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Antioxidant dysfunction: potential risk for neurotoxicity in ethylmalonic aciduria

  • Original Article
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Journal of Inherited Metabolic Disease

Abstract

Mitochondrial dysfunction and oxidative stress are central to the molecular basis of several human diseases associated with neuromuscular disabilities. We hypothesize that mitochondrial dysfunction also contributes to the neuromuscular symptoms observed in patients with ethylmalonic aciduria and homozygosity for ACADS c.625G>A-a common variant of the short-chain acyl-coenzyme A (CoA) dehydrogenase (SCAD) enzyme in the mitochondrial fatty acid oxidation pathway. This study sought to identify the specific factors that initiate cell dysfunction in these patients. We investigated fibroblast cultures from 10 patients with neuromuscular disabilities, elevated levels of ethylmalonic acid (EMA) (>50 mmol/mol creatinine), and ACADS c.625G>A homozygosity. Functional analyses, i.e., ACADS gene and protein expression as well as SCAD enzyme activity measurements, were performed together with a global nano liquid chromatography tandem mass spectroscopy (nano-LC-MS/MS)-based screening of the mitochondrial proteome in patient fibroblasts. Moreover, cell viability of patient fibroblasts exposed to menadione-induced oxidative stress was evaluated. Loss of SCAD function was detected in the patient group, most likely due to decreased ACADS gene expression and/or elimination of misfolded SCAD protein. Analysis of the mitochondrial proteome in patient fibroblasts identified a number of differentially expressed protein candidates, including reduced expression of the antioxidant superoxide dismutase 2 (SOD2). Additionally, patient fibroblasts demonstrated significantly higher sensitivity to oxidative stress than control fibroblasts. We propose that reduced mitochondrial antioxidant capacity is a potential risk factor for ACADS c.625G>A-associated ethylmalonic aciduria and that mitochondrial dysfunction contributes to the neurotoxicity observed in patients.

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Abbreviations

FCS:

Fetal calf serum

EMA:

Ethylmalonic acid

FAD:

Flavin adenine dinucleotide

SCAD:

Short-chain acyl-CoA dehydrogenase

SOD2:

Superoxide dismutase 2

PPAR:

Peroxisome proliferator activated receptor

ETF:

Electron transfer flavoprotein

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Acknowledgement

We thank all the clinicians and researchers from clinical departments around the world for providing DNA and cultured skin fibroblasts from patients and controls (Mike Gibson, OR, USA; Jerry Vockley, Pittsburgh, PA, USA; Orly Elpeleg, Jerusalem, Israel; Mary G. Ampola, Boston, MA, USA; Antonia Ribes, Barcelona, Spain; Dieter Matern, Rochester, NY, USA; Rhona Jack, Seattle, WA, USA; Tarja Linnankivi, Helsinki, Finland; Ina Knerr, Erlangen, Germany; Charles Roe, Dallas, TX, USA; Stanley Korman, Jerusalem, Israel; Sarah P. Young and David S. Millington, NC, USA). We also thank Christian Knudsen at the Department of Human Genetics for assisting with the cell culturing, and Dr. Kay Tanaka for providing the anti-SCAD antibodies. The work was supported by grants from the Danish Medical Research Council and Lundbeck Foundation, Denmark, as well as from a grant from the Physicians’ Services Incorporated Foundation of Ontario, Canada.

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  1. Christina B. Pedersen & Søren Vang

    Present address: Department of Molecular Medicine (MOMA), Aarhus University Hospital, Skejby, Brendstrupgaardsvej 100, 8200, Aarhus N, Denmark

Authors and Affiliations

  1. Research Unit for Molecular Medicine, Aarhus University Hospital, Skejby, Brendstrupgaardsvej 100, 8200, Aarhus N, Denmark

    Christina B. Pedersen, Søren Vang, Johan Palmfeldt, Margrethe Kjeldsen, Vibeke Stenbroen, Stinne P. Schmidt & Niels Gregersen

  2. Neurometabolic Research Laboratory, Department of Pediatrics, Hospital for Sick Children, University of Toronto, Toronto, Ontario, Canada

    Zarazuela Zolkipli & Ingrid Tein

  3. Lab. Genetic Metabolic Diseases, Department of Clinical Chemistry and Pediatrics, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    Ronald J. A. Wanders & Jos P. N. Ruiter

  4. Department of Clinical Genetics, Rigshospitalet, Copenhagen, Denmark

    Flemming Wibrand

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  1. Christina B. Pedersen
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  2. Zarazuela Zolkipli
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Corresponding author

Correspondence to Christina B. Pedersen.

Additional information

Communicated by: Matthias Baumgartner

References to electronic databases: Acyl-CoA Dehydrogenase, Short-chain, deficiency of: OMIM 201470; Ethylmalonic encephalopathy: OMIM 602473

Competing interest: None declared.

Electronic supplementary material

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Table S1

ETHE1 haplotypes in 10 c.625G>A homozygous EMA-patients (DOC 34 kb)

Table S2

Mitochondrial energy production (DOC 34.5 kb)

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Pedersen, C.B., Zolkipli, Z., Vang, S. et al. Antioxidant dysfunction: potential risk for neurotoxicity in ethylmalonic aciduria. J Inherit Metab Dis 33, 211–222 (2010). https://doi.org/10.1007/s10545-010-9086-6

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