Metilacija nestaničnoga DNA gena OCT3/4 i NANOG u ejakulatu bolesnika sa seminomom testisa

doctoral thesis

• Title (english): Cell Free DNA Methylation of OCT3/4 and NANOG Gene in Semen of Patients with Testicular Seminoma
• Author: Miroslav Tomić
• Mentor: Božo Krušlin (mentor)
• Mentor: Nino Sinčić (komentor)
• Committee member: Ana Katušić Bojanac (predsjednik povjerenstva)
• Committee member: Borislav Spajić (član povjerenstva)
• Committee member: Marijana Ćorić (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; Zagreb
• Defense date and country: 2022-09-09, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Clinical Cytology
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Urology
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences; Pathology
• Universal decimal classification (UDC): 616 - Pathology. Clinical medicine

Abstract

Tumori testisa iako rijetki tumori pogađaju mladu mušku populaciju u najproduktivnijoj dobi. Seminomi čine otprilike 50% svih dijagnosticiranih tumora testisa. Dijagnoza, terapija i praćenje uključuje invazivne postupke koji utječu na kvalitetu života svakog pacijenta. Klasični tumorski biomarkeri iz krvi imaju svoje mjesto u dijagnostici i praćenju pacijenata sa seminomom ali imaju i svoja ograničenja u senzitivnosti kao i specifičnosti.
Ejakulat, kao predstavnik tekućinskih biopsija, predstavlja neinvazivni izvor informacija za pacijente sa seminomom a izolacija slobodne nestanične DNA (cfDNA) iz ejakulata pacijenata sa seminomom potencijalni izvor biomarkera za probir i praćenje pacijenata sa seminomom.
U ovom istraživanju analizirali smo obrazac metilacije gena OCT3/4 i NANOG u ejakulatu pacijenata sa seminomom, preoperativno i postoperativno. Također analizirali smo i proteinsku ekspresiju istih gena u tkivu seminoma, GCNIS i okolnom tkivu sa očuvanom spermatogenezom.
Uočena je statistički značajna razlika proteinske ekspresije gena OCT3/4 i NANOG i GCNIS u odnosu na okolno tkivo s očuvanom spermatogenezom. Nismo dokazali statistički značajnu razliku u metilaciji gena OCT3/4 i NANOG između preoperativnih i postoperativnih ejakulata pacijenata sa seminomom testisa kao niti statistički značajnu razliku u odnosima korelacije između proteinske ekspresije i metilacije cfDNA gena.
Koliko je nama poznato, ovo je prvo istraživanje metilacije cfDNA OCT3/4 i NANOG gena iz ejakulata kod pacijenata sa seminomom testisa i može poslužiti kao važan izvor informacija za daljnja istraživanja ejakulata kao tekućinske biopsije za probir i praćenje pacijenata sa tumorima testisa.

Abstract (english)

Testicular tumors although rare tumors affect the young male population at the most productive age. Seminoma make up 50% of all diagnosed testicular tumors. Diagnosis, therapy and follow-up include invasive procedures that affect the quality of life of each patient. Classical tumor markers from the blood have their place in the diagnosis and monitoring of patients with seminoma, but they also have their limitations in sensitivity as well as specificity. Ejaculate, as a representative of liquid biopsies, is a non-invasive source of information for patients with seminoma and the isolation of cell free DNA (cfDNA) from the ejaculate of patients with seminoma is a potential source of markers for screening and monitoring patients with seminoma. In this study, we analyzed the methylation pattern of the OCT3/4 and NANOG genes in the ejaculate of patients with seminoma, preoperatively and postoperatively. We also analyzed the protein expression of the same genes in seminoma, GCNIS tissue and surrounding tissue with preserved spermatogenesis. A statistically significant difference in protein expression of OCT3/4 and NANOG and GCNIS was observed in relation to the surrounding tissue with preserved spermatogenesis. We did not prove a statistically significant difference in the methylation of the OCT3/4 and NANOG genes between preoperative and postoperative ejaculates of patients with seminoma, nor a statistically significant difference in the correlation relations between protein expression and methylation of cfDNA genes of the OCT3/4 and NANOG.
To the best of our knowledge, this is the first study of cfDNA methylation OCT3/4 and NANOG gene in ejaculate of patients with testicular seminoma and may serve as an important source of information for further ejaculate research as an ongoing biopsy for screening and monitoring of patients with testicular tumors.

• Keywords: Seminoma; Metilacija; Neoplazme testisa; Biomarkeri; tumor; Spermatogeneza; Nukleinske kiseline bez stanica; Biopsija
• Keywords (english): Seminoma; Methylation; Testicular Neoplasms; Biomarkers; Tumor; Spermatogenesis; Cell-Free Nucleic Acids; Biopsy
• Language: croatian
• URN:NBN: urn:nbn:hr:105:905393
• Project: Number: KK.01.1.1.01.0008; Title: Znanstveni centar izvrsnosti za reproduktivnu i regenerativnu medicinu – istraživanja novih platormi i potencijala; Leader: Slobodan Vukičević; Jurisdiction: eu; Funder: EK
• 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
• 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: 78 str.
• File origin: Born digital
• Access conditions: Open access
• Created on: 2022-10-25 07:53:38