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Urinary steroidomic profiles by LC-MS/MS to monitor classic 21-Hydroxylase deficiency.
J Steroid Biochem Mol Biol 2019; 198:105553JS

Abstract

21-hydroxylase deficiency, the most common enzyme defect associated with congenital adrenal hyperplasia (CAH) is characterized by an impairment of both aldosterone and cortisol biosynthesis. Close clinical and biological monitoring of Hydrocortisone (HC) and 9α-Fludrocortisone (FDR) replacement therapies is required to achieve an optimal treatment. As frequent and repeated reassessments of plasma steroids, 17-hydroxyprogesterone (17-OHP), androstenedione (Δ4-A) and testosterone (TESTO) is needed in childhood, urine steroid profiling could represent an interesting non-invasive alternative. We developed and validated a LC-MS/MS method for the measurement of 23-urinary mineralocorticoids, glucocorticoids and adrenal androgens. The usefulness of steroid profiling was investigated on single 08h00 am-collected spot urine for discriminating between 61 CAH patients and their age- and sex-matched controls. CAH patients were split into two groups according to their 08h00 am-plasma concentrations of 17-OHP: below (controlled patients, n = 26) and above 20 ng/mL (uncontrolled patients, n = 35). The lower limit of quantification and the wide analytical range allows to assay both free and total concentrations of the main urinary adreno-corticoids and their tetra-hydrometabolites. Extraction recoveries higher than 75% and intra-assay precision below 20% were found for most steroids. Urinary steroids upstream of the 21-hydroxylase defect were higher in uncontrolled CAH patients. Among CAH patients, plasma and urinary 17-OHP were closely correlated. As compared to controls, steroids downstream of the enzyme defect collapsed in CAH patients. This fall was more pronounced in controlled than in uncontrolled patients. Androgens (Δ4-A, TESTO and the sum etiocholanolone + androsterone) accumulated in uncontrolled CAH patients. A strong relationship was observed between plasma and urinary levels of androstenedione. Daily doses and urinary excretion of both FDR and HC were similar in both CAH groups. Urinary FDR was inversely related to the sodium-to-potassium ratio in urine. A partial least squares discriminant analysis model allowed to classify the patient's classes unaffected, controlled and un-controlled CAH patients based on urinary steroidomic profiles. Our LC-MS/MS method successfully established steroid profiling in urine and represents a useful and non-invasive tool for discriminating CAH patients according to treatment efficiency.

Authors+Show Affiliations

Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France; Service de Génétique Moléculaire, Pharmacogénétique et Hormonologie, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le Kremlin Bicêtre, F-94275, France. Electronic address: eric.pussard@aphp.fr.Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France; Service de Génétique Moléculaire, Pharmacogénétique et Hormonologie, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le Kremlin Bicêtre, F-94275, France.Département d'Endocrinologie Pédiatrique, Hôpital de Bicêtre, Hôpitaux Universitaires Paris Sud, Assistance Publique-Hôpitaux de Paris, Le Kremlin Bicêtre, F-94275, France; Centre de Référence des Maladies Rares du Développement Génital (DEVGEN), Le Kremlin Bicêtre, F-94275, France.Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France; Service de Génétique Moléculaire, Pharmacogénétique et Hormonologie, Hôpital Bicêtre, Assistance Publique-Hôpitaux de Paris, Le Kremlin Bicêtre, F-94275, France.Service de Biochimie, Assistance Publique-Hôpitaux de Paris, Hôpital Bicêtre, Le Kremlin Bicêtre, F-94275, France.Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France.Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France; Service d'Endocrinologie Pédiatrique, Hôpital Robert Debré, Assistance Publique Hôpitaux de Paris, Paris, F-75019, France; Université Paris Diderot, Sorbonne Paris Cité, Paris, F-75019, France.Inserm, U1185, Le Kremlin-Bicêtre, F-94276, France; Fac Med Paris-Sud, Univ. Paris-Sud, Université Paris Saclay, UMR-S 1185, Le Kremlin-Bicêtre, F-94276, France.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31778802

