Program Project: GENE MUTATION AND RESCUE IN HUMAN DIAPHRAGMATIC HERNIA

计划项目：人类膈疝的基因突变与挽救

• 批准号: 8515483
• 负责人: PATRICIA K DONAHOE
• 金额: $ 159.62万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8515483
• 关键词: Affect; Algorithms; Animal Model; Archives; Area; Bioinformatics; Biological; Biological Models; Birds; Boston; Candidate Disease Gene; Caring; Cell Line; Cells; Child; Childhood; Collaborations; Comorbidity; Congenital Abnormality; Congenital Heart Defects; Congenital diaphragmatic hernia; Copy Number Polymorphism; Custom; Cytogenetics; DNA; DNA Library; DNA Resequencing; DNA Sequence; Defect; Development; Diagnosis; Diaphragmatic Hernia; Doctor of Medicine; Doctor of Philosophy; Drosophila genus; Embryo; Etiology; Evolution; Family; Family member; Foundations; Funding; Future; Gene Dosage; Gene Expression; Gene Expression Profile; Gene Mutation; Gene-Modified; General Hospitals; Generations; Genes; Genetic; Genetic Screening; Genetic Variation; Genomics; Glean; Goals; Heart; Homologous Gene; Human; Immunohistochemistry; In Situ Hybridization; Individual; Infant; Institution; Institutional Review Boards; International; Interview; Knockout Mice; Laboratories; Libraries; Linkage Disequilibrium; Live Birth; Loss of Heterozygosity; Maps; Massachusetts; Medical; Metabolic; Methods; Microdissection; Middle East; Modeling; Molecular; Molecular Cytogenetics; Molecular Genetics; Morbidity - disease rate; Mus; Mutation; National Institute of Child Health and Human Development; Oligonucleotide Microarrays; Operative Surgical Procedures; Parents; Pathway interactions; Patients; Pediatric Hospitals; Phenotype; Postdoctoral Fellow; Proteins; Protocols documentation; RNA Interference; Recording of previous events; Recruitment Activity; Replacement Therapy; Research Infrastructure; Resolution; Respiratory Diaphragm; Sampling; Seminal; Siblings; Single Nucleotide Polymorphism; Site; Source; Stem cells; Survivors; Susceptibility Gene; Syndrome; Technology; Testing; Therapeutic; Tissues; Transgenic Organisms; Tretinoin; Urine; Validation; Variant; Virus; Work; base; cardiogenesis; cohort; comparative genomic hybridization; cost; design; dosage; exome sequencing; fly; follow-up; gene discovery; gene function; genetic linkage analysis; genetic pedigree; genome wide association study; genome-wide; high throughput screening; in utero; induced pluripotent stem cell; interest; kindred; member; mortality; mutant mouse model; next generation; next generation sequencing; novel; outreach; prevent; proband; programs; protein protein interaction; research study; small molecule; tool; trend; vector

