Introduction
Targeted molecular profiling combined with reproducible formalin-fixed paraffin-embedded (FFPE) EMB-based technology has the potential to support cardiac rejection diagnosis.
Objective
We aimed to develop and validate targeted gene expression diagnostic models of cardiac rejection and show their association with ISHLT pathological grades.
Method
We built a multicenter cohort of 591 FFPE-EMBs collected from four international centers between 2011 and 2021. Biopsies were graded according to the ISHLT working formulations, including 188 AMR cases, 289 ACR cases, and 114 non-rejection cases, randomly split in a derivation (n = 475) and a validation cohort (n = 116). Tissue gene expression was analyzed on FFPE-EMB using the Banff Human Organ Transplant gene set. Molecular classifiers for AMR and ACR were built using a supervised model. Association between molecular scores and pathology severity of rejection were analyzed in both derivation and validation set.
Results
The derivation included a total of 151 AMR (pAMR1H+: n = 46, pAMR1I+: n = 36, pAMR2-3: n = 69), 232 ACR (ACR 1R: n = 143, ACR 2-3R: n = 89) and 92 non-rejection cases. The validation set included a total of 37 AMR (pAMR1H+: n = 12, pAMR1I+: n = 15, pAMR2-3: n = 10), 57 ACR (ACR 1R: n = 31, ACR 2-3R: n = 26) and 22 non-rejection cases. Median AMR scores in AMR, ACR and non-rejection cases were 0.655 (IQR = 0.316), 0.217 (IQR = 0.254) and 0.140 (IQR = 0.209), respectively. Median ACR scores in ACR, AMR and non-rejection cases were 0.678 (IQR = 0.347), 0.263 (IQR = 0.364) and 0.302 (IQR = 270), respectively. AMR and ACR molecular scores were strongly associated with the pathology assessment of severity of rejection according to AMR and ACR international working formulations, respectively (derivation set: ACR: P for trend = 1.017E-46, AMR: P for trend = 2.911E-52; validation set: ACR: P for trend = 1.827E-13, AMR: P for trend = 5.174E-10, Fig. 1). Molecular AMR score was not associated with ACR severity; neither was the ACR molecular score with AMR severity.
Conclusion
Tissue-based molecular diagnostic system developed closely aligned with histological grading of cardiac allograft rejection, enhancing diagnostic precision, and offering a reliable companion tool for routine practice.
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