Congenital diaphragmatic hernia and other congenital diaphragmatic defects are associated with significant mortality and morbidity in neonates; however, the molecular basis of these developmental anomalies is unknown. In an analysis of E18.5 embryos derived from mice treated with N-ethyl-N-nitrosourea, we identified a mutation that causes pulmonary hypoplasia and abnormal diaphragmatic development. Fog2 (Zfpm2) maps within the recombinant interval carrying the N-ethyl-N-nitrosourea-induced mutation, and DNA sequencing of Fog2 identified a mutation in a splice donor site that generates an abnormal transcript encoding a truncated protein. Human autopsy cases with diaphragmatic defect and pulmonary hypoplasia were evaluated for mutations in FOG2. Sequence analysis revealed a de novo mutation resulting in a premature stop codon in a child who died on the first day of life secondary to severe bilateral pulmonary hypoplasia and an abnormally muscularized diaphragm. Using a phenotype-driven approach, we have established that Fog2 is required for normal diaphragm and lung development, a role that has not been previously appreciated. FOG2 is the first gene implicated in the pathogenesis of nonsyndromic human congenital diaphragmatic defects, and its necessity for pulmonary development validates the hypothesis that neonates with congenital diaphragmatic hernia may also have primary pulmonary developmental abnormalities.
Birth defects involving the diaphragm are as common and as serious as genetic disorders such as cystic fibrosis, yet the underlying causes of these defects are unknown. Most babies born with diaphragmatic defects have very small lungs, and many die in the newborn period with severe breathing difficulties. It is unknown whether the small lungs occur because these children have a diaphragmatic defect, or whether some patients might have a genetic abnormality that affects the development of both the lung and the diaphragm simultaneously.
In a screen of fetal mice carrying chemically induced genetic mutations, the authors found that a mutation in the gene Fog2 (Friend of gata 2), causes abnormal diaphragm development and small lungs. The lungs have a primary developmental abnormality that includes the specific loss of one lung lobe. Based on this result, the authors studied children affected with diaphragmatic abnormalities, and identified one human baby with a serious mutation in the human gene FOG2 who died at five hours of life with severe breathing difficulties, a diaphragmatic defect, and small lungs.
The authors have established that Fog2 is necessary for both diaphragm and lung development in mice and in humans. This is the first known cause of a nonsyndromic congenital diaphragmatic defect and establishes that some patients may have a primary developmental abnormality of both the lung and the diaphragm.