Basic Research in Cardiology

, 108:372 | Cite as

A distinct subgroup of cardiomyopathy patients characterized by transcriptionally active cardiotropic erythrovirus and altered cardiac gene expression

  • U. Kuhl
  • D. Lassner
  • A. Dorner
  • M. Rohde
  • F. Escher
  • B. Seeberg
  • E. Hertel
  • C. Tschope
  • C. Skurk
  • U. M. Gross
  • H.-P. Schultheiss
  • W. Poller
Original Contribution
First Online:
Received:
Revised:
Accepted:

Abstract

Recent studies have detected erythrovirus genomes in the hearts of cardiomyopathy and cardiac transplant patients. Assessment of the functional status of viruses may provide clinically important information beyond detection of the viral genomes. Here, we report transcriptional activation of cardiotropic erythrovirus to be associated with strongly altered myocardial gene expression in a distinct subgroup of cardiomyopathy patients. Endomyocardial biopsies (EMBs) from 415 consecutive cardiac erythrovirus (B19V)-positive patients with clinically suspected cardiomyopathy were screened for virus-encoded VP1/VP2 mRNA indicating transcriptional activation of the virus, and correlated with cardiac host gene expression patterns in transcriptionally active versus latent infections, and in virus-free control hearts. Transcriptional activity was detected in baseline biopsies of only 66/415 patients (15.9 %) harbouring erythrovirus. At the molecular level, significant differences between cardiac B19V-positive patients with transcriptionally active versus latent virus were revealed by expression profiling of EMBs. Importantly, latent B19V infection was indistinguishable from controls. Genes involved encode proteins of antiviral immune response, B19V receptor complex, and mitochondrial energy metabolism. Thus, functional mapping of erythrovirus allows definition of a subgroup of B19V-infected cardiomyopathy patients characterized by virus-encoded VP1/VP2 transcripts and anomalous host myocardial transcriptomes. Cardiac B19V reactivation from latency, as reported here for the first time, is a key factor required for erythrovirus to induce altered cardiac gene expression in a subgroup of cardiomyopathy patients. Virus genome detection is insufficient to assess pathogenic potential, but additional transcriptional mapping should be incorporated into future pathogenetic and therapeutic studies both in cardiology and transplantation medicine.

Keywords

Cardiomyopathy Erythrovirus Virus latency Virus reactivation 

Abbreviations

ADV

Adenovirus

ARB

Angiotensin receptor blocker

B19V

Erythrovirus/parvovirus B19

DCM

Dilated cardiomyopathy

EMB

Endomyocardial biopsy

EPC

Endothelial progenitor cell

ES

Mitral valve E-point to septal separation

EV

Enterovirus

FS

Fractional shortening

HHV-6

Human herpesvirus type 6

ICD

Imlantable cadioverter defibrillator

LVBB

Left ventricular branch block

LVEDD

Left ventricular enddiastolic diameter

LVEF

Left ventricular ejection fraction

LVESD

Left ventricular endsystolic diameter

MI

Myocardial infarction

PCR

Polymerase chain reaction

PM

Pacemaker

RT-QPCR

Reverse transcription quantitative PCR

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Notes

Acknowledgments

This work was supported by grants from the German Research Foundation (DFG) via Collaborative Research Centre SFB/TR 19 “Inflammatory Cardiomyopathy—Molecular Pathogenesis and Therapy”, and by the German Federal Ministry of Education and Research (BMBF) via KMU Innovative program (No. 616 0315296). For their excellent technical assistance we thank Ms. K. Winter, S. Ochmann, C. Seifert, M. Willner, and A. Kallel, Berlin, Germany.

Conflict of interest

None.

Supplementary material

395_2013_372_MOESM1_ESM.docx (316 kb)
Supplemental Figure 1 Panel A: Agarose gel analysis of PCR products obtained after various enzymatic pre-treatments of nucleic acids isolated from endomyocardial patient biopsies. Without DNAse digestion, a PCR product is expectedly generated from the single-stranded B19V genomic DNA contained in the biopsies (lanes 3-5). After extensive DNAse treatment of isolated nucleic acids, as used throughout the current study, no PCR could be obtained any more without reverse transcription (RT) before the PCR for VP1/VP2 sequence was conducted (lanes 6-8), indicating efficient removal of any residual viral genomic DNA. After the DNAs digestion, a PCR product was only generated if a RT reaction was conducted before the PCR (lanes 6-8, 11, 12). Panel B: In the RT-QPCR protocol used throughout this study for the quantification of viral sequences, there was also no signal after DNAse digestion without reverse transcription prior to quantitative PCR (lanes 2, 3, 5, 8). Quality controls as shown here in two patients (# 3512 for 2 biopsies, and #7806) were conducted for all patients investigated, and complete removal of viral genomic DNA by DNAse treatment before RT-qPCR was confirmed in all cases. The viral mRNA-derived quantitative signals as exemplified by lanes 4, 6, and 9 appear only after a reverse transcription (RT) before qPCR. (DOCX 823 kb)

