The Histone Deacetylase Inhibitor Vorinostat (SAHA) Increases the Susceptibility of Uninfected CD4 ⁺ T Cells to HIV by Increasing the Kinetics and Efficiency of Postentry Viral Events

Abstract

Latently infected cells remain a primary barrier to eradication of HIV-1. Over the past decade, a better understanding of the molecular mechanisms by which latency is established and maintained has led to the discovery of a number of compounds that selectively reactivate latent proviruses without inducing polyclonal T cell activation. Recently, the histone deacetylase (HDAC) inhibitor vorinostat has been demonstrated to induce HIV transcription from latently infected cells when administered to patients. While vorinostat will be given in the context of antiretroviral therapy (ART), infection of new cells by induced virus remains a clinical concern. Here, we demonstrate that vorinostat significantly increases the susceptibility of CD4؉ T cells to infection by HIV in a dose-and time-dependent manner that is independent of receptor and coreceptor usage. Vorinostat does not enhance viral fusion with cells but rather enhances the kinetics and efficiency of postentry viral events, including reverse transcription, nuclear import, and integration, and enhances viral production in a spreading-infection assay. Selective inhibition of the cytoplasmic class IIb HDAC6 with tubacin recapitulated the effect of vorinostat. These findings reveal a previously unknown cytoplasmic effect of HDAC inhibitors promoting productive infection of CD4 ؉ T cells that is distinct from their well-characterized effects on nuclear histone acetylation and long-terminal-repeat (LTR) transcription. Our results indicate that careful monitoring of patients and ART intensification are warranted during vorinostat treatment and indicate that HDAC inhibitors that selectively target nuclear class I HDACs could reactivate latent HIV without increasing the susceptibility of uninfected cells to HIV. IMPORTANCEHDAC inhibitors, particularly vorinostat, are currently being investigated clinically as part of a "shock-and-kill" strategy to purge latent reservoirs of HIV. We demonstrate here that vorinostat increases the susceptibility of uninfected CD4؉ T cells to infection with HIV, raising clinical concerns that vorinostat may reseed the viral reservoirs it is meant to purge, particularly under conditions of suboptimal drug exposure. We demonstrate that vorinostat acts following viral fusion and enhances the kinetics and efficiency of reverse transcription, nuclear import, and integration. The effect of vorinostat was recapitulated using the cytoplasmic histone deacetylase 6 (HDAC6) inhibitor tubacin, revealing a novel and previously unknown cytoplasmic mechanism of HDAC inhibitors on HIV replication that is distinct from their well-characterized effects of long-terminal-repeat (LTR)-driven gene expression. Moreover, our results suggest that treatment of patients with class I-specific HDAC inhibitors could induce latent viruses without increasing the susceptibility of uninfected cells to HIV.

In vitro effects of the small-molecule protein kinase C agonists on HIV latency reactivation

“…SAHA may successfully increase viral transcription, but fails to effectively enhance viral translation61. Moreover, SAHA treatment may increase susceptibility of uninfected CD4 T cell to HIV, and impair virus-specific CTLs6263. In comparison, synthetic analogues of protein kinase C (PKC) agonists represent HIV latency reversing agents (LRAs) of interest.…”

Histone Deacetylase Inhibitor Romidepsin InhibitsDe NovoHIV-1 Infections

“…Numerous in vitro, ex vivo, and clinical studies have shown that HDACi reactivate latent HIV infection (9, 12-16, 37, 38, 40-48). However, a theoretical concern is that HDACi-induced viral production in cART sanctuary sites could result in de novo HIV infections and reseeding of the latent reservoir (17,18). In the present study, we addressed this concern by comprehensively exploring the impact of romidepsin on HIV infection.…”

“…Recently, Lucera et al reported that supraphysiological concentrations (200 nM) of romidepsin increased the susceptibility of CD4 ϩ T cells to HIV infection (17). These data, as well as those of others (18), raise the concern that inducing latent HIV in anatomical compartments with suboptimal cART concentrations (19) could lead to the infection of new target cells and reseeding of the latent reservoir.…”

Autophagy Induction by Histone Deacetylase Inhibitors Inhibits HIV Type 1

“…However, a major clinical concern with this strategy is that, although administered in the context of cART, infection of new uninfected CD4 ϩ T cells and macrophages by induced virus during periods of viral activation from latency may occur. Indeed, a recent study has demonstrated that although vorinostat does not affect virus-CD4 ϩ T cell fusion, it increases the kinetics of post-entry events such as reverse transcription and integration thereby promoting productive infection of CD4 ϩ T cells and potentially reseeding the viral reservoirs being purged (4). Another important HIV reservoir is the macrophage.…”