Differences in plasma miRNA levels in inferior petrosal sinus samples of patients with ACTH-dependent Cushing’s syndrome

• DOI: 10.14341/probl12817
• Publication Date: 2022-01-12T09:11:47Z
• Authors: A. A. Malygina; Z. E. Belaya; A. G. Nikitin; P. A. Koshkin; I. I. Sitkin; A. M. Lapshina; P. M. Khandaeva; A. S. Lutsenko; D. A. Trukhina; G. A. Melnichenko
• Keywords: Adult; Male; 0303 health sciences; Middle Aged; Petrosal Sinus Sampling; 3. Good health; MicroRNAs; 03 medical and health sciences; Adrenocorticotropic Hormone; Case-Control Studies; Humans; Female; Cushing Syndrome; Aged

BACKGROUND : For the last decades microRNAs (miR) have proven themselves as novel biomarkers for various types of diseases. Identification specific circulating microRNA panel that differ patient with Cushing’s disease (CD) and ectopic ACTH syndrome (EAS) could improve diagnostic procedure. AIM to evaluate differences in miR levels plasma samples drained from inferior petrosal sinuses patients CD EAS. MATERIALS AND METHODS single-center, case-control study: we enrolled 24 ACTH-dependent (CS) requiring bilateral sinus sampling (BIPSS). Among them 12 subjects were confirmed (males=2, females=10; median age 46,5 [IR 33,8;53,5]) EAS (males=4, females=8, 54 38,75;60,75]). BIPSS was performed through a percutaneous approach. Once catheters properly placed, blood withdrawn simultaneously each peripheral vein. Plasma both centrifuged then stored at -80 C. MiRNA isolation carried out by an miRneasy Plasma/Serum Kit (Qiagen, Germany) on automatic QIAcube station according manufacturer protocol. To prevent degradation, added 1 unit RiboLock Rnase Inhibitor (Thermo Fisher Scientific, USA) per μL RNA solution. The concentration total aqueous solution evaluated NanoVue Plus spectrophotometer (GE Healthcare, USA). libraries prepared QIAseq miRNA Library following standard protocols. MiR expression analyzed sequencing Illumina NextSeq 500 (Illumina, RESULTS 108 miRNAs differently expressed (p <0,05) vs We divided these into 3 groups based significance results. first group consisted highest detected groups. Four included: miR-1203 downregulated — 36.74 (p=0,013), three other upregulated EAS: miR-383-3p 46.36 (p=0,01), miR-4290 6.84 (p=0,036), miR-6717-5p 4.49 (p=0,031). This miRs will be validated larger cohorts using RT-qPCR. CONCLUSION taken These need different methods order used potentially non-invasive differentiate CS.