"Despite the success of antiretroviral therapy (ART), HIV persists in long-lived immune cells as a latent reservoir that reactivates if treatment is interrupted. A major challenge in HIV cure research is identifying host factors required to maintain transcriptional competence of these latent viruses. Targeting such host dependencies represents a promising strategy for achieving durable viral suppression through “block-and- lock” approaches.
This pilot project investigates the Werner syndrome helicase (WRN), a host DNA repair protein, as a novel regulator of HIV-1 transcription. WRN resolves transcription-associated DNA structures and maintains genome stability under cellular stress. Prior studies suggest that HIV recruits WRN to support transcription, but whether WRN catalytic activity is functionally required remains unknown.
We will test whether WRN helicase activity is required for HIV transcriptional competence across different genomic contexts. Leveraging DC CFAR Core services, we will use well-defined J-Lat latency models to combine genetic knockdown, pharmacologic inhibition with a selective WRN inhibitor (HRO761), and rescue experiments with a helicase-deficient mutant to define WRN function. Transcriptional output will be measured using complementary assays including GFP reporter activity, RT-qPCR of viral transcripts, and p24 protein production under basal and stimulated conditions. In parallel, we will evaluate WRN dependency across J-Lat clones with distinct integration sites. To enhance physiological relevance, we will perform a focused validation in primary human CD4⁺ T cells to determine whether WRN inhibition similarly suppresses HIV transcription in a primary immune cell setting.
Together, these studies will provide the first direct test of WRN as a targetable host dependency factor and establish a mechanistic foundation for host-directed HIV cure strategies. The findings will align with NIH high-priority HIV/AIDS research areas and support future translational efforts toward durable viral silencing and improved health outcomes for people living with HIV.."
Project Summary provided by investigator.