MicroRNA signatures in cardiac biopsies and detection of allograft rejection

Andrea Di Francesco; Marny Fedrigo; Donato Santovito; Lucia Natarelli; Chiara Castellani; Fabio De Pascale; Giuseppe Toscano; Angela Fraiese; Giuseppe Feltrin; Elena Benazzi; Angela Nocco; Gaetano Thiene; Marialuisa Valente; Giorgio Valle; Andreas Schober; Gino Gerosa; Annalisa Angelini

doi:10.1016/j.healun.2018.06.010

MicroRNA signatures in cardiac biopsies and detection of allograft rejection

J Heart Lung Transplant. 2018 Nov;37(11):1329-1340. doi: 10.1016/j.healun.2018.06.010. Epub 2018 Jun 30.

Authors

Andrea Di Francesco¹, Marny Fedrigo², Donato Santovito³, Lucia Natarelli³, Chiara Castellani¹, Fabio De Pascale⁴, Giuseppe Toscano¹, Angela Fraiese¹, Giuseppe Feltrin¹, Elena Benazzi⁵, Angela Nocco⁵, Gaetano Thiene¹, Marialuisa Valente¹, Giorgio Valle⁴, Andreas Schober³, Gino Gerosa¹, Annalisa Angelini¹

Affiliations

¹ Department of Cardiac, Thoracic and Vascular Sciences, University of Padova, Padova, Italy.
² Department of Cardiac, Thoracic and Vascular Sciences, University of Padova, Padova, Italy. Electronic address: marny.fedrigo@unipd.it.
³ Institute for Cardiovascular Prevention, Ludwig-Maximillians University, Munich, Germany.
⁴ CRIBI Biotechnology Centre, University of Padova, Padova, Italy.
⁵ Organ and Tissue Transplantation Immunology, Fondazione IRCSS Ca' Granda Ospedale Maggiore Policlinico, Milano, Italy.

PMID: 30174164
DOI: 10.1016/j.healun.2018.06.010

Abstract

Background: Identification of heart transplant (HTx) rejection currently relies on immunohistology and immunohistochemistry. We aimed to identify specific sets of microRNAs (miRNAs) to characterize acute cellular rejection (ACR), antibody-mediated rejection (pAMR), and mixed rejection (MR) in monitoring formalin-fixed paraffin-embedded (FFPE) endomyocardial biopsies (EMBs) in HTx patients.

Methods: In this study we selected 33 adult HTx patients. For each, we chose the first positive EMB for study of each type of rejection. The next-generation sequencing (NGS) IonProton technique and reverse transcript quantitative polymerase chain reaction (RT-qPCR) analysis were performed on FFPE EMBs. Using logistic regression analysis we created unique miRNA signatures as predictive models of each rejection. In situ PCR was carried out on the same EMBs.

Results: We obtained >2,257 mature miRNAs from all the EMBs. The 3 types of rejection showed a different miRNA profile for each group. The logistic regression model formed by miRNAs 208a, 126-5p, and 135a-5p identified MR; that formed by miRNAs 27b-3p, 29b-3p, and 199a-3p identified ACR; and that formed by miRNAs 208a, 29b-3p, 135a-5p, and 144-3p identified pAMR. The expression of miRNAs on tissue, through in situ PCR, showed different expressions of the same miRNA in different rejections. miRNA 126-5p was expressed in endothelial cells in ACR but in cardiomyocytes in pAMR. In ACR, miRNA 29b-3p was significantly overexpressed and detected in fibroblasts, whereas in pAMR it was underexpressed and detected only in cardiomyocytes.

Conclusions: miRNA profiling on FFPE EMBs differentiates the 3 types of rejection. Localization of expression of miRNAs on tissue showed different expression of the same miRNA for different cells, suggesting different roles of the same miRNA in different rejections.

Keywords: HTx EMBs; NGS miRNA profiling; acute cellular rejection; antibody-mediated rejection; mixed rejection.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult
Aged
Biopsy
Female
Graft Rejection / genetics*
Graft Rejection / pathology
Heart Transplantation*
High-Throughput Nucleotide Sequencing
Humans
Immunoenzyme Techniques
Male
MicroRNAs / genetics*
Middle Aged
Myocardium / pathology*
Polymerase Chain Reaction
Transcriptome / genetics*

Substances

MicroRNAs