Lytic reactivation of EBV by HIV proteins and antiretroviral therapy with the heightened expression of MEF-2 oncogenes implicated in AIDS-related cancer

Despite effective antiretroviral therapy (ART), people living with HIV (PLWH) remain at a higher risk of developing AIDS-related cancers. The factors attributed to this increased risk are multifactorial, including long-term ART use, prolonged exposure to soluble HIV proteins, and drugs of abuse. Epstein-Barr virus (EBV) is a common driver of many malignancies, such as Burkitt’s lymphoma and Hodgkin’s lymphoma. While ART effectively suppresses HIV replication, it is not curative and prolonged use imposes health risks, including an accelerated course of cancers. Herein, we investigated the effects of various factors that can impact EBV latency in PLWH in cooperation with cellular oncoproteins. EBV negative and positive lymphoblastoid cell lines were stimulated with Biktarvy (a combination of Tenofovir, Emtricitabine, and Bictegravir), recombinant HIV proteins Nef and Rev, and/or drugs of abuse, including cocaine, METH, and morphine. We observed that simulation with Nef, Rev, or ART promoted the expression of EBV genes linked to lytic reactivation, such as BZLF1 and BMRF, as well as myocyte enhancer factor (MEF)-2 family oncogenes. MEF-2 observations were confirmed at the protein level and EBV gene changes were validated in samples from people with HIV/EBV. Enhancement of EBV lytic gene expression was confirmed by the DNA level changes and by assessing the protein products of BZLF1 and BMRF1. Drugs of abuse alone did not show any significant effects; however, METH combined with ART modestly promoted viral gene expression and suppressed anti-inflammatory cytokine production in a NF-KB dependent manner, a key signaling pathway for EBV reactivation. Indeed, NF-kB inhibition reversed the impact of HIV proteins and ART on EBV latent B cells. These findings establish a link between oncogenesis and persistent HIV and prolonged use of ART, highlighting the potential oncogenic risks for these HIV-associated factors.