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

• 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
• 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

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.

摘要 互惠基因组病(RGDS)涉及重复的完全相同基因组的微缺失和微复制 细分市场。RGD由非等位基因同源重组(Nahr)介导，共同存在于 神经发育障碍(NDD)和先天畸形最常见的复发遗传原因 在人类身上。鉴于RGDS的影响通常是在发育早期，这些疾病不成比例地 影响儿童，并往往导致终生残疾。分子后果基因的发现 因此，RGDS和构成这些疾病组成部分的基因将代表异常高的 机制研究和治疗靶向的优先靶点跨越孟德尔和复杂的光谱 精神错乱。我们的初步数据表明，在体外和体内的分子和计算集成 利用细胞和动物模型的基因组学方法可以识别与 RGDS和异常表型和网络失调的遗传驱动因素。在这些研究中，我们将 首先定义与微缺失和微缺失相关的基因表达谱和细胞表型 同源基因诱导的神经衍生物中最常见的8-12个RGD区的微复制 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点 最近开发的针对侧翼片段复制和模拟Nahr介导的机制 在人类中(目标1)。然后我们将寻找与RGD相关表型相关的特定基因 斑马鱼高通量驱动基因筛选(目标2)评估所有单个基因和成对分析 RGD区域内的相互作用。在目标3中，我们将尝试验证这些预测的驱动因素，并确定 它们对不同的神经细胞谱系和小鼠组织的影响。因此，这些研究将遵循一个框架 我们的研究团队此前曾用来确定非复发性微缺失综合征的基因驱动因素 和几个RGD地区，包括16p11.2 RGD，并应用创新的方法和技术来 使我们能够大规模地进行这些研究，并比较不同RGD的签名。在他们的结论中，Tehse 分析将定义人类最常见的RGD的分子特征，即驱动 这些信号的特定成分，它们的组织特异性，以及抢救最强者的能力 通过体外和体内剂量操作的签名。