Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing

Tobias B. Haack, Birgit Haberberger, Eva Maria Frisch, Thomas Wieland, Arcangela Iuso, Matteo Gorza, Valentina Strecker, Elisabeth Graf, Johannes A. Mayr, Ulrike Herberg, Julia B. Hennermann, Thomas Klopstock, Klaus A. Kuhn, Uwe Ahting, Wolfgang Sper, Ekkehard Wilichowski, Georg F. Hoffmann, Marketa Tesarova, Hana Hansikova, Jiri ZemanBarbara Plecko, Massimo Zeviani, Ilka Wittig, Tim M. Strom, Markus Schuelke, Peter Freisinger, Thomas Meitinger, Holger Prokisch

Research output: Contribution to journalArticle

Abstract

Background Next generation sequencing has become the core technology for gene discovery in rare inherited disorders. However, the interpretation of the numerous sequence variants identified remains challenging. We assessed the application of exome sequencing for diagnostics in complex I deficiency, a disease with vast genetic heterogeneity. Methods Ten unrelated individuals with complex I deficiency were selected for exome sequencing and sequential bioinformatic filtering. Cellular rescue experiments were performed to verify pathogenicity of novel disease alleles. Results The first filter criterion was 'Presence of known pathogenic complex I deficiency variants'. This revealed homozygous mutations in NDUFS3 and ACAD9 in two individuals. A second criterion was 'Presence of two novel potentially pathogenic variants in a structural gene of complex I', which discovered rare variants in NDUFS8 in two unrelated individuals and in NDUFB3 in a third. Expression of wild-type cDNA in mutant cell lines rescued complex I activity and assembly, thus providing a functional validation of their pathogenicity. Using the third criterion 'Presence of two potentially pathogenic variants in a gene encoding a mitochondrial protein', loss-of-function mutations in MTFMT were discovered in two patients. In three patients the molecular genetic correlate remained unclear and follow-up analysis is ongoing. Conclusion Appropriate in silico filtering of exome sequencing data, coupled with functional validation of new disease alleles, is effective in rapidly identifying disease-causative variants in known and new complex I associated disease genes.

Original languageEnglish
Pages (from-to)277-283
Number of pages7
JournalJournal of Medical Genetics
Volume49
Issue number4
DOIs
Publication statusPublished - Apr 2012

Fingerprint

Exome
Virulence
Alleles
Genes
Deficiency Diseases
Mutation
Genetic Heterogeneity
Mitochondrial Proteins
Genetic Association Studies
Computational Biology
Computer Simulation
Molecular Biology
Complementary DNA
Technology
Cell Line
Mitochondrial complex I deficiency

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Haack, T. B., Haberberger, B., Frisch, E. M., Wieland, T., Iuso, A., Gorza, M., ... Prokisch, H. (2012). Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing. Journal of Medical Genetics, 49(4), 277-283. https://doi.org/10.1136/jmedgenet-2012-100846

Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing. / Haack, Tobias B.; Haberberger, Birgit; Frisch, Eva Maria; Wieland, Thomas; Iuso, Arcangela; Gorza, Matteo; Strecker, Valentina; Graf, Elisabeth; Mayr, Johannes A.; Herberg, Ulrike; Hennermann, Julia B.; Klopstock, Thomas; Kuhn, Klaus A.; Ahting, Uwe; Sper, Wolfgang; Wilichowski, Ekkehard; Hoffmann, Georg F.; Tesarova, Marketa; Hansikova, Hana; Zeman, Jiri; Plecko, Barbara; Zeviani, Massimo; Wittig, Ilka; Strom, Tim M.; Schuelke, Markus; Freisinger, Peter; Meitinger, Thomas; Prokisch, Holger.

In: Journal of Medical Genetics, Vol. 49, No. 4, 04.2012, p. 277-283.

