Cooperative Agreement UG3CA290312

DEVELOPING RIBOSOME BIOGENESIS INHIBITORS FOR THE PREVENTION OF CANCER - SUMMARY TUMOR FORMA)ON AND CANCER CELL PROLIFERA)ON DEPEND ON ENHANCED RIBOSOME BIOGENESIS AND PROTEIN SYNTHESIS. RIBOSOMES ARE THE MOLECULAR MACHINES RESPONSIBLE FOR PROTEIN PRODUC)ON WITHIN CELLS. EMERGING DATA INDICATES THAT INHIBI)ON OF RIBOSOME FORMA)ON AND FUNC)ON CAN PREVENT CANCERS FROM FORMING IN CERTAIN CONTEXTS. FOR EXAMPLE, LIVER DISEASE PREDISPOSES PA)ENTS FOR LIVER CANCER, INCLUDING HEPATOCELLULAR CARCINOMA, AND THIS TRANSI)ON IS MARKED BY ENHANCED RIBOSOME BIOGENESIS. INHIBI)ON OF RIBOSOME FUNC)ON PREVENTS LIVER TUMOR FORMA)ON IN MOUSE MODELS. THUS, ADDI)ONAL COMPOUNDS THAT AAENUATE RIBOSOME PRODUC)ON AND AC)VITY MAY BE ABLE TO PREVENT THE FORMA)ON OF TUMORS. UNFORTUNATELY, PREVIOUSLY AVAILABLE METHODS USED TO STUDY RIBOSOME BIOGENESIS ARE LABORIOUS AND NOT EASILY SCALABLE. THIS HAS SIGNIFICANTLY HAMPERED THE DEVELOPMENT OF NEW RIBOSOME BIOGENESIS INHIBITORS. TO OVERCOME THIS CRI)CAL BARRIER, WE HAVE DEVELOPED AN INNOVA)VE PLADORM THAT WE CALL RIBO-SNAP, WHICH ALLOWS US TO ASSAY THE DYNAMICS OF RIBOSOME BIOGENESIS AT SINGLE CELL RESOLU)ON IN LIVING CELLS. USING THIS PLADORM, WE HAVE SUCCESSFULLY COMPLETED PILOT COMPOUND SCREENS AND IDEN)FIED NEW GENES AND SMALL MOLECULES THAT INHIBIT THE PROTEIN SYNTHESIS CAPACITY OF CANCER CELLS. IN PARALLEL, WE HAVE ALSO DEVELOPED A SECOND NOVEL ASSAY CALLED SIDBAIT, WHICH ALLOWS US TO ROBUSTLY AND RAPIDLY IDEN)FY COMPOUND TARGETS WITHIN CELLS. USING THESE APPROACHES, WE ARE IN A UNIQUE POSI)ON TO IDEN)FY AND DEVELOP RIBOSOME BIOGENESIS INHIBITORS AS EFFEC)VE AN)-CANCER AGENTS. WE NOW SEEK TO EXPAND OUR EFFORTS TO THE LARGER NCI NATURAL PRODUCT LIBRARY. THE PROPOSAL IS DIVIDED INTO UG3 AND UH3 SEC)ONS. UNDER AIM 1, WE WILL FURTHER OP)MIZE THE RIBO-SNAP PLADORM AND CONDUCT A SMALL PILOT SCREEN OF A SUBSET OF FRAC)ONS FROM THE 500K NATURAL PRODUCT LIBRARY. IN AIM 2, WE WILL USE SIDBAIT TO IDEN)FY THE TARGETS OF COMPOUNDS IDEN)FIED IN THE PILOT SCREEN OR OUR ONGOING SCREEN OF AN INDEPENDENT 350K COMPOUND LIBRARY. EXPERIMENTS DESCRIBED UNDER AIM 3 WILL DIRECTLY TEST THE EXTENT TO WHICH RIBOSOME BIOGENESIS INHIBITORS CAN PREVENT LIVER TUMOR FORMA)ON IN MOUSE MODELS. THESE THREE AIMS HAVE CLEAR MILESTONES AND WE AN)CIPATE THAT THE SUCCESSFUL COMPLE)ON OF THE UG3 PHASE WILL ALLOW THE PROJECT TO ADVANCE. DURING THE UH3 PHASE, THE LARGE OF SCREENING WILL EXPAND TO THE EN)RE 500K LIBRARY UNDER AIM 4. SIDBAIT WILL BE USED IN AIM 5 TO BEGIN TO DETERMINE THE MECHANISM OF AC)ON OF SPECIFIC COMPOUNDS. FINALLY, UNDER AIM 6, MOUSE MODELS WILL BE USED TO TEST WHETHER LEAD MOLECULE PREVENT CANCER PROGRESSION. THE SUCCESSFUL COMPLE)ON OF THE EN)RE PROJECT DESCRIBED IN THIS PROPOSAL WILL PROVIDE AN UNPARALLELED COLLEC)ON OF RIBOSOME BIOGENESIS INHIBITORS THAT CAN BE FURTHER DEVELOPED INTO NOVEL THERAPEU)CS FOR THE PREVEN)ON OF LIVER CANCER.