Somatic Mosaicism in the Brain of Tourette Syndrome

抽动秽语综合症大脑中的体细胞镶嵌

• 批准号: 9237729
• 负责人: FLORA M VACCARINO
• 金额: $ 23.23万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-10 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9237729
• 关键词: Affect; Autopsy; Basal Ganglia; Behavior; Biological Process; Blood; Blood specimen; Brain; Brain region; CRISPR/Cas technology; Cell Differentiation process; Cell Lineage; Cell Nucleus; Cell Survival; Cells; Cerebral cortex; Code; Conserved Sequence; Copy Number Polymorphism; Corpus striatum structure; Cortical Column; DNA; DNA Insertion Elements; DNA Sequence; DNA Transposable Elements; Data; Development; Disease; Dorsal; Embryonic Development; Engineering; Failure; Fibroblasts; Frequencies; Gene Expression; Genes; Genome; Genomic DNA; Genomic Segment; Genotype; Gilles de la Tourette syndrome; Health; Human; Human body; Individual; Informatics; Interneurons; Microglia; Mitotic; Molecular; Mosaicism; Mus; Mutation; Neurons; Nucleotides; Organoids; Pathogenesis; Patients; Phenotype; Population; Prefrontal Cortex; Process; Reporting; Retrotransposon; Sequence Analysis; Skin; Somatic Mutation; Sorting - Cell Movement; Techniques; Telencephalon; Testing; Tissues; Transgenic Mice; Variant; base; brain cell; brain tissue; cholinergic; digital; genetic risk factor; genetic variant; genome sequencing; genomic variation; hippocampal pyramidal neuron; induced pluripotent stem cell; insertion/deletion mutation; mind control; morphometry; mouse model; neuropsychiatric disorder; progenitor; protein function; targeted sequencing; transcriptome; whole genome; zygote

 DESCRIPTION (provided by applicant): Tourette Syndrome (TS) is a disorder of the developing telencephalon for which no significant causative genetic variant has yet emerged through the examination of blood samples. In this proposal we investigate whether somatic mutations might underlie in part the pathogenesis of TS. Existing evidence suggests that cells accumulate somatic mutations after the formation of the zygote, implying that cells of the human body do not have identical DNA sequence. Besides single nucleotide variation (SNV) and small insertion/deletions (InDels), cells can accumulate copy number variations (CNVs, i.e., duplications and deletions), insertions of transposable elements, inversions and translocations, all involving from few hundred to several millions of nucleotides. Somatic mosaicism arising in brain cells could explain the failure to discover consistent, replicable genetic risk factors in neuropsychiatric disorders like TS, and underlie at least in part the frequently observed variability between blood genotype and overall phenotype. There is no estimate of somatic mosaicism in either normal development or in disease. To test the hypothesis that somatic mutations might underlie the emergence of TS, in this proposal we will discover and quantify somatic genome variation in TS and normal control brains, followed by exploration of potential functional consequences of this variation. In Aim 1, we will perform using advanced sequencing techniques comprehensive discovery of lineage-specific and region-specific somatic genomic variations: SNVs, InDels, CNVs, retrotransposon insertions, inversion and translocations. The analysis will involve 20 TS brains and matched 20 normal control brains. Mosaic variants will be discovered and validated in prefrontal cortex (PFC), premotor cortex (PMC) and striatum (STR), three regions strongly implicated in TS, as well as in specific cell lineages isolated from these regions, including pyramidal neurons, medium spiny neurons, interneurons and microglial cells. In Aim 2, we will select 10 genomic variants, engineer them into iPSCs and in transgenic mice using CRISPR technologies, and characterize their impact on the molecular, tissue and behavior level. Together, these specific aims will provide the first estimate of somatic genomic variation (number, type, frequency) in the brain of TS and will yield hypotheses about their significance for brain development.

 描述（由申请人提供）：Tourette综合征（TS）是一种发育中的端脑疾病，通过血液样本检查尚未出现显著的致病遗传变异。在这个建议中，我们调查是否体细胞突变可能在TS的发病机制的一部分。现有证据表明，细胞在受精卵形成后积累体细胞突变，这意味着人体细胞不具有相同的DNA序列。除了单核苷酸变异（SNV）和小插入/缺失（InDel）之外，细胞可以积累拷贝数变异（CNV，即，复制和缺失）、转座因子的插入、倒位和易位，所有这些都涉及几百到几百万个核苷酸。脑细胞中出现的体细胞嵌合现象可以解释在TS等神经精神疾病中未能发现一致的、可复制的遗传风险因素，并且至少部分地解释了血液基因型和总体表型之间经常观察到的变异性。无论是在正常发育还是在疾病中，都没有体细胞嵌合现象的估计。为了验证体细胞突变可能是TS出现的基础这一假设，在本提案中，我们将发现并量化TS和正常对照大脑中的体细胞基因组变异，然后探索这种变异的潜在功能后果。在目标1中，我们将使用先进的测序技术全面发现谱系特异性和区域特异性的体细胞基因组变异：SNV，InDel，CNV，反转录转座子插入，倒位和易位。分析将涉及20个TS脑和匹配的20个正常对照脑。镶嵌变体将在前额叶皮层（PFC）、前运动皮层（PMC）和纹状体（STR）中被发现和验证，这三个区域与TS密切相关，以及在从这些区域分离的特定细胞谱系中，包括锥体神经元、中等多刺神经元、中间神经元和小胶质细胞。在目标2中，我们将选择10个基因组变体，使用CRISPR技术将它们改造成iPSC和转基因小鼠，并表征它们对分子，组织和行为水平的影响。总之，这些特定的目标将提供第一个估计的体细胞基因组变异（数量，类型，频率）在大脑中的TS，并会产生关于其意义的大脑发育的假设。