Research output: Contribution to journalArticle

Haack, TB, Haberberger, B, Frisch, EM, Wieland, T, Iuso, A, Gorza, M, Strecker, V, Graf, E, Mayr, JA, Herberg, U, Hennermann, JB, Klopstock, T, Kuhn, KA, Ahting, U, Sper, W, Wilichowski, E, Hoffmann, GF, Tesarova, M, Hansikova, H, Zeman, J, Plecko, B, Zeviani, M, Wittig, I, Strom, TM, Schuelke, M, Freisinger, P, Meitinger, T & Prokisch, H 2012, 'Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing', Journal of Medical Genetics, vol. 49, no. 4, pp. 277-283. https://doi.org/10.1136/jmedgenet-2012-100846
Haack, Tobias B. ; Haberberger, Birgit ; Frisch, Eva Maria ; Wieland, Thomas ; Iuso, Arcangela ; Gorza, Matteo ; Strecker, Valentina ; Graf, Elisabeth ; Mayr, Johannes A. ; Herberg, Ulrike ; Hennermann, Julia B. ; Klopstock, Thomas ; Kuhn, Klaus A. ; Ahting, Uwe ; Sper, Wolfgang ; Wilichowski, Ekkehard ; Hoffmann, Georg F. ; Tesarova, Marketa ; Hansikova, Hana ; Zeman, Jiri ; Plecko, Barbara ; Zeviani, Massimo ; Wittig, Ilka ; Strom, Tim M. ; Schuelke, Markus ; Freisinger, Peter ; Meitinger, Thomas ; Prokisch, Holger. / Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing. In: Journal of Medical Genetics. 2012 ; Vol. 49, No. 4. pp. 277-283.
@article{76456d10582c400cac0445ad4a002433,
title = "Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing",
abstract = "Background Next generation sequencing has become the core technology for gene discovery in rare inherited disorders. However, the interpretation of the numerous sequence variants identified remains challenging. We assessed the application of exome sequencing for diagnostics in complex I deficiency, a disease with vast genetic heterogeneity. Methods Ten unrelated individuals with complex I deficiency were selected for exome sequencing and sequential bioinformatic filtering. Cellular rescue experiments were performed to verify pathogenicity of novel disease alleles. Results The first filter criterion was 'Presence of known pathogenic complex I deficiency variants'. This revealed homozygous mutations in NDUFS3 and ACAD9 in two individuals. A second criterion was 'Presence of two novel potentially pathogenic variants in a structural gene of complex I', which discovered rare variants in NDUFS8 in two unrelated individuals and in NDUFB3 in a third. Expression of wild-type cDNA in mutant cell lines rescued complex I activity and assembly, thus providing a functional validation of their pathogenicity. Using the third criterion 'Presence of two potentially pathogenic variants in a gene encoding a mitochondrial protein', loss-of-function mutations in MTFMT were discovered in two patients. In three patients the molecular genetic correlate remained unclear and follow-up analysis is ongoing. Conclusion Appropriate in silico filtering of exome sequencing data, coupled with functional validation of new disease alleles, is effective in rapidly identifying disease-causative variants in known and new complex I associated disease genes.",
author = "Haack, {Tobias B.} and Birgit Haberberger and Frisch, {Eva Maria} and Thomas Wieland and Arcangela Iuso and Matteo Gorza and Valentina Strecker and Elisabeth Graf and Mayr, {Johannes A.} and Ulrike Herberg and Hennermann, {Julia B.} and Thomas Klopstock and Kuhn, {Klaus A.} and Uwe Ahting and Wolfgang Sper and Ekkehard Wilichowski and Hoffmann, {Georg F.} and Marketa Tesarova and Hana Hansikova and Jiri Zeman and Barbara Plecko and Massimo Zeviani and Ilka Wittig and Strom, {Tim M.} and Markus Schuelke and Peter Freisinger and Thomas Meitinger and Holger Prokisch",
year = "2012",
month = "4",
doi = "10.1136/jmedgenet-2012-100846",
language = "English",
volume = "49",
pages = "277--283",
journal = "Journal of Medical Genetics",
issn = "0022-2593",
publisher = "BMJ Publishing Group",
number = "4",

}

TY - JOUR

T1 - Molecular diagnosis in mitochondrial complex I deficiency using exome sequencing

AU - Haack, Tobias B.