References

  1. 1.
    Angelow A, Weitmann K, Schmidt M, Schwedler S, Vogt H, Havemann C, Staudt A, Felix SB, Stangl K, Klingel K, Kandolf R, Kuhl U, Lassner D, von Schlippenbach J, Schultheiss HP, Hoffmann W (2009) The German transregional collaborative research centre ‘Inflammatory Cardiomyopathy—Molecular Pathogenesis and Therapy’. Methods and baseline results from a 3-centre clinical study. Cardiology 113:222–230. doi: 10.1159/000203404 PubMedCrossRefGoogle Scholar
  2. 2.
    Bock CT, Klingel K, Kandolf R (2010) Human parvovirus B19-associated myocarditis. N Engl J Med 362:1248–1249. doi: 10.1056/NEJMc0911362 PubMedCrossRefGoogle Scholar
  3. 3.
    Bonsch C, Zuercher C, Lieby P, Kempf C, Ros C (2010) The globoside receptor triggers structural changes in the B19Virus capsid that facilitate virus internalization. J Virol 84:11737–11746. doi: 10.1128/JVI.01143-10 PubMedCrossRefGoogle Scholar
  4. 4.
    Bowles NE, Richardson PJ, Olsen EG, Archard LC (1986) Detection of Coxsackie-B-virus-specific RNA sequences in myocardial biopsy samples from patients with myocarditis and dilated cardiomyopathy. Lancet 1:1120–1123. doi: 10.1016/S0140-6736(86)91837-4 PubMedCrossRefGoogle Scholar
  5. 5.
    Breinholt JP, Moulik M, Dreyer WJ, Denfield SW, Kim JJ, Jefferies JL, Rossano JW, Gates CM, Clunie SK, Bowles KR, Kearney DL, Bowles NE, Towbin JA (2010) Viral epidemiologic shift in inflammatory heart disease: the increasing involvement of parvovirus B19 in the myocardium of pediatric cardiac transplant patients. J Heart Lung Transpl 29:739–746. doi: 10.1016/j.healun.2010.03.003 CrossRefGoogle Scholar
  6. 6.
    Brown KE, Anderson SM, Young NS (1993) Erythrocyte P antigen: cellular receptor for B19 parvovirus. Science 262:114–117. doi: 10.1126/science.8211117 PubMedCrossRefGoogle Scholar
  7. 7.
    Brown KE, Hibbs JR, Gallinella G, Anderson SM, Lehman ED, McCarthy P, Young NS (1994) Resistance to parvovirus B19 infection due to lack of virus receptor (erythrocyte P antigen). N Engl J Med 330:1192–1196. doi: 10.1056/NEJM199404283301704 PubMedCrossRefGoogle Scholar
  8. 8.
    De Regge N, Van Opdenbosch N, Nauwynck HJ, Efstathiou S, Favoreel HW (2010) Interferon alpha induces establishment of alphaherpesvirus latency in sensory neurons in vitro. PLoS One 5(9). doi:  10.1371/journal.pone.0013076
  9. 9.
    Donoso Mantke O, Nitsche A, Meyer R, Klingel K, Niedrig M (2004) Analysing myocardial tissue from explanted hearts of heart transplant recipients and multi-organ donors for the presence of parvovirus B19 DNA. J Clin Virol 31:32–39. doi: 10.1016/j.jcv.2003.12.013 PubMedCrossRefGoogle Scholar
  10. 10.
    Duechting A, Tschope C, Kaiser H, Lamkemeyer T, Tanaka N, Aberle S, Lang F, Torresi J, Kandolf R, Bock CT (2008) Human parvovirus B19 NS1 protein modulates inflammatory signaling by activation of STAT3/PIAS3 in human endothelial cells. J Virol 82:7942–7952. doi: 10.1128/JVI.00891-08 PubMedCrossRefGoogle Scholar
  11. 11.
    Gruhle S, Sauter M, Szalay G, Ettischer N, Kandolf R, Klingel K (2012) The prostacyclin agonist iloprost aggravates fibrosis and enhances viral replication in enteroviral myocarditis by modulation of ERK signaling and increase of iNOS expression. Basic Res Cardiol 107:287. doi: 10.1007/s00395-012-0287-z PubMedCrossRefGoogle Scholar
  12. 12.
