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Heteroplasmic mutant load differences in mitochondrial DNA-associated Leigh syndrome

  • Ji-Hoon Na
    Affiliations
    Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea
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  • Young-Mock Lee
    Correspondence
    Correspondence: Young-Mock Lee, MD, PhD, Department of Pediatrics, Yonsei University College of Medicine, Gangnam Severance Hospital, 211 Eonju-ro, Gangnam-gu, Seoul 135-720, Korea. Phone: +82-2-2019-3354, Fax: +82-2-2019-4881.
    Affiliations
    Department of Pediatrics, Yonsei University College of Medicine, Seoul, Korea

    Epilepsy Research Institute, Yonsei University College of Medicine, Seoul, Korea
    Search for articles by this author

      Highlights

      • Mutant load varies significantly in patients with mtDNA-associated Leigh Syndrome
      • MT-ATP6, MT-ND3, and MT-ND5 are the most common mutations
      • MT-ATP6 had a significantly higher mutant load than that of MT-ND3 and MT-ND5
      • Distribution of the heteroplasmic mutant load depends on the mutated gene

      Abstract

      Background

      Mitochondrial DNA (mtDNA)-associated Leigh syndrome is influenced by mutant pathogenicity and corresponding heteroplasmic loads; however, the manner in which heteroplasmic mutant load affects patient phenotypes and the relationship between mutant types and heteroplasmic mutant loads remain unknown.

      Objectives

      We aimed to investigate the distribution of the mutant load of different mtDNA mutations in a single-center cohort.

      Methods

      We used next-generation sequencing to confirm mtDNA mutations in 31 patients with Leigh syndrome. Subsequently, we counted the number of mtDNA reads to quantitatively analyze the heteroplasmic mutant load and categorize the patients according to the mtDNA mutations they harbored. Confirmed cases of mtDNA-associated Leigh syndrome were classified according to the mutations observed in 6 genes and 10 nucleotides.

      Results

      Of the 31 patients with Leigh syndrome, 27 harbored known pathogenic mutations. We discovered that MT-ATP6 was the most commonly mutated gene (n = 13 patients), followed by MT-ND3 (n = 7) and MT-ND5 (n = 4). MT-ATP6 had a significantly higher mutant load than MT-ND3 and MT-ND5 (p < 0.001, each). By contrast, MT-ND5 had a significantly lower mutant load than MT-ND3 (p = 0.007). Notably, the mutation loads varied significantly among patients carrying the MT-ATP6, MT-ND3, and MT-ND5 mutations.

      Conclusions

      Our study illustrated the heteroplasmic diversity and phenotypic expression threshold of mutated mitochondrial genes in mtDNA-associated Leigh syndrome. The results provide promising insights into the genotype–phenotype correlation in mtDNA-associated Leigh syndrome that are expected to guide the development of tailored treatments for Leigh syndrome.

