second messenger severe mental illness tool transcription factor valproate voltage

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项目摘要

DESCRIPTION (provided by applicant): This is a new R21 application to study in live human neurons the function of a bipolar disorder susceptibility gene encoding a neuronal calcium channel. Calcium ion channels are expressed in all excitable and many non-excitable cells to transduce electric signals at the cell membrane into chemical potentials of calcium influx. Calcium is a second messenger whose intracellular concentration controls many cellular functions including muscle contraction, neurotransmitter release, cell survival and growth. Calcium ion channels are important drug targets for hypertension and neuropathic pain. Coding mutations in the CACNA1C calcium ion channel gene cause a rare hereditary disorder called Timothy Syndrome, affecting the heart, the brain, and other organs. Bipolar disorder is a chronic and severe mental illness with 1-2% lifetime prevalence in population of the United States. Individuals with bipolar disorder suffer from recurrent episodes of mania and depression, and current bipolar treatments, such as lithium, are often ineffective in many patients. Treatment options for bipolar disorder are unlikely to advance substantially until the underlying molecular processes in pathogenic risk are better understood. There is a clear heritable risk in bipolar disorder, but it has been difficult to establish clear risk associations in any particular gene wit bipolar disorder. Large-scale genome-wide association studies in three independent cohorts and a large meta-analysis have revealed a strong and consistent association between bipolar disorder susceptibility and a genomic locus on chromosome 12 in the CACNA1C gene. Interestingly, the same genomic region was found to be associated with schizophrenia and major depression susceptibility last year, making it one of the most replicable and consistent associations in psychiatric genetics. These exciting genetic discoveries present unique opportunities to study how genomic variations in the CACNA1C gene may influence neuronal calcium channel activity. Recently, mouse and human fibroblasts have been directly converted to functional neuronal cells through ectopic expression of three transcription factors (ASCL1, MYT1L, BRN2). This technological breakthrough makes human-specific in vitro neuronal models possible. Transcription activator-like effector nuclease (TALEN) technology is a molecular engineering tool that recognizes unique sequences in a genomic locus, introduces site-specific changes in the native genome, and helps elucidate genotype-phenotype relationships in an isogenic background. We will use genome editing, together with a combination of gene expression, biochemical, and electrophysiological analyses, to reveal the functional role of intronic single nucleotide polymorphism (SNP) variations of CACNA1C in controlling L-type calcium channel activity in human induced neurons. Our work has the potential to provide novel insights into the molecular mechanism of a genetic risk locus for psychiatric illness and to validate a systematic approach to study the functional consequences of genetic variations in complex brain disorders.

描述（由申请人提供）：这是一种新的R21应用，用于研究活人神经元中的躁郁症敏感性基因编码神经元钙通道的功能。钙离子通道在所有令人兴奋的和许多不可驱行的细胞中表达，以将电信在细胞膜上转移到钙涌入的化学电位中。钙是第二个使者，其细胞内浓度控制许多细胞功能，包括肌肉收缩，神经递质释放，细胞存活和生长。钙离子通道是高血压和神经性疼痛的重要药物靶标。 CACNA1C钙离子通道基因中的编码突变引起一种罕见的遗传疾病，称为蒂莫西综合征，影响心脏，大脑和其他器官。躁郁症是一种慢性且严重的精神疾病，在美国人群中，终生患病率为1-2％。患有躁郁症的个体患有躁狂和抑郁症的复发发作，当前的双极治疗（例如锂）在许多患者中常常无效。躁郁症的治疗选择不可能实质性地进展，直到更好地理解致病风险中的基本分子过程。双相情感障碍有明显的遗传风险，但是很难在任何特定的基因双相情感障碍中建立明显的风险关联。在三个独立队列和大型荟萃分析中的大规模基因组关联研究表明，躁郁症的敏感性与CACNA1C基因中12号染色体的基因组基因座之间存在很强的一致性。有趣的是，去年发现相同的基因组区域与精神分裂症和重大抑郁易感性有关，使其成为精神遗传学中最可复制和一致的关联之一。这些令人兴奋的遗传发现提供了独特的机会来研究CACNA1C基因中基因组变异如何影响神经元通道活性。最近，小鼠和人的成纤维细胞已通过异位表达三个转录因子（ASCL1，MYT1L，BRN2）直接转化为功能性神经元细胞。这一技术突破使人类特异性的体外神经元模型成为可能。转录活化剂样效应子核酸酶（TALEN）技术是一种分子工程工具，可以识别基因组基因座中的独特序列，引入天然基因组中的位点特异性变化，并有助于阐明同基因性背景中的基因型 - 型 - 原型型关系。我们将使用基因组编辑以及基因表达，生化和电生理分析的组合来揭示CACNA1C内含子单核苷酸多态性（SNP）变化在控制人类诱导Neurons中L型钙通道活性中的功能作用。我们的工作有可能提供有关精神病遗传风险基因座的分子机制的新见解，并验证一种系统的方法来研究复杂脑疾病中遗传变异的功能后果。