mitoGENE:

High-throughput molecular diagnostics

State of the art

Diagnosis of mitochondrial disease patients is challenging given both clinical and genetic heterogeneity. Hence, in a major proportion of patients lack a molecular basis of the disease and new strategies to address this issue are urgently needed. Innovative high-throughput screening methods enable cost-efficient mutation screening of a large number of disease genes in large cohorts of patients. With high-throughput sequencing available, prior enrichment of the genomic region of interest remains a major hurdle. So far, two approaches have been suggested, enrichment by hybridization to a specific oligoarray and exon / gene specific amplification by PCR. Both strategies are focussed on a set of genes and need to be established for the disease of interest.

Aims of the project

The overall aim of this project is to provide comprehensive molecular diagnostics for mitochondrial disorders by using innovative and cutting-edge technology. This aim comprises three subgoals: (i) to increase the number of genes for which diagnostic testing is available at molecular genetics laboratories; (ii) to provide mutation screening for 100 candidate genes using an established high-throughput platform; (iii) to apply next-generation sequencing by developing mitochondrial disorder specific protocols. During the time frame of this project we will substantially increase the number of assays and develop new methods for molecular diagnostics.

Project management and contact addresses

Holger Prokisch, Dr.
Institut für Humangenetik
Technischen Universität München
Klinikum Rechts der Isar
Trogerstr. 32, 81675 München, Germany

Tel: (+49) 89 3187 2890
Fax: (+49) 89 3187 3297
E-mail: prokisch@helmholtz-muenchen.de

Publications:

  1. Haack TB, Danhauser K, Haberberger B, Hoser J, Strecker V, Boehm D, Uziel G, Lamantea E, Invernizzi F, Poulton J, Rolinski B, Iuso A, Biskup S, Schmidt T, Mewes HW, Wittig I, Meitinger T, Zeviani M, Prokisch H. (2010) Exome sequencing identifies ACAD9 mutations as a cause of complex I deficiency. Nat Genet. 42:1131-4.
  2. Gerards M, van den Bosch BJ, Danhauser K, Serre V, van Weeghel M, Wanders RJ, Nicolaes GA, Sluiter W, Schoonderwoerd K, Scholte HR, Prokisch H, Rötig A, de Coo IF, Smeets HJ. Riboflavin-responsive oxidative phosphorylation complex I deficiency caused by defective ACAD9: new function for an old gene. Brain. 134:210-9.
  3. Herzer M, Koch J, Prokisch H, Rodenburg R, Rauscher C, Radauer W, Forstner R, Pilz P, Rolinski B, Freisinger P, Mayr JA, Sperl W. (2010) Leigh disease with brainstem involvement in complex I deficiency due to assembly factor NDUFAF2 defect. Neuropediatrics. 41:30-4.
  4. O'Toole JF, Liu Y, Davis EE, Westlake CJ, Attanasio M, Otto EA, Seelow D, Nurnberg G, Becker C, Nuutinen M, Kärppä M, Ignatius J, Uusimaa J, Pakanen S, Jaakkola E, van den Heuvel LP, Fehrenbach H, Wiggins R, Goyal M, Zhou W, Wolf MT, Wise E, Helou J, Allen SJ, Murga-Zamalloa CA, Ashraf S, Chaki M, Heeringa S, Chernin G, Hoskins BE, Chaib H, Gleeson J, Kusakabe T, Suzuki T, Isaac RE, Quarmby LM, Tennant B, Fujioka H, Tuominen H, Hassinen I, Lohi H, van Houten JL, Rotig A, Sayer JA, Rolinski B, Freisinger P, Madhavan SM, Herzer M, Madignier F, Prokisch H, Nurnberg P, Jackson PK, Khanna H, Katsanis N, Hildebrandt F. (2010) Individuals with mutations in XPNPEP3, which encodes a mitochondrial protein, develop a nephronophthisis-like nephropathy. J Clin Invest. 120:791-802.
  5. Pham TT, Giesert F, Röthig A, Floss T, Kallnik M, Weindl K, Hölter SM, Ahting U, Prokisch H, Becker L, Klopstock T, Hrabé de Angelis M, Beyer K, Görner K, Kahle PJ, Vogt Weisenhorn DM, Wurst W. (2010) DJ-1-deficient mice show less TH-positive neurons in the ventral tegmental area and exhibit non-motoric behavioural impairments. Genes Brain Behav. 9:305-17
  6. Ghezzi D, Goffrini P, Uziel G, Horvath R, Klopstock T, Lochmüller H, D'Adamo P, Gasparini P, Strom TM, Prokisch H, Invernizzi F, Ferrero I, Zeviani M. (2009) SDHAF1,encoding a LYR complex-II specific assembly factor, is mutated in SDH-defective infantile leukoencephalopathy. Nat Genet. 4:654-6.
  7. Elstner M, Andreoli C, Klopstock T, Meitinger T, Prokisch H. (2009) The mitochondrial proteome database: MitoP2. Methods Enzymol. 457:3-20.
  8. Ahting U, Floss T, Uez N, Schneider-Lohmar I, Becker L, Kling E, Iuso A, Bender A, de Angelis MH, Gailus-Durner V, Fuchs H, Meitinger T, Wurst W, Prokisch H, Klopstock T. (2009) Neurological phenotype and reduced lifespan in heterozygous Tim23 knockout mice, the first mouse model of defective mitochondrial import. Biochim
    Biophys Acta. 1787:371-6.