      Keywords

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      References

        • Pfeffer G.
        • Chinnery P.F.
        Diagnosis and treatment of mitochondrial myopathies.
        Ann Med. 2013; 45: 4-16https://doi.org/10.3109/07853890.2011.605389
        • Dowling D.K.
        Evolutionary perspectives on the links between mitochondrial genotype and disease phenotype.
        Biochim Biophys Acta Gen Subj. 2014; 1840: 1393-1403https://doi.org/10.1016/j.bbagen.2013.11.013
        • Leigh D.
        Subacute necrotizing encephalomyelopathy in an infant.
        J Neurol Neurosurg Psychiatry. 1951; 14: 216-221https://doi.org/10.1136/jnnp.14.3.216
        • Rahman S.
        • Blok R.B.
        • Dahl H.H.
        • Danks D.M.
        • Kirby D.M.
        • Chow C.W.
        • et al.
        Leigh syndrome: clinical features and biochemical and DNA abnormalities.
        Ann Neurol. 1996; 39: 343-351https://doi.org/10.1002/ana.410390311
        • Marin S.E.
        • Mesterman R.
        • Robinson B.
        • Rodenburg R.J.
        • Smeitink J.
        • Tarnopolsky M.A.
        Leigh syndrome associated with mitochondrial complex I deficiency due to novel mutations in NDUFV1 and NDUFS2.
        Gene. 2013; 516: 162-167https://doi.org/10.1016/j.gene.2012.12.024
        • Björkman K.
        • Sofou K.
        • Darin N.
        • Holme E.
        • Kollberg G.
        • Asin-Cayuela J.
        • et al.
        Broad phenotypic variability in patients with complex I deficiency due to mutations in NDUFS1 and NDUFV1.
        Mitochondrion. 2015; 21: 33-40https://doi.org/10.1016/j.mito.2015.01.003
        • Han J.
        • Lee Y.-M.
        • Kim S.M.
        • Han S.Y.
        • Lee J.B.
        • Han S.-H.
        Ophthalmological manifestations in patients with Leigh syndrome.
        Br J Ophthalmol. 2015; 99: 528-535https://doi.org/10.1136/bjophthalmol-2014-305704
        • Monlleo-Neila L.
        • del Toro M.
        • Bornstein B.
        • Garcia-Arumi E.
        • Sarrias A.
        • Roig-Quilis M.
        • et al.
        Leigh syndrome and the mitochondrial m.13513G>A mutation: expanding the clinical spectrum.
        J Child Neurol. 2013; 28: 1531-1534https://doi.org/10.1177/0883073812460580
        • Ruiter E.M.
        • Siers M.H.
        • van den Elzen C.
        • van Engelen B.G.
        • Smeitink J.A.
        • Rodenburg R.J.
        • et al.
        The mitochondrial 13513G>A mutation is most frequent in Leigh syndrome combined with reduced complex I activity, optic atrophy and/or Wolff–Parkinson–White.
        Eur J Hum Genet. 2007; 15: 155-161https://doi.org/10.1038/sj.ejhg.5201735
        • Ortigoza-Escobar J.D.
        • Oyarzabal A.
        • Montero R.
        • Artuch R.
        • Jou C.
        • Jiménez C.
        • et al.
        Ndufs4 related Leigh syndrome: a case report and review of the literature.
        Mitochondrion. 2016; 28: 73-78https://doi.org/10.1016/j.mito.2016.04.001
        • Ruhoy I.S.
        • Saneto R.P.
        The genetics of Leigh syndrome and its implications for clinical practice and risk management.
        Appl Clin Genet. 2014; 7: 221-234https://doi.org/10.2147/TACG.S46176
        • Lake N.J.
        • Compton A.G.
        • Rahman S.
        • Thorburn D.R.
        Leigh syndrome: one disorder, more than 75 monogenic causes.
        Ann Neurol. 2016; 79: 190-203https://doi.org/10.1002/ana.24551
      1. Thorburn DR, Rahman J, Rahman S. Mitochondrial DNA-associated Leigh syndrome and NARP. In: Adam MP, Ardinger HH, Pagon RA, et al., editors. GeneReviews® [Internet]. Seattle: University of Washington; 1993–2022 [Updated 2017 Sep 28]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK1173/.

        • Wong L.-J.C.
        Next generation molecular diagnosis of mitochondrial disorders.
        Mitochondrion. 2013; 13: 379-387https://doi.org/10.1016/j.mito.2013.02.001
        • Ma Y.-Y.
        • Wu T.-F.
        • Liu Y.-P.
        • Wang Q.
        • Li X.Y.
        • Song J.Q.
        • et al.
        Heterogeneity of six children and their mothers with mitochondrial DNA 3243 A>G mutation.
        Mitochondrial DNA. 2013; 24: 297-302https://doi.org/10.3109/19401736.2012.760071
        • Ma Y.-Y.
        • Wu T.-F.
        • Liu Y.-P.
        • Wang Q.
        • Song J.Q.
        • Li X.Y.
        • et al.
        Genetic and biochemical findings in Chinese children with Leigh syndrome.
        J Clin Neurosci. 2013; 20: 1591-1594https://doi.org/10.1016/j.jocn.2013.03.034
        • Gould M.P.
        • Bosworth C.M.
        • McMahon S.
        • Grandhi S.
        • Grimberg B.T.
        • LaFramboise T.
        PCR-free enrichment of mitochondrial DNA from human blood and cell lines for high quality next-generation DNA sequencing.
        PLoS One. 2015; 10e0139253https://doi.org/10.1371/journal.pone.0139253
        • Craig D.W.
        • Nasser S.
        • Corbett R.
        • Chan S.K.
        • Murray L.
        • Legendre C.
        • et al.
        A somatic reference standard for cancer genome sequencing.
        Sci Rep. 2016; 624607https://doi.org/10.1038/srep24607
        • Huptas C.
        • Scherer S.
        • Wenning M.
        Optimized Illumina PCR-free library preparation for bacterial whole genome sequencing and analysis of factors influencing de novo assembly.
        BMC Res Notes. 2016; 9: 269https://doi.org/10.1186/s13104-016-2072-9
        • Palculict M.E.
        • Zhang V.W.
        • Wong L.-J.
        • Wang J.
        Comprehensive mitochondrial genome analysis by massively parallel sequencing.
        Methods Mol Biol. 2016; 1351: 3-17https://doi.org/10.1007/978-1-4939-3040-1_1
        • Preston J.L.
        • Royall A.E.
        • Randel M.A.
        • Sikkink K.L.
        • Phillips P.C.
        • Johnson E.A.
        High-specificity detection of rare alleles with paired-end low error sequencing (PELE-Seq).
        BMC Genomics. 2016; 17: 464https://doi.org/10.1186/s12864-016-2669-3
      2. Rahman S, Thorburn D. Nuclear gene-encoded Leigh syndrome spectrum overview. In: Adam MP, Ardinger HH, Pagon RA, et al., editors. GeneReviews® [Internet]. Seattle: University of Washington; 1993–2022 [Updated 2020 Jul 16]. Available from: https://www.ncbi.nlm.nih.gov/books/NBK320989/

