Kamata, S.; Taguchi, A.; Iuchi, H.; Ikeda, Y.; Maruyama, R.; Nakanishi, Y.; Sugi, T.; Okuma, Y.; Kobayashi, O.; Tomita, N.; Yoshimoto, D.; Wang, L.; Moritsugu, N.; Takahashi, C.; Tagami, M.; Matsunaga, H.; Okayama, T.; Manabe, R.-i.; Kiyotani, K.; Ikeo, K.; Okazaki, Y.; Kiyono, T.; Masuda, S.; Hamada, M.; Takeyama, H.; Kawana, K.

Abstract Human papillomavirus 18 (HPV18) preferentially infects cervical stem cell-like cells and is strongly associated with adenocarcinoma. However, the mechanisms underlying differentiation into cervical adenocarcinoma remain unclear due to the lack of appropriate experimental models. We aimed to establish a model of HPV18-associated cervical adenocarcinoma and elucidate its molecular and cellular differentiation mechanisms. HPV18 E6/E7 were introduced into induced pluripotent stem cell-derived reserve cell-like cells (iRCs) to generate tumor models. Spatial transcriptomics and single-cell multi-omics analyses were performed to integrate histological and molecular data. A distinct component (Gland_A) exhibited morphological and immunohistochemical features of cervical adenocarcinoma and was efficiently induced in iRC-18 tumors. Gland_A showed increased chromatin accessibility and elevated expression of FOXA1, FOXA2, and ALDH1A1. Analysis of clinical samples confirmed enrichment of ALDH1A1 in HPV-associated adenocarcinomas. This model recapitulates key features of HPV18-associated cervical adenocarcinoma and provides insights into its differentiation mechanisms.