A missense variant in the titin gene in Doberman pinscher dogs with familial dilated cardiomyopathy and sudden cardiac death

Abstract

The dog provides a large animal model of familial dilated cardiomyopathy for the study of important aspects of this common familial cardiovascular disease. We have previously demonstrated a form of canine dilated cardiomyopathy in the Doberman pinscher breed that is inherited as an autosomal dominant trait and is associated with a splice site variant in the pyruvate dehydrogenase kinase 4 (PDK4) gene, however, genetic heterogeneity exists in this species as well and not all affected dogs have the PDK4 variant. Whole genome sequencing of a family of Doberman pinchers with dilated cardiomyopathy and sudden cardiac death without the PDK4 variant was performed. A pathologic missense variant in the titin gene located in an immunoglobulin-like domain in the I-band spanning region of the molecule was identified and was highly associated with the disease (p < 0.0001). We demonstrate here the identification of a variant in the titin gene highly associated with the disease in this spontaneous canine model of dilated cardiomyopathy. This large animal model of familial dilated cardiomyopathy shares many similarities with the human disease including mode of inheritance, clinical presentation, genetic heterogeneity and a pathologic variant in the titin gene. The dog is an excellent model to improve our understanding of the genotypic phenotypic relationships, penetrance, expression and the pathophysiology of variants in the titin gene.

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References

  1. Anderson BR, Bogomolovas J, Labeit S, Granzier H (2013) Single molecule force spectroscopy on titin implicates immunoglobulin domain stability as a cardiac disease mechanism. J Biol Chem 288:5303–5315

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Bang ML, Centner T, Fornoff F, Geach AJ, Gotthardt M, McNabb M et al (2001) The complete gene sequence of titin, expression of an unusual approximately 700-kDa titin isoform, and its interaction with obscurin identify a novel Z-line to I-band linking system. Circ Res 89:1065–1072

    Article  CAS  PubMed  Google Scholar 

  3. Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30:2114–2120

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Calvert CA, Pickus CW, Jacobs GJ, Brown J (1997) Signalment, survival, and prognostic factors in Doberman pinschers with end-stage cardiomyopathy. J Vet Intern Med 11:323–326

    Article  CAS  PubMed  Google Scholar 

  5. Cheng Y, Hogarth KA, O’Sullivan ML, Regnier M, Pyle WG (2016) 2-Deoxyadenosine triphosphate restores the contractile function of cardiac myofibril from adult dogs with naturally occurring dilated cardiomyopathy. Am J Physiol Heart Circ Physiol 310:H80–H91

    Article  PubMed  Google Scholar 

  6. de Gonzalo-Calvo D, Quezada M, Campuzano O, Perez-Serra A, Broncano J, Ayala R et al (2017) Familial dilated cardiomyopathy: a multidisciplinary entity, from basic screening to novel circulating biomarkers. Int J Cardiol 228:870–880

    Article  PubMed  Google Scholar 

  7. DePristo MA, Banks E, Poplin RE, Garimella KV, Maguire JR, Hartl C et al (2011) A framework for variation discovery and genotyping using next-generation DNA sequencing data. Nat Genet 43:491–498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Franaszczyk M, Chmielweski P, Truszkowska G, Stawinski P, Michalak E, Rydzanicz M et al (2017) Titin truncating variants in dilated cardiomyopathy—prevalence and genotype-phenotype correlations. PLoS One 12:e0169007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Friedenberg SG, Meurs KM (2016) Genotype imputation in the domestic dog. Mamm Genome 27:485–494

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Gerull B, Gramlich M, Atherton J, McNabb M, Trombitas K, Sasse-Klaassen S et al (2002) Mutations of TTN, encoding the giant muscle filament titin, cause familial dilated cardiomyopathy. Nat Genet 30:201–204

    Article  CAS  PubMed  Google Scholar 

  11. Gerull B, Atherton J, Geupel A, Sasse-Klaasen S, Heuser A, Frenneaux M et al (2006) Identification of a novel frameshift mutation in the giant muscle filament titin in a large Australian family with dilated cardiomyopathy. J Mol Med 84:478–483

    Article  CAS  PubMed  Google Scholar 

  12. Gigli M, Begay RL, Morea G, Graw SL, Sinagra G, Taylor MRG et al (2016) A review of the giant protein titin in clinical molecular diagnostics of cardiomyopathies. Front Cardiovasc Med 3:21

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Granzier H, Labeit S (2002) Cardiac titin: an adjustable multi-functional spring. J Physiol 541:335–342

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Granzier HL, Labeit S (2006) The giant muscle protein titin is an adjustable molecular spring. Exerc Sport Sci Rev 34:50–53

