Urinary Proteomics Pilot Study for Biomarker Discovery and Diagnosis in Heart Failure with Reduced Ejection Fraction

PLoS One. 2016 Jun 16;11(6):e0157167. doi: 10.1371/journal.pone.0157167. eCollection 2016.

Abstract

Background: Biomarker discovery and new insights into the pathophysiology of heart failure with reduced ejection fraction (HFrEF) may emerge from recent advances in high-throughput urinary proteomics. This could lead to improved diagnosis, risk stratification and management of HFrEF.

Methods and results: Urine samples were analyzed by on-line capillary electrophoresis coupled to electrospray ionization micro time-of-flight mass spectrometry (CE-MS) to generate individual urinary proteome profiles. In an initial biomarker discovery cohort, analysis of urinary proteome profiles from 33 HFrEF patients and 29 age- and sex-matched individuals without HFrEF resulted in identification of 103 peptides that were significantly differentially excreted in HFrEF. These 103 peptides were used to establish the support vector machine-based HFrEF classifier HFrEF103. In a subsequent validation cohort, HFrEF103 very accurately (area under the curve, AUC = 0.972) discriminated between HFrEF patients (N = 94, sensitivity = 93.6%) and control individuals with and without impaired renal function and hypertension (N = 552, specificity = 92.9%). Interestingly, HFrEF103 showed low sensitivity (12.6%) in individuals with diastolic left ventricular dysfunction (N = 176). The HFrEF-related peptide biomarkers mainly included fragments of fibrillar type I and III collagen but also, e.g., of fibrinogen beta and alpha-1-antitrypsin.

Conclusion: CE-MS based urine proteome analysis served as a sensitive tool to determine a vast array of HFrEF-related urinary peptide biomarkers which might help improving our understanding and diagnosis of heart failure.

MeSH terms

  • Adult
  • Aged
  • Biomarkers / urine*
  • Female
  • Heart Failure / genetics
  • Heart Failure / pathology
  • Heart Failure / urine*
  • Humans
  • Male
  • Middle Aged
  • Peptide Fragments / genetics
  • Peptide Fragments / urine*
  • Proteomics*
  • Spectrometry, Mass, Electrospray Ionization
  • Stroke Volume
  • Support Vector Machine

Substances

  • Biomarkers
  • Peptide Fragments

Grants and funding

The funding organization (European Union; grants EU-MASCARA [HEALTH-2011.2.4.2-2] and HOMAGE [HEALTH-F7- 305507 HOMAGE] as well as the Muremester Laurits P Christensens Fund and the Kaptajnløjtnant Harald Jensen og Hustrus Fund had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript and only provided financial support. Harald Mischak, Thomas Koeck and Esther Nkuipou-Kenfack were employed by and received salary from Mosaiques Diagnostics GmbH. Being employed by Mosaiques Diagnostics GmbH did not influence study design, decision to publish or preparation of the manuscript. Urinary proteomic data collection and analysis was performed at Mosaiques Diagnostics GmbH following established standard operating procedures.