Abstract
Leigh syndrome is a subacute necrotising encephalomyopathy proven by post-mortem analysis of brain tissue showing spongiform lesions with vacuolation of the neuropil followed by demyelination, gliosis and capillary proliferation caused by mutations in one of over 75 different genes, including nuclear- and mitochondrial-encoded genes, most of which are associated with mitochondrial respiratory chain function. In this study, we report a patient with suspected Leigh syndrome presenting with seizures, ptosis, scoliosis, dystonia, symmetrical putaminal abnormalities and a lactate peak on brain MRS, but showing normal MRC enzymology in muscle and liver, thereby complicating the diagnosis. Whole exome sequencing uncovered compound heterozygous mutations in NADH dehydrogenase (ubiquinone) flavoprotein 1 gene (NDUFV1), c.1162+4A>C (NM_007103.3), resulting in skipping of exon 8, and c.640G>A, causing the amino acid substitution p.Glu214Lys, both of which have previously been reported in a patient with complex I deficiency. Patient fibroblasts showed a significant reduction in NDUFV1 protein expression, decreased complex CI and complex IV assembly and consequential reductions in the enzymatic activities of both complexes by 38% and 67%, respectively. The pathogenic effect of these variations was further confirmed by immunoblot analysis of subunits for MRC enzyme complexes in patient muscle, liver and fibroblast where we observed 90%, 60% and 95% reduction in complex CI, respectively. Together these studies highlight the importance of a comprehensive, multipronged approach to the laboratory evaluation of patients with suspected Leigh syndrome.
Competing interests: None declared
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Barrientos A, Fontanesi F, Diaz F (2009) Evaluation of the mitochondrial respiratory chain and oxidative phosphorylation system using polarography and spectrophotometric enzyme assays. Curr Protoc Hum Genet. Chapter 19, Unit19 3
Benit P, Chretien D, Kadhom N, DE Lonlay-Debeney P, Cormier-Daire V, Cabral A, Peudenier S, Rustin P, Munnich A, Rotig A (2001) Large-scale deletion and point mutations of the nuclear NDUFV1 and NDUFS1 genes in mitochondrial complex I deficiency. Am J Hum Genet 68:1344–1352
Bird MJ, Wijeyeratne XW, Komen JC, Laskowski A, Ryan MT, Thorburn DR, Frazier AE (2014) Neuronal and astrocyte dysfunction diverges from embryonic fibroblasts in the Ndufs4fky/fky mouse. Biosci Rep 34:e00151
Calvo SE, Tucker EJ, Compton AG, Kirby DM, Crawford G, Burtt NP, Rivas M, Guiducci C, Bruno DL, Goldberger OA, Redman MC, Wiltshire E, Wilson CJ, Altshuler D, Gabriel SB, Daly MJ, Thorburn DR, Mootha VK (2010) High-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency. Nat Genet 42:851–858
Frazier AE, Thorburn DR (2012) Biochemical analyses of the electron transport chain complexes by spectrophotometry. Methods Mol Biol 837:49–62
Hui J, Kirby DM, Thorburn DR, Boneh A (2006) Decreased activities of mitochondrial respiratory chain complexes in non-mitochondrial respiratory chain diseases. Dev Med Child Neurol 48:132–136
Lake NJ, Compton AG, Rahman S, Thorburn DR (2015) Leigh syndrome: one disorder, more than 75 monogenic causes. Ann Neurol. doi:10.1002/ana.24551
Lemire BD (2015) Glutathione metabolism links FOXRED1 to NADH:ubiquinone oxidoreductase (complex I) deficiency: a hypothesis. Mitochondrion 24:105–112
Leong DW, Komen JC, Hewitt CA, Arnaud E, Mckenzie M, Phipson B, Bahlo M, Laskowski A, Kinkel SA, Davey GM, Heath WR, Voss AK, Zahedi RP, Pitt JJ, Chrast R, Sickmann A, Ryan MT, Smyth GK, Thorburn DR, Scott HS (2012) Proteomic and metabolomic analyses of mitochondrial complex I-deficient mouse model generated by spontaneous B2 short interspersed nuclear element (SINE) insertion into NADH dehydrogenase (ubiquinone) Fe-S protein 4 (Ndufs4) gene. J Biol Chem 287:20652–20663
Mckenzie M, Lazarou M, Thorburn DR, Ryan MT (2007) Analysis of mitochondrial subunit assembly into respiratory chain complexes using Blue Native polyacrylamide gel electrophoresis. Anal Biochem 364:128–137
Menezes MJ, Guo Y, Zhang J, Riley LG, Cooper ST, Thorburn DR, Li J, Dong D, Li Z, Glessner J, Davis RL, Sue CM, Alexander SI, Arbuckle S, Kirwan P, Keating BJ, Xu X, Hakonarson H, Christodoulou J (2015) Mutation in mitochondrial ribosomal protein S7 (MRPS7) causes congenital sensorineural deafness, progressive hepatic and renal failure and lactic acidemia. Hum Mol Genet 24:2297–2307
Mimaki M, Wang X, Mckenzie M, Thorburn DR, Ryan MT (2012) Understanding mitochondrial complex I assembly in health and disease. Biochim Biophys Acta 1817:851–862
Ogilvie I, Kennaway NG, Shoubridge EA (2005) A molecular chaperone for mitochondrial complex I assembly is mutated in a progressive encephalopathy. J Clin Invest 115:2784–2792
