Molecular Assessment of Microcirculation Injury in Formalin-Fixed Human Cardiac Allograft Biopsies With Antibody-Mediated Rejection

Am J Transplant. 2017 Feb;17(2):496-505. doi: 10.1111/ajt.13956. Epub 2016 Aug 2.

Abstract

Precise diagnosis of antibody-mediated rejection (AMR) in cardiac allograft endomyocardial biopsies (EMBs) remains challenging. This study assessed molecular diagnostics in human EMBs with AMR. A set of 34 endothelial, natural killer cell and inflammatory genes was quantified in 106 formalin-fixed, paraffin-embedded EMBs classified according to 2013 International Society for Heart and Lung Transplantation (ISHLT) criteria. The gene set expression was compared between ISHLT diagnoses and correlated with donor-specific antibody (DSA), endothelial injury by electron microscopy (EM) and prognosis. Findings were validated in an independent set of 57 EMBs. In the training set (n = 106), AMR cases (n = 70) showed higher gene set expression than acute cellular rejection (ACR; n = 21, p < 0.001) and controls (n = 15, p < 0.0001). Anti-HLA DSA positivity was associated with higher gene set expression (p = 0.01). Endothelial injury by electron microscopy strongly correlated with gene set expression, specifically in AMR cases (r = 0.62, p = 0.002). Receiver operating characteristic curve analysis for diagnosing AMR showed greater accuracy with gene set expression (area under the curve [AUC] = 79.88) than with DSA (AUC = 70.47) and C4d (AUC = 70.71). In AMR patients (n = 17) with sequential biopsies, increasing gene set expression was associated with inferior prognosis (p = 0.034). These findings were confirmed in the validation set. In conclusion, biopsy-based molecular assessment of antibody-mediated microcirculation injury has the potential to improve diagnosis of AMR in human cardiac transplants.

Keywords: biopsy; clinical research/practice; heart transplantation/cardiology; molecular biology; pathology/histopathology; rejection: antibody-mediated (ABMR).

MeSH terms

  • Adult
  • Allografts
  • Biomarkers / analysis*
  • Biopsy
  • Female
  • Follow-Up Studies
  • Formaldehyde / chemistry*
  • Gene Expression Profiling
  • Graft Rejection / diagnosis*
  • Graft Rejection / etiology
  • Graft Survival
  • Heart Failure / surgery
  • Heart Transplantation / adverse effects*
  • Humans
  • Isoantibodies / immunology*
  • Killer Cells, Natural / immunology
  • Killer Cells, Natural / metabolism
  • Male
  • Microcirculation / genetics*
  • Middle Aged
  • Prognosis
  • Retrospective Studies
  • Risk Factors
  • Tissue Donors*

Substances

  • Biomarkers
  • Isoantibodies
  • Formaldehyde