CL_III_L: Clinical Medicine III. Lectures
Péter István Turai 1, 2, 3, Gábor Nyírő 3, Katalin Borka 4, Tamás Micsik 5, Attila Patócs 6, 7, 8, Péter Igaz 1, 2, 3
1 Department of Endocrinology, Faculty of Medicine, Semmelweis University, H-1083 Budapest, Hungary
2 Department of Internal Medicine and Oncology, Faculty of Medicine, Semmelweis University, H-1083 Budapest, Hungary
3 MTA-SE Molecular Medicine Research Group, Eötvös Loránd Research Network, H-1083 Budapest, Hungary
4 2nd Department of Pathology, Semmelweis University, H-1091 Budapest, Hungary
5 1st Department of Pathology and Experimental Cancer Research, Semmelweis University, H-1085 Budapest, Hungary
6 MTA-SE Hereditary Tumors Research Group, Eötvös Lóránd Research Network, H-1089 Budapest, Hungary
7 Department of Molecular Genetics, National Institute of Oncology, H-1022 Budapest, Hungary
8 Hungary Department of Laboratory Medicine, Faculty of Medicine, Semmelweis University, H-1089 Budapest, Hungary
Introduction. Most of the adrenocortical tumors are incidental clinical findings comprising 54% of benign adrenocortical adenomas (ACA) with excellent prognosis, rarely (0.7-2/million/year) however adrenocortical carcinoma (ACC) can be detected with a five-year survival of less than 30%. As the histological differentiation of ACA and ACC is difficult, molecular markers are of big clinicopathological relevance. Circular RNAs have been implicated as relevant factors in tumor pathogenesis, but their pathogenic relevance in adrenocortical tumors is unknown. There are also data supporting their utility as diagnostic markers of malignancy in various tumors. Based on their outstanding stability, circular RNAs can be investigated both in archived pathological samples and from blood.
Aim: to study the expression of circular RNAs in benign and malignant adrenocortical tumors.
Materials and methods. FFPE samples were selected 8-8-8 each from ACC, ACA, and normal adrenal cortex (NAC) and has been validated by pathologists. Total RNA isolation was performed with RecoverAll Total Nucleic Acid Kit for FFPE. Nanodrop 2000 and Qubit 4 were used to assess total RNA quantity. To maximize purification process for circular RNAs, we applied the RPAD method. For expression profiling of known and novel circular RNAs, MiSeq NGS platform was used. RNA-seq reads were mapped to Hg38 reference genome. CIRI2, CircExplorer2, AutoCirc were used for discovery of de novo circular RNAs.
Results. Circular RNAs were significantly differentially expressed between ACC versus ACA and ACC versus NAC cohorts. ACA versus NAC cohort did not differ significantly. Number of circular RNAs per sample by CircExplorer2 was 2700 out of which 1655 were known circular RNAs. CIRI2 found 2486 circular RNAs in total with 1679 known circular RNAs. AutoCirc total/known circular RNA ratio was 1346/578.
Conclusion. To the best of our knowledge, we are the first to describe the circular RNA patterns of adrenocortical tumors. We were also able to find novel circular RNAs in adrenocortical tissue and tumor. There is significant difference in expression of ACC versus ACA and ACC versus NAC cohorts. These expression changes need to be validated and bioinformatical analyses of novel circular RNAs for biological relevance are also warranted. These results might contribute to improved diagnosis of ACC.
Semmelweis University, Károly Rácz Doctoral School of Clinical Medicine