Gene Mutation and Rescue in Human Diaphragmatic Hernia

整体计划 摘要 我们的研究策略是“人类横隔疝的基因突变与挽救”项目 为了1)从基因组分析中确定先天性膈疝患者的基因变异，2)测试 在体外和体外以及动物模型中验证和功能的高优先级候选者，3)确定 通过整合已知的和新的候选基因定义的分子途径，以及4)选择分子 构思CDH产后治疗策略的目标。先天性巨结肠是一种先天缺陷，发生于 1/3000活产，死亡率和发病率很高，幸存者需要高科技驱动 第三或第四新生儿重症监护病房的介入性围产期护理。40%的CDH患者有 除CDH外，还有多个异常，但多达60%的异常是孤立的，有可能增加 如果能改善与之相关的肺发育不良导致其高发病率，则可存活下来。社会和 CDH的个人负担可望通过减轻患者和家庭的压力而减轻 相当高的发病率和死亡率，以及护理费用，都与这一出生缺陷相吻合。如果数量可观 肺分化可诱导发生在出生后的肺部，使用体外膜 氧合(ECMO)和积极的呼吸机管理可能会减少，从而保护脆弱的肺 死于继发性气压伤和支气管肺发育不良。 为了影响有益的变化，我们计划继续使用基因组方法，如整个Exome 测序(WES)、阵列比较基因组杂交(ACGH)和全基因组测序 (WGS)，并将不同的发现与来自发育中的横隔膜和肺的表达分析的数据相结合 小鼠模型中的组织，以及来自人类CDH患者的横隔膜活检。WES、WGS和aCGH 数据将在临床特征良好的独生子女队列中持续确定和分析 父母/孩子三人组，以及罕见的大家庭。这是P01的目标是利用这个不断增长的队列和我们的 一种分析方法来产生候选基因，这些基因将产生关于分子发病机制的重要线索 尤其是在将最高优先级的候选基因整合到分子通路之后。 项目一：基因组和基因表达分析，以发现人类CHD基因和途径。 项目II：小鼠模型将阐明CDH和相关的肺缺陷的遗传学并确定 临床上相关的靶点。 项目III：CDH患者基因变异的体外和选定动物模型的功能评估。

OVERALL PROGRAM ABSTRACT Our research strategy for the Program project “Gene Mutation and Rescue in Human Diaphragmatic Hernia” is to 1) identify gene variants in patients with Congenital Diaphragmatic Hernia from genomic analyses, 2) test high priority candidates for validation and function in vitro and ex vivo, and in animal models, 3) determine molecular pathways defined by integrating already known and new candidate genes, and 4) select molecular targets from which to conceive postnatal treatment strategies for CDH. CDH is a birth defect which occurs in 1/3000 live births, and has a high mortality and morbidity, with survivors requiring high technology-driven interventional perinatal care in tertiary or quarternary neonatal intensive care units. 40% of CDH patients have multiple anomalies in addition to CDH, but as many as 60% are isolated with the potential for increased survival if we can improve the associated lung hypoplasia responsible for its high morbidity. The societal and individual burden of CDH can hopefully be lessened by reducing the patient and family stresses caused by the considerable morbidity and mortality, as well as the cost of care, coincident with this birth defect. If substantial pulmonary differentiation can be induced to occur in the lung postnatally, the use of Extra Corporeal Membrane Oxygenation (ECMO) and aggressive ventilatory management may be reduced, thus protecting fragile lungs from secondary barotrauma and bronchopulmonary dysplasia. To affect a salutary change, we plan to continue to employ genomic methods such as Whole Exome Sequencing (WES), array Comparative Genomic Hybridization (aCGH), and Whole Genomic Sequencing (WGS), and to integrate variant findings with data from expression analyses of developing diaphragm and lung tissues in mouse models, and with diaphragm biopsies from human CDH patients. WES, WGS, and aCGH data will be continually ascertained and analyzed on a clinically well characterized cohort of singletons and parent/child trios, as well as rare extended families. It is the goal of this P01 to use this growing cohort and our analytic approach to generate candidate genes that will yield important clues as to the molecular pathogenesis of CDH, particularly after integrating the highest priority candidates into molecular pathways. PROJECT I: Genomic and gene expression analyses to discover human CHD genes and pathways. PROJECT II: Mouse models will elucidate genetics of CDH and associated pulmonary defects and identify clinically relevant targets. PROJECT III: Functional evaluation of CDH patient gene variants in vitro and in select animal models.