Genomic Risk Variants in Orofacial Clefting: Discovery and Functional Validation

摘要 口面部裂(OFCs，包括唇裂-CL、腭裂-CP或两者-CLP)是最常见的 人类的头面部异常，全世界大约每700个新生儿中就有一个受到影响，因此是 最常见的结构性出生缺陷。由于以下原因，离岸金融中心对公众健康有重大影响 相关的发病率和死亡率。患有OFC的普通儿童最初面临喂养困难，经历了 6次手术，住院30天，接受5年正畸治疗，并参与正在进行的 言语治疗，导致估计终身治疗总成本约为20万美元。此外，个人 患有强迫症的人有更高的心理健康问题发生率，在各个阶段的死亡率都更高。 生活风险和其他疾病(特别是乳腺癌、脑癌和结肠癌)的风险更高，以及婴儿更高 死亡率(特别是在获得医疗保健的机会可能有限的发展中国家)。离岸金融中心 病因复杂，由遗传变异、环境暴露及其相互作用引起。基因组 广泛的关联研究与测序结果相结合，已经识别出至少35个基因/区域 来自多个独立研究的全基因组意义，但这些结果只占 遗传力。随着全基因组测序(Wgs)的出现，罕见的变异研究正在涌现。 数据集，但罕见变体的功能验证对于生成链接这些变量所需的支持至关重要 OFC的基因/变异，以了解OFC背后的生物学，将关联信号转换为准确的 风险预测，并最终开发和/或改进治疗。这个新项目的总体目标是 协作项目是确定在我们的大型项目中确定的罕见风险变量的功能重要性 OFC家庭和控制的资源。这个新项目的拟议目标将有助于实现我们的总体目标 进球。我们将利用我们的OFC WGS资源(2,078个OFC病例/父母三人组)来发现新的基因组风险 具有创新分析的变体，强调单核苷酸变体(SNV)、结构变体(SVS)和 Indels。对于功能验证，这个新项目将利用 CRISPR/Cas9技术：杰克逊实验室的研究团队成员开发了一种新的 一种快速验证小鼠中假定的致病变异的方法，从而使哺乳动物能够使用 用于变异验证和对由此产生的裂隙表型进行更详细调查的系统。

ABSTRACT Orofacial clefts (OFCs, comprising cleft lip-CL, cleft palate-CP, or both-CLP) are the most common craniofacial anomalies in humans, affecting approximately 1 in 700 newborns worldwide, and are thus one of the most common structural birth defects. There are substantial public health impacts of OFCs, due to associated morbidity and mortality. An average child with an OFC initially faces feeding difficulties, undergoes 6 surgeries, spends 30 days in hospital, receives 5 years of orthodontic treatment, and participates in ongoing speech therapy, leading to an estimated total lifetime treatment cost of about $200,000. Further, individuals born with an OFC have an increased incidence of mental health problems, higher mortality rates at all stages of life and higher risk for other disorders (notably including breast, brain, and colon cancers), and higher infant mortality (particularly in developing countries where access to medical care may be limited). OFCs are etiologically complex, resulting from genetic variants, environmental exposures, and their interactions. Genome wide association studies coupled with sequencing results have identified at least 35 genes/regions achieving genome-wide significance from multiple independent studies but these results only account for a fraction of heritability. Rare variant studies are emerging with the availability of whole genome sequencing (WGS) datasets, but functional validation of rare variants is essential to generate the support necessary to link these genes/variants to OFCs, to understand the biology behind OFCs, to translate association signals into accurate risk predictions, and ultimately to develop and/or improve therapies. The overall goal of this new collaborative project is to identify the functional significance of rare risk variants identified in our large resource of OFC families and controls. The proposed aims of this new project will help achieve our overall goal. We will utilize our OFC WGS resources (2,078 OFC case/parent trios) to discover new genomic risk variants with innovative analyses emphasizing single nucleotide variants (SNVs), structural variants (SVs), and indels. For functional validation, this new project will take advantage of the high-efficiency and flexibility of CRISPR/Cas9 technology: research team members at the Jackson Laboratory have developed a novel approach to rapidly validate putative causative variants in the mouse, thus enabling the use of a mammalian system for both variant validation and for more detailed investigation of the resulting cleft phenotypes.