Pál Perge1, Henriett Butz2, Raffaele Pezzani3, Irina Bancos4, Zoltán Nagy1, Ábel Decmann1, Michaela Luconi5, Massimo Mannelli5, Edit I. Buzás6, Miklós Tóth1, Marco Boscaro3, Attila Patócs2,7, Peter Igaz1,6
1 2nd Department of Medicine, Semmelweis University, Budapest,
2 Molecular Medicine Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest,
3 Endocrinology Unit, Department of Medicine, University of Padua, Padova, Italy
4 Division of Endocrinology, Diabetes, Metabolism and Nutrition, Department of Internal Medicine, Mayo Clinic, , Rochester, MN USA
5 Department of Experimental and Clinical Biomedical Sciences, Endocrinology Unit, University of Florence, Florence, Italy
6 Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, Hungary
7 “Lendület-2013” Research Group, Hungarian Academy of Sciences and Semmelweis University, Budapest,
Introduction: Blood-borne microRNAs (miRNA) have been described in adrenocortical tumors, however there is no plasma marker for the preoperative diagnosis. Moreover, the expression of miRNAs in hormonally inactive and cortisol-producing tumors has not yet been evaluated.
Aims: Therefore the first objective of this study was to investigate the expression of exosomal miRNAs and their diagnostic utility in adrenocortical tumors. The second aim was to examine the expression of plasma exosomal miRNAs in patients suffering from non-functioning adrenocortical adenoma (NFA), cortisol-producing adrenocortical adenoma (CPA) and cortisol-producing adrenocortical carcinoma (CP-ACC).
Method: Exosomes were isolated either by performing ultracentrifugation or by applying Total Exosome Isolation Kit. Preoperative exosomal samples of 6 adrenocortical adenomas (ACA) and 6 adrenocortical carcinomas (ACC) were first screened by TaqMan Human Microarray A-cards, then two miRNAs (miR-101 and miR-483-5p) were validated in 18 ACAs and 16 ACCs by targeted RT-qPCR. Moreover, the expression of miR-22-3p, miR-27a-3p, miR-210-3p, miR-320b and miR-375 were analyzed in 13 NFA, 13 CPA and 9 CP-ACC samples.
Results: We found significant overexpression of miR-101 and miR-483-5p in ACC compared to ACA. Receiver operator characteristics analysis revealed miR-483-5p to have the highest diagnostic accuracy (AUC=0.965), the sensitivity and the specificity were 87.5 and 94.44, respectively. We observed significant overexpression of 3 miRNAs in both CPA and CP-ACC versus to NFA: miR-22-3p, miR-27a-3p and miR-320b. miR-320b was significantly overrepresented in CP-ACC compared to CPA. miR-210-3p was significantly overrepresented only in CP-ACC relative to NFA. Significant correlation was identified between circulating miRNA expression and urinary free cortisol concentrations for miR-22-3p, miR-27a-3p and miR-320b and cortisol level after low dose dexamethasone test (LDDT) for miR-22-3p and miR-320b. Furthermore, miR-27a-3p was significantly triggered by LDDT.
Conclusion: Exosomal miR-101 and miR-483-5p might be promising preoperative biomarkers of ACC. Moreover, exosomal miRNAs are dissimilarly expressed in hormonally inactive and cortisol-secreting adrenocortical neoplasms.
Clinical medicine, Hormonal regulations, Péter Igaz