Over 30,000 cases, the first prospective multi-center clinical study of methylation in China, CISCER® has good performance in cervical cancer triage

Preface

Cervical cancer (CC) is a major public health problem that seriously affects women's health worldwide and is the fourth leading cause of cancer death in women. With the advancement of vaccination, screening and treatment, the risk of cervical cancer has been significantly reduced in recent years. In the secondary prevention of cervical cancer, screening and triaging of abnormal screening results are increasingly important.

Current HPV screening strategies have limitations such as unnecessary follow-up, colposcopy referral and over-treatment due to transient HPV infection. The World Health Organization (WHO) recommends using HPV DNA testing combined with specific triage methods for primary screening of cervical cancer, and routine screening every 5-10 years. Compensating for the lack of low specificity of HPV testing, especially in non-HPV 16/18-positive patients, is through repeated cytology (LBC). Due to the limitations of cytology itself, it is necessary to explore better cervical cancer screening triage technologies and strategies.

Recently, a large-scale prospective multicenter clinical study led by Professor Li Lei of Peking Union Medical College Hospital and participated by Peking University International Hospital, Inner Mongolia Autonomous Region People's Hospital, Zhejiang University Second Hospital, Zhejiang University People's Hospital, Cangzhou Central Hospital and Beijing Origin Poly Bio-tec Co., Ltd(CISPOLY). was published in the journal Clinical Epigenetics (impact factor 4.8) - "Triage performance of PAX1m/JAM3m in opportunistic cervical cancer screening of non‒16/18 human papillomavirus‑positive women: a multicenter prospective study in China". The aim is to compare the accuracy of PAX1m/JAM3 methylation and traditional cytology in diagnosing CIN2+ and CIN3+ in non-HPV16/18 positive infection.

Research Methods

Sample source and flow chart: 30,084 women who underwent opportunistic screening for cervical cancer, 1,851 of whom had histopathology. The research flow chart is shown in Figure 1:

· Technical methods: All eligible participants underwent gynecological examination and collected cervical cytology. The remaining samples tested by hrHPV and cytology were used for PAX1/JAM3 gene methylation detection.

· Methylation result determination: According to the Ct values of PAX1 gene, JAM3 gene and internal reference gene (GAPDH), ΔCt was calculated (ΔCt=Ct detection gene-Ct internal reference gene). ΔCt PAX1 ≤ 6.6 or ΔCt JAM3 ≤10.0 was positive for methylation.

· Data analysis: The χ2 test was used for inter-group comparison. The receiver operating characteristic (ROC) curve was used to evaluate the AUCs of different detection methods in patients with < CIN3+ and CIN3+. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value of detecting CIN3+ were calculated with 95% confidence intervals. All tests were two-sided tests with P value < 0.05.

Study Results

1) PAX1/JAM3 gene methylation levels are significantly different in different levels of lesions (histopathology/cytopathology, as shown in Figure 2/3), and they increase with the increase in the severity of the lesions, showing its usefulness. Methylation has the ability to differentiate between lesions with different risk levels.

2) In the detection of CIN3+, the AUC area of PAX1m/JAM3 methylation was higher than that of cytology ≥ ASCUS (0.866 Vs 0.584, with a sensitivity and specificity of 84.8% and 88.5%, respectively, compared with 90.1% and 26.7% of cytology, indicating that methylation has a more balanced detection efficiency.

3) In the detection of CIN3+, cytological LBC ≥ASCUA or PAX1m/JAM3m (+) slightly increased the diagnostic sensitivity (98.0%) and referral rate (77.09%), but at the same time reduced the diagnostic specificity (24.8%). This suggests that methylation detection alone has the advantages of both high sensitivity and specificity.

4) Among all screening triage strategies, PAX1/JAM3 methylation will result in the lowest colposcopy referral (17.45%), compared to colposcopy referral due to cytology (74.66%), double gene methylation Significantly reduced the referral rate by 57.21%.

Conclusion and Outlook

The American Society for Colposcopy and Cervical Pathology (ASCCP), the Chinese Society for Colposcopy and Cervical Pathology (CSCCP) and other domestic and international guidelines recommend direct referral for colposcopy for HPV16/18 positives, while cytology is used for triage for non-16/18 positives. Individuals with LBC positive or ≥ASCUS are recommended to undergo colposcopy and pathological biopsy, while LBC negative patients are recommended to follow up. However, LBC results are easily affected by subjective factors, differences in the level of different pathologists, sampling, patient age and the lesion itself. Therefore, cervical cancer not infected with HPV 16/18 is the focus of future screening and treatment.

This study is the first and largest multicenter prospective trial in China on the efficacy of the new methylation technology in diversion. By comparing PAX1/JAM3 methylation with traditional cytology in non-16/18 positive women in the opportunistic screening population, it shows that methylation has better accuracy and more balanced detection efficacy. PAX1/JAM3 methylation for diversion will help find more patients with high-grade cervical lesions, and provides sufficient data support for its clinical application value and wide clinical application. It is a more promising new technology for cervical cancer diversion.

Clinical Epigenetics is an English academic journal focusing on the field of GENETICS & HEREDITY. It was founded in 2010 and published by BioMed Central Publisher with a publication cycle of 1 issue/year. The scope of this journal covers the fields of GENETICS & HEREDITY, and aims to timely, accurately and comprehensively report the experience, scientific research results, technological innovations, academic trends, etc. of GENETICS & HEREDITY workers at home and abroad in scientific research and other work in this field. The journal has been included in the SCIE database. In the latest upgraded version of the JCR partition table of the Chinese Academy of Sciences, the journal's partition information is the major discipline medicine zone 1, and the impact factor in 2023 is 4.8.