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Proteomic signature of circulating extracellular vesicles in dilated cardiomyopathy

Laboratory Investigation (2018) | Download Citation

Abstract

Dilated cardiomyopathy (DCM) remains a major cause of heart failure and carries a poor prognosis despite important advances in recent years. Better disease characterization using novel molecular techniques is needed to refine its progression. This study explored the proteomic signature of plasma-derived extracellular vesicles (EVs) obtained from DCM patients and healthy controls using size-exclusion chromatography (SEC). EV-enriched fractions were analyzed by liquid chromatography-mass spectrometry (LC-MS/MS). Raw data obtained from LC-MS/MS were analyzed against the Uniprot human database using MaxQuant software. Additional analyses using Perseus software were based on the Intensity-Based Absolute Quantification (iBAQ) values from MaxQuant analyses. A total of 90.07 ± 21 proteins (227 different proteins) in the DCM group and 96.52 ± 17.91 proteins (183 different proteins) in the control group were identified. A total of 176 proteins (74.6%) were shared by controls and DCM patients, whereas 51 proteins were exclusive for the DCM group and 7 proteins were exclusive for the control group. Fibrinogen (α, β and γ chain), serotransferrin, α-1-antitrypsin, and a variety of apolipoprotein family members (C-I, C-III, D, H or β-2-glycoprotein, and J or clusterin) were clustered in SEC-EVs derived from DCM patients relative to controls (p < 0.05). Regarding Gene Ontology analysis, response to stress and protein activation-related proteins were enriched in DCM-EVs compared with controls. Thus, the present study reports the distinct proteomic signature of circulating DCM-EVs compared with control-EVs. Furthermore, we confirm that SEC obtains highly purified EV fractions from peripheral blood samples for subsequent use in determining disease-specific proteomic signatures.

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Acknowledgements

We specifically acknowledge Dr. Francisco Sánchez-Madrid (Servicio de Inmunología, Hospital Universitario de la Princesa, IIS-IP, UAM; Cell-Cell Communication Laboratory, CNIC) and Dr. María Yáñez-Mó (Unidad de Investigación, Hospital Sta Cristina, IIS-IP; Departamento Biología Molecular/CBM-SO, UAM) for kindly gifting antibodies against CD9, CD63 and CD81. We also thank Beatriz González, Roser Cabanes, Margarita Rodríguez, Carmen Rivas, Nuria Benito and Alba Ros for invaluable work in the Heart Failure Clinic; Eduard Sabido and Eva Borràs from the UPF/CRG Proteomics Unit for proteomic data processing and analysis; and Hernando del Portillo (ICREA Research Professor at ISGLOBAL-IGTP) for access to the NTA instrument.

Funding

This work was supported by grants from the Ministerio de Economía, Industria y Competitividad (SAF2014-59892-R), Fundació La MARATÓ de TV3 (201405/10, 201502, 201516), Societat Catalana de Cardiologia, Generalitat de Catalunya (SGR 2014, CERCA Programme) and the Fundació Bancària La Caixa. This work was also funded by the Red de Terapia Celular—TerCel (RD16/0011/0006), CIBER Cardiovascular—(CB16/11/00403), and Fondo de Investigación Sanitaria, Instituto de Salud Carlos III (FIS PI14/01682) as part of the Plan Nacional de I+D+I cofounded by ISCIII-Sudirección General de Evaluación y el Fondo Europeo de Desarrollo Regional (FEDER).

Author information

Author notes

  1. These authors contributed equally: Santiago Roura, Ana Gámez-Valero.

Affiliations

  1. Heart Failure and Cardiac Regeneration (ICREC) Research Program, Health Science Research Institute Germans Trias i Pujol (IGTP), Badalona, Spain

    • Santiago Roura
    • , Carolina Gálvez-Montón
    •  & Antoni Bayes-Genis
  2. Center of Regenerative Medicine in Barcelona, Barcelona, Spain

    • Santiago Roura
  3. CIBERCV, Instituto de Salud Carlos III, Madrid, Spain

    • Santiago Roura
    • , Josep Lupón
    • , Carolina Gálvez-Montón
    •  & Antoni Bayes-Genis
  4. REMAR-IVECAT Group, Germans Trias i Pujol Research Institute (IGTP), Can Ruti Campus, Badalona, Spain

    • Ana Gámez-Valero
    •  & Francesc E. Borràs
  5. Department of Pathology, Hospital Universitari and Health Sciences Research Institute Germans Trias i Pujol, Universitat Autònoma de Badalona, Badalona, Spain

    • Ana Gámez-Valero
  6. Cardiology Service, HUGTiP, Badalona, Spain

    • Josep Lupón
    •  & Antoni Bayes-Genis
  7. Department of Medicine, Universitat Autònoma de Badalona, Badalona, Spain

    • Josep Lupón
    •  & Antoni Bayes-Genis
  8. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain

    • Francesc E. Borràs
  9. Nephrology Service, Hospital Universitari Germans Trias i Pujol, Badalona, Spain

    • Francesc E. Borràs

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Conflict of interest

The authors declare that they have no conflict of interest.

Corresponding authors

Correspondence to Francesc E. Borràs or Antoni Bayes-Genis.

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DOI

https://doi.org/10.1038/s41374-018-0044-5