ABSTRACT
OBJECTIVE: The lifetime risk of metabolic diseases in offspring of women with gestational diabetes mellitus (GDM) depends, at least in part, on the impact of glycemic fetal programming. To quantify this impact, we have developed and validated a unique mass spectrometry method to measure the percentage of glycated hemoglobin in cord blood. RESEARCH DESIGN AND METHODS: This case-control study includes 37 GDM women and 30 pregnant women with normal glucose tolerance (NGT). RESULTS: Glycation of the α-chain (Glα) was higher in neonates from GDM (2.32 vs. 2.20%, P < 0.01). Glα strongly correlated with maternal A1C measured at delivery in the overall cohort (r = 0.67, P < 0.0001) as well as in each group (GDM: r = 0.66, P < 0.0001; NGT: r = 0.50, P = 0.01). CONCLUSIONS: Thus, Glα may reflect hyperglycemic exposure during the last weeks of fetal development. Future studies will confirm Glα is a predictive biomarker of prenatally programmed lifetime metabolic health and disease.
Subject(s)
Diabetes, Gestational/blood , Fetal Hemoglobin/metabolism , Glycated Hemoglobin/metabolism , Hyperglycemia/blood , Metabolic Diseases/epidemiology , Prenatal Exposure Delayed Effects/blood , Adult , Blood Glucose/metabolism , Case-Control Studies , Female , Fetal Blood/metabolism , Fetal Development , Glycosylation , Humans , Infant, Newborn , Male , Metabolic Diseases/etiology , Pregnancy , Prenatal Exposure Delayed Effects/diagnosis , Young AdultABSTRACT
Fabry disease is a lysosomal storage disorder caused by deficiency of α-galactosidase A, resulting in glycosphingolipid accumulation in organs and tissues, including plasma and urine. Two disease-specific Fabry biomarkers have been identified and quantified in plasma and urine: globotriaosylceramide (Gb(3)) and globotriaosylsphingosine (lyso-Gb(3)). The search continues for biomarkers that might be reliable indicators of disease severity and response to treatment. The main objective of this study was to target other urinary biomarkers using a time-of-flight mass spectrometry metabolomic approach. Urinary metabolites of 63 untreated Fabry patients and 59 controls were analyzed. A multivariate statistical analysis performed on a subset of male samples revealed seven novel Fabry biomarkers in urine, all lyso-Gb(3) analogues having modified sphingosine moieties. The empirical formulas of the sphingosine modifications were determined by exact mass measurements (- C(2)H(4), - C(2)H(4) + O, - H(2), - H(2) + O, + O, + H(2)O(2), + H(2)O(3)). We evaluated the relative concentration of lyso-Gb(3) and its seven analogues by measuring area counts for each analogue in all Fabry patients. All samples were normalized to creatinine. We found higher concentrations for males with Fabry disease compared to females. None of these biomarkers were detected in controls. To our knowledge, this is the first time that lyso-Gb(3)-related Fabry disease biomarkers are detected in urine.
Subject(s)
Fabry Disease/metabolism , Fabry Disease/urine , Glycolipids/urine , Metabolomics/methods , Sphingolipids/urine , Adolescent , Adult , Aged , Biomarkers/metabolism , Biomarkers/urine , Child , Child, Preschool , Creatine/metabolism , Creatine/urine , Data Mining , Female , Glycolipids/metabolism , Humans , Infant , Male , Middle Aged , Multivariate Analysis , Spectrometry, Mass, Electrospray Ionization/methods , Sphingolipids/metabolism , Young AdultABSTRACT
The characterization of cord blood hemoglobin at the molecular level is a daunting challenge because hemoglobin F (HbF) and hemoglobin A (HbA) coexist in neonatal blood. We developed and validated a method using electrospray time-of-flight mass spectrometry (ES-TOF-MS) that measures, in a single analysis, relative levels of glycated and acetylated hemoglobin and allows the calculation of relative proportions of HbA, HbF(0), and HbF(1) in cord blood. Specific sections of acquired spectra were deconvoluted using a maximum entropy-based approach to true mass scale spectra. Mass precisions were less than 3 ppm with similar accuracies. Intra-interday precisions for α- and γ-chain glycation levels were 2.10%/3.72% and 2.75%/6.79%, respectively. The linearity of the α-chain glycation response was excellent (r(2) = 0.9990). We performed sample analysis on 39 cord blood specimens and found that the glycated α- and γ-chain levels were 2.27 ± 0.21% and 2.38 ± 0.29%, respectively, while the acetylated (G)γ and (A)γ-chain levels were 8.48 ± 0.53% and 7.14 ± 0.74%, respectively. We observed three types of HbF distinguishable by the intensities of γ-chain variants. Two-thirds of cord blood specimens were classified as HbF(I) with an intensity ratio (G)γ/(A)γ of 1.90 ± 0.12. For HbF(II) type (10/39 neonates), the intensity ratio of (G)γ/(A)γ was 3.71 ± 0.28. For three neonates with HbF(III), no (A)γ-chain was detected.
