Olwal, C. O.; Rathore, U.; Makanani, S.; Kaushal, P.; Ashley, I. A.; Ummadi, M. R.; Appiah, V.; Djomkam Zune, A. L.; Blanc, S.; Winters, D.; Delgado, Y.; Muthoka, K.; Fabius, J. M.; Eckhardt, M.; Kaake, R. M.; Su, M.; Fregoso, O. I.; Hultquist, J. F.; Orango, E. O.; Swaney, D. L.; Kyei, G. B.; Krogan, N. J.; Quashie, P. K.; Bediako, Y.; Bouhaddou, M.

Persistent infection with human papillomavirus (HPV) is the primary cause of cervical cancer worldwide. Notably, women co-infected with HPV and human immunodeficiency virus type 1 (HIV-1) have a six-fold higher lifetime risk of developing cervical cancer compared to those without HIV, even when adhering to antiretroviral therapy (ART) and achieving T-cell reconstitution. While chronic HIV-1 infection is known to cause inflammation, how paracrine signals from immune cells alter signaling in cervical cells remain poorly understood. To address this, we conducted global transcriptomics analysis on cervical swabs from Kenyan women with HPV, stratified by HIV-1 and cancer status. Strikingly, women with HIV-1 showed cancer-like gene expression patterns in non-cancerous cervical epithelial cells. Complementary global mass spectrometry (MS) proteomics of cervical cells exposed to the secretome of HIV-1 infected primary CD4+ T-cells revealed altered expression of proteins in MAPK, PI3K-AKT, and beta-catenin signaling pathways. Integrative network analyses of transcriptomic and proteomic datasets revealed that HIV-1 altered gene expression in key pathways known to drive cervical cancer, including genes commonly mutated in patients without HIV-1. Notably, IRS-1, a key PI3K-AKT pathway activator, was found to be consistently upregulated in both participant samples and cell culture models, as were interferon-stimulated genes. Phosphoproteomics MS analysis confirmed PI3K-AKT pathway activation in cervical cells exposed to conditioned media from HIV-1-infected T-cells. Together, our findings uncover how HIV-1 reshapes cervical cell signaling via paracrine mechanisms and highlights the PI3K pathway as a potential therapeutic target in HIV-associated cervical cancer.