        • Naess K.
        • Freyer C.
        • Bruhn H.
        • Wibom R.
        • Malm G.
        • Nennesmo I.
        • et al.
        MtDNA mutations are a common cause of severe disease phenotypes in children with Leigh syndrome.
        Biochim Biophys Acta. 2009; 1787: 484-490https://doi.org/10.1016/j.bbabio.2008.11.014
        • Ogawa E.
        • Shimura M.
        • Fushimi T.
        • Tajika M.
        • Ichimoto K.
        • Matsunaga A.
        • et al.
        Clinical validity of biochemical and molecular analysis in diagnosing Leigh syndrome: a study of 106 Japanese patients.
        J Inherit Metab Dis. 2017; 40: 685-693https://doi.org/10.1007/s10545-017-0042-6
        • Ng Y.S.
        • Lax N.Z.
        • Maddison P.
        • Alston C.L.
        • Blakely E.L.
        • Hepplewhite P.D.
        • et al.
        MT-ND5 mutation exhibits highly variable neurological manifestations at low mutant load.
        EBioMedicine. 2018; 30: 86-93https://doi.org/10.1016/j.ebiom.2018.02.010
        • Sofou K.
        • de Coo I.F.M.
        • Ostergaard E.
        • Isohanni P.
        • Naess K.
        • De Meirleir L.
        • et al.
        Phenotype-genotype correlations in Leigh syndrome: new insights from a multicentre study of 96 patients.
        J Med Genet. 2018; 55: 21-27https://doi.org/10.1136/jmedgenet-2017-104891
        • Wei Y.
        • Cui L.
        • Peng B.
        Mitochondrial DNA mutations in late-onset Leigh syndrome.
        J Neurol. 2018; 265: 2388-2395https://doi.org/10.1007/s00415-018-9014-5
        • Tatuch Y.
        • Christodoulou J.
        • Feigenbaum A.
        • Clarke J.T.
        • Wherret J.
        • Smith C.
        • et al.
        Heteroplasmic mtDNA mutation (T----G) at 8993 can cause Leigh disease when the percentage of abnormal mtDNA is high.
        Am J Hum Genet. 1992; 50 (PMID: 1550128): 852-858
        • White S.L.
        • Collins V.R.
        • Wolfe R.
        • Cleary M.A.
        • Shanske S.
        • DiMauro S.
        • et al.
        Genetic counseling and prenatal diagnosis for the mitochondrial DNA mutations at nucleotide 8993.
        Am J Hum Genet. 1999; 65: 474-482https://doi.org/10.1086/302488
        • Stendel C.
        • Neuhofer C.
        • Floride E.
        • Yuqing S.
        • Ganetzky R.D.
        • Park J.
        • et al.
        Delineating MT-ATP6-associated disease: From isolated neuropathy to early-onset neurodegeneration.
        Neurol Genet. 2020; 6: e393https://doi.org/10.1212/NXG.0000000000000393
      3. Stenton SL, Zou Y, Cheng H, Liu Z, Wang J, Shen D, et al. Leigh syndrome: a study of 209 patients at the Beijing Children's Hospital. Ann Neurol. 2022 [Epub ahead of print]. https://doi.org/10.1002/ana.26313.

        • Na J.-H.
        • Lee M.J.
        • Lee C.H.
        • Lee Y.-M.
        Association between epilepsy and Leigh syndrome with MT-ND3 mutation, particularly the m.10191T>C point mutation.
        Front Neurol. 2021; 12752467https://doi.org/10.3389/fneur.2021.752467