    Guan W, Cheng F, Yoto Y, Kleiboeker S, Wong S, Zhi N, Pintel DJ, Qiu J (2008) Block to the production of full-length B19Virus transcripts by internal polyadenylation is overcome by replication of the viral genome. J Virol 82:9951–9963. doi: 10.1128/JVI.01162-08 PubMedCrossRefGoogle Scholar
  13. 13.
    Guan W, Wong S, Zhi N, Qiu J (2009) The genome of human parvovirus B19 can replicate in nonpermissive cells with the help of adenovirus genes and produces infectious virus. J Virol 83:9541–9553. doi: 10.1128/JVI.00702-09 PubMedCrossRefGoogle Scholar
  14. 14.
    Kuethe F, Lindner J, Matschke K, Wenzel JJ, Norja P, Ploetze K, Schaal S, Kamvissi V, Bornstein SR, Schwanebeck U, Modrow S (2009) Prevalence of parvovirus B19 and human bocavirus DNA in the heart of patients with no evidence of dilated cardiomyopathy or myocarditis. Clin Infect Dis 49:1660–1666. doi: 10.1086/648074 PubMedCrossRefGoogle Scholar
  15. 15.
    Kuhl U, Lassner D, Pauschinger M, Gross UM, Seeberg B, Noutsias M, Poller W, Schultheiss HP (2008) Prevalence of erythrovirus genotypes in the myocardium of patients with dilated cardiomyopathy. J Med Virol 80:1243–1251. doi: 10.1002/jmv.21187 PubMedCrossRefGoogle Scholar
  16. 16.
    Kuhl U, Lassner D, von Schlippenbach J, Poller W, Schultheiss HP (2012) Interferon-beta improves survival in enterovirus-associated cardiomyopathy. J Am Coll Cardiol 60:1295–1296. doi: 10.1016/j.jacc.2012.06.026 PubMedCrossRefGoogle Scholar
  17. 17.
    Kuhl U, Pauschinger M, Bock T, Klingel K, Schwimmbeck CP, Seeberg B, Krautwurm L, Poller W, Schultheiss HP, Kandolf R (2003) Parvovirus B19 infection mimicking acute myocardial infarction. Circulation 108:945–950. doi: 10.1161/01.CIR.0000085168.02782.2C PubMedCrossRefGoogle Scholar
  18. 18.
    Kühl U, Pauschinger M, Noutsias M, Seeberg B, Bock T, Lassner D, Poller W, Kandolf R, Schultheiss H-P (2005) High prevalence of viral genomes and multiple viral infections in the myocardium of adults with “Idiopathic” left ventricular dysfunction. Circulation 111:887–893. doi: 10.1161/01.CIR.0000155616.07901.35 PubMedCrossRefGoogle Scholar
  19. 19.
    Kuhl U, Pauschinger M, Schwimmbeck PL, Seeberg B, Lober C, Noutsias M, Poller W, Schultheiss HP (2003) Interferon-beta treatment eliminates cardiotropic viruses and improves left ventricular function in patients with myocardial persistence of viral genomes and left ventricular dysfunction. Circulation 107:2793–2798. doi: 10.1161/01.CIR.0000072766.67150.51 PubMedCrossRefGoogle Scholar
  20. 20.
    Kuhl U, Rohde M, Lassner D, Gross UM, Escher F, Schultheiss HP (2012) miRNA as activity markers in Parvo B19 associated heart disease. Herz 37:637–643. doi: 10.1007/s00059-012-3656-3 PubMedCrossRefGoogle Scholar
  21. 21.
    Moulik M, Breinholt JP, Dreyer WJ, Kearney DL, Price JF, Clunie SK, Moffett BS, Kim JJ, Rossano JW, Jefferies JL, Bowles KR, O’Brian Smith E, Bowles NE, Denfield SW, Towbin JA (2010) Viral endomyocardial infection is an independent predictor and potentially treatable risk factor for graft loss and coronary vasculopathy in pediatric cardiac transplant recipients. J Am Coll Cardiol 56:582–592. doi: 10.1016/j.jacc.2010.02.060 PubMedCrossRefGoogle Scholar
  22. 22.
