ESPN 51th Annual Meeting

ESPN 2018


 
MICRORNA PROFILING IN PEDIATRIC KIDNEY TRANSPLANTATION: A TOOL TO FIND NOVEL BIOMARKERS OF GRAFT REJECTION
ANDREA CARRARO 1 SUSANNA NEGRISOLO 1 GIULIA FREGONESE 1 EMANUELE VIANELLO 1 PIERA DE GASPARI 2 PIERGIORGIO GAMBA 3 GIORGIO VALLE 4 ENRICO VIDAL 1 LUISA MURER 1

1- HOSPITAL-UNIVERSITY OF PADUA, PEDIATRIC NEPHROLOGY, DIALYSIS AND TRANSPLANT UNIT, DEPARTMENT OF WOMEN’S AND CHILDREN’S HEALTH, PADUA, ITALY
2- HOSPITAL-UNIVERSITY OF PADUA, NEUROLOGICAL CLINIC, PADUA, ITALY
3- HOSPITAL-UNIVERSITY OF PADUA, PEDIATRIC SURGERY, WOMEN’S AND CHILDREN’S HEALTH DEPARTMENT, PADUA, ITALY
4- UNIVERSITY OF PADUA, CRIBI BIOTECHNOLOGY CENTRE, PADUA, ITALY
 
Introduction:

In the last years, the study of novel non-invasive biomarkers has led to the study of micro-RNAs (miRNAs) as possible markers useful to prevent kidney rejection. miRNAs are short non coding RNA sequences involved in different physiological process that might play a key role in kidney rejection. Therefore, it would be useful to check the expression of miRNAs in renal tissue and in biological fluids. In this perspective we performed a miRNA profiling analysis in protocol biopsies of pediatric transplanted patients enrolled in our center.

Material and methods:

Using commercial kits, miRNAs fraction was extracted at one year post transplantation from kidney tissue of 30 patients. miRNAs quality and concentration were estimated by Agilent Bioanalyzer 2100 and Qubit Fluorometer. The samples with a good RIN (Rate Integrity Number) were selected. 10 patients samples with a normal histology (Banff I) and 10 patients samples with acute/chronic Kidney rejection (Banff II-III-IV) were processed. We used a miRNA enrichment kit for miRNA-seq (Illumina platform) that succeed to analyze small RNA species and generate sequencing libraries starting from 1 ng of material.

Results:

The tissues miRNAs showed a concentration range between 6.42 and 30.8 ng/µl and an adequate RIN (6.5 - 8.5). The miRNA-seq analysis showed about 1092 different miRNAs. Statistical analysis revealed a significant over-expression of five miRNAs in patients with rejection: hsa-miR-142-5p, hsa-miR-142-3p, hsa-miR-106b-3p, hsa-miR-101-3p, hsa-miR-185-5p.

Conclusions:

These results showed an overexpression of 5 miRNAs in biopsies of pediatric patients with a subclinical kidney rejection. Particularly, hsa-miR-142-5p and hsa-miR-142-3p have been reported before to be associated to acute kidney rejection in transplanted adults, on the contrary hsa-miR-106b-3p, hsa-miR-101-3p, hsa-miR-185-5p not. A prospective study will be performed to correlate these data with those that will be obtained from biological fluids. The final aim is to ameliorate the monitoring and prolong the graft survival.