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Molecular analysis of 51 unrelated pedigrees with late-onset multiple acyl-CoA dehydrogenation deficiency (MADD) in southern China confirmed the most common ETFDH mutation and high carrier frequency of c.250G>A

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Abstract

Multiple acyl-CoA dehydrogenation deficiency (MADD) is an autosomal recessive disease affecting amino acid, fatty acid, and choline metabolisms and is a common genetic defect responsible for lipid storage myopathy. Most forms of MADD are caused by a deficiency of electron transfer flavoprotein (ETF) or ETF dehydrogenase (ETFDH). However, its molecular feature has not been found uniformly in previous reports of Chinese patients. A large cohort of 56 late-onset MADD patients from 51 unrelated pedigrees in southern China was recruited to investigate a clear correlation between clinical phenotype and molecular genetic basis. All exons of ETFA, ETFB, and ETFDH, including the intron–exon boundaries, and 5′ and 3′ untranslated regions were directly sequenced. ETFDH deficiencies affected 94.1% (48/51) of the pedigrees. ETFDH-c.250G>A is the most common mutation, representing a high allelic frequency of 83.3% (80/96). Carrier frequency of c.250G>A is estimated to be 1.35% (7/520) in the normal population. A significant reduced expression of ETFDH was identified in the muscle of ETFDH-deficient patients. ETFDH deficiency is a major cause of riboflavin-responsive MADD in southern China, and c.250G>A is an important mutation that could be employed as a fast and reliable screening method.

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Acknowledgments

The authors sincerely thank the participants for their help and willingness to participate in this study and the anonymous reviewers for helping to improve this manuscript.

Funding/support

This work was supported by a key program of scientific research of Fujian Medical University (2009D064), a program for Innovative Research Teams in Science (FMU-RT002), Fuzhou, and a grant from Huashan Hospital for the special professorship of Fudan University, Shanghai (to Wu ZY), China.

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The authors declare that they have no conflicts of interest.

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Authors and Affiliations

  1. Department of Neurology and Institute of Neurology, Huashan Hospital, Institutes of Brain Science and State Key Laboratory of Medical Neurobiology, Shanghai Medical College, Fudan University, 12 Wulumuqi Zhong Road, Shanghai, 200040, China

    Zhi-Qiang Wang & Zhi-Ying Wu

  2. Department of Neurology and Institute of Neurology, First Affiliated Hospital, Fujian Medical University, 20 Chazhong Road, Fuzhou, 350005, China

    Zhi-Qiang Wang, Xue-Jiao Chen, Shen-Xing Murong, Ning Wang & Zhi-Ying Wu

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  1. Zhi-Qiang Wang
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  2. Xue-Jiao Chen
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  3. Shen-Xing Murong
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  4. Ning Wang
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Corresponding author

Correspondence to Zhi-Ying Wu.

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Supplementary Fig. 1

Muscle biopsy of a patient with MADD showing common histological and electron microscopy findings. a Hematoxylin and eosin stain showed moderate to severe vacuolar myopathy. b ATPase stain (pH 4.6) revealed that the vacuolar change was most prominent in type-I fibers. c ORO stain showed lipid accumulation in type-I fibers. d There was a marked increase in the amount of lipid deposited within the myofibers. A′, B′, C′, and D′ represented the boxed region in A, B, C, D at a higher magnification (JPEG 1315 kb)

Supplementary Fig. 2

Four known mutations including c.250G>A (p.Ala84Thr), c.524G>A (p.Arg175His), c.770A>G (p.Tyr257Cys), and c.1395T>G (p.Tyr465X) in ETFDH. The upper panel for each chromatogram depicts the normal sequence, whereas the lower panel represents mutated sequence (JPEG 1251 kb)

Supplementary Fig. 3

Primer mismatch PCR-RFLP analysis of c.770A>G, c.998A>G, and c.1395T>G mutations. Left: Analysis of the c.770A>G mutation. After cleavage with RsaI, the allele with c.770A>G was cut into 156-bp and 20-bp fragments, whereas the normal allele was not cut. 1–3 Patients with compound heterozygous mutations of c.[250G>A]+[770A>G], c.[524G>A]+[770A>G], c.[770A>G]+[1254_1257del], respectively. 4-5 Normal controls. Middle: Analysis of the c.998A>G mutation. After cleavage with MaeII (or TaiI), the normal allele was cut into 106-bp and 24-bp fragments, whereas the allele with c.998A>G was not cut. 1-2 Two patients with compound heterozygous mutations of c.[250G>A]+[998A>G]. 3–5 Normal controls. Right: Analysis of the c.1395T>G mutation. After cleavage with Csp68III, the normal allele was cut into 115-bp and 20-bp fragments, whereas the allele with c.1395T>G was not cut. 1 Patient with a single heterozygous of c.[1395T>G]+[?]. 2–5 Normal controls. M DL2000 marker (JPEG 383 kb)

Supplementary Table

Primers designed for ETFA, ETFB, ETFDH, and PCR conditions (DOC 78 kb)

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Wang, ZQ., Chen, XJ., Murong, SX. et al. Molecular analysis of 51 unrelated pedigrees with late-onset multiple acyl-CoA dehydrogenation deficiency (MADD) in southern China confirmed the most common ETFDH mutation and high carrier frequency of c.250G>A. J Mol Med 89, 569–576 (2011). https://doi.org/10.1007/s00109-011-0725-7

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  • DOI: https://doi.org/10.1007/s00109-011-0725-7

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