Single-Cell Profiling of HDAC Inhibitor-Induced EBV Lytic Heterogeneity Defines Abortive and Refractory States in B
Lauren E Haynes1, Ashley P Barry1, Micah A Luftig1,2
1Department of Molecular Genetics and Microbiology, Duke University School of Medicine, Duke Center for Virology, Durham, NC, USA.
Biorxiv : the Preprint Server for Biology
|November 24, 2025
Summary
Histone deacetylase (HDAC) inhibitors show promise for Epstein-Barr virus (EBV)-associated cancers by inducing lytic reactivation. However, host factors like NF-κB signaling can impede this process, impacting therapeutic strategies.
Area of Science:
- Oncology
- Virology
- Molecular Biology
Background:
- Epstein-Barr virus (EBV) is linked to several cancers, including lymphomas and carcinomas.
- EBV-positive tumors often exhibit latent gene expression, posing challenges for conventional therapies.
- Lytic reactivation therapies using HDAC inhibitors are being explored but require deeper mechanistic understanding.
Purpose of the Study:
- To investigate the effects of pan-HDAC inhibitor (panobinostat) and class I HDAC inhibitor (nanatinostat) on EBV-positive cancer cell lines.
- To characterize the impact of these inhibitors on cell growth, survival, and EBV lytic reactivation.
- To elucidate host factors that may prevent successful EBV lytic reactivation.
Main Methods:
- Treatment of four EBV-positive cell lines (BL, immunoblastic lymphoma, LCLs) with panobinostat and nanatinostat.
- Single-cell approaches to analyze EBV lytic gene expression, cell cycle, and apoptosis.
- Single-cell RNA sequencing on panobinostat-treated P3HR1-ZHT BL cells to identify host factor roles.
Main Results:
- All EBV-positive cell lines showed sensitivity to HDAC inhibitors, with unique response profiles.
- Successful EBV lytic reactivation occurred in a small fraction of cells across models.
- Abortive lytic cells upregulated NF-κB-dependent genes and immune signaling pathways, including CD137/CD137L.
Conclusions:
- HDAC inhibitors induce EBV lytic reactivation, but efficacy varies among cancer models.
- Host factors, particularly NF-κB and immune signaling pathways, play a role in limiting successful lytic reactivation.
- Findings offer insights for developing novel oncolytic therapies targeting EBV-associated malignancies.
