Neonatal Fellow Cincinnati Children's Hospital Cincinnati Children's Hospital Medical Center Cincinnati, Ohio, United States
Background: Congenital Diaphragmatic hernia (CDH) is characterized by a defect in the diaphragm leading to protrusion of abdominal contents into the thoracic cavity during fetal lung development. CDH results in the development of severe pulmonary hypertension (PH) and lung hypoplasia. Despite recent advances in treatment methods, morbidity and mortality of CDH remain high. The detailed mechanisms of CDH pathogenesis are yet to be understood despite the increasing interests in the community. The understanding of lung metabolism during the CDH development in utero could help us discern the changes in the CDH pathogenesis.
Objective: The present study aimed to analyze the differences in the metabolic profile of the fetal lungs with CDH compared with control developed fetal lungs.
Design/Methods: Timed-pregnant dams received Nitrofen or vehicle (olive oil) on E9. All fetal lungs exposed to Nitrofen or vehicle control were harvested at day E21 by C-section and then processed for metabolomics analysis using nuclear magnetic resonance (NMR) spectroscopy. The three groups analyzed were Nitrofen CDH (NCDH), Nitrofen control (NC), and vehicle control (VC). A total of 64 metabolites were quantified and before statistical analysis,
Results: The multivariate analysis identified forty-two metabolites that were different between the three groups. The two main components on the PLS-DA comparing NCDH with VC accounted for 75.6%. The highest VIP-score (>2) metabolites were lactate, glutamate, and adenosine 5′-triphosphate (ATP) (Figure1). Fetal CDH lungs have changes related to oxidative stress (metabolism of nicotinate, nicotinamide, ascorbate, aldarate, and glutathione), nucleotide synthesis (pyrimidine metabolism, aminoacyl-t-RNA biosynthesis, amino sugar, and nucleotide sugar metabolism), amino acid metabolism (glycine, serine, threonine, alanine, aspartate, glutamate, glutamine, histidine, arginine, proline metabolism), glycerophospholipid metabolism and glucose metabolism (pyruvate, TCA cycle metabolism and starch and sucrose metabolism)(Figure2). Conclusion(s): This work provides new insights into the molecular mechanisms behind the CDH pathophysiology and can lead to explore novel treatments in CDH.
C- ANOVA followed by least significant difference post hoc tests.
D- Three groups PCA showed a good separation
E- PCA loading plots (NC-NCDH-VC)
F- PLS-DA showed a well-defined separation between groups.
G- 5 components cross-validation
H- Metabolites variable importance in projection (VIP) in PLS-DA
A-The metabolome view shows all matched pathways from the pathway topology analysis.
B- Most relevant pathways with FDR p<0.05 from the pathway enrichment analysis and pathway impact values >0.1
Authors/Institutions: Maria del Mar Romero Lopez, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States; Marc Oria, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States; Miki Watanabe-Chailand, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States; Maria Florencia Varela, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States; Lindsey Romick-Rosendale, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States; Jose L. Peiro, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, United States