INTRAGRAFT miRNA SIGNATURES DIFFERENTIATE REJECTION FROM NON-REJECTION PHENOTYPES IN KIDNEY ALLOGRAFTS

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INTRAGRAFT miRNA SIGNATURES DIFFERENTIATE REJECTION FROM NON-REJECTION PHENOTYPES IN KIDNEY ALLOGRAFTS

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Co-author 1

Neva Bezeljak neva.bezeljak@kclj.si University Medical Center Ljubljana Department of Nephrology Ljubljana Slovenia - University of Ljubljana Medical Faculty Ljubljana Slovenia

Co-author 2

Nika Kojc nika.kojc@mf.uni-lj.si Medical Faculty, University of Ljubljana Institute of Pathology Ljubljana Slovenia -

Co-author 3

Živa Pipan Tkalec Ziva.PIPAN-TKALEC@mf.uni-lj.si Medical Faculty, University of Ljubljana Institute of Pathology Ljubljana Slovenia -

Co-author 4

Miha Arnol miha.arnol@mf.uni-lj.si University Medical Center Ljubljana Department of Nephrology Ljubljana Slovenia - University of Ljubljana Medical Faculty Ljubljana Slovenia

Co-author 5

Emanuela Boštjančič emanuela.bostjancic@mf.uni-lj.si Medical Faculty, University of Ljubljana Institute of Pathology Ljubljana Slovenia -

Co-author 6

Željka Večerić-Haler zeljka.vecerichaler@kclj.si University Medical Center Ljubljana Department of Nephrology Ljubljana Slovenia * University of Ljubljana Medical Faculty Ljubljana Slovenia

Co-author 7

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Introduction

MicroRNAs (miRNAs) are small non-coding RNAs regulating immune activation and fibrosis and have been implicated in various forms of kidney allograft injury. However, few studies have combined clinical, histological, and molecular confirmation of diagnosis when assessing intragraft miRNA expression. This study aimed to evaluate the expression of five selected miRNAs (miR-29b-3p, miR-142-3p, miR-142-5p, miR-155-5p, and miR-223-3p) across distinct phenotypes of allograft injury, including rejection, BK virus-associated nephropathy, and acute tubular necrosis.

Methods

We analyzed kidney allograft biopsies from patients with antibody-mediated rejection (n = 26), T-cell-mediated rejection (n = 14), BK virus–associated nephropathy (n = 15), acute tubular necrosis (n = 8), and normal controls (n = 20). All diagnoses were established using a tripartite diagnostic approach integrating clinical evolution, histopathology according to Banff 2019 criteria, and molecular transcript analysis (Molecular Microscope® Diagnostic System, MMDx), ensuring maximal diagnostic certainty. Quantitative real-time PCR was used for miRNA profiling, and expression data were correlated with clinicopathological parameters and molecular injury signatures.

Results

Rejection phenotypes showed the highest expression of immune-related miRNAs (miR-155-5p, miR-142-5p, miR-223-3p), whereas miR-29b-3p was down-regulated in both rejection and BK virus-associated nephropathy, reflecting fibrotic remodeling. A combined multifactorial model of the analyzed miRNAs significantly improved discrimination between rejection and non-rejection phenotypes. Among individual candidates, miR-223-3p showed the strongest discriminatory power between rejection and acute tubular necrosis, indicating its potential as a complementary molecular biomarker.

Conclusion

This study provides the first integrated analysis of intragraft miRNA expression linking clinical, histological, and molecularly verified diagnoses. The five-miRNA panel, selected based on prior experimental and translational data, reliably distinguishes between rejection and other injury patterns. The findings support the incorporation of multifactorial miRNA models into diagnostic workflows to refine phenotypic classification and improve post-transplant monitoring.

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