Epstein-Barr virus nuclear antigen 2 in transcription regulation

Epstein-Barr virus (EBV) remains a significant burden to global health. This orally transmitted virus is associated with a spectrum of diseases, including nasopharyngeal carcinoma, a type of head and neck cancer, oral hairy leukoplakia, infectious mononucleosis, and certain B cell malignancies, and about 10% of gastric cancers. Despite its substantial health impact, there are currently no commercial vaccines or targeted therapies for EBV infection or EBV-associated diseases, and the mechanisms underlying EBV's diverse pathogenic effects remain largely elusive. A critical aspect of EBV's oncogenic potential lies in its ability to transform primary B cells into immortalized lymphoblastoid cell lines (LCLs), a process central to EBV-associated tumorigenesis. EBNA2, an EBV encoded nuclear antigen, plays a pivotal role in this transformation by acting as a master regulator of both viral and host gene expression. EBNA2's pathogenesis is intrinsically linked to its transcriptional regulation activity. This protein regulates the expression of hundreds of viral and host genes. However, the mechanisms through which EBNA2 regulates gene expression remain elusive. In this proposal, we aim to elucidate the mechanisms by which EBNA2 regulates gene expression and its contribution to B cell transformation and EBV pathogenesis. We will investigate this from three key aspects: (1) Identification of EBNA2 co-factors involved in EBNA2- mediated transcriptional regulation. EBNA2 regulates gene expression through largely unidentified co-factors. Dissecting these crucial co-factors will not only enhance our understanding of EBNA2's regulatory mechanisms but also identify potential targets for treating EBV-associated diseases. (2) Determine the mechanism through which EBNA2 regulates genome organization and its impact on B cell transformation and gene expression. Our preliminary data indicate that EBNA2 globally alters host genome structure. Given the critical role of genome structure in gene transcription regulation, we hypothesize that EBNA2 manipulates genome organization to regulate gene expression. We will investigate the mechanisms through which EBNA2 regulates genome organization and its effect on gene expression and B cell transformation. (3) Single-cell analysis of EBNA2 in transcriptional regulation. LCLs are heterogeneous cell populations. Consequently, studying the EBNA2 regulome using bulk RNA-seq from LCLs may obscure crucial information about differential responses to EBNA2 among distinct cell subpopulations. To gain a more comprehensive understanding of EBNA2's impact on transcriptional regulation across diverse cell populations in LCLs and during EBV-mediated B cell transformation, we will employ single-cell RNA-seq and ATAC-seq techniques. In summary, this proposal will comprehensively evaluate the role of EBNA2 in transcriptional regulation. This research addresses critical knowledge gaps in EBV biology, potentially leading to new targeted therapies and preventive strategies against EBV-associated malignancies. Project Number: 1R01DE035120-01 | Fiscal Year: 2025 | NIH Institute/Center: National Institute of Dental and Craniofacial Research (NIDCR) | Principal Investigator: Chong Wang | Institution: UNIVERSITY OF MINNESOTA, MINNEAPOLIS, MN | Award Amount: $450,462 | Activity Code: R01 | Study Section: Viral Dynamics and Transmission Study Section [VDT] View on NIH RePORTER: https://reporter.nih.gov/project-details/11204387