1fe54d932b1fc100198898dc3a24af76 ijms-26-11598-v2.pdf b22ee750692cd634c4689c9062476ab264cabb99 ijms-26-11598-v2.pdf 8d78b3f5e04abb341b7003ca4c0a24b35b95d39869e958cb33a113229ef64a40 ijms-26-11598-v2.pdf Title: Mapping of Determinants of Urinary Sex Steroid Metabolites During Late Pregnancy: Results from Two Spanish Cohorts Subject: Steroid hormones (SHs), including sex steroids and corticosteroids, are crucial for a healthy pregnancy. We aimed to comprehensively characterize the maternal SH metabolome in late pregnancy and identify clinical, lifestyle, and sociodemographic determinants influencing SH metabolism with a replication in an independent cohort. Urinary SH metabolites were analyzed in 1221 third-trimester pregnant women (aged 28 to 37 years) from two Spanish cohorts, BiSC (2018–2021, n = 721) and INMA-Sabadell (2004–2006, n = 500), using targeted UHPLC-MS/MS. We quantified 50 SH metabolites, resulting in 13 hormone groups, 9 sulfate/glucuronide ratios, and 17 estimated steroid enzymatic activities across steroidogenesis pathways. We applied elastic net regression to identify determinants, and multivariable linear regression models to estimate variance explained. Among the 47 and 28 determinants from BiSC and INMA-Sabadell, respectively, 10 determinant-SH metabolome pairs showed statistically significant associations (p < 0.05), supporting robust replication. Maternal BMI was the main determinant linked to higher corticosteroid and androgen metabolites. Higher physical activity was associated with lower glucocorticoids and progestogen metabolites, while older maternal age was related with lower levels of androgen and corticosteroid metabolites. Tobacco exposure in the first trimester predicted higher levels of cortisol metabolites. Latin American women had lower cortolone levels compared with Spanish women. Parity, dietary fat intake, sleep, alcohol intake, and sex of the fetus contributed to smaller variations in different SHs. This dual-cohort analysis provides the most detailed and replicated evidence to date of how clinical, lifestyle, and sociodemographic factors shape the maternal SH metabolome during late pregnancy. Keywords: steroidomics; maternal metabolome; metabolic yprofiling; urinary steroid prolife; phase II metabolism; conjugated metabolites Author: Emily P. Laveriano-Santos, Estelle Renard-Dausset, Mariona Bustamante, Dolors Pelegri, Zoraida García-Ruiz, Marina Ruiz-Rivera, Marta Cosin-Tomas, Elisa Llurba-Olive, Maria Dolores Gomez-Roig, Noemi Haro, Óscar J. Pozo, Payam Dadvand, Martine Vrijheid and Léa Maitre Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25; modified using OpenPDF 1.4.2 CreationDate: Wed Dec 3 07:14:42 2025 CET ModDate: Thu Dec 4 04:43:25 2025 CET Custom Metadata: yes Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 30 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 3830553 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- PDCZKO+VnURWPalladioL Type 1 Custom yes yes yes 175 0 ZESVVA+URWPalladioL-Bold Type 1 Custom yes yes yes 176 0 VXLEHZ+URWPalladioL-Roma Type 1 Custom yes yes yes 177 0 XYXRRC+VnURWPalladioL-Bold Type 1 Custom yes yes yes 178 0 ZWBQKP+URWPalladioL-Ital Type 1 Custom yes yes yes 179 0 NXQZWG+EURM10 Type 1 Builtin yes yes yes 219 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 215 0 UWJUYN+CMSY10 Type 1 Builtin yes yes yes 270 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 248 0 URYLRZ+URWPalladioL-BoldItal Type 1 Custom yes yes yes 281 0 XIRVNA+EURB10 Type 1 Builtin yes yes yes 291 0 GNPBMJ+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 302 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 303 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 319 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 337 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 499 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 641 0 GOOENK+TimesNewRoman TrueType WinAnsi yes yes no 642 0 GNPBMH+PalatinoLinotype TrueType WinAnsi yes yes no 1061 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2026-01-23 03:08:42 CET RepresentationInformation: ijms-26-11598-v2.