Persistence of a herpesvirus - in both the host and population - requires a balance between latent and lytic phases. This balance is also important for herpesvirus-mediated pathogenesis, including EBV-related diseases. EBV provides a powerful system for studying human herpesvirus latency (and lytic activation), due to its ability to establish latency in vitro. While latent EBV can be (re-)activated into the lytic phase by chemicals or immunoglobulin crosslinking, not every EBV-infected B cell is susceptible to lytic activation. This refractory state, while crucial for viral persistence, negatively impacts success f viral oncolytic therapy for EBV-cancers. Unfortunately, determinants of susceptibility to lytic activation signals are poorly understood. To address this critical issue at the single cell level, e pioneered a method for separating lytic and refractory B cells, and made the fundamental discovery that cellular STAT3, a transcription factor overactive in many human cancers, regulates susceptibility to lytic activation. We propose to test the hypothesis that STAT3 does this by employing cellular DNA-binding protein SZF1 to recruit the transcriptional co-repressor KAP1 to simultaneously repress multiple EBV lytic genes, thereby promoting the refractory state.
In Aim 1, we will identify and validate SZF1-binding sites on the EBV genome in infected cells, using ChIP-exo to achieve single nucleotide resolution.
In Aim 2, we will elucidate the mechanism of SZF1-mediated transcriptional repression of EBV lytic genes - using STAT3, SZF1, and KAP1 mutants, as well as purified lytic and refractory cells. The proposed research is expected to significantly impact our understanding of 1) how 2 major host transcriptional mechanisms (mediated via STAT3 and KAP1) regulate expression of EBV lytic genes to affect susceptibility to lytic activation, and thereby viral persistence, 2) how STAT3, SZF1, and KAP1 function may be exogenously manipulated to increase the number of lytic cells, and 3) how KAP1 repressor identifies targets in a genomic context - promoting discovery of cellular targets and physiologic functions of SZF1. Importantly, by providing tools to devise approaches (e.g., small molecules) to enhance lytic activation, our work promises to cripple viral persistence, and improve effectiveness of therapies for EBV-associated cancers.

Public Health Relevance

A fundamental issue in herpesvirus research is understanding what controls whether virus will be active or will, instead, lie quiescent within infected cells. This balance between active and quiescent states directly and profoundly impacts viral persistence and disease - as well as efficacy of treatments to control those diseases, particularly viral cancers. Our research is focused on understanding the contribution of human, host cells to regulating the active/quiescent balance. To do this, we use the best available herpesvirus model relevant to both active and quiescent states - Epstein-Barr virus (EBV), which is present in most humans. This research will provide the knowledge and tools needed to therapeutically target host-herpesvirus interactions to eliminate the virus, and improve therapies for EBV-associated cancers.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
1R01AI113134-01
Application #
8764258
Study Section
Virology - A Study Section (VIRA)
Program Officer
Beisel, Christopher E
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2014
Total Cost
Indirect Cost
Name
State University New York Stony Brook
Department
Pediatrics
Type
Schools of Medicine
DUNS #
City
Stony Brook
State
NY
Country
United States
Zip Code
11794