Background: The development of new non-invasive markers for prostate cancer (PC) diagnosis, prognosis, and management is an important issue that needs to be addressed to decrease PC mortality. Small extracellular vesicles (SEVs) secreted by prostate gland or prostate cancer cells into the plasma are considered next-generation diagnostic tools because their chemical composition might reflect the PC development. The population of plasma vesicles is extremely heterogeneous. The study aimed to explore a new approach for prostate-derived SEV isolation followed by vesicular miRNA analysis. Methods: We used superparamagnetic particles functionalized by five types of DNA-aptamers binding the surface markers of prostate cells. Specificity of binding was assayed by AuNP-aptasensor. Prostate-derived SEVs were isolated from the plasma of 36 PC patients and 18 healthy donors and used for the assessment of twelve PC-associated miRNAs. The amplification ratio (amp-ratio) value was obtained for all pairs of miRNAs, and the diagnostic significance of these parameters was evaluated. Results: The multi-ligand binding approach doubled the efficiency of prostate-derived SEVs’ isolation and made it possible to purify a sufficient amount of vesicular RNA. The neighbor clusterization, using three pairs of microRNAs (miR-205/miR-375, miR-26b/miR375, and miR-20a/miR-375), allowed us to distinguish PC patients and donors with sensitivity—94%, specificity—76%, and accuracy—87%. Moreover, the amp-ratios of other miRNAs pairs reflected such parameters as plasma PSA level, prostate volume, and Gleason score of PC. Conclusions: Multi-ligand isolation of prostate-derived vesicles followed by vesicular miRNA analysis is a promising method for PC diagnosis and monitoring.

PUB were compared to the postsurgical pathological specimen or follow-up for non-operated tumors. ResultsFrom November 2016 to February 2022, 34 patients were included. Based on surgical specimen (n=23) or follow up (n=11, including 4 metastasis), the final diagnoses were: 11 sarcomas and 23 non-sarcomas including 22 LM and one inflammatory myofibroblastic tumor [JS1] . The median followup was 12 months (IQR: 6-37). The diagnostic accuracy of M-PUB and MCGH-PUB were 94% and 100%. The sensitivity, specificity and Negative Predictive Value of MCGH-PUB were 100%, 100% and 100%. A high GI was significantly associated with malignancy (p<0•001). Genomic analyses allowed correct malignancy upgrade for four tumors after suspicious microscopic examination. There was no PUB complication and no dissemination on the biopsy track. Conclusion MCGH-PUB is safe and accurate to discriminate pre operatively benign tumors from uterine sarcoma.

Cervical cancer is one of the most common gynecological malignancies and it is preventable through the yearly diagnosis and management of pre-cancerous cervical disease. The profile of miRNA expression in cervical epithelium cells is altered with cervical dysplasia development and further progression. The NOVAprep-miR-CERVIX is a new approach for the assessment of cervical dysplasia through the analysis of six marker miRNAs. This study aims to evaluate theperformance and diagnostic potency of the new method. Cytological smears from 226 women (NILM, n.114; HSIL, n.112) were included in the study. A VPH test was performed with RealBest DNAHPV HR screen Kit, six marker miRNAs (miR-21, -29b, -145, -451a, -1246, -1290) were assayed using NOVAprep-miR-CERVIX kit. Obtained data were analyzed using the Delta Ct method and random forest machine learning algorithm. The results of the quantitative analysis of six microRNAs were expressed as a miR-CERVIX parameter, which ranged from 0 to 1, where “0” corresponded to the healthy cervical epithelium, while “1” corresponded to high-grade squamous intraepithelial dysplasia. The average value of miR-CERVIX differed in groups of NILM and HSIL samples (0.34 vs. 0.72; p < 0.000005). An estimation of miR-CERVIX allowed for the differentiation between healthy and pre-cancerous samples with sensitivity of 0.79 and specificity of 0.79, as well as to confirm HSIL with specificity of 0.98. Interestingly, the HSIL group included HPV(+) and HPV(−) samples, which were statistically significantly different in terms of miR-CERVIX value. Analysis of CC-associated miRNAs in material of cervical smear might serve as an additional method for the evaluation of cervical dysplasia severity.

Introduction. Testicular germ cell tumor is a relatively rare disease. Its high social significance is due to the fact that this pathology occurs in young patients. The standard schemes of polychemotherapy determine the potential possibility of effective treatment for most of the patients even with an advanced disease. Several circulating markers (alpha-fetoprotein, human chorionic gonadotropin and lactate dehydrogenase) are being used for therapy monitoring, but the low diagnostic specificity of these molecules determines the need to develop new approaches. Over the past years, circulating microRNA, for instance miR-371a-3p, appeared to be promising marker for testicular germ cell tumor monitoring. However, to develop and to implement in practice the microRNA-based diagnostic technologies, it’s necessarily to understand the features of the microRNA expression alterations specific for different histological types of testicular germ cell tumor.The study objective – to evaluate changes in the expression of several potential marker microRNA molecules (miR-302/ miR-367, miR-371/miR-373) in testicular germ cell tumor samples of various histological types.Materials and methods. Testicular germ cell tumor samples (n = 61), including seminomas, embryonic carcinomas, post-pubertal teratomas, yolk sac tumors, chorioncarcinomas, and corresponding normal tissue samples (n = 61) were included in the study. The analysis of selected miRNA expression was performed by reverse transcription and polymerase chain reaction.Results. We identified the changes in the expression profile of the miR-302/miR-367 cluster typical for semines, embryonic carcinomas, post-pubertal teratomas, yolk sac tumors and chorioncarcinomas, as well as changes in the expression profile of the miR-371/miR-373 cluster, universal for all histotypes except chorioncarcinomas. Inhibition of miR-10b and miR-145 expression in semines, embryonic carcinomas, and post-pubertal teratomas was demonstrated.Conclusion. Activation of miR-302b, miR-302d, miR-371a expression and inhibition of miR-10b, miR-145 expression in the tissue of the most common variants of testicular germ cell tumor is a characteristic feature of these tumors. The detected changes are significant and can lead to corresponding changes in the profile of circulating microRNAs.