Abstract 3021: Epigenetic signatures of virus-associated cervical cancer in women living with HIV
Authors: Zheng Yinan , Silas Olugbenga , Musa Jonah , Qu Yishu , Gao Tao , Kim Kyeezu , Joyce Brian , Wang Jun , Gursel Demirkan , Fatimah Abdulkareem , Imade Godwin , Akanmu Alani , Wei Jian-Jun , Kocherginsky Masha , Kim Kwang-Youn , Wehbe Firas , Achenbach Chad , Anorlu Rose , Simon Melissa , Sagay Atiene , Ogunsola Folasade , Murphy Robert , Hou Lifang 
Published: April 2023

Abstract
Background: Low- and middle-income countries are facing a high health burden of cervical cancer (CC). The situation is worsened by a high prevalence of human immunodeficiency virus (HIV). We aim to identify epigenetic signatures to help understand the virus-associated pathways underlying CC development, which is fundamental to developing effective CC screening tools or therapeutic approaches for women living with HIV.
Methods: We recruited a total of 365 Nigerian women with mean age of 52 (239 HIV+ and 126 HIV-; 98 without CC, 106 with cervical lesions, and 161 with CC). DNA methylation profiling in cervical tissue samples was performed using Illumina EPIC array covering over 860K methylation sites. The epigenetic signatures were identified among HIV+ women through epigenome-wide association study comparing CC vs. CC-free (Bonferroni adjusted p &lt;0.05), which were further validated by comparing: 1) pairs of tumor and adjacent normal samples; and 2) pairs of cervical lesions and adjacent normal samples. The comparisons were adjusted for age, BMI, education, employment, parity, study site, and technical batch variables. We then compared the identified signatures between HIV+ and HIV- among CC women to delineate the role of HIV. We constructed a methylation risk score (MRS) using the signatures, and we performed receiver operating characteristic (ROC) analysis to evaluate the performance of distinguishing across cancer statuses (CC, cervical lesions, CC-free).
Results: We identified 46 differentially methylated markers (p-value ranging from 5.7e-8 to 2.4e-19) in HIV+ CC women. The effect sizes among the 46 markers in the paired tumor-normal analysis and the paired lesion-normal analysis were highly correlated with the CC vs. CC-free analysis (r=0.99 and r=0.95, respectively) with smaller magnitudes (1.8 and 4.3 times smaller, respectively). Gene ontology and Reactome pathway enrichment analysis revealed that these 46 markers were enriched in genes involving activation of PI3K/AKT/mTOR signaling network (e.g., RPTOR, HDAC3, MAPKAP1), which plays a crucial role in virus/host crosstalk and virus-induced carcinogenesis. The PI3K/AKT/mTOR signaling cascade suppressors, including gene PRDM8, were silenced by promoter hypermethylation among HIV+ CC women (p=4.3e-12). These epigenetic changes, however, were not observed in CC women without HIV. In an independent dataset, MRS was the lowest in women without CC (2.3±2.1), followed by women with cervical lesions (4.9±1.1), and the highest in CC women (9.6±3.9) (p-trend &lt; 2e-16). The MRS achieved areas under the ROC curve = 0.93 and 0.88 in distinguishing CC and cervical lesions from CC-free among HIV+ women, respectively.
Conclusion: HIV may communicate with cervical cells and promote CC through epigenetic activation of PI3K/AKT/mTOR signaling pathway. Our epigenetic signatures may serve as novel biomarkers for CC early detection and treatment for women living with HIV.
Citation Format: Yinan Zheng, Olugbenga Silas, Jonah Musa, Yishu Qu, Tao Gao, Kyeezu Kim, Brian Joyce, Jun Wang, Demirkan Gursel, Abdulkareem Fatimah, Godwin Imade, Alani Akanmu, Jian-Jun Wei, Masha Kocherginsky, Kwang-Youn Kim, Firas Wehbe, Chad Achenbach, Rose Anorlu, Melissa Simon, Atiene Sagay, Folasade Ogunsola, Robert Murphy, Lifang Hou. Epigenetic signatures of virus-associated cervical cancer in women living with HIV [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3021.