Molecular markers for diagnosis and prognosis in cervical neoplasia

For a long time practice in detection, diagnosis and therapy of cervical neoplasia has been based on classic clinicopathologic parameters such as cytomorphology of cervical smears, histology of cervical biopsies and/or radical hysterectomy specimens, fi ndings at rectovaginal examinati- on etc. In contrast to malignancies such as breast cancer, where molecular markers such as estrogen receptor expression and c-erbB-2 amplifi cati on have greatly impacted on clinical practice, comparable molecular markers have not been implemented in screening, diagnosis and treatment of cervical neoplasia. High risk HPV (hr-HPV) DNA detecti on in cervical smears for populati onbased screening might be regarded as the excepti on to this rule, but its implementati on is still a matt er of debate. Aim of this thesis was to explore possible opportuniti es of a variety of molecular markers for implementati on in screening, diagnosis and treatment of cervical neoplasia. Populati on-based screening has reduced the incidence of cervical cancer signifi cantly in the Western world. However, both the methodology for primary screening as well as the diagnostic work-up, treatment and follow-up of pati ents with (pre)malignant cervical neoplasia are characterized by signifi cant fl aws, that leave ample room for improvement. A primary screening test with a higher sensiti vity and specifi-city than the currently used Pap smear e.g. primary hr- HPV testi ng in combinati on with a triage test for hr-HPV positi ve women may increase efficacy of current populati on-based screening for cervical cancer. A test based on detecti- on of DNA methylati on appears to be an att racti ve tool to use for cervical neoplasia screening, since gene promoter methylati on is an early event in cervical carcinogenesis. So far, no methylation markers however have been shown to be useful for cervical neoplasia screening, especially due to low sensiti vity for high-grade premalignant lesions. Recently, we reported 13 possible cervical cancer specifi c methylated biomarkers identi fi ed by pharmacological unmasking of cervical cancer cell lines as determined by global gene expression microarrays in combinati on with large-genome computati onal screening [1]. In chapter 2 of this thesis an in-depth analysis of the methylati on patt erns of these 13 candidate genes in cervical neoplasia was performed and their diagnostic relevance determined. Five of 13 gene promoters (C13ORF18, CCNA1, TFPI2, C1ORF166 and NPTX1) were found to be more frequently methylated in frozen cervical cancer compared to normal cervix specimens. Quanti tati ve methylati on analysis for these 5 markers revealed that CCNA1 and C13ORF18 methylati on were both present in 68/97 cervical scrapings from cervical cancer pati ents and in only 5 and 3 scrapings, respectively, from 103 healthy controls (p