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2026-01-22 09:57:35 CET Size: 3830553 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 1300 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: ViewerPreferences: HideToolbar: false HideMenubar: false HideWindowUI: false FitWindow: true CenterWindow: false DisplayDocTitle: false NonFullScreenPageMode: UseNone Direction: L2R ViewArea: CropBox ViewClip: CropBox PrintArea: CropBox PageClip: CropBox PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Item: Title: Results Children: Item: Title: Participants’ Characteristics Item: Title: Identification of SH Metabolites Item: Title: Main Determinant Selection of the SH Metabolome (BiSC): ExWAS and ENET Analyses Item: Title: Determinants of Steroid Hormones: Relative Importance Analysis in the BiSC Item: Title: External Replication Analysis (INMA-Sabadell) Item: Title: Discussion Children: Item: Title: A New Window into Maternal Steroid Metabolism Item: Title: Clinical Factors Are Consistently Associated with SH-Level Differences Item: Title: Lifestyle Behaviors: Physical Activity, Sleep, Smoking, Alcohol, and Diet Are Determinants of SH Levels Item: Title: Maternal Age and Ethnicity as Sociodemographic Determinants Associated with SH Metabolism Item: Title: Genetic: SNPs from Steroidogenesis Enzymes Item: Title: Fetal Determinants: Sex Item: Title: Strengths, Limitations, and Future Directions Item: Title: Materials and Methods Children: Item: Title: Study Participants and Study Overview Item: Title: Urine Collection and Steroid Targeted Metabolome Profiling Item: Title: Determinants Children: Item: Title: Sociodemographic and Clinical Parameters Item: Title: Fetal Sex and Growth Item: Title: Genetic Item: Title: Mental Health Item: Title: Lifestyle Item: Title: Statistical Analysis Children: Item: Title: Data Preparation Item: Title: Primary Analysis—BiSC Item: Title: Replication Analysis—INMA-Sabadell Cohort Item: Title: Conclusions Item: Title: References Info: Title: Mapping of Determinants of Urinary Sex Steroid Metabolites During Late Pregnancy: Results from Two Spanish Cohorts Author: Emily P. Laveriano-Santos, Estelle Renard-Dausset, Mariona Bustamante, Dolors Pelegri, Zoraida García-Ruiz, Marina Ruiz-Rivera, Marta Cosin-Tomas, Elisa Llurba-Olive, Maria Dolores Gomez-Roig, Noemi Haro, Óscar J. Pozo, Payam Dadvand, Martine Vrijheid and Léa Maitre Subject: Steroid hormones (SHs), including sex steroids and corticosteroids, are crucial for a healthy pregnancy. We aimed to comprehensively characterize the maternal SH metabolome in late pregnancy and identify clinical, lifestyle, and sociodemographic determinants influencing SH metabolism with a replication in an independent cohort. Urinary SH metabolites were analyzed in 1221 third-trimester pregnant women (aged 28 to 37 years) from two Spanish cohorts, BiSC (2018–2021, n = 721) and INMA-Sabadell (2004–2006, n = 500), using targeted UHPLC-MS/MS. We quantified 50 SH metabolites, resulting in 13 hormone groups, 9 sulfate/glucuronide ratios, and 17 estimated steroid enzymatic activities across steroidogenesis pathways. We applied elastic net regression to identify determinants, and multivariable linear regression models to estimate variance explained. Among the 47 and 28 determinants from BiSC and INMA-Sabadell, respectively, 10 determinant-SH metabolome pairs showed statistically significant associations (p < 0.05), supporting robust replication. Maternal BMI was the main determinant linked to higher corticosteroid and androgen metabolites. Higher physical activity was associated with lower glucocorticoids and progestogen metabolites, while older maternal age was related with lower levels of androgen and corticosteroid metabolites. Tobacco exposure in the first trimester predicted higher levels of cortisol metabolites. Latin American women had lower cortolone levels compared with Spanish women. Parity, dietary fat intake, sleep, alcohol intake, and sex of the fetus contributed to smaller variations in different SHs. This dual-cohort analysis provides the most detailed and replicated evidence to date of how clinical, lifestyle, and sociodemographic factors shape the maternal SH metabolome during late pregnancy. 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