Abstract
Organic acid (OAD) and fatty acid oxidation disorders (FAOD) are inborn errors of metabolism often presenting with life-threatening metabolic decompensation followed by (irreversible) organ failure, and even death during catabolic state. Most of these diseases are considered as treatable, and metabolic decompensations can be avoided by early diagnosis and start of therapy. Confirmation of suspected diagnosis currently relies on enzymatic and mutation analyses and in vitro loading of palmitic acid in human skin fibroblast cultures. Furthermore, in some cases potentially life-threatening in vivo loading or fasting tests are still performed. In this study, we established a standardized in vitro loading test in peripheral blood mononuclear cells (PBMC) that allows reliable biochemical confirmation of a suspected diagnosis within 1 week. Patients with confirmed diagnosis of short-, medium-, very-long-chain, and long-chain 3-hydroxyacyl-CoA dehydrogenase deficiencies, methylmalonic, propionic, isovaleric acidurias, and glutaric aciduria type I were included in the study. PBMC, isolated from heparinized venous blood samples of these individuals were incubated for 5 days with palmitic acid or 2-oxoadipic acid (glutaric aciduria type I), respectively, and quantitative acylcarnitine profiling was subsequently performed in supernatants using electrospray ionization tandem mass spectrometry. All patients were clearly identified, including those with mild biochemical phenotypes who, in particular, are at risk to be missed under balanced metabolic conditions. In glutaric aciduria type I, the same results were also obtained using lymphoblasts. In conclusion, our assay allows biochemical confirmation of a number of FAOD and OAD and could easily be implemented into the confirmatory diagnostic work-up.
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Abbreviations
- DBS:
-
dried blood spots
- FAOD:
-
fatty acid oxidation disorders
- GA-I:
-
glutaric aciduria type I
- GC/MS:
-
gas chromatography/mass spectrometry
- IL-2:
-
interleukin-2
- IVA:
-
isovaleric aciduria
- LCHAD(D):
-
long-chain 3-hydroxyacyl-CoA dehydrogenase [EC 1.1.1.211] (deficiency)
- MCAD(D):
-
medium-chain acyl-CoA dehydrogenase [EC 1.3.99.3] (deficiency)
- MMA:
-
methylmalonic aciduria
- MS/MS:
-
electrospray ionisation-tandem mass spectrometry
- OAD:
-
organic acid disorders
- PA:
-
propionic aciduria
- PBMC:
-
peripheral blood mononuclear cells
- SCAD(D):
-
short-chain acyl-CoA dehydrogenase [EC 1.3.99.2] (deficiency)
- VLCAD(D):
-
very long-chain acyl-CoA dehydrogenase [EC 1.3.99.13] (deficiency)
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Acknowledgements
The authors are grateful to S. Exner-Camps and M. Herm for excellent technical support.
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This study was supported by a grant from the Medical Faculty of the University of Heidelberg to A. S.-B. (# F.206611) and by a grant from the German Federal Ministry of Education and Science (BMBF # 01GM0305; project # 8) to S. K.
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Schulze-Bergkamen, A., Okun, J., Spiekerkötter, U. et al. Quantitative Acylcarnitine Profiling in Peripheral Blood Mononuclear Cells Using In Vitro Loading With Palmitic and 2-Oxoadipic Acids: Biochemical Confirmation of Fatty Acid Oxidation and Organic Acid Disorders. Pediatr Res 58, 873–880 (2005). https://doi.org/10.1203/01.PDR.0000181378.98593.3E
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DOI: https://doi.org/10.1203/01.PDR.0000181378.98593.3E
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