    Munakata Y, Saito-Ito T, Kumura-Ishii K, Huang J, Kodera T, Ishii T, Hirabayashi Y, Koyanagi Y, Sasaki T (2005) Ku80 autoantigen as a cellular coreceptor for human parvovirus B19 infection. Blood 106:3449–3456. doi: 10.1182/blood-2005-02-0536 PubMedCrossRefGoogle Scholar
  23. 23.
    Norja P, Hokynar K, Aaltonen LM, Chen R, Ranki A, Partio EK, Kiviluoto O, Davidkin I, Leivo T, Eis-Hubinger AM, Schneider B, Fischer HP, Tolba R, Vapalahti O, Vaheri A, Soderlund-Venermo M, Hedman K (2006) Bioportfolio: lifelong persistence of variant and prototypic erythrovirus DNA genomes in human tissue. Proc Natl Acad Sci USA 103:7450–7453. doi: 10.1073/pnas.0602259103 PubMedCrossRefGoogle Scholar
  24. 24.
    Noutsias M, Rohde M, Block A, Klippert K, Lettau O, Blunert K, Hummel M, Kuhl U, Lehmkuhl H, Hetzer R, Rauch U, Poller W, Pauschinger M, Schultheiss HP, Volk HD, Kotsch K (2008) Preamplification techniques for real-time RT-PCR analyses of endomyocardial biopsies. BMC Mol Biol 9:3. doi: 10.1186/1471-2199-9-3 PubMedCrossRefGoogle Scholar
  25. 25.
    Noutsias M, Rohde M, Goldner K, Block A, Blunert K, Hemaidan L, Hummel M, Blohm JH, Lassner D, Kuhl U, Schultheiss HP, Volk HD, Kotsch K (2011) Expression of functional T-cell markers and T-cell receptor Vbeta repertoire in endomyocardial biopsies from patients presenting with acute myocarditis and dilated cardiomyopathy. Eur J Heart Fail 13:611–618. doi: 10.1093/eurjhf/hfr014 PubMedCrossRefGoogle Scholar
  26. 26.
    Pauschinger M, Bowles NE, Fuentes-Garcia FJ, Pham V, Kuhl U, Schwimmbeck PL, Schultheiss HP, Towbin JA (1999) Detection of adenoviral genome in the myocardium of adult patients with idiopathic left ventricular dysfunction. Circulation 99:1348–1354. doi: 10.1161/01.CIR.99.10.1348 PubMedCrossRefGoogle Scholar
  27. 27.
    Piccoli C, Quarato G, Ripoli M, D’Aprile A, Scrima R, Cela O, Boffoli D, Moradpour D, Capitanio N (2009) HCV infection induces mitochondrial bioenergetic unbalance: causes and effects. Biochim Biophys Acta 1787:539–546. doi: 10.1016/j.bbabio.2008.11.00 PubMedCrossRefGoogle Scholar
  28. 28.
    Pozzuto T, von Kietzell K, Bock T, Schmidt-Lucke C, Poller W, Zobel T, Lassner D, Zeichhardt H, Weger S, Fechner H (2011) Transactivation of human parvovirus B19 gene expression in endothelial cells by adenoviral helper functions. Virology 411:50–64. doi: 10.1016/j.virol.2010.12.019 PubMedCrossRefGoogle Scholar
  29. 29.
    Schenk T, Enders M, Pollak S, Hahn R, Huzly D (2009) High prevalence of human parvovirus B19 DNA in myocardial autopsy samples from subjects without myocarditis or dilative cardiomyopathy. J Clin Microbiol 47:106–110. doi: 10.1128/JCM.01672-08 PubMedCrossRefGoogle Scholar
  30. 30.
    Schmidt-Lucke C, Spillmann F, Bock T, Kuhl U, Van Linthout S, Schultheiss HP, Tschope C (2010) Interferon beta modulates endothelial damage in patients with cardiac persistence of human parvovirus b19 infection. J Infect Dis 201:936–945. doi: 10.1086/650700 PubMedCrossRefGoogle Scholar
  31. 31.
    Servant-Delmas A, Lefrere JJ, Morinet F, Pillet S (2010) Advances in human B19 erythrovirus biology. J Virol 84:9658–9665. doi: 10.1128/JVI.00684-10 PubMedCrossRefGoogle Scholar
  32. 32.
    Vallbracht KB, Schwimmbeck PL, Kuhl U, Rauch U, Seeberg B, Schultheiss HP (2005) Differential aspects of endothelial function of the coronary microcirculation considering myocardial virus persistence, endothelial activation, and myocardial leukocyte infiltrates. Circulation 111:1784–1791. doi: 10.1161/01.CIR.0000160863.30496.9B PubMedCrossRefGoogle Scholar
