The Role of FAM19A4 and Hsa-mir-124 Methylation in Predicting Prognosis of Untreated Cervical Intraepithelial Neoplasia 2 (CIN 2)

Sponsor

University Medical Centre Maribor (Other)

Overall Status

Recruiting

CT.gov ID

NCT05624827

Collaborator

(none)

100

Enrollment

Location

Arm

Anticipated Duration (Months)

3.1

Patients Per Site Per Month

Study Details

Study Description

Brief Summary

High-risk precancerous cervical lesions are divided into stage 2 and 3 cervical intraepithelial neoplasia (CIN 2 and 3). CIN 3 represents a direct pre-stage of invasive cancer, has a high rate of progression and a high degree of agreement with the final histological diagnosis. In CIN 2 lesions, the rate of agreement with the final histological diagnosis is lower and the rate of spontaneous regression is higher. Due to the higher rate of regression and possible complications after excisional treatment, conservative active monitoring can be considered in selected young CIN 2 patients. A recent meta-analysis reported a high rate of spontaneous clinical regression of CIN 2, particularly in women under 30 years old. There are currently no prospectively validated prognostic biomarkers to determine which CIN 2 will progress to higher grade and which will regress to lower grade of change. Recent research has studied HPV methylation and microbiome analysis as biomarkers. A number of studies have shown that host cell DNA methylation levels in cervical scrapes increase with underlying cervical disease severity and are highest in cervical cancer. DNA methylation involves the covalent binding of a methyl group to the 5´ position of a cytosine molecule in CpG dinucleotides. Besides global hypomethylation, the overall loss of methylation during carcinogenesis, resulting in chromosomal instability, and the silencing of tumour suppressor genes by local hypermethylation of CpG-rich promoter regions contribute to cancer development. Gene promoter methylation can be easily accessed by sensitive, quantitative methylation-specific PCR providing an objective test outcome. The aim of this study was to determine the effect of the methylation rate of two suppressor genes- FAM19A4 and hsa-mir-124 on the rate of CIN 2 regression, persistence or progression in women younger than 36 years (≤35 years old).

Condition or Disease	Intervention/Treatment	Phase
Cervical Intraepithelial Neoplasia Grade 2 DNA Methylation	Diagnostic Test: Testing DNA methylation test for predicting prognosis of untreated CIN 2	N/A

Study Design

Study Type:

Interventional

Anticipated Enrollment :

100 participants

Allocation:

N/A

Intervention Model:

Single Group Assignment

Masking:

None (Open Label)

Primary Purpose:

Diagnostic

Official Title:

The Role of FAM19A4 and Hsa-mir-124 Methylation in Predicting Prognosis of Untreated Cervical Intraepithelial Neoplasia 2 (CIN 2)

Actual Study Start Date :

Sep 1, 2021

Anticipated Primary Completion Date :

Sep 1, 2023

Anticipated Study Completion Date :

May 1, 2024

Arms and Interventions

Arm	Intervention/Treatment
Experimental: Women with CIN 2 and under 36 years old We will analyze patients under 36 years old (≤ 35) with histological confirmed CIN 2 lesions and without any additional risk factors.	Diagnostic Test: Testing DNA methylation test for predicting prognosis of untreated CIN 2 After being diagnosed with CIN 2, patients will first be contacted by telephone and invited to participate in the study. If patients agree to participate in the research, they will sign a consent to participate in the research. After that, we will perform a colposcopy and take a cervical swab for analysis with the QIAsure Methylation Test Kit (Qiagen, Gaithersburg, USA), which will determine the methylation of tumor suppressor genes FAM19A4 and has-mir-124. Patients will complete a questionnaire. The total duration of tracking in both groups will be two years. The QIAsure Methylation Test will be performed to analyze methylation. It is a methylation-specific PCR test that detects hypermethylation of the tumor promoter suppressor genes FAM19A4 and has-mir-124. The samples on which we will use this test are bisulfite-converted DNA obtained by triage test for high-risk HPV - Hybrid Capture 2 HPV DNA Test (hc2, Qiagen, Gaithersburg, USA).

Arm

Intervention/Treatment

Experimental: Women with CIN 2 and under 36 years old

We will analyze patients under 36 years old (≤ 35) with histological confirmed CIN 2 lesions and without any additional risk factors.

Diagnostic Test: Testing DNA methylation test for predicting prognosis of untreated CIN 2

After being diagnosed with CIN 2, patients will first be contacted by telephone and invited to participate in the study. If patients agree to participate in the research, they will sign a consent to participate in the research. After that, we will perform a colposcopy and take a cervical swab for analysis with the QIAsure Methylation Test Kit (Qiagen, Gaithersburg, USA), which will determine the methylation of tumor suppressor genes FAM19A4 and has-mir-124. Patients will complete a questionnaire. The total duration of tracking in both groups will be two years. The QIAsure Methylation Test will be performed to analyze methylation. It is a methylation-specific PCR test that detects hypermethylation of the tumor promoter suppressor genes FAM19A4 and has-mir-124. The samples on which we will use this test are bisulfite-converted DNA obtained by triage test for high-risk HPV - Hybrid Capture 2 HPV DNA Test (hc2, Qiagen, Gaithersburg, USA).

Outcome Measures

Primary Outcome Measures

Impact of the degree of methylation on progression of CIN 2 [two years]
The primary clinical outcome will be the effect of the degree of methylation on the progression of CIN 2 to CIN 3+ (CIN 3 and cervical cancer).

Secondary Outcome Measures

Rate of progression of CIN 2 [two years]
The secondary clinical outcome will be the rate of progression of CIN 2 to CIN 3+, the rate of clinical decline to normal histological picture (˂ CIN 1) and the rate of persistence of precancerous change after two years (CIN 2 or persistent CIN 1).

Eligibility Criteria

Criteria

Ages Eligible for Study:

20 Years to 36 Years

Sexes Eligible for Study:

Female

Accepts Healthy Volunteers:

Yes

Inclusion Criteria:

Histologically confirmed CIN 2 (with biopsy of colposcopically suspicious changes in the cervix)
Age under 30 years
Satisfactory colposcopy (transformation zone fully visible)
Size of change below 75% of transformation zone
The change in the ectocervix is fully visible
Age 30-35 years, if the patient is non-smoker and the change in the cervix does not exceed 50% of the area of the transformation zone
Signing an informed consent to participate in the survey
Willingness to perform inspections every 6 months

Exclusion Criteria:

Age 36 years or older
Unsatisfactory colposcopy (transformation zone not fully visible)
Size of change exceeds 75% of the transformation zone
The change in the ectocervix is not completely visible
Age 30-35 years for smokers or if the change exceeds 50% of the area of the transformation zone
Suspicted glandular precancerous changes
Histologically verified CIN 2 with cytological changes of glandular cells
Colposcopically suspected invasive disease
Histologically verified CIN 2 and histologically verified AIS
Histologically verified CIN 2 and histologically verified invasive cancer elsewhere in the cervix
Refusal to sign participation in the survey
Unwillingness to perform control examinations
Weakness of an immune system
Cervical conization performed in the past
Treatment with local immunomodulators