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Principal Investigator: Laura Murray-Nerger
Organization: BRIGHAM AND WOMEN'S HOSPITAL
Fiscal Year: 2024
Award: $76,984
Funding agency: National Institute of Allergy and Infectious Diseases
PROJECT SUMMARY/ABSTRACT
Epstein-Barr virus (EBV) is spread through saliva, infects oropharyngeal tissues including the tonsillar epithelium,
and establishes life-long latency in the B-cell compartment in over 95% of the global adult human population.
The oral transmission of EBV can result in infectious mononucleosis and can lead to several B-cell and epithelial
cancers, including Burkitt lymphoma. This frequently presents as craniofacial and nasopharyngeal carcinomas.
EBV uses both latent and lytic phases of its replication cycle to colonize the oropharynx and tonsils. Reactivation
from latency is closely tied to EBV pathogenicity; however, the mechanisms that maintain latency and regulate
reactivation in tonsillar memory B-cells and in EBV-associated diseases remain incompletely understood.
Because EBV can evade immune detection while latent, most currently available therapies cannot harness the
presence of the latent EBV genome. Developing a detailed understanding of the switch from latency to lytic
reactivation may lay the foundation for lytic induction therapeutic strategies. One of the challenges to
understanding the EBV lytic switch is to define the host factors necessary for maintaining EBV latency and how
these factors are circumvented by EBV to allow reactivation. To begin to address this question, a human
CRISPR/Cas9 screen was performed in Burkitt B-cells that were originally derived from a craniofacial biopsy. It
showed that knockout of the human nuclear protein splicing factor proline and glutamine rich (SFPQ) strongly
induces EBV lytic reactivation. SFPQ binds both DNA and RNA and is known to regulate both transcription and
RNA splicing. Thus, SFPQ is poised as a master regulator of human and viral gene expression and viral genomic
conformation during EBV latency. The goal of this study is to test the hypothesis that SFPQ suppresses
EBV lytic reactivation in both DNA- and RNA- dependent manners and that EBV relieves this suppression
by redistributing SFPQ to paraspeckles. Aim 1 is to determine the mechanism by which SFPQ represses EBV
lytic reactivation. Aim 2 is to define the factors that mediate SFPQ subnuclear redistribution and function upon
lytic reactivation. Molecular virology, transcriptomic, genomic, and microscopy approaches will be integrated to
test this hypothesis. Understanding how SFPQ regulates the EBV and host genomes during latency and how
SFPQ responds during lytic reactivation will contribute to a fundamental knowledge gap in how EBV subverts
host factors to colonize oropharyngeal tissues and the B-cell compartment. This study may lay the foundation
for lytic induction therapeutic strategies. The Brigham and Women’s Hospital at Harvard Medical School will
provide an environment rich in both physical and intellectual resources for completion of this training.
Furthermore, working with Dr. Gewurz in the collaborative atmosphere of his lab will enhance training in multi-
disciplinary approaches to study EBV-host interactions. Altogether, this is an excellent environment for scientific
training for development towards running an independent research program.
Terms: <21+ years old><3-D><3-Dimensional><3D><AIDS Virus><Acquired Immune Deficiency Syndrome Virus><Acquired Immunodeficiency Syndrome Virus><Address><Adult><Adult Human><Architecture><B blood cells><B cell><B cells><B-Cells><B-Lymphocytes><B-cell><Basal Transcription Factor><Basal transcription factor genes><Binding><Biology><Biopsy><Body Tissues><Burkitt Herpesvirus><Burkitt Lymphoma><Burkitt Lymphoma Virus><Burkitt Tumor><Burkitt's Lymphoma/Leukemia><Burkitt's Type Small Non-Cleaved Cell Lymphoma><Business-Friendly Atmosphere><CRISPR editing screen><CRISPR screen><CRISPR-based screen><CRISPR/Cas9 screen><Cancers><Carcinoma><Cell Body><Cell Compartmentation><Cell Compartmentations><Cells><DNA><DNA Binding><DNA Binding Interaction><DNA bound><DNA-Binding Proteins><Deoxyribonucleic Acid><Detection><Development><Disease><Disorder><Down-Regulation><EB virus><EBV><EBV Infections><EBV latency><EBV-associated disease><Engineering / Architecture><Environment><Epithelial Cells><Epithelial cancer><Epithelium><Epstein Barr Virus><Epstein-Barr Virus