THE IDENTIFICATION OF CLINICALLY RELEVANT METABOLIC INHIBITORS TO TREAT GAMMAHERPESVIRUS INFECTIONS - PROJECT ABSTRACT SIGNIFICANCE: IT IS ESTIMATED THAT ~15% OF ALL CANCERS ARE CAUSED BY ONCOGENIC VIRUS INFECTIONS. TWO OF THE TOP SEVEN HUMAN ONCOGENIC VIRUSES ARE MEMBERS OF THE GAMMAHERPESVIRUS FAMILY: EPSTEIN BARR VIRUS (EBV) AND KAPOSI'S SARCOMA HERPESVIRUS (KSHV). MURINE HERPESVIRUS 68 (MHV-68), A MOUSE GAMMAHERPESVIRUS WHICH SHARES SIGNIFICANT HOMOLOGY WITH KSHV AND EBV, IS A MODEL SYSTEM TO STUDY ONCOGENIC GAMMAHERPESVIRUS INFECTIONS. THIS PROPOSAL WILL FOCUS ON THE ALTERATION OF HOST CELL METABOLISM BY MHV-68 DURING LYTIC INFECTION, AN OFTEN-OVERLOOKED ASPECT OF THE GAMMAHERPESVIRUS LIFE CYCLE. WE WILL ELUCIDATE WHICH HOST CELL METABOLISM GENES ARE INDUCED DURING ONCOGENIC GAMMAHERPESVIRUS LYTIC INFECTION AND IDENTIFY CLINICALLY RELEVANT ANTI- METABOLISM DRUGS THAT CAN BLOCK GAMMAHERPESVIRUS PRODUCTION. INDUCTION OF THE LYTIC CYCLE ALLOWS THE ESTABLISHMENT OF A LIFELONG PERSISTENT INFECTION BY CONTINUOUSLY SPREADING VIRUS WITHIN THE SAME HOST OR TO NEW HOSTS. THE LYTIC CYCLE ALSO HAS IMPLICATIONS IN SEEDING NEW TUMORS. UNDERSTANDING THE LYTIC CYCLE'S ROLE IN ALTERING HOST CELL METABOLISM CAN PROVIDE US WITH NEW THERAPEUTIC MECHANISMS TO BLOCK LYTIC REPLICATION AND VIRUS SPREAD. HYPOTHESIS: MHV-68 ALTERATION OF HOST CELL METABOLISM IS NECESSARY FOR LYTIC REPLICATION AND BLOCKING THESE PATHWAYS WITH CLINICALLY RELEVANT ANTI-METABOLISM DRUGS CAN THERAPEUTICALLY TREAT GAMMAHERPESVIRUS INFECTIONS. METHODS: FIRST, UNDERGRADUATE RESEARCHERS WILL PERFORM RNA-SEQ ANALYSIS OF MOCK VS MHV-68 INFECTED CELLS AND DETERMINE WHICH HOST CELL METABOLISM GENES ARE INDUCED DURING INFECTION. SECOND, STUDENTS WILL IDENTIFY CLINICALLY RELEVANT METABOLIC INHIBITORS THAT CAN BLOCK MHV-68 PRODUCTION IN VITRO. COMMON EXPERIMENTAL METHODS WILL INCLUDE DRUG TOXICITY ASSAYS, WHICH MEASURE CELL DEATH AND PROLIFERATION, AND VIRAL PRODUCTION (TITER) ASSESSMENT USING TRADITIONAL PLAQUE ASSAYS. THIRD, STUDENTS WILL IDENTIFY WHICH STEP(S) OF THE VIRAL LIFE CYCLE IS BLOCKED BY THESE METABOLIC INHIBITORS BY USING REAL-TIME RT-PCR TO DETECT PERTURBATIONS IN VIRAL GENE TRANSCRIPTION THROUGHOUT THE VIRAL REPLICATION CYCLE. TEACHING AIMS: THE EXPERIMENTS IN THIS PROPOSAL WILL BE PERFORMED BY UNDERGRADUATES AND HAVE THE POTENTIAL TO SPARK SCIENTIFIC CURIOSITY. THIS PROPOSAL WILL EXPOSE STUDENTS TO GENUINE RESEARCH EXPERIENCES, HELP PREPARE THEM FOR GRADUATE SCHOOL, GENERATE EXCITEMENT ABOUT THE SCIENTIFIC PROCESS, AND INCREASE OUR NATION'S BIOMEDICAL WORKFORCE. IMPLICATIONS: THIS RESEARCH IS TRANSFORMATIVE BY FOCUSING ON HOW ALTERATION OF HOST CELL METABOLISM DURING LYTIC REPLICATION OF GAMMAHERPESVIRUSES CONTRIBUTES TO VIRION PRODUCTION AND THEREFORE CANCER FORMATION. ADDITIONALLY, INHIBITION OF METABOLIC PATHWAYS WITH CLINICALLY RELEVANT METABOLIC INHIBITORS CAN POTENTIALLY BE USED CHEMOTHERAPEUTICALLY TO TREAT VIRAL INDUCED TUMORS.