Metilacija nestaničnoga DNA gena RASSF1A u krvi i ejakulatu kao epigenetički biomarker u pacijenata sa seminomom testisa

doctoral thesis

• Title (english): Methylation of cell-free DNA of the RASSF1A gene in blood and semen as an epigenetic biomarker in patients with testicular seminoma
• Author: Dora Raos
• Mentor: Davor Ježek (mentor)
• Mentor: Nino Sinčić (komentor)
• Committee member: Floriana Jakuš (predsjednik povjerenstva)
• Committee member: Ana Katušić Bojanac (član povjerenstva)
• Committee member: Monika Ulamec (član povjerenstva)
• Committee member: Božo Krušlin (član povjerenstva)
• Committee member: Nikola Knežević (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; Zagreb
• Defense date and country: 2022-01-12, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Urology
• Universal decimal classification (UDC): 616 - Pathology. Clinical medicine

Abstract

Seminomi (SE) su tumori testisa koji pogađaju mladu mušku populaciju. Dijagnoza i liječenje SE predstavljaju invazivne postupake. Serumski biomarkeri koriste se u dijagnostici SE, međutim imaju ograničenu specifičnost i osjetljivost. Slobodni nestanični DNA (cfDNA) iz tjelesnih tekućina predstavlja neinvazivni postupak za probir i praćenje SE pacijenata. Cilj rada je utvrditi metilaciju cfDNA gena RASSF1A u krvi i ejakulatu bolesnika sa SE te njezin potencijal kao biomarkera. Pirosekvenciranjem, istražili smo metilacijski obrazac gena supresor tumora RASSF1A u cfDNA iz krvi i ejakulata pacijenata sa SE jer je njegova aberantna metilacija već uočena kod genomskoga DNA (gDNA) iz SE tkiva. RASSF1A u gDNA iz SE bio je statistički značajno hipometiliran u odnosu na okolno tkivo. Hipometilirani RASSF1A detektiran je i u predoperativnim uzorcima krvi što znači da je metilacija RASSF1A iz gDNA tumora odražena u cfDNA iz krvi. Međutim, u predoperativnim uzorcima ejakulata RASSF1A je bio hipermetiliran. Iz toga slijedi da krv predstavlja bolji izvor SE biomarkera od ejakulata. Analizom metilacije svakog pojedinom CpG dinukleotida zaključeno je da status prosječne metilacije RASSF1A ima veću statističku značajnost od metilacije RASSF1A na pojedinim CpG dinukleotidima. Ovim istraživanjem dobiveni su vrijedni rezultati u kontekstu tekućinskih biopsija kao izvora za probir i praćenje SE pacijenata.

Abstract (english)

Seminoma (SE) is the most frequent type of testicular tumour, affecting predominantly young men. Diagnosis and treatment of SE represent invasive procedures. In SE diagnostics, serum biomarkers represent a valuable tool. However, they have limited specificity and sensitivity. Cell-free DNA (cfDNA) from liquid biopsies represents a non-invasive procedure for SE screening and monitoring. The aim of this study was to determine the methylation of gene RASSF1A in cfDNA from blood and ejaculate of SE patients, as well as its potential as an SE biomarker. By pyrosequencing, we investigated the methylation of the RASSF1A tumor suppressor gene in cfDNA from the blood and ejaculate of SE patients, as well as in gDNA from SE tissue. RASSF1A in gDNA from SE was significantly hypomethylated regarding surrounding tissue. Hypomethylated RASSF1A was also detected in preoperative blood samples, while in preoperative ejaculate samples RASSF1A was hypermethylated. Analysis of the methylation of each CpG dinucleotide concluded that the status of the average methylation of RASSF1A has a higher statistical significance than the methylation of RASSF1A of individual CpG dinucleotides. This research yielded valuable results in the context of liquid biopsies as a source for screening and monitoring SE patients

• Keywords: Seminom; Tumor testisa; Geni; Ejakulat; Biomarkeri
• Keywords (english): Seminom; Testicular tumor; Genes; Ejaculate; Biomarkers
• Language: croatian
• URN:NBN: urn:nbn:hr:105:812948
• Project: Number: IP-2016-06-3692; Title: Epigenetički biomarkeri u krvi i ejakulatu bolesnika sa seminomom testisa; Acronym: epiSem; Leader: Davor Ježek; Jurisdiction: Croatia; Funder: HRZZ; Funding stream: IP
• Project: Number: KK.01.1.1.01.0008; Title: Regenerativna i reproduktivna medicina - istraživanje novih platformi i potencijala; Leader: Slobodan Vukičević; Jurisdiction: eu; Funder: EK
• 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: 90 str.
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-04-14 09:26:51