    Article  PubMed  Google Scholar 

  15. Granzier HL, Radke MH, Peng J, Westermann D, Nelson OL, Rost K, King NMP et al (2009) Truncation of titin’s elastic PEVK region leads to cardiomyopathy with diastolic dysfunction. Circ Res 105:557–564

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Hensley MT, Tang J, Woodruff K, Defrancesco T, Tou S, Williams CM et al (2017) Intracoronary allogeneic cardiosphere-derived stem cells are safe for use in dogs with dilated cardiomyopathy. J Cell Mol Med 21:1503–1512

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Herman DS, Lam L, Taylor MRG, Wang L, Teekakirikul P, Christodoulou D et al (2012) Truncations of titin causing dilated cardiomyopathy. N Engl J Med 366:619–628

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Hershberger RE, Hedges DJ, Morales A (2013) Dilated cardiomyopathy: the complexity of a diverse genetic architecture. Nat Rev Cardiol 10:531–547

    Article  CAS  PubMed  Google Scholar 

  19. Hinson JT, Chopra A, Nafissi N, Polacheck WJ, Benson CC, Swist S et al (2015) Titin mutations in iPS cells define sarcomere insufficiency as a cause of dilated cardiomyopathy. Science 349:982–986

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Itoh-Satoh M, Hayashi T, Nishi H, Koga Y, Arimura T, Koyanagi T et al (2002) Titin mutations as the molecular basis for dilated cardiomyopathy. Biochem Biophys Res Commun 291:385–393

    Article  CAS  PubMed  Google Scholar 

  21. Kimura A (2016) Molecular genetics and pathogenesis of cardiomyopathy. J Hum Genet 61:41–50

    Article  CAS  PubMed  Google Scholar 

  22. Labeit S, Lahmers S, Burkart C, Fong C, McNabb M, Witt S et al (2006) Expression of distinct classes of titin isoforms in striated and smooth muscles by alternative splicing, and their conserved interaction with filamins. J Mol Biol 362:664–681

    Article  CAS  PubMed  Google Scholar 

  23. Laddach A, Gautel M, Fraternali F (2017) TITINdb-a computational tool to assess titin’s role as a disease gene. Bioinformatics 33:3482–3485

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Lahmers S, Wu Y, Call DR, Labeit S, Granzier H (2004) Developmental control of titin isoform expression and passive stiffness in fetal and neonatal myocardium. Circ Res 94:505–513

    Article  CAS  PubMed  Google Scholar 

  25. LeWinter MM, Granzier HL (2014) Cardiac titin and heart disease. J Cardiovasc Pharmacol 63:207–212

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25:1754–1760

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Lindblad-Toh K, Wade CM, Mikkelson TS, Karlsson EK, Jaffe DB, Kamal M et al (2005) Genome sequence, comparative analysis and haplotype structure of the domestic dog. Nature 438:803–819

    Article  CAS  Google Scholar 

  28. Linke WA (2018) Titin gene and protein functions in passive and active muscle. Annu Rev Physiol 80:389–411

    Article  CAS  PubMed  Google Scholar 

  29. Martin MW, Stafford Johnson MJ, Strehlau G, King JN (2010) Canine dilated cardiomyopathy: a retrospective study of prognostic findings in 367 clinical cases. J Small Anim Pract 51:428–436

    Article  CAS  PubMed  Google Scholar 

  30. Mausberg TB, Wess G, Simak J, Keller L, Drogemuller M, Drogemuller C et al (2011) A locus on chromosome 5 is associated with dilated cardiomyopathy in Doberman pinschers. PLoS One 6:e20042

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. McKenna A, Hanna M, Banks E, Sivachenko A, Cibulskis K, Kernytsky A et al (2010) The Genome Analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res 20:1297–1303

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. McLaren W, Gil L, Hunt SE, Riat HS, Ritchie GRS, Thormann A et al (2016) The ensembl variant effect predictor. Genome Biol 17:122

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Meurs KM, Fox PR, Norgard M, Spier AW, Lamb A, Koplitz SL et al (2007) A prospective genetic evaluation of familial dilated cardiomyopathy in the Doberman pinscher. J Vet Intern Med 21:1016–1020

    Article  PubMed  Google Scholar 

  34. Meurs KM, Hendrix KP, Norgard MM (2008) Molecular evaluation of five cardiac genes in Doberman pinschers with dilated cardiomyopathy. Am J Vet Res 69:1050–1053

    Article  PubMed  Google Scholar 

  35. Meurs KM, Lahmers S, Keene BW, White SN, Oyama MA, Maucell E et al (2012) A splice site mutation in a gene encoding for PDK4, a mitochondrial protein, is associated with the development of dilated cardiomyopathy in the Doberman pinscher. Hum Genet 131:1319–1325

