Three novel epigenetic-modifying compounds identified as HIV latency-reversing agents in Ghana

Human Immunodeficiency Virus (HIV) persists despite antiretroviral therapy (ART) due to latent viral reservoirs that remain undetected by the immune system. A promising cure approach involves using latency-reversing agents (LRAs) to reactivate latent proviruses, enabling immune-mediated clearance. Because HIV latency is regulated by epigenetic modifications of chromatin surrounding the integrated virus, we hypothesized that epigenetic-modifying compounds could function as effective LRAs. We screened 150 such compounds using the J.LAT 10.6 cell line, which harbors a single integrated HIV-GFP reporter. Reactivation was assessed by GFP expression via flow cytometry. Positive hits were screened across multiple J.LAT clones and a primary CD4⁺ T cell latency model, with lead candidates evaluated ex vivo in CD4⁺ T cells isolated from four ART-suppressed individuals (viral load <50 copies/mL). HIV-1 mRNA levels were quantified by RT-qPCR. Screening identified 15 positive hits, of which five (MC1568, Abexinostat, Pracinostat, EPZ2015666, and CXC6258-HCL) induced 20–91% GFP-positive cells. MC1568 showed consistent activity across J.LAT clones (47–91% GFP⁺). In the primary T cell model, three compounds (MC1568, Abexinostat, and Pracinostat) were validated as lead LRAs. Ex vivo, these compounds significantly increased intracellular HIV-1 mRNA expression (6–19-fold) in CD4⁺ T cells from all four participants. We identified three promising LRAs (MC1568, Abexinostat, and Pracinostat) that robustly reactivated latent HIV both in vitro and ex vivo. These findings warrant preclinical evaluation in animal models as potential components of HIV cure strategies. Notably, this is the first study in Ghana to establish a medium- to high-throughput HIV latency reversal screening platform.