TRANSCRIPTIONAL CONTROL OF THE ERYTHROPOIETIN GENE

ERYTHROPOIETIN (EPO) INDUCES ERYTHROID PROGENITOR CELLS TO DIFFERENTIATE INTO ERYTHROCYTES. RECOMBINANT EPO IS APPROVED FOR TREATING ANEMIA IN PATIENTS WITH CHRONIC RENAL FAILURE, AND IS BEING EVALUATED IN ANEMIAS ASSOCIATED WITH CHEMOTHERAPY, AIDS AND SURGERY. HOWEVER, THE USE OF RECOMBINANT EPO AS A PHARMACEUTICAL SUFFERS FROM SEVERAL LIMITATIONS, EG. THE HIGH COST OF MANUFACTURE, TECHNICAL DIFFICULTIES ASSOCIATED WITH REPRODUCIBLE LARGE SCALE PURIFICATION, PROTEIN STORAGE AND THE NEED FOR ADMINISTRATION BY INJECTION. SUCH PROBLEMS COULD BE CIRCUMVENTED BY DEVELOPING PHARMACEUTICAL COMPOUNDS WHICH CAN REPLACE RECOMBINANT EPO THERAPY BY TRANSCRIPTIONAL ACTIVATION OF THE ENDOGENOUS EPO GENE. ONCOGENE SCIENCE HAS DEVELOPED A UNIQUE ROBOTIC TECHNOLOGY TO IDENTIFY MOLECULES WHICH ACT AS SPECIFIC MODULATORS OF TRANSCRIPTION. THE CELL-BASED SCREEN CAN ANALYZE MORE THAN 2,000 COMPOUNDS PER WEEK AGAINST MULTIPLE GENES. PRELIMINARY EXPERIMENTS INDICATE THAT THE EPO EXPRESSION UNIT IS COMPLEX, PREVENTING THE ROUTINE CONSTRUCTION OF A REGULATED PROMOTER/REPORTER VECTOR. IN PHASE I, WE WILL CONSTRUCT VARIOUS PROMOTER/REPORTER PLASMIDS TO IDENTIFY TRANSCRIPTIONAL ELEMENTS WHICH REGULATE EXPRESSION OF EPO IN RESPONSE TO ANOXIA. WE WILL SCREEN THE APPROPRIATE CONSTRUCT IN PHASE II TO IDENTIFY NOVEL COMPOUNDS WITH THE GOAL TO DEVELOP PHARMACEUTICAL AGENTS WHICH STIMULATE TRANSCRIPTION OF THE EPO GENE.