The potential role of TP63 regulating ZC3H13-mediated HLA-A m6A methylation modification in HR-HPV persistent infection patients

Abstract Objective The aim of this study is to investigate the short-term persistent HPV infection and natural prognosis in patients with IA1 cervical cancer after CKC, and to explore the mechanism of persistent HPV infection leading to cervical lesions based on database analysis. Methods This is a prospective observation cohort study which enrolled the stage IA1 patients who select to receive CKC as the treatment modality in a single center from January 24, 2018, to June 9, 2022. The primary outcome was the persistent infection status and remession rates within two years after the CKC. In addition, the relevant mechanism was explored based on database analysis. The Cancer Genome Atlas (TCGA) database is the source of RNA sequencing data of cervical cancer patients. The gene Expression omnibus (GEO) database was used as the validation set to verify the expression of TP63 mRNA in the process from normal cervical to precancerous lesions, and the difference of TP63 between cancerous lesions and paracarcinoma was verified by Western blot. The limma package of R software, Kaplan-Meier survival curve and Log-rank test were used to screen the genes related to m6A methylation modification affecting the prognosis of cervical cancer. spearman correlation analysis was used to verify the correlation between genes, and Timer2.0 immune database was used to analyze the correlation between the expression level of key genes and the level of immune infiltration. JASPAR and SRAMP open access databases were used to verify the relevant RNA sequence binding sites and m6A methylation modification sites. Results A total of 98 eligible patients were included and the main types of HPV was as following: HPV 16 (76.19%,64/84), HPV 58 (10.71%,9/84), HPV 33 (9.52%,8/84), and unknown type 14.28% (14/98). The HR-HPV negative rates of the whole cohort at 3, 6, 9, 12, 15, 18, 21, and 24 months after CKC were 76% (19/25), 80.95% (34/42), 76.47%(39/51), 76.92%(40/52), 75.93%(41/54), 76.36%(42/55), 75.44%(43/57), and 74.58% (44/59), which was stable within 2 years after surgery. Based on the database analysis, this study proposes the relevant mechanism hypothesis of HPV persistent infection and difficult to clear: TP63 acts on the promoter of ZC3H13 to induce its expression and promote the m6A methylation modification of HLA-A mRNA 3’UTR, which leads to the accelerated degradation of HLA-A mRNA and further inhibits the antigen presentation of HPV viral proteins, leading to HPV escape from CD8 + T cell killing. Conclusion The negative rate of HR-HPV remained stable within 2 years after the CKC.Standardized follow-up after conization is very important for patients with stage IA1 cervical cancer.This study elucidates the mechanism of TP63-ZC3H13-HLA-A axis and provides A therapeutic target for HPV clearance and prevention of recurrence in patients with stage IA cervical cancer after conization.