  33. 33.
    Vallbracht KB, Schwimmbeck PL, Kuhl U, Seeberg B, Schultheiss HP (2004) Endothelium-dependent flow-mediated vasodilation of systemic arteries is impaired in patients with myocardial virus persistence. Circulation 110:2938–2945. doi: 10.1161/01.CIR.0000146891.31481.CF PubMedCrossRefGoogle Scholar
  34. 34.
    Weigel-Kelley KA, Yoder MC, Srivastava A (2003) Alpha5beta1 integrin as a cellular coreceptor for human parvovirus B19: requirement of functional activation of beta1 integrin for viral entry. Blood 102:3927–3933. doi: 10.1182/blood-2003-05-1522 PubMedCrossRefGoogle Scholar
  35. 35.
    Wessely R, Klingel K, Santana LF, Dalton N, Hongo M, Jonathan Lederer W, Kandolf R, Knowlton KU (1998) Transgenic expression of replication-restricted enteroviral genomes in heart muscle induces defective excitation-contraction coupling and dilated cardiomyopathy. J Clin Invest 102:1444–1453. doi: 10.1172/JCI1972 PubMedCrossRefGoogle Scholar
  36. 36.
    Wittchen F, Suckau L, Witt H, Skurk C, Lassner D, Fechner H, Sipo I, Ungethum U, Ruiz P, Pauschinger M, Tschope C, Rauch U, Kuhl U, Schultheiss HP, Poller W (2007) Genomic expression profiling of human inflammatory cardiomyopathy (DCMi) suggests novel therapeutic targets. J Mol Med 85:257–271. doi: 10.1007/s00109-006-0122-9 PubMedCrossRefGoogle Scholar
  37. 37.
    Xiong D, Yajima T, Lim BK, Stenbit A, Dublin A, Dalton ND, Summers-Torres D, Molkentin JD, Duplain H, Wessely R, Chen J, Knowlton KU (2007) Inducible cardiac-restricted expression of enteroviral protease 2A is sufficient to induce dilated cardiomyopathy. Circulation 115:94–102. doi: 10.1161/CIRCULATIONAHA.106.631093 PubMedCrossRefGoogle Scholar
  38. 38.
    Xu J, Nie HG, Zhang XD, Tian Y, Yu B (2011) Down-regulated energy metabolism genes associated with mitochondria oxidative phosphorylation and fatty acid metabolism in viral cardiomyopathy mouse heart. Mol Biol Rep 38:4007–4013. doi: 10.1007/s11033-010-0519-y PubMedCrossRefGoogle Scholar
  39. 39.
    Yilmaz A, Mahrholdt H, Athanasiadis A, Vogelsberg H, Meinhardt G, Voehringer M, Kispert EM, Deluigi C, Baccouche H, Spodarev E, Klingel K, Kandolf R, Sechtem U (2008) Coronary vasospasm as the underlying cause for chest pain in patients with PVB19 myocarditis. Heart 94:1456–1463. doi: 10.1136/hrt.2007.131383 PubMedCrossRefGoogle Scholar
  40. 40.
    Zakrzewska K, Cortivo R, Tonello C, Panfilo S, Abatangelo G, Giuggioli D, Ferri C, Corcioli F, Azzi A (2005) Human parvovirus B19 experimental infection in human fibroblasts and endothelial cells cultures. Virus Res 114:1–5. doi: 10.1016/j.virusres.2005.05.003 PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • U. Kuhl
    • 1
  • D. Lassner
    • 2
  • A. Dorner
    • 1
  • M. Rohde
    • 2
  • F. Escher
    • 1
  • B. Seeberg
    • 1
  • E. Hertel
    • 1
  • C. Tschope
    • 1
  • C. Skurk
    • 1
  • U. M. Gross
    • 2
  • H.-P. Schultheiss
    • 1
  • W. Poller
    • 1
  1. 1.Department of Cardiology and Pneumology, Charité Centrum 11 (Cardiovascular Medicine)Charité-Universitätsmedizin BerlinBerlinGermany
  2. 2.Institut Kardiale Diagnostik und Therapie (IKDT)BerlinGermany

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