Clinical trial NCT04687267: Conservative Management of CIN2 Lesions and Biomarkers Evaluation
Official title: Gestione Conservativa di Lesioni CIN2 e Valutazione di Biomarcatori Indicativi di Regressione
Study type: OBSERVATIONAL
Overall status: COMPLETED
Conditions: CIN2
Keywords: biomarkers
HPV
methylation
clinical outcome
Lead sponsor: Istituto Oncologico Veneto IRCCS
Sponsor class: OTHER
Collaborators: name: Azienda ULSS 3 Serenissima
name: Azienda Ulss 6 EUGANEA
name: Azienda Ulss 2 Marca Trevigiana
name: Azienda Ulss 9 Scaligera
name: Regione Veneto
First posted: 2020-12-29
Enrollment: 319
Sex: FEMALE
Minimum age (years): 25
Maximum age (years): 45
Standard ages: ADULT
Accepts healthy volunteers: no
Observational model: COHORT
Time perspective: PROSPECTIVE
Sampling method: NON_PROBABILITY_SAMPLE
Study population: Women attending organized population-based cervical cancer screening with a histological CIN2 lesion, meeting the selection criteria and consenting to participate.

Brief summary: Prospective study including women aged 25-45 years, adherent to the cervical screening program of four different centers of the Veneto region, with a diagnosis of CIN2 lesion. After enrollment according to predefined criteria, and informed consent to participate, the CIN2 lesions are managed by follow-up; cases with progressive lesions will be treated immediately, cases with CIN2 persistence for more than 12 months will be treated as well. Viral, molecular and immunocytochemical biomarkers will be studied, and evaluated in relation to the clinical outcome.

Detailed description: Women aged 25-45 years, adherent to the organized population-based cervical screening program, with a histological diagnosis of CIN2 and fulfilling the inclusion criteria will be invited to participate to the study, previously providing specific information; in case of acceptance, informed consent is signed.

STUDY PROTOCOL:

The adherent women will attend periodical control visits:

* every 6 months up to 24 months, with performance of: pap test (PT) and colposcopy (with biopsy in case of visible alterations);
* at 6 and 12 months control visit: a liquid-based sample of cervical cells will be collected for the biomarkers' analyses.

BIOMARKERS:

1. - HPV search and partial HPV16/18 genotyping, by cobas 4800 high-risk HPV assay (Roche); PCR with MY09/MY11 consensus primers and full genotyping by restriction fragment length analysis, plus PCR with beta-globin primers (in-house);
2. - methylation analysis of the cellular genes FAM194A and hsa-mir124-2, by methylation-specific quantitative PCR test (qMSP - QIAsure methylation test, Qiagen);
3. - methylation analysis of the L1 and L2 viral genes of HPV types 16 and 18, by pyrosequencing;
4. - immunocytochemical analysis for p16INK4A/Ki67 proteins (dual stain), by p16INK4A/Ki67 immunocytochemical analysis by CINtec Plus kit (Roche).

Eligibility criteria: Inclusion Criteria:

* age range 25-45 years,
* CIN2 lesions located in the exocervix and completely visible at colposcopy.

Exclusion Criteria:

* age \>45 years;
* history of previous high-grade lesions;
* squamo-columnar junction not completely visible (type 3);
* cytology with suspect or indicative for invasive lesion;
* lesions exclusively located in the endocervix;
* lesions located in the exocervix but not completely visible;
* pregnancy

Primary outcomes:
measure: Rate of spontaneous regression of CIN2 lesions
timeFrame: Through study completion, an average of 2 years.
measure: CIN2 clinical outcome by HPV genotype
timeFrame: Through study completion, an average of 2 years.
measure: CIN2 clinical outcome by DNA methylation
timeFrame: Through study completion, an average of 2 years.
measure: CIN2 clinical outcome by p16/ki67 protein expression
timeFrame: Through study completion, an average of 2 years.
Secondary outcomes:
measure: Adhesion to CIN2 conservative management.
timeFrame: 2 years
Other outcomes:
Locations:
city: Padova
country: Italy
facility: Azienda ULSS 6 Euganea
state: PD
city: Treviso
country: Italy
facility: Azienda ULSS 2 Marca Trevigiana
state: TV
city: Mestre
country: Italy
facility: Azienda ULSS 3 Serenissima
state: VE
city: Verona
country: Italy
facility: Azienda ULSS 9 Scaligera
state: VR
Study officials:
affiliation: Azienda ULSS 3 Serenissima
name: Tiziano Maggino, MD
role: PRINCIPAL_INVESTIGATOR
References:
citation: Del Mistro A, Matteucci M, Insacco EA, Onnis G, Da Re F, Baboci L, Zorzi M, Minucci D. Long-Term Clinical Outcome after Treatment for High-Grade Cervical Lesions: A Retrospective Monoinstitutional Cohort Study. Biomed Res Int. 2015;2015:984528. doi: 10.1155/2015/984528. Epub 2015 Jun 9.
citation: Del Mistro A, Frayle H, Rizzi M, Fantin G, Ferro A, Angeletti PM, Giorgi Rossi P, Altobelli E. Methylation analysis and HPV genotyping of self-collected cervical samples from women not responding to screening invitation and review of the literature. PLoS One. 2017 Mar 6;12(3):e0172226. doi: 10.1371/journal.pone.0172226. eCollection 2017.
citation: Ordi J, Sagasta A, Munmany M, Rodriguez-Carunchio L, Torne A, del Pino M. Usefulness of p16/Ki67 immunostaining in the triage of women referred to colposcopy. Cancer Cytopathol. 2014 Mar;122(3):227-35. doi: 10.1002/cncy.21366.
citation: Gillio-Tos A, Fiano V, Grasso C, Trevisan M, Gori S, Mongia A, De Marco L, Ronco G; New Technologies for Cervical Cancer Screening (NTCC) Working Group. Assessment of viral methylation levels for high risk HPV types by newly designed consensus primers PCR and pyrosequencing. PLoS One. 2018 Mar 26;13(3):e0194619. doi: 10.1371/journal.pone.0194619. eCollection 2018.