Multiomics analysis of benzodiazepine-mediated epigenetic reprogramming in HIV-1 infected hMDMs

About This Grant

Despite the success of antiretroviral therapy (ART), HIV remains incurable due to the persistence of viral reservoirs, particularly in the central nervous system (CNS). Myeloid cells—macrophages and microglia— serve as long-lived HIV reservoirs in the CNS and are implicated in neuroinflammation and HIV-associated neurocognitive disorders (HAND). Unlike CD4+ T cells, infected myeloid cells exhibit a state we define as semi‗quiescence, in which transcription from the HIV-1 promoter persists, but viral protein production is minimal. This persistent yet attenuated activity complicates efforts to eradicate the virus from the CNS. Our preliminary data show that ART-treated human monocyte-derived macrophages (hMDMs) maintain stable proviral levels over time, with reduced p24 Gag protein expression but active transcription. Chromatin immunoprecipitation (ChIP-qPCR) analysis of these cells revealed euchromatin markers at the viral LTR, consistent with transcriptionally active but translationally restricted infection. Strikingly, treatment with benzodiazepines (BDZs)—commonly prescribed to people living with HIV—reactivates viral protein production in these cells, suggesting they may act as latency reversal agents (LRAs) in the CNS. BDZs appear to target RUNX1, a transcription factor that interacts with the HIV-1 LTR, and may disrupt epigenetic control of viral persistence. We hypothesize that HIV-infected myeloid cells adopt a unique global epigenetic signature early in infection, orchestrated in part by RUNX1 and HIV Tat, which modulates effector function and maintains semi-quiescence. BDZs may override this regulation, reactivating latent virus and worsening neuroinflammatory outcomes. We aim to: 1) define the global epigenetic landscape of HIV-infected hMDMs under ART and characterize how RUNX1 and Tat occupancy correlates with gene expression and viral activity and 2) determine how BDZ exposure alters the global epigenetic state of infected and uninfected hMDMs, testing the hypothesis that BDZs promote viral reactivation via a euchromatic shift. This research will uncover new mechanisms of HIV persistence in myeloid reservoirs and inform safer therapeutic strategies for PLWH, especially those at risk for HAND.