REV-ERBα/β Targeting Transcriptionally Reprograms HIV-Infected CD4 ⁺ T-Cells for Increased Viral Reactivation but Limited Virion Spread

The circadian clock repressors REV-ERBα/β control rhythmic gene expression and inhibit HIV-1 transcription. Whether REV-ERBα/β modulate the HIV-1 replication cycle beyond transcription remains unknown. Here, we demonstrate that memory CD4 ⁺ T-cells predominantly express the REV-ERBβ isoform ex vivo and that T-cell receptor (TCR) triggering downregulates both REV-ERBα/β expression, with levels of REV-ERBα/β mRNA being lower in ART-treated people with HIV (PWH) receiving antiretroviral therapy (ART) compared to people without HIV (PWoH) before/after TCR triggering. In single-round infection, the REV-ERBα/β antagonist SR8278 facilitated HIV-1 reverse transcription, integration, and intracellular HIV-p24 expression, but limited virion release. Moreover, SR8278 downregulated CCR5 mRNA expression, inhibited R5-tropic HIV-1 replication in vitro, and limited viral outgrowth in CD4 ⁺ T-cells from ART-treated PWH. Finally, genome-wide RNA-sequencing and functional validations revealed HIV-1 restriction/dependency factors that represent novel putative REV-ERBα/β targets. Thus, pharmacological inhibition of REV-ERBα/β uniquely combines a latency reversal activity with the inhibition of progeny virion spread.