Conservative Management of CIN2 Lesions and Biomarkers Evaluation

Gestione Conservativa di Lesioni CIN2 e Valutazione di Biomarcatori Indicativi di Regressione

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.

Study Overview

• Status: Completed
• Conditions: CIN2

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).

• Study Type: Observational
• Enrollment (Actual): 319

Contacts and Locations

Study Locations

• Italy
  • PD
    • Padova, PD, Italy
      • Azienda ULSS 6 Euganea
  • TV
    • Treviso, TV, Italy
      • Azienda ULSS 2 Marca Trevigiana
  • VE
    • Mestre, VE, Italy
      • Azienda Ulss 3 Serenissima
  • VR
    • Verona, VR, Italy
      • Azienda ULSS 9 Scaligera

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 23 years to 43 years (Adult)
• Accepts Healthy Volunteers: No

• 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.

Description

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

Study Plan

How is the study designed?

Design Details

• Observational Models: Cohort
• Time Perspectives: Prospective

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Rate of spontaneous regression of CIN2 lesions	Eligible women will not be treated at diagnosis, but periodically followed-up. Treatment will be provided for progressive lesions and lesions persisting more than 12 months. The rates of lesion regression will be calculated: number of lesions regressed to CIN1 or normal / total number of cases.	Through study completion, an average of 2 years.
CIN2 clinical outcome by HPV genotype	Rate of CIN2 regression will be calculated in relation to positivity for HPV16 vs positivity for other high-risk types (number of regressed HPV16-related CIN2 lesions / total number of HPV16-related CIN2 vs number of regressed non-HPV16-related CIN2 lesions / total number of non-HPV16-related CIN2 lesions).	Through study completion, an average of 2 years.
CIN2 clinical outcome by DNA methylation	Rate of CIN2 regression will be calculated in relation to DNA methylation of cellular and viral genes (number of regressed hypermethylated CIN2 lesions / total number of CIN2 lesions with valid result for each gene analyzed).	Through study completion, an average of 2 years.
CIN2 clinical outcome by p16/ki67 protein expression	Rate of CIN2 regression will be calculated in relation to positivity for p16/ki67 expression (number of regressed p16/ki67-positive CIN2 lesions / total number of CIN2 lesions with valid result for p16/ki67 expression).	Through study completion, an average of 2 years.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Adhesion to CIN2 conservative management.	The rate of eligible women consenting to participate to the study will be calculated (enrolled women / eligible women)	2 years

Collaborators and Investigators

• Sponsor: Istituto Oncologico Veneto IRCCS
• Collaborators: Azienda Ulss 2 Marca Trevigiana; Azienda ULSS 3 Serenissima; Regione Veneto; Azienda Ulss 6 EUGANEA; Azienda Ulss 9 Scaligera
• Investigators: Principal Investigator: Tiziano Maggino, MD, Azienda Ulss 3 Serenissima

Publications and helpful links

General Publications

• 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.
• 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.
• 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.
• 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.

Study record dates

Study Major Dates

• Study Start (Actual): April 15, 2019
• Primary Completion (Actual): October 31, 2021
• Study Completion (Actual): December 31, 2022

Study Registration Dates

• First Submitted: December 10, 2020
• First Submitted That Met QC Criteria: December 23, 2020
• First Posted (Actual): December 29, 2020

Study Record Updates

• Last Update Posted (Actual): September 28, 2023
• Last Update Submitted That Met QC Criteria: September 27, 2023
• Last Verified: September 1, 2023

More Information

Terms related to this study

• Keywords: biomarkers; HPV; clinical outcome; methylation
• Other Study ID Numbers: CIN2-RSFR

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No