OLIGONUCLEOTIDES AS INHIBITORS OF PAPILLOMAVIRUS

THE PURPOSE OF THIS PROJECT IS TO TEST THE FEASIBILITY AND TECHNICAL MERIT OF USING ANTISENSE OLIGONUCLEOTIDES AS INHIBITORS OF PAPILLOMAVIRUS GENE FUNCTION. BOVINE PAPILLOMAVIRUS TYPE-1 (BPV-1) HAS BEEN SELECTED AS THE MODEL SYSTEM BECAUSE IT IS THE ONLY IN VITRO SYSTEM WHERE THE PAPILLOMAVIRUS GENOME IS MAINTAINED AS AN EPISOME AS IT IS IN MOST NATURALLY OCCURRING LESIONS. EARLY GENE 2 (E2) HAS BEEN SELECTED AS THE TARGET GENE FOR TWO MAJOR REASONS: 1) E2 PLAYS A CENTRAL ROLE IN REGULATION OF PAPILLOMAVIRUS TRANSCRIPTION. THE E2 TRANSREGULATION OF VIRAL TRANSCRIPTION IS A COMMON FEATURE AMONG ALL PAPILLOMAVIRUSES ANALYZED, BOTH ANIMAL AND HUMAN; 2) A SIMPLE ASSAY EXISTS TO QUALITATIVELY MEASURE E2 GENE FUNCTION THUS ALLOWING QUANTITATIVE MEASUREMENT OF INHIBITION OF E2 EXPRESSION BY ANTISENSE COMPOUNDS. IN THE INITIAL PHASE OF THIS PROGRAM, THE FEASIBILITY OF THIS APPROACH WILL BE TESTED BY DESIGNING, SYNTHESIZING AND TESTING ANTISENSE OLIGONUCLEOTIDES AGAINST E2 IN A CELL CULTURE BASED ASSAY. THIS ASSAY WILL NOT ONLY ALLOW QUANTITATIVE MEASUREMENT OF E2 EXPRESSION BUT ALSO ALLOW THE ANALYSIS OF THE EFFECTS OF INHIBITION OF E2 EXPRESSION ON THE ABILITY OF BPV-1 TO MAINTAIN AND REPLICATE ITS GENOME IN VIRALLY TRANSFORMED CELLS.