Abstract
Mitochondrial fatty acid oxidation is a vital biochemical process for energy metabolism. Among the known fatty-acid metabolism disorders, very-long-chain acyl-CoA dehydrogenase (VLCAD) deficiency and medium-chain acyl-CoA dehydrogenase (MCAD) deficiency count among the most frequent. Both are potentially very serious diseases as they carry a risk of severe neurological post-crisis sequelae, and even sudden death. Diagnosis relies on plasma acylcarnitine profile analysis and urine organic acid analysis, followed by genetic testing to confirm diagnosis. However, in some cases, it is crucial to run a specific diagnostic assay for enzyme activity, which is generally performed in leukocytes or fibroblasts. The aim of this study was to address this need, first by developing a MCAD and VLCAD enzyme activity-specific diagnostic assay in fibroblasts (by measuring the reaction products, i.e. enoyl-CoA) via a rapid LC–MS/MS-based technique, and then by testing MCAD-deficient patients (n = 6), VLCAD-deficient patients (n = 10), and control patients (n = 12). MCAD activity was significantly different in the MCAD-deficiency (MCADD) group (mean = 0.07 nmol C8:1 formed/min/mg protein) compared to the control group (mean = 0.36 nmol C8:1 formed/min/mg protein). All MCADD patients showed less than 35% residual MCAD activity. VLCAD activity was significantly decreased in the VLCADD group (mean = 0.06 nmol C16:1 formed/min/mg protein) compared to the control group (mean = 0.86 nmol C16:1 formed/min/mg protein, respectively). All VLCADD patients showed less than 35% residual VLCAD activity. This technique allowed also to confirm that a novel ACADVL gene mutation (c.1400T>C) is responsible for a defective VLCAD activity (residual activity at 10%).
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Abbreviations
- ETF:
-
Electron Transfer Flavoprotein
- FA:
-
Fatty acids
- FAD:
-
Flavin adenine dinucleotide
- HPLC:
-
High-performance liquid chromatography
- MCAD:
-
Medium-chain acyl-CoA dehydrogenase
- MCADD:
-
MCAD deficiency
- MES:
-
2-(N-morpholino)ethanesulfonic acid
- OMIM:
-
Online Mendelian Inheritance in Man
- VLCAD:
-
Very-long-chain acyl-CoA dehydrogenase
- VLCADD:
-
VLCAD deficiency
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The authors thank A.T.T. (Auvergne Traduction Technique®) for proofreading the manuscript.
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Communicated by: Cornelis Jakobs
Appendices
Take-Home Message
The reader will learn about the usefulness of MCAD and VLCAD activity measurement and the performance of the method developed in fibroblasts.
Conflict of Interest
Damien Bouvier, Christine Vianey-Saban, Séverine Ruet, and Cécile Acquaviva declare that they have no conflict of interest.
Compliance with ethics guidelines: Informed consent was obtained from all patients for being included in the study.
Authors Contribution
Damien Bouvier and Severine Ruet have performed technical part of the study.
Damien Bouvier, Christine Vianey-Saban, Severine Ruet, and Cécile Acquaviva have performed analysis and interpretation of the data.
Damien Bouvier drafted the article.
Christine Vianey-Saban and Cécile Acquaviva revised the article.
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© 2016 Society for the Study of Inborn Errors of Metabolism (SSIEM)
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Bouvier, D., Vianey-Saban, C., Ruet, S., Acquaviva, C. (2016). Development of a Tandem Mass Spectrometry Method for Rapid Measurement of Medium- and Very-Long-Chain Acyl-CoA Dehydrogenase Activity in Fibroblasts. In: Morava, E., Baumgartner, M., Patterson, M., Rahman, S., Zschocke, J., Peters, V. (eds) JIMD Reports, Volume 35. JIMD Reports, vol 35. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2016_22
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DOI: https://doi.org/10.1007/8904_2016_22
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