THE INFLUENCE OF TARGET PROTEIN HALF-LIFE ON THE EFFECTIVENESS OF ANTISENSE OLIGONUCLEOTIDE ANALOG-MEDIATED BIOLOGIC RESPONSES

Citation
Dg. Spiller et al., THE INFLUENCE OF TARGET PROTEIN HALF-LIFE ON THE EFFECTIVENESS OF ANTISENSE OLIGONUCLEOTIDE ANALOG-MEDIATED BIOLOGIC RESPONSES, Antisense & nucleic acid drug development, 8(4), 1998, pp. 281-293
Citations number
31
Categorie Soggetti
Biology,"Medicine, Research & Experimental","Biothechnology & Applied Migrobiology
ISSN journal
10872906
Volume
8
Issue
4
Year of publication
1998
Pages
281 - 293
Database
ISI
SICI code
1087-2906(1998)8:4<281:TIOTPH>2.0.ZU;2-0
Abstract
During the course of a study aimed at improving antisense oligodeoxynu cleotide-mediated ex vivo bone marrow purging of patients suffering fr om chronic myeloid leukemia (CML), the properties of a number of antis ense structures intended to reduce the expression of c-myc, mutant p53 , and bcr-abl mRNAs and proteins were examined. The majority of the an tisense oligodeoxynucleotides were designed to be capable of directing ribonuclease H (RNase H) cleavage of their target mRNAs, Streptolysin O (SLO) reversible permeabilization was used to deliver the oligodeox ynucleotides into the CML line KYO-1, We found that the efficiency and specificity of antisense oligonucleotide-induced reductions of target protein expression depended on target protein half-life, the oligonuc leotide structure, and the specific sequence within the target mRNA, T ransient reductions of c-myc mRNA and protein were achieved with a chi meric methylphosphonate-phosphodiester oligodeoxynucleotide antisense to the initiation codon, but cell proliferation was unaffected. In con trast, a chimeric oligodeoxynucleotide of similar structure targeted t o an alternative site in the coding region of c-myc mRNA reduced targe t mRNA and protein levels for over 24 hours and halted cell proliferat ion. Chimeric methylphosphonate-phosphodiester oligodeoxynucleotide an tisense to a point mutation in KYO-1 p53 mRNA efficiently reduced targ et mRNA expression, but only small, transient reductions in p53 protei n expression were observed. However, a chimeric methylphosphonate-phos phorothioate oligodeoxynucleotide targeted to the same site reduced p5 3 protein to 30% of control levels over a 48-hour period. BCR-ABL prot ein expression was unaffected by chimeric oligodeoxynucleotides target ed to the breakpoint in bcr-abl mRNA, even when mRNA levels at early t imes were substantially reduced.