Rapid Identification and Validation of Human Craniofacial Development Genes

人类颅面发育基因的快速鉴定和验证

• 批准号: 9267961
• 负责人: RICHARD L MAAS
• 金额: $ 57.45万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-15 至 2019-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9267961
• 关键词: Affect; Alleles; Animal Model; Behavior; Boston; Candidate Disease Gene; Clinical Investigator; Collection; Communities; Congenital Abnormality; Consanguinity; DNA Sequence; Data; Data Set; Development; Disease; Enhancers; Ensure; Etiology; Face; FaceBase; Family; Family member; Fishes; Gene Expression; Gene Expression Profiling; Gene Targeting; Gene Transfer Techniques; Genes; Genetic; Genomic DNA; Genomics; Grant; Hospitals; Human; Incidence; Individual; Mus; Mutagenesis; Mutate; Mutation; Patient Recruitments; Patients; Pediatric Hospitals; Regulator Genes; Research; Research Infrastructure; Research Personnel; Resources; Saudi Arabia; Sequence Analysis; Specialist; Technology; TimeLine; Validation; Variant; Work; Zebrafish; base; biobank; biomedical referral center; comparative genomic hybridization; craniofacial; craniofacial complex; craniofacial development; cranium; developmental disease; exome; gene discovery; genome integrity; genome wide association study; knock-down; mouse model; mutant; new technology; novel; proband; public health relevance; success; whole genome

DESCRIPTION (provided by applicant): The advent of new genomic sequencing technologies has made the task of gene discovery in human developmental disorders highly efficient. Simultaneously, advances in gene targeting in model organisms, specifically in zebrafish, have made semi-high throughput validation and analysis of human candidate genes feasible, including those responsible for craniofacial disorders. This application for a new spoke project in FaceBase 2 will take advantage of this convergence of new technologies to identify and functionally validate approximately two dozen genes involved in novel aspects of human craniofacial development. Specifically, we will take advantage of already ascertained collections of craniofacial dysmorphoses from Boston Children's Hospital (BCH) and from King Faisal Specialist Hospital and Research Center (KFSHRC) in Saudi Arabia, where the high incidence of consanguinity makes autozygosity mapping and the identification of recessive causal loci highly feasible. We will extend the work of FaceBase beyond its current focus on disorders of palatal development by including a relatively wide range of craniofacial disorders that involve other components of the craniofacial complex. In addition, use of resources already compiled by FaceBase, including detailed gene expression data in mouse and zebrafish, enhancer analyses, and genome wide association studies, in combination with the present data and publicly available datasets, will further facilitate the functional annotation of these newly validated gene. To provide valuable deliverable resources to other FaceBase investigators and to the community at large, we will pursue three Specific Aims. In Aim 1, we will ascertain and recruit patients with a wide range of craniofacial dysmorphoses of likely monogenic etiology. These patients will not only be identified at the BCH and KFSHRC referral centers, but also solicited from other clinical investigators and potentially even the FaceBase Biorepository. In Aim 2, patients will be prioritized for further study based on the genetic likelihood of identifying a caual variant. We will then perform whole exome and in some cases whole genome sequence (WES/WGS) analysis, on the proband and potentially other family members, using aCGH to ensure genomic integrity and autozygosity mapping where applicable. An existing state-of-the-art computational pipeline will be used to derive a limited set of potentially causal DNA sequence variants and candidate genes. Lastly, in Aim 3, in cases where causation cannot be readily established from known function and expression data, we will seek additional independent confirmatory cases and, in parallel, employ a rapid analysis strategy consisting of high-throughput gene expression analysis, morpholino knockdown, and mutagenesis and transgenesis to prepare GOF and LOF alleles. The results will be forwarded to the FaceBase 2 Coordinating Center, with the key deliverables to the community being a validated gene list of human craniofacial developmental regulatory genes and a set of corresponding zebrafish mutants that can be widely shared for further detailed study.

描述（由申请人提供）：新的基因组测序技术的出现使在人类发育障碍中发现基因发现的任务非常有效。同时，模型生物（尤其是斑马鱼的基因靶向）的进展使人对人类候选基因的半高吞吐量验证和分析可行，包括负责颅面疾病的基因。此申请在新的讲话项目中 Facebase 2将利用这种新技术的这种融合来识别并在功能上验证大约二十二个基因参与人类颅面发育的新方面。具体来说，我们将利用已经确定的来自波士顿儿童医院（BCH）的颅面畸形收集，以及位于沙特阿拉伯的Faisal国王专家医院和研究中心（KFSHRC）的收集，在那里，高管的发病率很高，使自身的自动化映射和衰退的标识非常高。我们将通过包括涉及颅面复合物的其他成分的相对较广泛的颅面疾病，将面碱的工作扩展到其当前对帕拉塔尔发育障碍的关注。此外，使用面库已经编制的资源，包括小鼠和斑马鱼中的详细基因表达数据，增强子分析和基因组广泛的关联研究，以及当前数据和公开可用的数据集，将进一步促进这些新验证的基因的功能注释。为了向其他面部调查人员和整个社区提供宝贵的可交付资源，我们将追求三个具体目标。在AIM 1中，我们将确定并招募患有广泛的颅面畸形患者，可能是单基因病因。这些患者不仅将在BCH和KFSHRC转介中心鉴定，而且还可以从其他临床研究人员征求任何临床研究者，甚至可能是Facebase BioreSository的征求者。在AIM 2中，将根据鉴定CAUAL变异的遗传可能性来对患者进行进一步研究。然后，我们将对所有基因组序列（WES/WGS）分析进行整个外显子组和可能的其他家庭成员分析，并使用ACGH来确保在适用的情况下确保基因组完整性和自动合格映射。现有的最新计算管道将用于得出一组有限的潜在因果DNA序列变体和候选基因。最后，在目标3中，如果无法从已知功能和表达数据中轻易建立因果关系，我们将寻求其他独立的验证性案例，并同时采用由高通量基因表达分析，morpholino敲低以及诱变和转基因组成的快速分析策略来制备GOF和LOF。结果将转发到面底2协调中心，关键的可交付成果是人类颅面发育监管基因的经过验证的基因清单，以及一组相应的斑马鱼突变体，可以广泛共享，以进行进一步的详细研究。