AU - Haberberger, Birgit

AU - Frisch, Eva Maria

AU - Wieland, Thomas

AU - Iuso, Arcangela

AU - Gorza, Matteo

AU - Strecker, Valentina

AU - Graf, Elisabeth

AU - Mayr, Johannes A.

AU - Herberg, Ulrike

AU - Hennermann, Julia B.

AU - Klopstock, Thomas

AU - Kuhn, Klaus A.

AU - Ahting, Uwe

AU - Sper, Wolfgang

AU - Wilichowski, Ekkehard

AU - Hoffmann, Georg F.

AU - Tesarova, Marketa

AU - Hansikova, Hana

AU - Zeman, Jiri

AU - Plecko, Barbara

AU - Zeviani, Massimo

AU - Wittig, Ilka

AU - Strom, Tim M.

AU - Schuelke, Markus

AU - Freisinger, Peter

AU - Meitinger, Thomas

AU - Prokisch, Holger

PY - 2012/4

Y1 - 2012/4

N2 - Background Next generation sequencing has become the core technology for gene discovery in rare inherited disorders. However, the interpretation of the numerous sequence variants identified remains challenging. We assessed the application of exome sequencing for diagnostics in complex I deficiency, a disease with vast genetic heterogeneity. Methods Ten unrelated individuals with complex I deficiency were selected for exome sequencing and sequential bioinformatic filtering. Cellular rescue experiments were performed to verify pathogenicity of novel disease alleles. Results The first filter criterion was 'Presence of known pathogenic complex I deficiency variants'. This revealed homozygous mutations in NDUFS3 and ACAD9 in two individuals. A second criterion was 'Presence of two novel potentially pathogenic variants in a structural gene of complex I', which discovered rare variants in NDUFS8 in two unrelated individuals and in NDUFB3 in a third. Expression of wild-type cDNA in mutant cell lines rescued complex I activity and assembly, thus providing a functional validation of their pathogenicity. Using the third criterion 'Presence of two potentially pathogenic variants in a gene encoding a mitochondrial protein', loss-of-function mutations in MTFMT were discovered in two patients. In three patients the molecular genetic correlate remained unclear and follow-up analysis is ongoing. Conclusion Appropriate in silico filtering of exome sequencing data, coupled with functional validation of new disease alleles, is effective in rapidly identifying disease-causative variants in known and new complex I associated disease genes.

AB - Background Next generation sequencing has become the core technology for gene discovery in rare inherited disorders. However, the interpretation of the numerous sequence variants identified remains challenging. We assessed the application of exome sequencing for diagnostics in complex I deficiency, a disease with vast genetic heterogeneity. Methods Ten unrelated individuals with complex I deficiency were selected for exome sequencing and sequential bioinformatic filtering. Cellular rescue experiments were performed to verify pathogenicity of novel disease alleles. Results The first filter criterion was 'Presence of known pathogenic complex I deficiency variants'. This revealed homozygous mutations in NDUFS3 and ACAD9 in two individuals. A second criterion was 'Presence of two novel potentially pathogenic variants in a structural gene of complex I', which discovered rare variants in NDUFS8 in two unrelated individuals and in NDUFB3 in a third. Expression of wild-type cDNA in mutant cell lines rescued complex I activity and assembly, thus providing a functional validation of their pathogenicity. Using the third criterion 'Presence of two potentially pathogenic variants in a gene encoding a mitochondrial protein', loss-of-function mutations in MTFMT were discovered in two patients. In three patients the molecular genetic correlate remained unclear and follow-up analysis is ongoing. Conclusion Appropriate in silico filtering of exome sequencing data, coupled with functional validation of new disease alleles, is effective in rapidly identifying disease-causative variants in known and new complex I associated disease genes.

UR - http://www.scopus.com/inward/record.url?scp=84864082138&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84864082138&partnerID=8YFLogxK

U2 - 10.1136/jmedgenet-2012-100846

DO - 10.1136/jmedgenet-2012-100846

M3 - Article

C2 - 22499348

AN - SCOPUS:84864082138

VL - 49

SP - 277

EP - 283

JO - Journal of Medical Genetics

JF - Journal of Medical Genetics

SN - 0022-2593

IS - 4

ER -