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

基本信息

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
项目类别：
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

项目摘要

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）：先天性膈疝 (CDH) 是一种常见的（1:2500 活产）高影响（50% 死亡率，高发病率）出生缺陷，具有表型和遗传复杂性，为了揭示其病因，我们正在将新颖和传统的研究策略结合起来，研究最大的 CDH 人类患者群体之一。总的来说，我们采用分子遗传学、分子细胞遗传学、基因组、发育、生物信息学和干细胞策略以及多种模式生物的使用。我们采用的策略可以作为模板来识别通路中的基因，这些通路不仅负责 CDH，而且还负责其他先天性异常。我们工作的基础是为确定和招募来自两个学术三级儿科外科中心（麻省总医院 (MGH) 和波士顿儿童医院 (CHB)）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、外科、以及基因诊断和管理。研究人员对 CDH 患者和家属进行全面访谈，并由资深遗传学家仔细进行表型分析；使用适用于婴儿和儿童的敏感方案从每个先证者和尽可能多的家庭成员中获取生物样本，并储存起来用于拟议的迭代遗传研究。当多条查询趋同时，选择候选基因进行测序 [即，在阵列比较基因组杂交 (aCGH) 或单核苷酸多态性 (SNP) 阵列上检测到拷贝数变异 (CNV)、在连锁分析中检测到共享区间内的基因在属于多重 CDH 家族的受影响者中，表达基因开发原始膈肌和优先考虑多个候选变体的生物信息学算法通过上述平台和具有隔膜缺陷的动物模型的注释检测到]。随后在多种动物模型中评估最有希望的候选者的功能意义。可用测序能力的发展降低了成本，这为检查多基因或多基因性打开了大门，我们现在知道这些基因或多基因性将导致 CDH 等出生缺陷表型。鉴于我们的遗传伙伴关系在国内和国际上的影响力和声誉，以及 CDH 患者杰出父母提供的自愿支持网络的力量，该研究已超出我们两个机构的范围，以确定许多重要的多重亲属，包括近亲亲属来自美国和国际的非近亲家庭。相关性：本研究的首要目标是将这些互补方法检测到的多个基因汇聚成令人信服的分子途径，这将揭示途径中的多基因缺陷，并指出预防或减轻这种致命或病态先天异常的潜在治疗范例。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。