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

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An Erratum to this article was published on 26 November 2013

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.

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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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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.

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Authors and Affiliations

  1. Department of Cardiology and Pneumology, Charité Centrum 11 (Cardiovascular Medicine), Charité-Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200, Berlin, Germany

    U. Kuhl, A. Dorner, F. Escher, B. Seeberg, E. Hertel, C. Tschope, C. Skurk, H.-P. Schultheiss & W. Poller

  2. Institut Kardiale Diagnostik und Therapie (IKDT), Berlin, Germany

    D. Lassner, M. Rohde & U. M. Gross

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  1. U. Kuhl
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  2. D. Lassner
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  3. A. Dorner
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  4. M. Rohde
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  5. F. Escher
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  6. B. Seeberg
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  7. E. Hertel
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  8. C. Tschope
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  9. C. Skurk
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  10. U. M. Gross
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  11. H.-P. Schultheiss
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  12. W. Poller
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Corresponding author

Correspondence to W. Poller.

Additional information

U. Kühl, D. Lassner, H.-P. Schultheiss and W. Poller are contributing equally to this paper.

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395_2013_372_MOESM1_ESM.docx

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)

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Kuhl, U., Lassner, D., Dorner, A. et al. A distinct subgroup of cardiomyopathy patients characterized by transcriptionally active cardiotropic erythrovirus and altered cardiac gene expression. Basic Res Cardiol 108, 372 (2013). https://doi.org/10.1007/s00395-013-0372-y

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