Study on the Potential Biomarkers of Maternal Urine Metabolomics for Fetus with Congenital Heart Diseases Based on Modified Gas Chromatograph-Mass Spectrometer

Abstract

Background. There has been significant research on the genetic and environmental factors of congenital heart defects (CHDs), but few causes of teratogenicity, especially teratogenic mechanisms, can be clearly identified. Metabolomics has a potential advantage in researching the relationship between external factors and CHD. Objective. To find and identify the urinary potential biomarkers of pregnancy (including in the second and third trimesters) for fetuses with CHD based on modified gas chromatograph-mass spectrometer (GC-MS), which could reveal the possibility of high-risk factors for CHD and lay the foundation for early intervention, treatment, and prevention. Methods. Using a case-control design, we measured the urinary potential biomarkers of maternal urine metabolomics based on GC-MS in a population-based sample of women whose infants were diagnosed with CHD (70 case subjects) or were healthy (70 control subjects). SIMCA-P 13.0 software, principal component analysis (PCA), orthogonal partial least squares-discriminant analysis (OPLS-DA), Wilcoxon-Mann-Whitney test, and logistics regression were used to find significant potential biomarkers. Result. The 3D score graph of the OPLS-DA showed that the CHD and control groups were fully separated. The fitting parameters were R2x=0.78 and R2y=0.69, and the forecast rate was Q2=0.61, indicating a high forecast ability. According to the ranking of VIPs from the OPLS-DA models, we found 34 potential metabolic markers with a VIP > 1, and after two pairwise rank sum tests, we found 20 significant potential biomarkers, which were further used in multifactor logistic regressions. Significant substances, including 4-hydroxybenzeneacetic acid (OR=4.74, 95% CI: 1.06-21.06), 5-trimethylsilyloxy-n-valeric acid (OR=15.78, 95% CI: 2.33-106.67), propanedioic acid (OR=5.37, 95% CI: 1.87-15.45), hydracrylic acid (OR=6.23, 95% CI: 1.07-36.21), and uric acid (OR=5.23, 95% CI: 1.23-22.32), were associated with CHD. Conclusion. The major potential biomarkers in maternal urine associated with CHD were 4-hydroxybenzeneacetic acid, 5-trimethylsilyloxy-n-valeric acid, propanedioic acid, hydracrylic acid, and uric acid, respectively. These results indicated that the short chain fatty acids (SCFAs) and aromatic amino acid metabolism may be relevant with CHD.

Analysis of Biomarkers for Congenital Heart Disease Based on Maternal Amniotic Fluid Metabolomics

“…Graca et al ( 16 , 17 ) did not find changes in proline and uric acid levels in AF when using MRI to detect fetal malformations, including CHD. However, the changes in uric acid were consistent with the urine biomarker reported by Xie et al ( 19 ), as shown in Table 1 . Our results using AF were not consistent with those obtained using the maternal serum in early pregnancy reported by Bahado-Singh et al ( 18 ) and our own results in maternal serum.…”

“…Several metabolomics studies have focused on fetal CHD, and we summarized these studies as shown in Table 1. In these studies, the types of samples used were maternal serum, maternal urine and amniotic fluid (AF), and the methods used were direct injection/liquid chromatography and tandem mass spectrometry (DI/LC-MS/MS), tandem mass spectrometry (MS/MS), gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) (16)(17)(18)(19)(20). However, studies of fetal CHD based on amniotic fluid metabolomics remains limited due to the availability of AF.…”

Biomarkers for Isolated Congenital Heart Disease Based on Maternal Amniotic Fluid Metabolomics analysis

“…In terms of metabolomic methods, a total of 2472 metabolites were identi ed using the UHPLC-QTOF-MS untargeted metabolomics detection in this study. Many new metabolites were found compared to previous studies, which mostly used NMR or GC-TOF-MS methods, could only detect hundreds of metabolites [15][16][17][18] . UHPLC increasingly displacing conventional high performance liquid chromatography 23 LC-MS is the main workhorse of metabolomics owing to its high degree of analytical sensitivity and speci city when measuring diverse chemistry in complex biological samples 24 .…”

Metabolomics Signatures and Subsequent Maternal Health among Mothers with a Congenital Heart Defect-Affected Pregnancy

“…Metabolites 2022, 12, 100 2 of 16 This is particularly important, as disrupted metabolism related to the affected birth could impact long-term maternal health. Women with pregnancies affected by CHD have been reported with altered metabolic profiles and were associated with dysregulated metabolic pathways [7][8][9][10]. These markers were often utilized as predictive markers for fetuses with congenital heart defects.…”