DESCRIPTION (provided by applicant): In the quest to unravel the etiology of Congenital Diaphragmatic Hernia (CDH), a prevalent (1:2500 live births) high impact (50% mortality with high morbidity) birth defect with phenotypic and genetic complexity, we are combining novel and traditional investigative strategies to one of the largest cohort of human patients with CDH. Collectively, we are employing molecular genetic, molecular cytogenetic, genomic, developmental, bioinformatic, and stem cell strategies along with the use of multiple model organisms. The strategies that we employ can serve as a template for identifying genes in pathways responsible not only for CDH, but for additional congenital anomalies, as well. The foundation of our work is the Infrastructure developed for ascertaining and recruiting almost all patients with CDH from two academic tertiary pediatric surgical centers, Massachusetts General Hospital (MGH) and Children's Hospital Boston (CHB), each offering state of the art medical, surgical, and genetic diagnosis, and management. Patients and families with CDH are interviewed comprehensively by study staff and carefully phenotyped by a senior geneticist; biological samples are obtained using sensitive protocols adapted for infants and children from each proband and from as many family members as possible, and are banked for proposed iterative genetic studies. Gene candidates are selected for sequencing when there is convergence from multiple lines of inquiry [i.e., copy number variations (CNVs) detected on array Comparative Genomic Hybridization (aCGH) or Single Nucleotide Polymorphism (SNP) arrays, genes in shared intervals detected on linkage analysis in affected belonging to multiplex CDH families, expressed genes in the developing primordial diaphragm, and bioinformatic algorithms that prioritize multiple candidate variants detected by the aforementioned platforms and from annotation of animal models with diaphragm defects]. The functional significance of the most promising candidates is subsequently assessed in multiple animal models. The evolution of available sequencing capabilities to lower costs opens the door to examine the multiple genes or polygeneity that we now know will contribute to a birth defect phenotype such as CDH. Given the national and International outreach and reputation of our genetic partnerships and the strength of the voluntary support network provided by the exceptional parents of patients with CDH, the study has extended beyond our two institutions to ascertain a number of seminal multiplex kindreds, including both consanguineous and non-consanguineous families, from the U.S. and internationally. RELEVANCE: The overarching goal of this study is to converge the multiple genes detected by these complementary approaches into cogent molecular pathways that will reveal polygeneic defects in the pathways and point toward potential treatment paradigms to prevent or alleviate this mortal or morbid congenital anomaly. Further, the methods devised for this study of CDH may have broader implications across other congenital anomalies.

描述(由申请人提供)：为了揭开先天性横隔疝(CDH)的病因，CDH是一种普遍的(1：2500活产)高影响(50%死亡率和高发病率)出生缺陷，具有表型和遗传复杂性，我们正在结合新的和传统的研究策略来研究人类CDH患者中最大的队列之一。总的来说，我们正在使用分子遗传学、分子细胞遗传学、基因组学、发育学、生物信息学和干细胞策略，同时使用多种模式生物。我们采用的策略可以作为一个模板，用于识别不仅导致CDH，而且还导致其他先天性异常的通路中的基因。我们工作的基础是为确定和招募来自马萨诸塞州综合医院(MGH)和波士顿儿童医院(CHB)这两个学术三级儿科外科中心的几乎所有CDH患者而开发的基础设施，每个中心都提供最先进的医疗、手术和基因诊断和管理。研究人员对CDH患者和家属进行了全面的访谈，并由一名资深遗传学家进行了仔细的表型鉴定；生物样本是根据每个先证者和尽可能多的家庭成员适用于婴儿和儿童的敏感方案获得的，并被储存起来用于拟议的迭代遗传学研究。当从多个查询行[即，在阵列比较基因组杂交(ACGH)或单核苷酸多态(SNP)阵列上检测到的拷贝数变异(CNV)，在受影响的CDH家族的连锁分析中检测到的共享间隔中的基因，在多个CDH家族中表达的基因， 开发原始隔膜，以及优先处理多个候选变体的生物信息学算法 由上述平台检测，并从具有横隔膜缺陷的动物模型的注释中检测到]。随后在多个动物模型中对最有希望的候选基因的功能意义进行了评估。现有测序能力的演变，以降低成本，打开了检查多基因或多基因的大门，我们现在知道这些基因或多基因将导致出生缺陷表型，如CDH。鉴于我们的遗传伙伴关系在国内和国际上的推广和声誉，以及CDH患者特殊父母提供的自愿支持网络的力量，这项研究已经扩展到我们的两个机构，以确定来自美国和国际的许多精液多重亲属，包括血缘和非血缘家庭。 相关性：这项研究的首要目标是将这些互补方法检测到的多个基因融合到令人信服的分子途径中，揭示这些途径中的多基因缺陷，并指出潜在的治疗方案，以预防或减轻这种致命或病态的先天性异常。此外，为CDH研究设计的方法可能对其他先天性异常具有更广泛的影响。