Določanje mikro RNA v urinu bolnikov z nefroblastomom

Lavrič, Špela (author), Trebušak Podkrajšek, Katarina (mentor), Tesovnik, Tine (comentor)

Wilmsov tumor ali nefroblastom je najpogostejše maligno obolenje ledvic v pediatrični populaciji. V večini primerov je nastanek tumorja posledica patogenih različic genov, vključenih v proces celične diferenciacije in posttranskripcijskega uravnavanja genov. Najpogostejši vzrok za nastanek tumorja so patološke različice genov CTNNB1, WTX, TP53, MYCN, SIX1, SIX2, AMER1, DROSHA, DICER 1, DGCR8, XPO5 in TARBP2. V manjši meri je nastanek tumorja posledica prirojenih genetskih sindromov, kot so Beckwith-Wiedemann, Denys-Drash, sindrom WAGR in sindrom Perlman, ali patoloških različic v zarodnih celicah oziroma dednih sprememb, ki povečujejo tveganje za razvoj rakavih bolezni. Pomembno vlogo pri nastanku in razvoju različnih vrst rakavih bolezni, vključno z Wilmsovim tumorjem, imajo molekule miRNA. Gre za skupino majhnih nekodirajočih molekul RNA, ki se po sintezi ne prepišejo v proteine, temveč imajo v celici pomembno vlogo v posttranskripcijski regulaciji izražanja genov. Spremenjen profil izražanja miRNA lahko vodi v nastanek bolezni, zato so miRNA v zadnjem obdobju predmet številnih študij, ki raziskujejo njihovo uporabnost v diagnostiki in zdravljenju, predvsem pri nekaterih vrstah rakavih bolezni. Namen magistrske naloge je bil opredeliti profil izražanja miRNA v urinu bolnikov z Wilmsovim tumorjem z metodo sekvenciranja naslednje generacije, ter prepoznati miRNA, ki bi predstavljale potencialne biološke označevalce za neinvazivno zgodnje odkrivanje bolezni in spremljanje uspešnosti zdravljenja. V raziskavo smo vključili 18 preiskovancev, 9 bolnikov z Wilmsovim tumorjem in 9 zdravih preiskovancev, starih od 1 do 12 let. Opredelili smo nivoje miRNA v urinu preiskovancev s tumorjem pred operacijo, po operaciji in pri zdravih preiskovancih, ter primerjali razlike v nivojih miRNA med različnimi skupinami. V vzorcih urina preiskovancev s tumorjem smo zaznali spremenjen nivo miR-16-5p, miR-181b-5p, miR-20a-5p, miR-196-5p in miR-184, za katere predhodne raziskave navajajo spremenjeno izražanje v tumorskem tkivu oziroma serumu bolnikov z Wilmsovim tumorjem. Izsledki naše raziskave kažejo na potencial določenih miRNA v urinu za zgodnje in neinvazivno odkrivanje bolezni. V prihodnje bodo potrebne dodatne raziskave, ki bodo na večjem številu vzorcev potrdile te ugotovitve in dodatno osvetlile vlogo različno izraženih miRNA pri nastanku in razvoju te bolezni ter njihovo morebitno diagnostično uporabnost.

• Language: Slovenian
• Keywords: Wilmsov tumor, mikro RNA, spremembe nivojev miRNA, sekvenciranje naslednje generacije
• Work type: Master's thesis/paper
• Organization: FFA - Faculty of Pharmacy
• Year: 2025
• PID: 20.500.12556/RUL-171881
• Publication date in RUL: 04.09.2025

Secondary language

• Language: English
• Title: Urinary micro RNA determination in nephroblastoma patients

Abstract

Wilms tumor or nephroblastoma is the most common renal malignancy in the pediatric population. In most cases, tumor development is the result of pathological variants of genes involved in the process of cell differentiation or post-transcriptional regulation of genes, for example in the genes CTNNB1, WTX, TP53, MYCN, SIX1, SIX2, AMER1, DROSHA, DICER. 1, DGCR8, XPO5 and TARBP2. In some instances, WT is linked to several congenital abnormalities or syndromes, such as Beckwith-Wiedemann, Denys-Drash, WAGR, and Perlman Syndrome, as well as hereditary genetic changes in genes that elevate the risk of developing cancer. miRNA molecules, which are small, noncoding RNAs, play a crucial role in the post-transcriptional regulation of gene expression and are significant in the development of various types of cancerous diseases. In recent years, studies have focused on identifying differentially expressed miRNAs and their use in diagnosis and treatment of cancer. The master's thesis aimed to define the miRNA expression profile in the urine of patients with Wilms tumor using next-generation sequencing and to identify miRNAs that could serve as potential biomarkers for non-invasive early disease detection and monitoring of treatment efficacy. We included samples from 18 children aged from 1 to 12, 9 of them diagnosed with Wilms tumor, and 9 healhy individuals. We analysed the expression of miRNAs in the urine of patients with tumor before surgery, after surgery and in healthy individuals, and compared the differences in miRNA expression between the different groups. In the urine samples of patients with tumors, we detected altered expression of certain miRNAs that have been reported in studies to show differential expression in tumor tissue or serum of patients with WT. These include miR-16-5p, miR-181b-5p, miR-20a-5p, miR-196-5p, and miR-184. In the future, further studies are required to confirm our findings in a larger number of urine samples and define the role of the most frequently dysregulated miRNAs in the development and progression of Wilms tumor, as well as their potential use in diagnostics.

• Keywords: Wilms tumor, micro RNA, alteration of miRNA expression, next-generation sequencing