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Principal Investigator: Italo Tempera
Organization: WISTAR INSTITUTE
Fiscal Year: 2024
Award: $457,575
Funding agency: National Institute of Allergy and Infectious Diseases
Epstein-Barr Virus (EBV) reprograms host cell gene expression and metabolism during the establishment of latency and the immortalization of B-lymphocytes. The regulatory mechanisms coordinating this reprogramming with EBV latency reflect important events in viral oncogenesis, yet remain poorly understood. We have found that key viral regulators of EBV latency, including the major tegument protein BNRF1 and the EBV Nuclear Antigen EBNA1 coordinate key aspects of purine metabolism during establishment of latency and immortalization of primary B-cells. In this R01, we focus on how EBV reprograms purine metabolic gene expression, and how purine metabolites contribute directly to EBV tumorigenesis. One clue to this coordinate regulation is provided by the viral-encoded tegument protein BNRF1 that shares extensive structural similarity to the purine biosynthetic enzyme FGARAT (also called PFAS) and functions in viral chromatin assembly during primary infection. Orthologues of BNRF1 are found in all gamma herpesviruses, including KSHV ORF75, and share the common function of disarming components of the PML-nuclear body (PML-NB) and its anti-viral functions. We have previously shown that BNRF1 interacts with the histone H3.3 chaperone DAXX and displaces its interaction with the ATP-dependent SNF2-like helicase ATRX to enable selective expression of latency-specific viral genes during primary infection. However, it has not yet been shown how the viral FGARAT homology domain is linked to cellular purine biosynthesis and/or signaling. Using metabolomics mass spectrometry, we provide new preliminary data indicating that the purine biosynthetic pathway is among the most significantly perturbed by EBV during B-cell immortalization. Integrating gene expression (RNA-Seq), chromatin accessibility (ATAC-Seq), and EBNA1-DNA binding to host chromosome (ChIP-Seq), we identified cellular metabolic genes, including adenine deaminase (ADA), adenosine kinase 4 (AK4), and purinergic receptors P2RY8 and P2RX5 as direct targets of EBNA1 transcriptional regulation during EBV immortalization. We now propose to investigate the mechanisms by which EBV senses and reprograms purine metabolism and how purinergic signaling regulates establishment of EBV latency and host cell transformation. We will test the central hypothesis that EBV coordinately regulates cellular purine metabolism with viral and cellular gene expression during the B-cell immortalization process, and that purinergic signaling is critical for viral latency and oncogenesis. Specifically, we will investigate how EBV regulates expression of purine metabolic genes during primary infection (Aim 1), elucidate how purine metabolism impacts the establishment of EBV latency (Aim 2), and investigate the role of purine metabolism and signaling in B-cell immortalization, immune signaling, and EBV-induced tumorigenesis (Aim 3). These studies will advance our understanding of basic mechanisms coordinating gene expression with metabolism and identify new targets for therapeutic intervention in viral latency and viral-associated cancers.
