A systematic test of the relation of ASD heterogeneity to synaptic function

ASD 异质性与突触功能关系的系统测试

基本信息

批准号：
7842915
负责人：
ROBERT C MALENKA
金额：
$ 89.8万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2011-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7842915
关键词：
Accounting Acute Address Affect Autistic Disorder Biological Biological Assay Brain Candidate Disease Gene Cell Survival Cells Classification Clinical Cognition Disorders Communication Communities Complex Copy Number Polymorphism Data Effectiveness Electroporation Exhibits Funding Gene Deletion Gene Duplication Gene Mutation Gene Silencing Generations Genes Genetic Goals Heritability Heterogeneity Hippocampus (Brain)Homologous Gene Impairment Injection of therapeutic agent Laboratories Libraries Measures Molecular Morphology Mus Mutation Neurobiology Neurons Optics Other Genetics Pathogenesis Pathway interactions Patients Phenotype Physiological Process Proteins Publications Reagent Role Slice Subfamily lentivirinae Symptoms Synapses Techniques Testing Therapeutic Intervention Translating Variant Virus autism spectrum disorder base clinical phenotype combinatorial density design gain of function in utero in vivo knock-down neuron development overexpression public health relevance response small hairpin RNA synaptic function synaptogenesis tool tool development vector web site

项目摘要

DESCRIPTION (provided by applicant): Although autism spectrum disorders (ASDs) are highly heritable, ASDs are heterogeneous, and no single genetic cause contributes to ASDs in a large proportion of patients. Instead, heterogeneous genetic changes, including many single gene mutations and copy-number variations (CNVs) are found in ASDs. Thus, a key question is whether different genetic changes contribute to ASDs via multiple, independent, pathogenic pathways, or whether the various genetic changes in ASDs converge onto a single pathogenic pathway. Several independent mutations in genes encoding synaptic proteins, such as neurexin-1, neuroligins, and SHANK3, suggested that ASDs may generally involve an impairment of synaptic communication between neurons. However, most of the other genetic changes observed in ASDs have no known effect on synapses in fact, have no known effect on any brain function. Thus, the major goal of the present proposal is to conduct a large scale, systematic analysis of the synaptic effects of genetic changes in ASDs. The approach will be to over express (to mimic gene duplications) or knock down (to mimic gene inactivations) mRNAs corresponding to 81 ASD candidate genes, and to test the effect of these manipulations on synapses using standardized assays. Cell viability, neuronal development, synapse density and synapse function will be assessed in cultured mouse neurons using optical and electro-physiological assays that are well established in the PI's laboratories. Genes that were found to affect neuronal development, synapse formation, or synapse function in cultured neurons will be studied by the same manipulations in vivo after stereotaxic injection of lentiviruses into the mouse hippocampus, or after in utero electroporation. Changes in synapse function and plasticity will then be examined in acute slices from these mice using standard electrophysiological techniques well established in the PI's laboratories. All results from this project will be posted on a dedicated public website, and all reagents generated will be made readily available to the scientific community. The results of this project will provide a standardized reference point for the function of ASD candidate genes, and provide an initial test of the hypothesis that despite their clinical and genetic heterogenity, ASDs involve a common, if diverse, pathway acting on synaptic communication in the brain. PUBLIC HEALTH RELEVANCE: Autism spectrum disorders are known to be clinically and genetically heterogeneous, but it is unclear whether these two types of heterogeneity are related, and how specifically the various genetic changes affect brain function. This project will address these issues by studying the changes in neuron-to-neuron communication caused by the genes associated with autism.

描述（由申请人提供）：尽管自闭症谱系障碍（ASDS）是高度遗传的，但ASD是异质的，并且没有任何遗传原因在很大一部分患者中会造成ASD的贡献。取而代之的是，在ASD中发现了异质遗传变化，包括许多单个基因突变和拷贝数变化（CNV）。因此，一个关键的问题是，不同的遗传变化是否通过多个，独立的，致病的途径导致ASD，或者ASDS中的各种遗传变化是否会融合到单个致病途径上。编码突触蛋白的基因中的几个独立突变，例如Neurexin-1，Neuroligins和Shank3，表明ASD通常涉及神经元之间突触通信的损害。但是，在ASD中观察到的大多数其他遗传变化实际上对突触没有已知影响，对任何大脑功能尚无影响。因此，本提案的主要目标是对ASDS遗传变化的突触作用进行大规模的系统分析。该方法将是过度表达（模仿基因的重复）或敲低（模仿基因失活的）mRNA，与81个ASD候选基因相对应，并使用标准化测定法测试这些操纵对突触的影响。细胞活力，神经元发育，突触密度和突触功能将在PI实验室中良好确定的光学和电生物学测定法中评估在培养的小鼠神经元中。发现培养神经元中神经元发育，突触形成或突触功能的基因将通过体内的同一操作在体内进行体内进行研究，而慢病毒将慢病毒注射到小鼠海马中，或者在子宫型电穿孔中。然后，将使用PI实验室中确定的标准电生理技术在这些小鼠的急性切片中检查突触功能和可塑性的变化。该项目的所有结果都将发布在专用的公共网站上，并且生成的所有试剂将易于为科学界提供。该项目的结果将为ASD候选基因的功能提供标准化的参考点，并提供了以下假设的初步检验：尽管具有临床和遗传异质性，但ASD涉及一种对大脑突触通信作用的常见途径。公共卫生相关性：已知自闭症谱系在临床和遗传上是异质性的，但目前尚不清楚这两种异质性是否相关，以及各种遗传变化如何影响大脑功能。该项目将通过研究与自闭症相关的基因引起的神经元间通信的变化来解决这些问题。