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.
OVERALL PROGRAM NARRATIVE: Congenital Diaphragmatic Hernia (CDH) is a prevalent (1:3000 live births) high impact (50% mortality with high morbidity) birth defect with phenotypic and genetic complexity and heterogeneity. The aim of this Program Project 'Gene Mutation and Rescue in Human Congenital Diaphragmatic Hernia' is to 1) identify gene variants in patients with Congenital Diaphragmatic Hernia from genomic sequencing and structural variant 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. 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. Steps that we employ herein can serve ultimately as a template to address other polygenic congenital anomalies and treatment strategies conceived for CDH can have broader applications in the care of children with the bronchopulmonary dysplasia of prematurity.
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