Please visit our lab website: gewurzlab.bwh.harvard.edu
Our research focuses on EBV-mediated oncogenesis. We use CRISPR genetic and proteomic techniques to interrogate how EBV reprograms key B-cell growth, survival, metabolic and immune-evasion pathways.
EBV is an oncogenic gamma-herpesvirus that persistently infects >95% of adults worldwide. EBV is associated with multiple human malignancies, with nearly 200,000 cancers attributable to EBV each year worldwide. These include Burkitt lymphoma, Hodgkin lymphoma, HIV-associated lymphomas, post-transplant lymphoproliferative diseases, and immune-senescence-associated lymphomas. EBV is also causally associated with epithelial malignancies, including nasopharyngeal carcinoma and gastric carcinoma. EBV's association with cancer is an outgrowth of its relationship with host cells. Upon infection of primary B lymphocytes, EBV enters a state of viral latency, but is hardly quiescent. Rather, EBV expresses oncogenic membrane proteins, transcription factors and microRNAs that efficiently transform resting B-lymphocytes into rapidly growing lymphoblasts. In vitro, EBV converts primary resting B lymphocytes into immortalized lymphoblastoid cell lines (LCLs), which provide an excellent tissue culture model for studies of EBV latency proteins and lymphoproliferative disorders.
Current research includes the following areas:
1) CRISPR/Cas9 screens to identify novel synthetic lethal targets in EBV-transformed B-cells. We are using CRISPR-based genetic approaches to identify key oncogenic strategies that support EBV-driven immortalized B-cell growth, including how the virus evades tumor repressor responses. These vulnerabilities may be exploited to rationally target the growth and survival of EBV-transformed B-cells.
2) EBV LMP1-MEDIATED NF-KB PATHWAY ACTIVATION
The EBV oncoprotein Latent Membrane Protein 1 (LMP1) mimics CD40 signaling to potently activate the canonical and non-canonical NF-kB, MAP kinase, and IRF7 pathways. LMP1-mediated NF-kB activation is essential for EBV-mediated B-cell transformation, and for the growth and survival of transformed cells. We use genetic and proteomic approaches to study LMP1 signaling from the assembly of plasma membrane signalosomes through the activation of NF-kB transcription factor complexes. Likewise, we use next-generation sequencing-based technologies, including ChIP-seq. We are using CRISPR/Cas9, RNAi, ChiP-seq, and mass-spectrometry-based proteomic approaches to characterize how LMP1 constitutively activates NF-kB pathways. We are also comparing LMP1 and CD40-mediated NF-kB target gene regulation in B-cells.
3) QUANTITATIVE TEMPORAL VIROMIC ANALYSIS OF EBV LYTIC INFECTION
We are using whole cell mass spectrometry-based approaches to probe the EBV/host relationship during B-cell transformation versus lytic replication. This unbiased, systematic approach promises identify novel viral strategies central to EBV biology, including EBV subversion of host growth and survival pathways, metabolic pathways, innate and adaptive immune pathways. Likewise, we are identifying whole proteome changes that accompany EBV-mediated primary B-cell transformation.
4) NOVEL PRIMARY HUMAN IMMUNODEFICIENCIES WITH EBV SUSCEPTIBILITY
We are conducting whole exome and immunologic studies oF individuals with persistent active EBV infection and EBV-associated B-cell malignancy. These studies promise to identify key aspects of the host immune response required to control EBV lytic and latent infection.