Influenza della variabilità preanalitica sul profilo di espressione dei microRNA plasmatici

Résumé fourni par la source

INTRODUCTION: MicroRNAs (miRNAs) represent potential non-invasive circulating biomarkers for oncological diseases like the myelodysplastic syndrome (MDS). Although the effect of biological variability on miRNA-expression profile is well documented, limited data are available on the role of preanalytical variability, both relevant in the prospective of an imminent clinical application of these molecules. The aim of this study was to investigate the impact of time and temperature storage in platelet-poor plasma samples (PPP) on miRNA-expression profile.METHODS: K2-EDTA plasma from 12 patients with MDS and 12 healthy donors (HD) were collected and centrifuged to obtain PPP within 3 hours (h). PPP aliquots were stored before miRNA extraction as follows: A = 4 °C for 30 minutes; B = 4 °C for 24 h; C = room temperature (RT) for 24 h; D = -20 °C for 10 days. miRNA libraries were prepared using QIAseq miRNA Library Kit (Qiagen), sequenced in the Next-generation sequencing platform NextSeq550 (Illumina) and analyzed using CLC workbench (Qiagen).RESULTS: Plasma miRNA-expression profile was affected the most by the C condition (RT for 24 h) followed by B; A and D showed no significant differences. 16 miRNAs were constantly differentially expressed in MDS compared to HD in A, B, C and D (13 upregulated, 3 downregulated in MDS).DISCUSSION: Our results highlight that the miRNA extraction immediately after PPP separation or PPP long-term storage at -20 °C represents the best preanalytical conditions for an unbiased comprehensive miRNA-expression profile analysis using NGS. The standardization and statement about preanalytical conditions should be always recommended in both miRNA research studies and in their clinical translation.

Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.