RETROVIRUS MODEL--EVUALUATE ANTI-SENSE PROBE THERAPY

VIRAL OR PLASMID CONSTRUCTS CONTAINING A HUMAN GROWTH HORMONE (HGH) GENE POSITIONED DOWNSTREAM FROM AN AUTHENTIC VIRAL OR HOST TRANSCRIPTIONAL CONTROL SEQUENCE SHOULD PROVIDE A CONVENIENT SYSTEM IN WHICH TO RAPIDLY AND QUANTITATIVELY EVALUATE ANTISENSE OLIGONUCLEOTIDES DIRECTED TOWARD LEADER RNA. PHASE I WORK WILL USE A PROVIRAL VECTOR DERIVED FROM A MODIFIED ROUS SARCOMA VIRUS CONTAINING THE HGH GENE SUBSTITUTED FOR THE SRC GENE. VIRAL INFECTIONS WILL BE ESTABLISHED IN CELL CULTURE USING THIS VECTOR, AND THE EFFECTS OF ANTISENSE OLIGONUCLEOTIDES ON RNA PROCESSING WILL BE MONITORED BY ASSAYING FOR HGH GENE EXPRESSION. OLIGONUCLEOTIDES OF VARIOUS LENGTHS, CONTAINING EITHER NORMAL PHOSPHODIESTER OR METHYLPHOSPHONATE LINKAGES, WILL BEEVALUATED FOR ANTISENSE ACTIVITY. THE ADDITION OF TERMINAL ACRIDINE RESIDUES TO THE OLIGONUCLEOTIDES WILL ALSO BE EVALUATED FOR THEIR EFFECT ON ANTISENSE ACTIVITY. DEMONSTRATION OF THE EFFICACY OF THIS APPROACH WILL LEAD TO A PHASE II EFFORT IN WHICH A HUMAN IMMUNODEFICIENCY VIRUS(HIV)-DIRECTED HGH GENE EXPRESSION SYSTEM WILL BE CONSTRUCTED FOR EVALUATION OF ANTISENSE PROBES PRIOR TO USE IN AN HIV-INFECTED CELL LINE.