Infections><Epstein-Barr Virus associated disease><Epstein-Barr Virus latency><Epstein-Barr pathogenesis><Epstein-Barr viral infections><Equilibrium><Foundations><Functional RNA><Gastric Carcinoma><Gene Expression><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genetic Transcription><Genome><Genomic Segment><Genomics><Germinoblastic Sarcoma><Germinoblastoma><Glandular Fever><Gln><Glutamine><Goals><HHV-4><HHV4><HIV><Histones><Hodgkin Disease><Hodgkin Disorder><Hodgkin lymphoma><Hodgkin's><Hodgkin's Lymphoma><Hodgkin's disease><Hodgkins lymphoma><Hospitals><Host Factor><Host Factor Protein><Human><Human Herpesvirus 4><Human Immunodeficiency Viruses><Immune Evasion><Infectious Mononucleosis><Infectious Mononucleosis Virus><Integration Host Factors><Knock-out><Knockout><Knowledge><L-Glutamine><L-Proline><LAV-HTLV-III><Location><Lymphadenopathy-Associated Virus><Lymphoma><Lytic><Lytic Cycle><Lytic Infection><Lytic Phase><Lytic Virus><Malignant Epithelial Neoplasms><Malignant Epithelial Tumors><Malignant Lymphogranuloma><Malignant Lymphoma><Malignant Nasopharyngeal Neoplasm><Malignant Nasopharyngeal Tumor><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Nasopharynx><Malignant neoplasm of nasopharynx><Mediating><Membrane><Memory B Cell><Memory B-Lymphocyte><Methods><Microscopy><Modern Man><Molecular Configuration><Molecular Conformation><Molecular Interaction><Molecular Stereochemistry><Molecular Virology><Nasopharyngeal Cancer><Nasopharyngeal Carcinoma><Nasopharynx><Nasopharynx Cancer><Nasopharynx Carcinoma><Nervous System Diseases><Nervous System Disorder><Neurologic Disorders><Neurological Disorders><Non-Coding><Non-Coding RNA><Non-Polyadenylated RNA><Non-translated RNA><Noncoding RNA><Nontranslated RNA><Nuclear Protein><Oral><Oropharyngeal><Oropharynx><Oropharynxs><Phase><Population><Proline><Protein Splicing><Proteins><Q Levoglutamide><Q. Levoglutamide><RNA><RNA Binding><RNA Expression><RNA Gene Products><RNA Splicing><RNA bound><RNA-Binding Proteins><Repression><Research><Research Resources><Resources><Reticulolymphosarcoma><Rhinopharynx><Ribonucleic Acid><Role><Running><Saliva><Salivary><Site><Splicing><Stomach Carcinoma><Structure><Testing><Therapeutic><Tissues><Tonsil><Tonsillar Tissue><Training><Transcription><Transcription Factor Proto-Oncogene><Transcription Regulation><Transcription factor genes><Transcriptional Control><Transcriptional Regulation><Transmission><Untranslated RNA><Viral Diseases><Viral Genes><Viral Genome><Viral Latency><Virus Diseases><Virus Latency><Virus-HIV><Woman><Work><adulthood><balance><balance function><business-friendly environment><clustered regularly interspaced short palindromic repeats screen><collaborative atmosphere><collaborative environment><conformation><conformational><conformational state><conformationally><conformations><craniofacial><craniofacies><developmental><epithelial carcinoma><gamma-herpesvirus><gammaherpesvirus><genome segment><genomic region><immune evasive><infection mouth><insight><interactive atmosphere><interactive environment><interdisciplinary approach><interdisciplinary atmosphere><interdisciplinary environment><lab atmosphere><laboratory atmosphere><latency/reactivation><live cell image><live cell imaging><live cellular image><live cellular imaging><lytic gene expression><lytic replication><lytic viral replication><lytic virus replication><malignancy><medical college><medical schools><membrane structure><mononucleosis><multidisciplinary approach><nasopharnygeal><neoplasm/cancer><neurological disease><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapy approaches><new treatment approach><new treatment strategy><noncoding><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapy approach><oral infection><oral infectious><oral pharyngeal><pathogen><pathogenesis of Epstein-Barr virus><pathogenesis of Epstein-Barr virus infection><pathogenic virus><peer-group atmosphere><peer-group environment><post-transplant><post-transplantation><posttransplant><posttransplantation><programs><promoter><promotor><reactivation from latency><research atmosphere><school of medicine><science atmosphere><science oriented atmosphere><scientific atmosphere><social role><three dimensional><tonsillar><transcription factor><transcriptomics><transmission process><viral genomics><viral infection><viral pathogen><virus genome><virus genomics><virus host interaction><virus infection><virus pathogen><virus-induced disease><γ-herpesvirus>