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
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.