Amniotic fluid metabolic fingerprinting indicated metabolites which may play a role in the pathogenesis of foetal Down syndrome — a preliminary report

Objectives: Down syndrome is the most common human chromosomal aberration. It is commonly known that it is a genetic-based disease, but still, pathomechanisms which lead to observed disorders have not been explained. The objective of thisstudy was to determine the metabolic fingerprinting of the amniotic fluid women carrying foetuses with Down syndrome (DS).Material and methods: The study and control groups consisted of women who underwent routine amniocentesis betweenthe 15th and 18th week of gestation. After analysis of the karyotyping results, 13 women with foetal DS were chosen. Forthe control group, 13 healthy patients with uncomplicated pregnancies who delivered healthy newborns at term wasselected. Amniotic fluid was analyzed using liquid chromatography combined with high resolution mass spectrometry.Results: In the amniotic fluid of women with foetal DS compared to patients with healthy foetuses, we reported significantdifferences in the level of four metabolites: methylhistidine, hexanoylcarnitine, diacetylspermine and p-cresol sulfate whichmay be connected with improper development of nervous system and muscles. We detected bacterial metabolite, whichsupport the latest thesis about non-sterile intrauterine environment.Conclusions: Based on our findings, we hypothesise that differences in the level of four metabolites in the amniotic fluidmay play role in the pathogenesis of DS. Defining their potential as biochemical pathogenic factors of DS requires furtherinvestigation of the biological pathways involving in the foetal development

Maternal plasma and amniotic fluid sphingolipids profiling in fetal Down syndrome.

INTRODUCTION:Sphingolipids can be potentially involved in the formation of the central and peripheral nervous systems, which are particularly connected with the pathogenesis of Down syndrome. The aim of the study was to determine the concentration of selected sphingolipids in the plasma and amniotic fluid of pregnant patients with fetal Down syndrome. MATERIAL AND METHODS:Out of 190 amniocentesis we had 10 patients with confirmed Down syndrome. For the purpose of our control we chose 14 women without confirmed chromosomal aberration. To assess the concentration of 11 sphingolipids in the blood plasma and amniotic fluid we used an ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometry (UHPLC/MS/MS). RESULTS:We showed a significant increase in the concentration of 2 ceramides, C22-Cer and C24:1-Cer, in the plasma of women with fetal Down syndrome. Furthermore we showed a decrease in the concentration of 7 ceramides--C16-Cer, C18-Cer, C18:1-Cer, C20-Cer, C22-Cer, C24:1-Cer, and C24-Cer--in the amniotic fluid of women with fetal Down syndrome. We created ROC curves for all significant sphingolipids in maternal plasma, which set the threshold values and allowed for predicting the likelihood of Down syndrome in the fetus with specific sensitivity and specificity. We demonstrated a significantly higher risk of Down syndrome when the plasma concentration of C22-Cer > 12.66 ng/100 ul (sens. 0.9, sp. 0.79, P value = 0.0007) and C24:1-Cer > 33,19 ng/100 ul (sens. 0.6, sp. 0.86, P value = 0.0194). CONCLUSION:On the basis of our findings, it seems that the sphingolipids may play a role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on a larger group of patients

Maternal Plasma and Amniotic Fluid Chemokines Screening in Fetal Down Syndrome

Objective. Chemokines exert different inflammatory responses which can potentially be related to certain fetal chromosomal abnormalities. The aim of the study was to determine the concentration of selected chemokines in plasma and amniotic fluid of women with fetal Down syndrome. Method. Out of 171 amniocentesis, we had 7 patients with confirmed fetal Down syndrome (15th–18th weeks of gestation). For the purpose of our control, we chose 14 women without confirmed chromosomal aberration. To assess the concentration of chemokines in the blood plasma and amniotic fluid, we used a protein macroarray, which allows the simultaneous determination of 40 chemokines per sample. Results. We showed significant decrease in the concentration of 4 chemokines, HCC-4, IL-28A, IL-31, and MCP-2, and increase in the concentration of CXCL7 (NAP-2) in plasma of women with fetal Down syndrome. Furthermore, we showed decrease in concentration of 3 chemokines, ITAC, MCP-3, MIF, and increase in concentration of 4 chemokines, IP-10, MPIF-1, CXCL7, and 6Ckine, in amniotic fluid of women with fetal Down syndrome. Conclusion. On the basis of our findings, our hypothesis is that the chemokines may play role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on larger group of patients

Maternal plasma angiogenic and inflammatory factor profiling in foetal Down syndrome

<div><p>Objective and design</p><p>Angiogenic factors are proteins that are related to certain foetal chromosomal abnormalities. The aim of this study was to determine the concentration of 60 angiogenic factors in the plasma of women with offspring possessing trisomy 21/Down syndrome (DS).</p><p>Method</p><p>After analysing karyotyping results, we selected 20 patients with foetuses possessing DS, and for the control group, we selected 28 healthy patients with uncomplicated pregnancies who delivered healthy newborns at term (i.e., 15–18 weeks of gestation). To assess the concentration of proteins in the blood plasma, we used a protein macroarray which enabled simultaneous determination of 60 angiogenic factors per sample.</p><p>Results</p><p>We observed a statistically significant increase in the concentration of these five angiogenic and inflammatory factors: TGFb1 (p = 0.039), angiostatin (p = 0.0142), I-309 (p = 0.0476), TGFb3 (p = 0.0395), and VEGF-D (p = 0.0173)—compared to concentrations in patients with healthy foetuses.</p><p>Conclusion</p><p>Our findings suggest that angiogenic factors may play role in DS pathogenesis.</p></div

Concentrations of angiogenic factors in maternal plasma.

<p>Concentrations of angiogenic factors in maternal plasma.</p

The list of quantified sphingolipids with the limit of detection.

<p>The list of quantified sphingolipids with the limit of detection.</p

Maternal plasma and amniotic fluid chemokines screening

Objective. Chemokines exert different inflammatory responses which can potentially be related to certain fetal chromosomal abnormalities. The aim of the study was to determine the concentration of selected chemokines in plasma and amniotic fluid of women with fetal Down syndrome. Method. Out of 171 amniocentesis, we had 7 patients with confirmed fetal Down syndrome (15th-18th weeks of gestation). For the purpose of our control, we chose 14 women without confirmed chromosomal aberration. To assess the concentration of chemokines in the blood plasma and amniotic fluid, we used a protein macroarray, which allows the simultaneous determination of 40 chemokines per sample. Results. We showed significant decrease in the concentration of 4 chemokines, HCC-4, IL-28A, IL-31, and MCP-2, and increase in the concentration of CXCL7 (NAP-2) in plasma of women with fetal Down syndrome. Furthermore, we showed decrease in concentration of 3 chemokines, ITAC, MCP-3, MIF, and increase in concentration of 4 chemokines, IP-10, MPIF-1, CXCL7, and 6Ckine, in amniotic fluid of women with fetal Down syndrome. Conclusion. On the basis of our findings, our hypothesis is that the chemokines may play role in the pathogenesis of Down syndrome. Defining their potential as biochemical markers of Down syndrome requires further investigation on larger group of patients