Molecular mechanisms and genetic drivers of reciprocal genomic disorders

相互基因组疾病的分子机制和遗传驱动因素

基本信息

批准号：
9982392
负责人：
MICHAEL E TALKOWSKI
金额：
$ 69.6万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9982392
关键词：
16p11.2 22q11.2 Ablation Affect Animal Model Architecture Biological Models CRISPR/Cas technology CRKL gene Cell Line Cell model Child Clustered Regularly Interspaced Short Palindromic Repeats Companions Complex Computer Models Critical Pathways DNA Sequence Alteration Data Defect Development Diagnosis Disease Disease model Dose Engineering Expression Profiling Gene Combinations Gene Expression Gene Expression Profile Genes Genetic Genetic Transcription Genome Genomic Segment Genomic approach Genomics Goals Grant Human In Vitro Individual Kidney Mediating Modeling Molecular Molecular Computations Molecular Profiling Mus Nature Neurodevelopmental Disorder Neurons Pathogenicity Pathway interactions Phenotype Population Prevention Process Recurrence Reproducibility Resources Specificity Syndrome Technology Testing Therapeutic Tissues Validation Zebrafish base common treatment comorbidity congenital anomaly disability dosage gene discovery genome editing homologous recombination improved in vivo in vivo Model induced pluripotent stem cell innovation insight microdeletion molecular phenotype mouse model nerve stem cell neuropsychiatry novel relating to nervous system screening therapeutic target trait transcriptome

项目摘要

ABSTRACT Reciprocal genomic disorders (RGDs) involve recurrent microdeletion and microduplicaton of identical genomic segments. RGDs are mediated by non-allelic homologous recombination (NAHR) and are collectively among the most common recurrent genetic causes of neurodevelopmental disorders (NDD) and congenital anomalies in humans. Given that the impact of RGDs is usually early in development, these disorders disproportionately affect children and often result in lifelong disabilities. Discovery of the genes the molecular consequences of RGDs and the genes that underlie components of these disorders would therefore represent exceptionally high priority targets for mechanistic studies and therapeutic targeting across a spectrum of Mendelian and complex disorders. Our Preliminary Data suggest that an integrated in vitro and in vivo molecular and computational genomics approach using cellular and animal modeling can identify molecular signatures associated with RGDs and the genetic drivers of aberrant phenotypes and dysregulated networks. In these studies, we will first define the gene expression profiles and cellular phenotypes associated with microdeletion and microduplication of the 8-12 most prevalent RGD regions in neural derivatives from isogenic induced pluripotent stem cell (iPSC) lines. We will accomplish this using a CRISPR/Cas9 genome editing approach we recently developed that targets the flanking segmental duplications and mimics NAHR-mediated mechanisms in humans (Aim 1). We will then seek the specific genes associated with RGD-associated phenotypes using high-throughput driver gene screening in zebrafish (Aim 2) to evaluate all individual genes and pairwise interactions within RGD regions. In Aim 3 we will then seek to validate these predicted drivers and determine their impact in diverse neuronal lineages and across mouse tissues. These studies will thus follow a framework our investigative team has previously used to identify genetic drivers of non-recurrent microdeletion syndromes and several RGD regions, including 16p11.2 RGD, and apply innovative approaches and technologies to enable us to conduct these studies at scale and compare signatures across RGDs. At their conclusion, tehse analyses will define the molecular signatures of the most common RGDs in humans, the genes that drive specific components of these signatures, their tissue specificity, and the capacity to rescue the strongest signatures through dosage manipulation in vitro and in vivo.

抽象的相互基因组疾病（RGD）涉及相同基因组的反复微缺失和微重复段。 RGD 由非等位基因同源重组 (NAHR) 介导，并且统称为神经发育障碍 (NDD) 和先天性异常最常见的复发性遗传原因在人类中。鉴于 RGD 的影响通常发生在发育早期，这些疾病不成比例地影响儿童并常常导致终身残疾。基因的发现及其分子后果因此，RGD 和构成这些疾病成分的基因将代表异常高的水平。跨孟德尔和复杂谱系的机制研究和治疗靶向的优先目标失调。我们的初步数据表明，集成的体外和体内分子和计算使用细胞和动物模型的基因组学方法可以识别与 RGD 以及异常表型和失调网络的遗传驱动因素。在这些研究中，我们将首先定义与微缺失相关的基因表达谱和细胞表型等基因诱导的神经衍生物中 8-12 个最常见的 RGD 区域的微复制多能干细胞 (iPSC) 系。我们将使用 CRISPR/Cas9 基因组编辑方法来实现这一目标最近开发的针对侧翼节段重复并模仿 NAHR 介导的机制人类（目标 1）。然后，我们将使用以下方法寻找与 RGD 相关表型相关的特定基因：对斑马鱼进行高通量驱动基因筛选（目标 2），以评估所有个体基因和成对基因 RGD 区域内的相互作用。在目标 3 中，我们将寻求验证这些预测的驱动因素并确定它们对不同神经元谱系和小鼠组织的影响。因此，这些研究将遵循一个框架我们的研究团队之前曾用于识别非复发性微缺失综合征的遗传驱动因素和几个 RGD 区域，包括 16p11.2 RGD，并应用创新方法和技术使我们能够大规模地进行这些研究并比较 RGD 之间的特征。在他们的结论中，tese 分析将定义人类最常见 RGD 的分子特征，即驱动基因这些特征的具体组成部分、它们的组织特异性以及拯救最强者的能力通过体外和体内剂量操作来识别特征。