Metilacija nestanične DNA gena RASSF1A i PRSS21 u krvi i ejakulatu bolesnika s neseminomskim tumorima zametnih stanica testisa

doctoral thesis

• Title (english): Cell Free DNA Methylation of RASSF1A and PRSS21 Genes in Blood and Semen of Patients with Nonseminomatous Testicular Germ Cell Tumors
• Author: Jure Krasić
• Mentor: Nino Sinčić (mentor)
• Mentor: Tomislav Kuliš (komentor)
• Committee member: Floriana Jakuš (predsjednik povjerenstva)
• Committee member: Davor Ježek (član povjerenstva)
• Committee member: Monika Ulamec (član povjerenstva)
• Committee member: Ana Katušić Bojanac (član povjerenstva)
• Committee member: Frane Paić (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; Zagreb
• Defense date and country: 2022-10-17, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Basic Medical Sciences; Cytology, Histology and Embryology
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Urology
• Universal decimal classification (UDC): 616 - Pathology. Clinical medicine

Abstract

Tumori zametnih stanica testisa (TZST) su najčešća neoplazija koja pogađa mlađu mušku populaciju. Dijele se na seminome i neseminome (NSE). Potvrda dijagnoze TZST neizostavno uključuje radikalnu orhidektomiju te je pouzdana neinvazivna dijagnostička metoda potrebna za raniju dijagnostiku. Analiza slobodnocirkulirajuće nestanične DNA (cfDNA) iz tjelesnih tekućina je perspektivan neinvazivan postupak za dijagnozu onkoloških pacijenata. Metilacija genomske DNA (gDNA) gena RASSF1A i PRSS21 iz tkiva TZST-a se pokazala perspektivnim biomarkerom. Cilj rada je utvrditi potencijal metilacije cfDNA gena RASSF1A i PRSS21 iz krvi i ejakulata kao biomarkera za identifikaciju pacijenata s NSE. Pirosekvenciranjem je istražena stopa metilacije cfDNA promotorske regije gena RASSF1A i PRSS21 iz krvi i ejakulata pacijenata s potvrđenom dijagnozom NSE i zdravih dobrovoljaca bez prijašnje dijagnoze TZST-a te gDNA iz tumorskog tkiva i okolnog zdravog tkiva pacijenata s NSE. Hipermetilirana gDNA RASSF1A bila je detektirana u tkivu NSE naspram okolnog zdravog tkiva. U gena PRSS21 nije detektirana razlika u stupnju metilacije gDNA. Kod pacijenata s NSE detektirana je hipermetilacija cfDNA gena RASSF1A u krvi, dok je u ejakulatu detektirana hipermetilacija cfDNA gena PRSS21. Metilacija cfDNA gena RASSF1A i PRSS21 je pokazala višu osjetljivost i specifičnost od biomarkera koji su trenutno u kliničkoj upotrebi. Ovim istraživanjem je potvrđen potencijal metilacije cfDNA gena RASSF1A u krvi te je otkriven potencijal metilacije cfDNA gena PRSS21 u ejakulatu kao dijagnostičkih biomarkera pacijenata s NSE.

Abstract (english)

Testicular germ cell tumors (TGCT) are the most common malignancy among young males. TGCT are subdivided into seminomas and nonseminomas (NSE). Confirmation of a TGCT diagnosis always includes radical orchidectomy, making a reliable noninvasive diagnostic method required for earlier diagnosis. Analysis of circulating cell-free DNA (cfDNA) from body liquids is a perspective noninvasive procedure for diagnosis of oncological patients. Genomic DNA methylation (gDNA) of RASSF1A and PRSS21 genes in the tissue of TGCT has been shown as a potential biomarker. The aim of this study was to assess the potential of cfDNA methylation of genes RASSF1A and PRSS21 in the blood and ejaculate as biomarkers for patients with NSE. Pyrosequencing was used to analyze cfDNA methylation of genes RASSF1A and PRSS21 in blood and ejaculate of patients with a confirmed diagnosis of NSE and healthy donors with no prior diagnosis of TGCT as well as in gDNA from the tumor tissue and the surrounding healthy tissue of patients with NSE. RASSF1A was hypermethylated in gDNA of tumor tissue. No difference in gDNA methylation levels was found in PRSS21. In NSE patients hypermethylated RASSF1A was detected in cfDNA isolated from the blood, while hypermethylated PRSS21 was detected in cfDNA isolated from the ejaculate. Methylation of RASSF1A and PRSS21 cfDNA shows higher sensitivity and specificity than the currently used clinical biomarkers. This study confirms the potential of methylation of RASSF1A in cfDNA from blood and discovers the methylation of PRSS21 in cfDNA from the ejaculate as diagnostic biomarkers of patients with NSE.

• Keywords: Tumor zametnih stanica testisa; DNK metilacija; Orhiektomija; Seminoma; Biomarkeri; Genomika
• Keywords (english): Testicular Germ Cell Tumor; DNA Methylation; Orchiectomy; Seminoma; Biomarkers; Genomics
• Language: croatian
• URN:NBN: urn:nbn:hr:105:873523
• Project: Number: KK.01.1.1.01.0008; Title: Reproduktivna i regenerativna medicina - istraživanje novih platformi i potencijala; Leader: Ana Katušić Bojanac; Jurisdiction: eu; Funder: MZO
• Study programme: Title: Biomedicine and Health Sciences; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstvo (doktor/doktorica znanosti, područje biomedicine i zdravstvo)
• Type of resource: Text
• Extent: 81 str.
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2024-10-18
• Created on: 2022-11-25 12:39:59