Functional Dissection of CNVs in Neurodevelopmental Traits

神经发育特征中 CNV 的功能剖析

基本信息

批准号：
10366987
负责人：
Erica Ellen Davis
金额：
$ 55.39万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-03-01 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10366987
关键词：
16p11.2 19p13.1 Affect Animal Model Area Attention Biological Biological Assay Biology Cartilage Cells Collaborations Complex Copy Number Polymorphism Data Data Set Disease Disease susceptibility Dissection Equilibrium Face Funding Gene Combinations Gene Dosage Gene Expression Profile Genes Genetic Genetic Epistasis Genetic Transcription Genomics Histone Deacetylase Human Human Pathology Individual Knowledge Maps Measures Methods Modeling Molecular Mus Mutation Neurodevelopmental Disorder Neurons Pathogenicity Pathology Pathway Analysis Pathway interactions Pharmaceutical Preparations Phenocopy Phenotype Physiological Probability Property Protocols documentation RORA gene Rattus Repressor Proteins Role Safety Syndrome Testing Therapeutic Therapeutic Intervention Transcript Transcription Repressor Transcriptional Regulation Translating Validation Work Zebrafish biological systems burden of illness cell type combinatorial craniofacial craniofacial development dosage drug candidate drug development drug discovery drug repurposing efficacy testing genetic architecture genetic corepressor genomic data genomic locus in vivo inhibitor/antagonist neurodevelopment novel phenotypic data safety testing therapeutic candidate therapeutic development tool trait transcriptome transcriptomics

项目摘要

Understanding the contribution of copy number variants (CNVs) to human pathology remains challenging, in part because of the complex and varied influence of loci within them. In some cases, a single dosage-sensitive locus drives pathology. In other examples, multiple genes contribute to phenotypes through additive effects and through complex genetic interactions. In the previous funding period, we focused on deletion syndromes and reciprocal CNVs and asked whether we could develop tools to aid the identification of driver genes. Progress in that work has informed several areas of pathology. For example, the identification of SIN3B as a driver of a non-recurrent 230 kb deletion on 19p13.1 that causes a complex neurodevelopmental phenotype reinforced the role of the Sin3/HDAC transcriptional corepressor complex in neurodevelopment and facilitated the identification of Mendelian mutations that phenocopy the CNV. In the case of the 17q24.2 CNV, we learned about the synthetic CNV phenotype likely caused by dosage imbalance of both BPTF and PSMD12. Finally, modeling of multiple phenotypes associated with the 16p11.2 CNV showed how: (a) key neurodevelopmental and craniofacial pathologies were conserved from humans, to mice, to rats, to zebrafish; (b) demonstrated discrete epistatic interactions between loci; and (c) generated a host of surrogate model organisms in which to perform both pathway analysis and drug discovery. Overall, the genetic and functional dissection of CNV exemplars has underscored three observations: (a) not all genes in a CNV are dosage sensitive (suspected, but now supported by experimental evidence); (b) some genes are dosage sensitive in one direction (gain, loss, but not both); and (c) non-driver genes can participate in distinct genetic interactions. The knowledge and experimental tools gained affords us the opportunity to expand our ambition. We still do not understand how intra-CNV combinatorial dosage influences phenotype, and whether the lessons learned from prior work are broadly generalizable. Moreover, given the non-trivial contribution of CNVs to disease burden, it is crucial that we pay attention to the development of therapeutic avenues. We propose three Aims: (1) We will dissect a unique set of CNVs, in which the phenotype is driven only by duplication to identify triplosensitive genes and ask whether disease susceptibility is driven by a threshold effect as a means to understanding why only a subset of human genes are dosage sensitive. (2) We will harness our existing zebrafish models to characterize the molecular properties of epistatic effects using the neuronal and facial cartilage phenotypes of the 16p11.2 CNV and ask whether we can detect epistasis at the level of transcriptional regulation. (3) We will combine our extensive repertoire of zebrafish models of CNV drivers with the Connectivity Map (CMap) Touchstone dataset and a newly developed safety/efficacy protocol to identify candidate therapeutics. Together, our studies will refine the genetic architecture of genomic disorders, inform the molecular basic of genetic interactions, and chart a systematic path toward therapeutics for neurodevelopmental traits.

了解拷贝数变体（CNV）对人类病理学的贡献仍然具有挑战性部分原因是基因座在其中的复杂和多样化的影响。在某些情况下，单剂量敏感基因座驱动病理学。在其他示例中，多个基因通过添加效应有助于表型并通过复杂的遗传相互作用。在上一个资金期间，我们专注于缺失综合症和相互的CNV，并询问我们是否可以开发工具来帮助识别驱动器基因。这项工作的进展为病理的几个领域提供了信息。例如，识别sin3b为一个 19p13.1上非旋转230 kb删除的驱动器，该驱动会导致复杂的神经发育表型加强了SIN3/HDAC转录核心复合物在神经发育中的作用并促进鉴定Mendelian突变，即CNV。对于17q24.2 CNV，我们学会了关于BPTF和PSMD12剂量不平衡引起的合成CNV表型。最后，与16p11.2 CNV相关的多种表型的建模显示：（a）关键神经发育从人到小鼠到大鼠，再到斑马鱼，颅面病理是保守的。（b）证明基因座之间的离散上任相互作用；（c）产生了许多替代模型生物同时执行途径分析和药物发现。总体而言，CNV的遗传和功能解剖示例强调了三个观察结果：（a）CNV中的所有基因并非剂量敏感（可疑，，但是现在得到实验证据的支持）；（b）某些基因在一个方向上对剂量敏感（增益，损失，但不是两者）；（c）非驱动器基因可以参与不同的遗传相互作用。知识和获得的实验工具为我们提供了扩大野心的机会。我们仍然不明白如何 CNV内组合剂量会影响表型，以及从先前工作中学到的经验教训是否是广泛的可推广。而且，鉴于CNV对疾病负担的非平凡贡献，至关重要的是我们关注治疗途径的发展。我们提出三个目标：（1）我们将剖析独特的CNV集，其中表型仅由重复驱动以识别三度敏感基因和询问疾病的易感性是否是由阈值效应驱动的，以此作为理解为什么只有人类基因的子集对剂量敏感。（2）我们将利用现有的斑马鱼模型来表征使用16p11.2的神经元和面部软骨表型的上皮作用的分子特性 CNV并询问我们是否可以在转录调控水平上检测上毒。（3）我们将结合我们的 CNV驱动程序的斑马鱼模型与连接图（CMAP）Touchstone数据集的广泛曲目以及新开发的安全/功效协议，以识别候选治疗疗法。一起，我们的研究将完善基因组疾病的遗传结构，告知遗传相互作用的分子基础，并为神经发育特征的治疗学途径绘制了系统的途径。