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One mutation, three phenotypes: novel metabolic insights on MELAS, MIDD and myopathy caused by the m.3243A > G mutation

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Abstract

Introduction

The m.3243A > G mitochondrial DNA mutation is one of the most common mitochondrial disease-causing mutations, with a carrier rate as high as 1:400. This point mutation affects the MT-TL1 gene, ultimately affecting the oxidative phosphorylation system and the cell’s energy production. Strikingly, the m.3243A > G mutation is associated with different phenotypes, including mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS), maternally inherited diabetes and deafness (MIDD) and myopathy.

Objectives

We investigated urine metabolomes of MELAS, MIDD and myopathy patients in order to identify affected metabolic pathways and possible treatment options.

Methods

A multiplatform metabolomics approach was used to comprehensively analyze the metabolome and compare metabolic profiles of different phenotypes caused by the m.3243A > G mutation. Our analytical array consisted of NMR spectroscopy, LC-MS/MS and GC-TOF-MS.

Results

The investigation revealed phenotypic specific metabolic perturbations, as well as metabolic similarities between the different phenotypes. We show that glucose metabolism is highly disturbed in the MIDD phenotype, but not in MELAS or myopathy, remodeled fatty acid oxidation is characteristic of the MELAS patients, while one-carbon metabolism is strongly modified in both MELAS and MIDD, but not in the myopathy group. Lastly we identified increased creatine in the urine of the myopathy patients, but not in MELAS or MIDD.

Conclusion

We conclude by giving novel insight on the phenotypes of the m.3243A > G mutation from a metabolomics point of view. Directives are also given for future investigations that could lead to better treatment options for patients suffering from this debilitating disease.

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Abbreviations

2-HG:

2-Hydroxyglutaric acid

ADP:

Adenosine diphosphate

ANOVA:

Analysis of variance

ATP:

Adenosine triphosphate

AUC:

Area under the curve

BCAA:

Branched-chain amino acid

Complex V:

ATP synthase

FAD:

Oxidized flavin adenine dinucleotide

FADH2 :

Reduced flavin adenine dinucleotide

FAS:

Fatty acids synthesis

GC–TOF–MS:

Gas chromatography time-of-flight mass spectrometry

KSS:

Kearns–Sayre syndrome

LC–MS/MS:

Liquid chromatography tandem mass spectrometry

LS:

Leigh syndrome

MCFA:

Medium chain fatty acids

MD:

Mitochondrial disease

MELAS:

Mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes

MERRF:

Myoclonus epilepsy with ragged red fibers

MIDD:

Maternally inherited diabetes and deafness

MS:

Mass spectrometry

mtDNA:

Mitochondrial DNA

mTOR:

mammalian target of rapamycin

MT-TL1:

Mitochondria-encoded tRNA leucine 1

NAD+ :

Oxidized nicotine amide adenine dinucleotide

NADH:

Reduced nicotine amide adenine dinucleotide

NMR:

Proton nuclear magnetic resonance spectroscopy

OXPHOS:

Oxidative phosphorylation

PCA:

Principal component analysis

PCr:

Phosphocreatine

PEO:

progressive external ophthalmoplegia

ROC:

Receiver operating characteristic

SCFA:

Short chain fatty acids

SI:

Supplementary information

TCA:

Tricarboxylic acid

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Acknowledgements

We would like to thank Prof. L.J. Mienie from the North-West University, Potchefstroom, South Africa, for valuable help with the interpretation of the metabolic data. The research was supported by a grant from the Technology Innovation Agency (TIA, Grant Number Metabol. 01), South Africa. The research facilities were provided by the National Metabolomics Platform, North-West University, Potchefstroom Campus, Potchefstroom, South Africa.

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Authors and Affiliations

  1. Mitochondria Research Laboratory, Human Metabolomics, North-West University, Potchefstroom, South Africa

    Karien Esterhuizen, J. Zander Lindeque, Shayne Mason, Francois H. van der Westhuizen & Roan Louw

  2. Department of Pediatrics, Radboud Center for Mitochondrial Medicine, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands

    Richard J. Rodenburg, Paul de Laat, Jan A. M. Smeitink & Mirian C. H. Janssen

  3. Department of Internal Medicine, Radboud Center for Mitochondrial Medicine, Radboud University Nijmegen Medical Center, Nijmegen, The Netherlands

    Mirian C. H. Janssen

  4. Human Metabolomics, Faculty of Natural and Agricultural Sciences, North-West University, Potchefstroom Campus, Private Bag X6001, Potchefstroom, South Africa

    Roan Louw

Authors
  1. Karien Esterhuizen
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  2. J. Zander Lindeque
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  3. Shayne Mason
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  4. Francois H. van der Westhuizen
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  5. Richard J. Rodenburg
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  7. Jan A. M. Smeitink
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Contributions

RL, MJ, ZL, FVDW and JS planned and oversaw the study. KE, ZL and SM performed all analytical work. Data mining was performed by RL, ZL, SM and KE. The manuscript was written by KE, RL and ZL, assisted by all the authors.

Corresponding author

Correspondence to Roan Louw.

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Conflict of interest

The authors declare no competing interests. Jan Smeitink is the founding CEO of Khondrion.

Ethical approval

The study complied with all Institutional guidelines and terms of the Declaration of Helsinki of 1975 (as revised in 2013) for investigation of human participants. Ethical approval was obtained from the Health Research Ethics Committee (HREC) of the North-West University, South Africa (NWU-00170-13-S1) and the Ethics Committee of the Nijmegen-Arnhem region of the Netherlands (2010/183; NL32683.091.10). All the patients, as well as the controls, provided written informed consent for their urine samples to be used for these research purposes.

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Esterhuizen, K., Lindeque, J.Z., Mason, S. et al. One mutation, three phenotypes: novel metabolic insights on MELAS, MIDD and myopathy caused by the m.3243A > G mutation. Metabolomics 17, 10 (2021). https://doi.org/10.1007/s11306-020-01769-w

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