P Alcaide1, J Krijt2, P Ruiz-Sala1, P Ješina2, M Ugarte1, V Kožich2, B Merinero3. 1. Centro de Diagnóstico de Enfermedades Moleculares, Facultad de Ciencias, Universidad Autónoma Madrid, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), 28049 Madrid, Spain. 2. Institute of Inherited Metabolic Disorders, Charles University in Prague, 1st Faculty of Medicine and General University Hospital in Prague, Ke Karlovu 2, 128 08 Praha 2, Czech Republic. 3. Centro de Diagnóstico de Enfermedades Moleculares, Facultad de Ciencias, Universidad Autónoma Madrid, Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), 28049 Madrid, Spain. Electronic address: bmerinero@cbm.csic.es.
Abstract
BACKGROUND: Cystathionine β-synthase (CBS) is released into plasma from organs expressing this enzyme. Decreased plasma CBS activity has been demonstrated in CBS-deficient patients with 16 different genotypes. The aim of this study was to determine plasma CBS activity in patients carrying 11 additional genotypes using two LC-MS/MS methods. Patients and methods CBS activity was measured in EDTA or heparin plasma using either a previously described or a newly developed LC-MS/MS method optimized for analysis of the reaction product, 3,3-(2)H2-cystathionine, as its butyl ester derivative. We analyzed plasma samples from 26 CBS-deficient patients with known genotypes and 57 controls. RESULTS: We developed a new LC-MS/MS method for simple and sensitive determination of CBS activity. Plasma CBS activity was low (i.e., 0.001-0.036 of the multiples of median control values, MoM) in patients homozygous for the prevalent Hispanic mutation c.572C>T (p.T191M) but was highly elevated (2.95 MoM) in a single patient homozygous for the c.1330G>A (p.D444N) mutation. Patients with the remaining nine genotypes exhibited decreased activities (0.00-0.22 MoM), which did not overlap with the controls (0.29-2.10 MoM). CONCLUSIONS: The determination of CBS activity in plasma is a rapid and non-invasive procedure for detecting a subgroup of CBS-deficient patients with distinct genotypes.
BACKGROUND: Cystathionine β-synthase (CBS) is released into plasma from organs expressing this enzyme. Decreased plasma CBS activity has been demonstrated in CBS-deficientpatients with 16 different genotypes. The aim of this study was to determine plasma CBS activity in patients carrying 11 additional genotypes using two LC-MS/MS methods. Patients and methods CBS activity was measured in EDTA or heparin plasma using either a previously described or a newly developed LC-MS/MS method optimized for analysis of the reaction product, 3,3-(2)H2-cystathionine, as its butyl ester derivative. We analyzed plasma samples from 26 CBS-deficientpatients with known genotypes and 57 controls. RESULTS: We developed a new LC-MS/MS method for simple and sensitive determination of CBS activity. Plasma CBS activity was low (i.e., 0.001-0.036 of the multiples of median control values, MoM) in patients homozygous for the prevalent Hispanic mutation c.572C>T (p.T191M) but was highly elevated (2.95 MoM) in a single patient homozygous for the c.1330G>A (p.D444N) mutation. Patients with the remaining nine genotypes exhibited decreased activities (0.00-0.22 MoM), which did not overlap with the controls (0.29-2.10 MoM). CONCLUSIONS: The determination of CBS activity in plasma is a rapid and non-invasive procedure for detecting a subgroup of CBS-deficientpatients with distinct genotypes.
Authors: Andrew A M Morris; Viktor Kožich; Saikat Santra; Generoso Andria; Tawfeg I M Ben-Omran; Anupam B Chakrapani; Ellen Crushell; Mick J Henderson; Michel Hochuli; Martina Huemer; Miriam C H Janssen; Francois Maillot; Philip D Mayne; Jenny McNulty; Tara M Morrison; Helene Ogier; Siobhan O'Sullivan; Markéta Pavlíková; Isabel Tavares de Almeida; Allyson Terry; Sufin Yap; Henk J Blom; Kimberly A Chapman Journal: J Inherit Metab Dis Date: 2016-10-24 Impact factor: 4.982
Authors: Song Sun; Jochen Weile; Marta Verby; Yingzhou Wu; Yang Wang; Atina G Cote; Iosifina Fotiadou; Julia Kitaygorodsky; Marc Vidal; Jasper Rine; Pavel Ješina; Viktor Kožich; Frederick P Roth Journal: Genome Med Date: 2020-01-30 Impact factor: 11.117
Authors: Viktor Kožich; Jitka Sokolová; Andrew A M Morris; Markéta Pavlíková; Florian Gleich; Stefan Kölker; Jakub Krijt; Carlo Dionisi-Vici; Matthias R Baumgartner; Henk J Blom; Martina Huemer Journal: J Inherit Metab Dis Date: 2020-12-28 Impact factor: 4.982