Citation

Pussard, Eric, et al. "Urinary Steroidomic Profiles By LC-MS/MS to Monitor Classic 21-Hydroxylase Deficiency." The Journal of Steroid Biochemistry and Molecular Biology, vol. 198, 2019, p. 105553.
Pussard E, Travers S, Bouvattier C, et al. Urinary steroidomic profiles by LC-MS/MS to monitor classic 21-Hydroxylase deficiency. J Steroid Biochem Mol Biol. 2019;198:105553.
Pussard, E., Travers, S., Bouvattier, C., Xue, Q. Y., Cosson, C., Viengchareun, S., ... Lombès, M. (2019). Urinary steroidomic profiles by LC-MS/MS to monitor classic 21-Hydroxylase deficiency. The Journal of Steroid Biochemistry and Molecular Biology, 198, p. 105553. doi:10.1016/j.jsbmb.2019.105553.
Pussard E, et al. Urinary Steroidomic Profiles By LC-MS/MS to Monitor Classic 21-Hydroxylase Deficiency. J Steroid Biochem Mol Biol. 2019 Nov 26;198:105553. PubMed PMID: 31778802.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Urinary steroidomic profiles by LC-MS/MS to monitor classic 21-Hydroxylase deficiency. AU - Pussard,Eric, AU - Travers,Simon, AU - Bouvattier,Claire, AU - Xue,Qiong-Yao, AU - Cosson,Claudine, AU - Viengchareun,Say, AU - Martinerie,Laetitia, AU - Lombès,Marc, Y1 - 2019/11/26/ PY - 2019/07/05/received PY - 2019/11/21/revised PY - 2019/11/24/accepted PY - 2019/11/30/pubmed PY - 2019/11/30/medline PY - 2019/11/29/entrez KW - Congenital adrenal hyperplasia KW - Fludrocortisone KW - Hydrocortisone KW - LC-MS/MS KW - Replacement therapy monitoring KW - Urinary steroidomic profile SP - 105553 EP - 105553 JF - The Journal of steroid biochemistry and molecular biology JO - J. Steroid Biochem. Mol. Biol. VL - 198 N2 - 21-hydroxylase deficiency, the most common enzyme defect associated with congenital adrenal hyperplasia (CAH) is characterized by an impairment of both aldosterone and cortisol biosynthesis. Close clinical and biological monitoring of Hydrocortisone (HC) and 9α-Fludrocortisone (FDR) replacement therapies is required to achieve an optimal treatment. As frequent and repeated reassessments of plasma steroids, 17-hydroxyprogesterone (17-OHP), androstenedione (Δ4-A) and testosterone (TESTO) is needed in childhood, urine steroid profiling could represent an interesting non-invasive alternative. We developed and validated a LC-MS/MS method for the measurement of 23-urinary mineralocorticoids, glucocorticoids and adrenal androgens. The usefulness of steroid profiling was investigated on single 08h00 am-collected spot urine for discriminating between 61 CAH patients and their age- and sex-matched controls. CAH patients were split into two groups according to their 08h00 am-plasma concentrations of 17-OHP: below (controlled patients, n = 26) and above 20 ng/mL (uncontrolled patients, n = 35). The lower limit of quantification and the wide analytical range allows to assay both free and total concentrations of the main urinary adreno-corticoids and their tetra-hydrometabolites. Extraction recoveries higher than 75% and intra-assay precision below 20% were found for most steroids. Urinary steroids upstream of the 21-hydroxylase defect were higher in uncontrolled CAH patients. Among CAH patients, plasma and urinary 17-OHP were closely correlated. As compared to controls, steroids downstream of the enzyme defect collapsed in CAH patients. This fall was more pronounced in controlled than in uncontrolled patients. Androgens (Δ4-A, TESTO and the sum etiocholanolone + androsterone) accumulated in uncontrolled CAH patients. A strong relationship was observed between plasma and urinary levels of androstenedione. Daily doses and urinary excretion of both FDR and HC were similar in both CAH groups. Urinary FDR was inversely related to the sodium-to-potassium ratio in urine. A partial least squares discriminant analysis model allowed to classify the patient's classes unaffected, controlled and un-controlled CAH patients based on urinary steroidomic profiles. Our LC-MS/MS method successfully established steroid profiling in urine and represents a useful and non-invasive tool for discriminating CAH patients according to treatment efficiency. SN - 1879-1220 UR - https://www.unboundmedicine.com/medline/citation/31778802/Urinary_steroidomic_profiles_by_LC-MS/MS_to_monitor_classic_21-Hydroxylase_deficiency L2 - https://linkinghub.elsevier.com/retrieve/pii/S0960-0760(19)30406-6 DB - PRIME DP - Unbound Medicine ER -