ESPN 53rd Annual Meeting

ESPN 2021


 
VESICULAR miRNAs AS NEW FRONTIER IN THE PREVENTION OF RENAL SUBCLINICAL REJECTION
Andrea Carrao 1 Diana Marzenta 1 Emanuele Vianello 2 Benedetta Bussolati 3 Federica Collino 4 Piera De Gaspari 1 Loris Bertoldi 5 Giuseppe Benvenuto 5 Davide Meneghesso 2 Mattia Parolin 2 Germana Longo 2 Luisa Murer 1 Elisa Benetti 2 Susanna Negrisolo 1

1- LABORATORY OF IMMUNOPATHOLOGY AND MOLECULAR BIOLOGY OF THE KIDNEY, INSTITUTE OF PEDIATRIC RESEARCH CITTà DELLA SPERANZA, DEPARTMENT OF WOMEN’S AND CHILDREN’S HEALTH, PADUA UNIVERSITY HOSPITAL, PADUA, ITALY
2- PEDIATRIC NEPHROLOGY DIALYSIS AND TRANSPLANT UNIT, DEPARTMENT OF WOMEN’S AND CHILDREN’S HEALTH, PADUA UNIVERSITY HOSPITAL, PADUA, ITALY
3- UNIVERSITY OF TURIN, DEPARTMENT OF MOLECULAR BIOTECHNOLOGY AND HEALTH SCIENCES, ITALY
4- MAJOR GENERAL HOSPITAL OF MILAN, CA GRANDA FOUNDATION IRCCS, LABORATORY OF TRANSLATIONAL RESEARCH IN PEDIATRIC NEPHRO-UROLOGY, ITALY
5- BMR-GENOMICS, PADUA, ITALY
 
Introduction:

The study of novel biomarkers useful to prevent subclinical rejection has grown up in the last decades. Among the various candidates emerge the microRNAs (miRNAs), short non-coding sequences of RNA (21 – 23 nt) involved in different post-transcriptional gene pathways. Their presence is confirmed in different types of tissues and biological fluids, such as urine. Due to their high degradability, miRNAs are often carry by extracellular vesicles (EV), lipid nano-bound particles release from all types of cells. Particularly the study of urinary extracellular vesicles (UEVs) and their miRNAs content could be helpful to reflect directly the condition of the kidney. For this reason, we try to identify a miRNAs profile from UEVs samples useful to prevent subclinical kidney rejection in pediatric transplanted patients.

Material and methods:

The miRNAs were extracted from UEVs of 20 pediatric transplanted patients, with a stable condition or kidney rejection at one-year post-transplantation. The UEVs were isolated from urine samples by the ultracentrifugation method. The vesicles concentration and size were identifying by electron microscopy and scattering analysis (Nanosight 3000). The extracted miRNAs were enriched before sequencing with the Illumina sequencer.

Results:

The miRNAs sequencing shows about 522 different miRNAs and 48 of them were differentially expressed between patients with subclinical rejection or stable condition. UEVs characterization show a concentration of 2,79 x 1011 – 9,56 x 1011 particles with a size diameter of about 197 ± 7 nm. The miRNAs concentration was between 197 – 907 pg/µl.

Conclusions:

These results emphasize the relevance of vesicles miRNAs as useful biomarkers lead to the evaluation and prevention of subclinical kidney rejection in pediatric transplanted patients.