Terms: <1H-Purin-6-amine><ATAC sequencing><ATAC-seq><ATACseq><ATRX><ATRX gene><Adenine><Adenosine><Adenosine Kinase><Anabolism><Assay for Transposase-Accessible Chromatin using sequencing><B Cell Proliferation><B blood cells><B cell><B cell immortalization><B cells><B lymphocyte immortalization><B-Cells><B-Lymphocytes><B-cell><BING2><Burkitt Herpesvirus><Burkitt Lymphoma Virus><Cancers><Cell Body><Cell Communication and Signaling><Cell Signaling><Cells><Cellular biology><ChIP Sequencing><ChIP-seq><ChIPseq><Chaperone><Chromatin><Chromatin Assembly><Chromatin Modeling><Chromosomes><Complex><DAXX><DAXX gene><DNA Binding><DNA Binding Interaction><DNA Helicases><DNA Unwinding Proteins><DNA bound><DNA unwinding enzyme><Data><Deaminase><Death Associated Protein 6><EB virus><EBV><EBV Infections><EBV Nuclear Antigens><EBV latency><EBV malignancies><EBV(+) malignancy><EBV-Related Malignancy><EBV-associated cancers><EBV-associated disease><EBV-associated malignancy><EBV-positive cancer><EBV-positive malignancy><EBV-positive malignant tumor><EBV-related cancer><Enzyme Gene><Enzymes><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Epstein Barr Virus><Epstein Barr Virus Malignancy><Epstein Barr Virus associated cancers><Epstein Barr Virus positive cancer><Epstein-Barr Virus Infections><Epstein-Barr Virus Nuclear Antigens><Epstein-Barr Virus associated disease><Epstein-Barr Virus associated malignancy><Epstein-Barr Virus latency><Epstein-Barr Virus-Related Malignancy><Epstein-Barr Virus-Related Malignant Neoplasm><Epstein-Barr viral infections><Event><G Protein-Complex Receptor><G Protein-Coupled Receptor Genes><G-Protein-Coupled Receptors><GPCR><Gene Expression><Genes><Genome><HHV-4><HHV-8><HHV4><HHV8><Histone H3.3><Human Herpesvirus 4><Human Herpesvirus 8><Immune><Immune signaling><Immunes><Infectious Mononucleosis Virus><Inflammatory><Intermediary Metabolism><Intracellular Communication and Signaling><KSHV><Kaposi Sarcoma-Associated Herpes Virus><Kaposi Sarcoma-Associated Herpesvirus><Kaposi sarcoma associated virus><Kaposi sarcoma herpes virus><Kaposi's sarcoma (KS)-associated herpesvirus><Link><Lymphomagenesis><Lytic Virus><Malignant Neoplasms><Malignant Tumor><Mass Photometry/Spectrum Analysis><Mass Spectrometry><Mass Spectroscopy><Mass Spectrum><Mass Spectrum Analyses><Mass Spectrum Analysis><Metabolic><Metabolic Control><Metabolic Processes><Metabolism><Molecular Chaperones><Nuclear><Oncogenesis><PFAS><Pathway interactions><Play><Poly-fluoroalkyl substances><Primary Infection><Process><Proteins><Purine Metabolism Pathway><Purine Receptors><Purinergic Receptors><Purines><Purinoceptor><RNA Seq><RNA sequencing><RNAseq><Regulation><Role><Signal Transduction><Signal Transduction Systems><Signaling><Testing><Therapeutic Intervention><Transcription Activation><Transcription Regulation><Transcriptional Activation><Transcriptional Control><Transcriptional Regulation><Viral><Viral Activity><Viral Diseases><Viral Function><Viral Genes><Viral Latency><Viral Physiology><Virus><Virus Diseases><Virus Latency><Virus-HHV8><Virus-Related Malignancy><Virus-Related Malignant Neoplasm><Vitamin B4><alpha thalassemia mental retardation X-linked gene><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><biological signal transduction><biosynthesis><cell biology><cell transformation><chromatin immunoprecipitation-sequencing><epigenetically><gamma-herpesvirus><gammaherpesvirus><helicase><inhibitor><intervention therapy><kaposi's sarcoma herpesvirus><kaposi's sarcoma-associated human herpesvirus><malignancy><metabolism measurement><metabolomics><metabonomics><mouse model><murine model><neoplasm/cancer><new drug target><new druggable target><new pharmacotherapy target><new therapeutic target><new therapy target><novel><novel drug target><novel druggable target><novel pharmacotherapy target><novel therapeutic target><novel therapy target><pathway><perfluorinated alkyl substances><perfluoroalkyl substances><perfluoroalkylated substances><polyfluorinated alkyl substances><polyfluoroalkyl substances><programs><purine metabolism><selective expression><selectively expressed><social role><transcriptional reprogramming><transcriptome sequencing><transcriptomic sequencing><transformed cells><tumor><tumorigenesis><viral associated cancer><viral associated malignancy><viral associated malignant neoplasm><viral induced cancer><viral induced malignancy><viral induced malignant neoplasm><viral infection><viral related cancer><viral related malignancy><viral related malignant neoplasm><virus associated cancer><virus associated malignancy><virus associated malignant neoplasm><virus induced cancer><virus induced malignancy><virus induced malignant neoplasm><virus infection><virus related cancer><virus-induced disease><α-thalassemia mental retardation X-linked gene><γ-herpesvirus>