Rahman S, Blok RB, Dahl HH, Danks DM, Kirby DM, Chow CW, Christodoulou J, Thorburn DR (1996) Leigh syndrome: clinical features and biochemical and DNA abnormalities. Ann Neurol 39:343–351
Riley LG, Menezes MJ, Rudinger-Thirion J, Duff R, de Lonlay P, Rotig A, Tchan MC, Davis M, Cooper ST, Christodoulou J (2013) Phenotypic variability and identification of novel YARS2 mutations in YARS2 mitochondrial myopathy, lactic acidosis and sideroblastic anaemia. Orphanet J Rare Dis 8:193
Schuelke M, Smeitink J, Mariman E, Loeffen J, Plecko B, Trijbels F, Stockler-Ipsiroglu S, van den Heuvel L (1999) Mutant NDUFV1 subunit of mitochondrial complex I causes leukodystrophy and myoclonic epilepsy. Nat Genet 21:260–261
Suthammarak W, Morgan PG, Sedensky MM (2010) Mutations in mitochondrial complex III uniquely affect complex I in Caenorhabditis elegans. J Biol Chem 285:40724–40731
Tang G, Gutierrez Rios P, Kuo SH, Akman HO, Rosoklija G, Tanji K, Dwork A, Schon EA, Dimauro S, Goldman J, Sulzer D (2013) Mitochondrial abnormalities in temporal lobe of autistic brain. Neurobiol Dis 54:349–361
Thorburn DR, Chow CW, Kirby DM (2004) Respiratory chain enzyme analysis in muscle and liver. Mitochondrion 4:363–375
Tuppen HAL, Hogan VE, He L, Blakely EL, Worgan L, Al-Dosary M, Saretzki G, Alston CL, Morris AA, Clarke M, Jones S, Devlin AM, Mansour S, Chrzanowska-Lightowlers ZMA, Thorburn DR, Mcfarland R, Taylor RW (2010) The p.M292T NDUFS2 mutation causes complex I-deficient Leigh syndrome in multiple families. Brain 133:2952–2963
Ugalde C, Janssen RJ, van den Heuvel LP, Smeitink JA, Nijtmans LG (2004) Differences in assembly or stability of complex I and other mitochondrial OXPHOS complexes in inherited complex I deficiency. Hum Mol Genet 13:659–667
Wenchich L, Drahota Z, Honzik T, Hansikova H, Tesarova M, Zeman J, Houstek J (2003) Polarographic evaluation of mitochondrial enzymes activity in isolated mitochondria and in permeabilized human muscle cells with inherited mitochondrial defects. Physiol Res 52:781–788
Acknowledgements
This research was supported by Australian NHMRC grant 1026891 to J.C., an NHMRC Principal Research Fellowship to D.R.T., an Australian Mitochondrial Disease Foundation (AMDF) PhD Scholarship to M.N. and a Research Grant from the Shenzhen Municipal Government of China (NO.CXB201108250094A) to X.X. We would also like to acknowledge the Institutional Development Funds to H.H. Finally, we also gratefully acknowledge donations to J.C. by the Crane and Perkins families.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Additional information
Communicated by: Shamima Rahman, FRCP, FRCPCH, PhD
Electronic Supplementary Material
Below is the link to the electronic supplementary material.
Appendices
Compliance with Ethics Guidelines
All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000. Informed consent was obtained from all patients included in the study.
Synopsis
A definitive genetic diagnosis was provided by whole exome sequencing and functional studies in a patient with late-onset childhood Leigh syndrome and normal muscle and liver mitochondrial RC enzyme activities.
Conflicts of Interest
Michael Nafisinia, Yiran Guo, Xiao Dang, Jiankang Li, Yulan Chen, Jianguo Zhang, Nicole J Lake, Wendy A Gold, Lisa G Riley, David R Thorburn, Brendan Keating, Xun Xu, Hakon Hakonarson and John Christodoulou declare that they have no conflict of interest.
Details of Contributions of Individual Authors
Michael Nafisinia: design and implementation of structural and functional studies, data analysis and manuscript preparation
Yiran Guo, Xiao Dang, Jiankang Li, Yulan Chen, Jianguo Zhang, Brendan Keating, Xun Xu, Hakon Hakonarson: performance of whole exome sequencing, implementation of bioinformatics, data analysis and manuscript preparation
Wendy Gold, Lisa Riley: contribution to design of structural and functional studies, data analysis and manuscript preparation
Nicole J. Lake: implementation of structural and functional studies
David Thorburn: oversight of spectrophotometric assays, data analysis and manuscript preparation
John Christodoulou: overall oversight of the research project, clinical interface, data analysis and manuscript preparation
Rights and permissions
Copyright information
© 2016 SSIEM and Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Nafisinia, M. et al. (2016). Whole Exome Sequencing Identifies the Genetic Basis of Late-Onset Leigh Syndrome in a Patient with MRI but Little Biochemical Evidence of a Mitochondrial Disorder. In: Morava, E., Baumgartner, M., Patterson, M., Rahman, S., Zschocke, J., Peters, V. (eds) JIMD Reports, Volume 32. JIMD Reports, vol 32. Springer, Berlin, Heidelberg. https://doi.org/10.1007/8904_2016_541
Download citation
DOI: https://doi.org/10.1007/8904_2016_541
Received:
Revised:
Accepted:
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-662-54384-9
Online ISBN: 978-3-662-54385-6
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)