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 神经氧蛋白1（NRXN1）的杂合缺失，一种突触前细胞粘附分子，与 神经精神疾病和精神病。 NRXN1是高度剪接的，最近是这些作用 已经提出了在神经元同一性中的剪接同工型。但是，尚未表征NRXN1同工型 在人类神经元中。 NRXN1缺失显示的动物模型在行为和神经元活动中定义。然而， 动物模型无法在其他风险的情况下概括众多患者特定缺失的影响 患者基因组中的变体；这些效果在一起可能会影响NRXN1的可变渗透率 删除，会议会议一组各种临床诊断。使用来自稀有队列的HIPSC-神经元 四名携带NRXN1缺失的精神病患者，该提案将调查这些建议的随意贡献 兴奋性HIPSC-神经元中的基因表达，替代剪接和神经元表型的缺失。那里 是三个目的：第一个是识别NRXN1删除HIPSC-NERORS中替代剪接的差异， 其次是恢复同工型缺陷并操纵HIPSC-神经元中的NRXN1表达，第三个是 建立hipsc-神经元中NRXN1缺失的功能意义。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NRXN1同工型的差异表达，这些表达 对于适当的神经元活性和形态至关重要。我们包括强大的初步数据，证明 我们的提议的可行性；此外，我们已经观察到NRXN1 HIPSC-Neurons表达 野生型和删除的等位基因变体。我们希望这些实验将提高我们对 神经精神疾病的遗传风险的分子机制。