Program Project: GENE MUTATION AND RESCUE IN HUMAN DIAPHRAGMATIC HERNIA

描述（由申请人提供）：为了阐明先天性膈疝（CDH）的病因，CDH是一种具有表型和遗传复杂性的流行（1：2500活产）高影响（50%死亡率和高发病率）出生缺陷，我们将新型和传统研究策略结合到最大的CDH人类患者队列中。总的来说，我们正在采用分子遗传学，分子细胞遗传学，基因组学，发育，生物信息学和干细胞策略沿着使用多种模式生物。我们采用的策略可以作为一个模板，用于识别不仅负责CDH，而且还负责其他先天性异常的途径中的基因。 我们工作的基础是为确定和招募几乎所有来自两个学术三级儿科外科中心的CDH患者而开发的基础设施，这两个中心是马萨诸塞州总医院（MGH）和波士顿儿童医院（CHB），每个中心都提供最先进的医疗，手术和遗传诊断和管理。CDH患者和家庭由研究人员进行全面访谈，并由高级遗传学家进行仔细的表型分析;使用适用于每个先证者和尽可能多的家庭成员的婴儿和儿童的敏感方案获得生物样本，并储存用于拟议的迭代遗传研究。当从多条查询线中存在收敛时，选择基因候选物用于测序[即，比较基因组杂交（aCGH）或单核苷酸多态性（SNP）阵列上检测到的拷贝数变异（CNV），多重CDH家族中受影响者连锁分析中检测到的共享间隔中的基因， 开发原始隔膜，以及优先考虑多个候选变体的生物信息学算法 通过上述平台和从具有隔膜缺陷的动物模型的注释检测]。随后在多个动物模型中评估最有希望的候选物的功能意义。 现有测序能力的发展为降低成本打开了大门，以检查我们现在知道将导致出生缺陷表型（如CDH）的多基因或多核苷酸。 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患者的特殊父母提供的自愿支持网络的力量，该研究已经超出了我们的两个机构，以确定一些精液的多重激酶，包括美国和国际上的近亲和非近亲家庭。 相关性：本研究的总体目标是将这些互补方法检测到的多个基因汇聚到令人信服的分子通路中，这将揭示通路中的多基因缺陷，并指向潜在的治疗范例，以预防或减轻这种致命或病态的先天性异常。此外，为这项CDH研究设计的方法可能对其他先天性异常具有更广泛的意义。

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.