Investigating role of neurexin-1 mutation in autism using human induced neurons

使用人类诱导神经元研究 neurexin-1 突变在自闭症中的作用

• 批准号: 8594881
• 负责人: Changhui Pak
• 金额: $ 4.92万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-30 至 2016-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8594881
• 关键词: Accounting; Address; Affect; Autistic Disorder; Behavior; Biological Assay; Brain; Cell Adhesion Molecules; Cell Line; Cells; Child; Communication; Complex; Coupled; Data; Defect; Development; Diagnosis; Disease; Environment; Etiology; Event; Excitatory Synapse; Exhibits; Frequencies; Functional disorder; Gene Expression; Gene Targeting; Generations; Genes; Genetic; Human; Human Genetics; Image Analysis; Knock-out; Lead; Life; Maintenance; Measurement; Measures; Mediating; Messenger RNA; Molecular; Monitor; Mutate; Mutation; N-Methyl-D-Aspartate Receptors; Neurobiology; Neurodevelopmental Disorder; Neurons; Pathogenesis; Patients; Phenotype; Physiologic pulse; Prosencephalon; Protocols documentation; Research; Reverse Transcriptase Polymerase Chain Reaction; Role; Schizophrenia; Social Interaction; Staining method; Stains; Symptoms; Synapses; Synapsins; Synaptic Transmission; Synaptic Vesicles; Testing; Training; autism spectrum disorder; base; career; density; effective therapy; embryonic stem cell; excitatory neuron; genome-wide; homologous recombination; human embryonic stem cell; immunoreactivity; loss of function mutation; mouse model; multidisciplinary; mutant; neuron development; neuropsychiatry; neurotransmitter release; new therapeutic target; novel therapeutics; postsynaptic; presynaptic; public health relevance; receptor; recombinase; research study; response; stem cell biology; synaptic function; synaptogenesis; treatment strategy; uptake

DESCRIPTION (provided by applicant): The object of this proposal is to enhance our limited understanding of the pathophysiology of autism spectrum disorders (ASDs), which is a devastating neurodevelopmental condition affecting 1 in 88 US children. This will be accomplished by studying human neurons induced from human embryonic stem cells (ESCs) that are conditionally knock out for neurexin-1 (Nrxn1), a gene that is frequently mutated in patients with ASDs. The synaptic role of Nrxn1 and its functional contribution to autism in human neuronal context has not been studied, which makes this study unique. Preliminary data indicates the feasibility of this approach, whereby ESCs conditionally mutant for Nrxn1 locus has been generated. Using these cells, human excitatory neurons will be generated whereby Nrxn1 gene will be conditionally knocked out during neuronal development. Based on the hypothesis that Nrxn1 is required for proper synaptic function in human neurons, I will compare synapse development and function (synaptic transmission) in control and Nrxn1 mutant human neurons. Nrxn1 function in synapse development will be monitored by morphological studies examining synapse density and size as well as synaptic vesicle cycling. Using, electrophysiolgical assays, I will assess both pre and postsynaptic functions of Nrxn1 in mediating synaptic transmission at human excitatory synapses. In total, these experiments attempt to determine the mechanism by which mutated Nrxn1 influence human synaptic transmission and as a consequence, contribute to autism, using a combination of experimental approaches that assess synaptic function. !

描述（由申请人提供）：该提案的目的是增强我们对自闭症谱系障碍（ASDS）病理生理学的有限​​理解，这是一种毁灭性的神经发育状况，影响了88名美国儿童中的1个。这将通过研究由人类胚胎干细胞（ESC）诱导的人类神经元来实现，这些神经元有条件地将神经毒素1（NRXN1）敲定，这是一种经常在ASDS患者中突变的基因。尚未研究NRXN1的突触作用及其对自闭症的功能贡献，这使得这项研究与众不同。初步数据表明了这种方法的可行性，从而生成了NRXN1基因座的有条件的ESC突变体。使用这些细胞，将产生人类兴奋性神经元，从而在神经元发育过程中有条件地淘汰NRXN1基因。基于以下假设：NRXN1是人类神经元中适当突触功能所必需的，我将比较对照和NRXN1突变体神经元中突触的发育和功能（突触传播）。 NRXN1突触发育中的功能将通过检查突触密度和大小以及突触囊泡循环的形态学研究来监测。使用电生理测定，我将评估NRXN1在人类兴奋性突触中介导突触传播中NRXN1的前后功能。总的来说，这些实验试图确定突变的NRXN1影响人类突触传播的机制，因此，使用评估突触功能的实验方法的组合，从而有助于自闭症。呢