    Article  CAS  PubMed  Google Scholar 

  36. Morales A, Hershberger RE (2015) The rationale and timing of molecular genetic testing for dilated cardiomyopathy. Can J Cardiol 31:1309–1312

    Article  PubMed  PubMed Central  Google Scholar 

  37. Neiva-Sousa M, Almeida-Coelho J, Falcao-Pires I, Leite-Moreira AF (2015) Titin mutations: the fall of Goliath. Heart Fail Rev 20:579–588

    Article  CAS  PubMed  Google Scholar 

  38. Ottenheijm CA, Hooljiman P, DeChene ET, Stienen GJ, Beggs AH, Granzier H (2010) Altered myofilament function depresses force generation in patients with nebulin-based nemaline myopathy (NEM2). J Struct Biol 170:334–343

    Article  CAS  PubMed  Google Scholar 

  39. Owczarek-Lipska M, Mausberg TB, Stephenson H, Dukes-McEwan H, Wess G, Leeb T (2013) A 16-bp deletion in the canine PDK4 gene is not associated with dilated cardiomyopathy in a European cohort of Doberman pinschers. Anim Genet 44:239

    Article  CAS  PubMed  Google Scholar 

  40. Pires DE, Ascher DB, Blundell TL (2014a) DUET: a server for predicting effects of mutations on protein stability using an integrated computational approach. Nucleic Acids Res 42:W314–W319

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Pires DE, Ascher DB, Blundell TL (2014b) mCSM: predicting the effects of mutations in proteins using graph-based signatures. Bioinformatics 30:335–342

    Article  CAS  PubMed  Google Scholar 

  42. Schafer S, de Marvao A, Adami E, Fiedler LR, Ng B, Khin E et al (2017) Titin-truncating variants affect heart function in disease cohorts and the general population. Nat Genet 49:46–53

    Article  CAS  Google Scholar 

  43. Sen-Chowdhry S, Syrris P, McKenna WJ (2005) Genetics of right ventricular cardiomyopathy. J Cardiovasc Electrophysiol 16:927–935

    Article  PubMed  Google Scholar 

  44. Taylor M, Graw S, Sinagra G, Barnes C, Slavov D, Brun F et al (2011) Genetic variation in titin in ARVC-overlap syndromes. Circulation 124:876–885

    Article  PubMed  PubMed Central  Google Scholar 

  45. Thomas WP, Gaber CE, Jacobs GJ, Kaplan PM, Lombard CW, Moise NS et al (1993) Recommendations for standards in transthoracic two-dimensional echocardiography in the dog and cat. J Vet Intern Med 7:247–252

    Article  CAS  PubMed  Google Scholar 

  46. Towbin JA (2014) Inherited cardiomyopathies. Circ J 78:2347–2356

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  47. Van der Auwera GA, Carneiro MO, Hartl C, Poplin R, del Angel G, Levy-Moonshine A et al (2013) From FastQ data to high confidence variant calls: the Genome Analysis Toolkit best practices pipeline. Curr Protoc Bioinformatics 43:1110–1133

    Google Scholar 

  48. Vikhorev PG, Smoktunowicz N, Munster AB, Copeland O, Kostin S, Montgiraud C et al (2017) Abnormal contractility in human heart myofibrils from patients with dilated cardiomyopathy due to mutations in TTN and contractile protein genes. Sci Rep 7:14829

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Vilella AJ, Severin J, Ureta-Vidal A, Heng L, Durbin R, Birney E (2009) EnsemblCompara GeneTrees: complete, duplication-aware phylogenetic trees in vertebrates. Genome Res 19:327–335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Warren CM, Krzesinski PR, Greaser ML (2003) Vertical agarose gel electrophoresis and electroblotting of high-molecular-weight proteins. Electrophoresis 24:1695–1702

    Article  CAS  PubMed  Google Scholar 

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Funding

Research reported in this publication was supported by the Heart Lung and Blood Institute of the National Institutes of Health under award number R35HL144998.

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Correspondence to Kathryn M. Meurs.

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On behalf of all authors, the corresponding author states that there is no conflict of interest. North Carolina State University offers the canine titin variant test as a risk assement tool for dogs.

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Meurs, K.M., Friedenberg, S.G., Kolb, J. et al. A missense variant in the titin gene in Doberman pinscher dogs with familial dilated cardiomyopathy and sudden cardiac death. Hum Genet 138, 515–524 (2019). https://doi.org/10.1007/s00439-019-01973-2

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