Gene methylation of CADM1 and MAL identified as a biomarker of high grade anal intraepithelial neoplasia

Samuel Phillips,Fengyi Jin,Monica Molano,Kahli Cassells,Dorothy A Machalek,Suzanne M Garland,Jennifer M Roberts,Sepehr N Tabrizi,Alyssa M Cornall,Richard J Hillman,I Mary Poynten,David J Templeton,Andrew E Grulich,Gerald L Murray

doi:10.1038/s41598-022-07258-5

Abstract

Human papillomavirus (HPV) is detected in up to 96% of anal squamous cell cancers, where screening programs needed. However, the best methodology is still undetermined. Host DNA methylation markers CADM1, MAL and miR124 have been identified in cervical disease, but not anal disease. Anal swabs varying by disease grade were assessed for DNA methylation of CADM1, MAL and miR124-2. Each marker was compared across disease grades, stratified by HPV and HIV status. Receiver operating characteristic curves identified the predictive value of significant gene candidates. CADM1 methylation was significantly higher in high-grade squamous intraepithelial lesions (HSIL) compared with low-grade (LSIL) (p = 0.005) or normal (p < 0.001) samples with 67.2% correctly identified as HSIL. MAL methylation was significantly (p = 0.002) increased in HSIL compared with LSIL in HIV positive participants with 79.8% correctly indicated as HSIL. Gene miR124-2, showed no difference between disease grades. Biomarkers with established diagnostic value in cervical disease have limited utility in the prediction of anal disease, with CADM1 identified as a marker with screening potential in a gay and bisexual men (GBM) population and MAL in HIV positive GBM population. New markers specific to the anal mucosa are required to improve triage of high-risk individuals.

Highlights

cell adhesion molecule 1 (CADM1) Cell adhesion molecule 1 MAL T-lymphocyte maturation associated protein miR124 Micro RNA 124 ACTB Beta actin gene cervical intraepithelial neoplasia (CIN) Cervical intraepithelial neoplasia
This study aimed to examine the performance of previously identified cervical cancer methylation markers (CADM1, MAL, and miR124-2) in anal high-grade squamous intraepithelial lesions (HSIL), using samples collected as part of the Study of the Prevention of Anal Cancer (SPANC)[5]
Among HIV positive individuals only, gene MAL had significantly less gene methylation in men whose highest grade of disease was LSIL compared with both normal histology and HSIL

Summary

Introduction

CADM1 Cell adhesion molecule 1 MAL T-lymphocyte maturation associated protein miR124 Micro RNA 124 ACTB Beta actin gene CIN Cervical intraepithelial neoplasia. Vaccination against high-risk HPV (HRHPV) genotypes that cause the majority of anal cancers, when administered prior to HPV exposure, are highly effective in reducing the incidence of the precursor lesion, anal high-grade squamous intraepithelial lesions (HSIL)[7]. Other high-risk populations with increased rates of ASCC include women and heterosexual men with HIV infection (12-fold higher), women with previous lower genital tract disease related to HPV, and transplant r ecipients[5,10–12]. A recent meta-analysis evaluated the performance of anal cytology in women and men and identified an overall sensitivity and specificity of atypical squamous cells of undetermined significance or higher positive cytology for the detection of AIN2 or higher at 77.3% and 55.5%, r espectively[14]. The application of HRHPV DNA detection as a primary test for anal screening in high-risk populations may be limited, due to the high prevalence of anal HRHPV and the presence of multiple HPV types, resulting in high sensitivity but poor s pecificity[14,17–19]

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