Characterization of hiPSC-neurons from psychosis patients with neurexin-1 deletions

Neurexin-1 缺失精神病患者 hiPSC 神经元的表征

• 批准号: 9328489
• 负责人: Erin Flaherty
• 金额: $ 4.39万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-04 至 2020-04-03
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328489
• 关键词: Affect; Alternative Splicing; Animal Model; Behavior; Bipolar Disorder; Cell Adhesion Molecules; Cell Line; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Data; Delusions; Engineering; Exhibits; Exons; Foundations; Gene Expression; Genetic; Genetic Risk; Genetic Transcription; Genome; Goals; Hallucinations; Heritability; Human; Individual; Link; Mediating; Messenger RNA; Modeling; Molecular; Morphology; Mus; Mutation; Neurodevelopmental Disorder; Neurons; Patients; Pattern; Penetrance; Phenotype; Population; Prefrontal Cortex; Protein Isoforms; Psychotic Disorders; RNA Splicing; Risk; Role; Schizophrenia; Symptoms; Synaptic Transmission; Testing; Time; Tissue-Specific Gene Expression; Transcript; Up-Regulation; Work; autism spectrum disorder; base; clinical Diagnosis; cohort; differential expression; excitatory neuron; experience; experimental study; genetic risk factor; genetic variant; improved; induced pluripotent stem cell; neuropsychiatric disorder; neuropsychiatric symptom; neurotransmitter release; novel; nuclease; presynaptic; risk variant; small molecule; synaptic function; targeted sequencing

Project Summary Heterozygous deletion of neurexin-1 (NRXN1), a presynaptic cell adhesion molecule, is strongly associated with neuropsychiatric disorders and psychosis. NRXN1 is highly alternatively spliced and recently a role for these splice isoforms in neuronal identity has been suggested; however, NRXN1 isoforms have not been characterized in human neurons. Animal models of NRXN1 deletion display deficits in behavior and neuronal activity. However, animal models cannot recapitulate the effect of myriad patient-specific deletions in the context of other risk variants within a patient’s genome; together, these effects likely influence the variable penetrance of NRXN1 deletions, which confer a diverse set of clinical diagnoses. Using hiPSC-neurons derived from a rare cohort of four psychosis patients carrying NRXN1 deletions, this proposal will investigate the casual contribution of these deletions on gene expression, alternative splicing and neuronal phenotypes in excitatory hiPSC-neurons. There are three aims: the first is identify differences in alternative splicing in NRXN1 deletion hiPSC-neurons, the second is to restore isoform deficiencies and manipulate NRXN1 expression in hiPSC-neurons, and the third is to establish the functional significance of NRXN1 deletion in hiPSC-neurons. My hypothesis is that excitatory hiPSC-neurons derived from four psychosis patients will exhibit differential expression of NRXN1 isoforms which are crucial for proper neuronal activity and morphology. We include strong preliminary data demonstrating the feasibility of our proposal; moreover, we have already observed that NRXN1 hiPSC-neurons express both wildtype and deleted allele variants. Our hope is that these experiments will improve our understanding of the molecular mechanisms underlying genetic risk for neuropsychiatric disorders.

项目摘要 突触前细胞粘附分子neurexin-1（NRXN 1）的杂合缺失与 神经精神障碍和精神病。NRXN 1是高度选择性剪接的，最近在这些细胞中起作用。 已经提出了神经元身份中的剪接异构体;然而，NRXN 1异构体尚未被表征 在人类神经元中。NRXN 1缺失的动物模型显示行为和神经元活性的缺陷。然而，在这方面， 动物模型不能概括大量患者特异性缺失在其他风险背景下的作用， 患者基因组内的变异;这些效应可能共同影响NRXN 1的可变突变率。 缺失，这赋予了一组不同的临床诊断。使用来自一个罕见的hiPSC-神经元的队列， 四名携带NRXN 1缺失的精神病患者，这项提案将调查这些患者的偶然贡献。 在兴奋性hiPSC神经元中基因表达、选择性剪接和神经元表型的缺失。那里 有三个目的：第一个是确定NRXN 1缺失hiPSC神经元中选择性剪接的差异， 第二是恢复同种型缺陷并操纵hiPSC神经元中的NRXN 1表达，第三是 建立NRXN 1缺失在hiPSC神经元中的功能意义。我的假设是兴奋性 来自四名精神病患者的hiPSC-神经元将表现出NRXN 1同种型的差异表达， 对正常的神经元活动和形态至关重要。我们包括强有力的初步数据证明， 此外，我们已经观察到，NRXN 1 hiPSC神经元表达 野生型和缺失的等位基因变体。我们希望这些实验能够提高我们对 神经精神疾病遗传风险的分子机制