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Open AccessArticle

Identification of a Diagnostic Set of Endomyocardial Biopsy microRNAs for Acute Cellular Rejection Diagnostics in Patients after Heart Transplantation Using Next-Generation Sequencing

1
Department of Cardiovascular Diseases, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Pekařská 53, 65691Brno, Czech Republic
2
Central European Institute of Technology, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic
3
Department of Pathological Physiology, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic
4
Department of Pathology, St. Anne’s University Hospital and Faculty of Medicine, Masaryk University, Pekařská 53, 65691 Brno, Czech Republic
5
Centre of Cardiovascular Surgery and Organ Transplantation, Pekařská 53, 65691 Brno, Czech Republic
*
Authors to whom correspondence should be addressed.
Both authors contributed equally to the manuscript (co-equal first authors).
Cells 2019, 8(11), 1400; https://doi.org/10.3390/cells8111400
Received: 20 October 2019 / Revised: 3 November 2019 / Accepted: 4 November 2019 / Published: 6 November 2019
(This article belongs to the Special Issue microRNA as Biomarker)
Introduction: Acute cellular rejection (ACR) of heart allografts represents the most common reason for graft failure. Endomyocardial biopsies (EMB) are still subject to substantial interobserver variability. Novel biomarkers enabling precise ACR diagnostics may decrease interobserver variability. We aimed to identify a specific subset of microRNAs reflecting the presence of ACR. Patients and Methods: Monocentric retrospective study. A total of 38 patients with the anamnesis of ACR were identified and for each patient three consecutive samples of EMB (with, prior and after ACR) were collected. Sixteen trios were used for next-generation sequencing (exploratory cohort); the resting 22 trios were used for validation with qRT-PCR (validation cohort). Statistical analysis was performed using R software. Results: The analysis of the exploration cohort provided the total of 11 miRNAs that were altered during ACR, the three of which (miR-144, miR-589 and miR-182) were further validated in the validation cohort. Using the levels of all 11 miRNAs and principal component analysis, an ACR score was created with the specificity of 91% and sensitivity of 68% for detecting the presence of ACR in the EMB sample. Conclusion: We identified a set of microRNAs altered in endomyocardial biopsies during ACR and using their relative levels we created a diagnostic score that can be used for ACR diagnosis.
Keywords: microRNA; endomyocardial biopsy; heart transplantation; acute cellular rejection; diagnostics microRNA; endomyocardial biopsy; heart transplantation; acute cellular rejection; diagnostics
MDPI and ACS Style

Nováková, T.; Macháčková, T.; Novák, J.; Hude, P.; Godava, J.; Žampachová, V.; Oppelt, J.; Zlámal, F.; Němec, P.; Bedáňová, H.; Slabý, O.; Bienertová-Vašků, J.; Špinarová, L.; Krejčí, J. Identification of a Diagnostic Set of Endomyocardial Biopsy microRNAs for Acute Cellular Rejection Diagnostics in Patients after Heart Transplantation Using Next-Generation Sequencing. Cells 2019, 8, 1400.

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