Second-generation antisense oligonucleotides: structure—activity relationships and the design of improved signal-transduction inhibitors

Altmann, Karl Heinz; Fabbro, Doriano; Dean, Nicholas M.; Geiger, Thomas; Monia, Brett P.; Müller, Matthias; Nicklin, Paul

doi:10.1042/bst0240630

Cited by 104 publications

(95 citation statements)

References 8 publications

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“…To do this, 22 oligonucleotides designed to hybridize to multiple sites on the human Bcl-x L mRNA (including the 5' untranslated region, coding region, and 3' untranslated region) were synthesized as 20-mer 2'-O-methoxyethyl chimeric oligonucleotides (Altmann et al, 1996) and tested for their ability to inhibit Bcl-x L expression. ISIS 16009 was selected as the Bcl-x L inhibitor for this study because it greatly inhibited Bcl-x L mRNA expression ( Figure 2).…”

Section: Selection Of a Bcl-x L Antisense Oligonucleotide Inhibitormentioning

confidence: 99%

Inhibition of Bcl-xL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli

et al. 1999

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The epidermis is continually exposed to harmful mutagens that have the potential to cause DNA damage. To protect the skin from accumulating mutated cells, keratinocytes have developed a highly regulated mechanism of eliminating damaged cells through apoptosis. Bclx L is a well-described cell survival protein that when overexpressed in skin can protect keratinocytes from UV radiation-induced apoptosis. To begin to unravel the complex mechanisms that keratinocytes use to survive, we wanted to characterize the role of endogenous Bcl-x L in protecting cells from death. In this study, we describe the development and characterization of an antisense inhibitor to Bcl-x L . We show that this inhibitor reduces Bcl-x L RNA and protein in a concentration-dependent, sequence-speci®c manner. Furthermore, treatment of keratinocytes and epithelial cells with this inhibitor sensitizes these cells to UV-B radiation and cisplatinum treatment-induced apoptosis. Thus, these results o er direct evidence that Bcl-x L is critical in the protection of skin and epithelial cells from apoptosis and provide a basis for the role of Bcl-x L in keratinocyte and epithelial cell survival.

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Section: Selection Of a Bcl-x L Antisense Oligonucleotide Inhibitormentioning

confidence: 99%

Inhibition of Bcl-xL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli

et al. 1999

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“…Additional modifications of oligonucleotides have focused on increasing further the stability of these macromolecules and increasing target hybridization affinity. The primary modifcations currently being explored involve the 2' position on the ribose and the linkages of the nucleotide backbone (2,9). The 2' position modifications involve alkylation and halogenation ; the primary intent of these modifications is to protect a phosphodiester backbone from nuclease degradation.…”

Section: Antisense Therapeuticsmentioning

confidence: 99%

Synthetic Oligonucleotides: The Development of Antisense Therapeutics

Monteith

Levin

1999

Toxicol Pathol

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Antisense therapeutics using synthetic oligodeoxynucleotides (ODNs) Antisense therapeutics are species specific because they are based on the genomic code. Although sequences may be relatively conserved among species, typical target regions for antisense therapeutics will differ by at least several bases between species. Therefore, to define pharmacologic effects, species-specific homologues of the human target must be identified and tested in the appropriate animal model. One exception is the use of human

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“…Further modifications in the sugar moiety have been engineered into these ASOs to generate a variety of second-generation structures that include 2 0 -Omethoxyethyl-substituted (2 0 -MOE) ribonucleosides. 16 These oligonucleotides have increased RNA binding activity, superior nuclease resistance, greater potency and selective inhibition of mRNA/ protein expression but less toxicity than the phosphorothioate-only structures. 16 ISIS 13650 is an example of a 2 0 -MOE ASO targeted against Raf-1 and has the same base sequence as that of ISIS 5132.…”

mentioning

confidence: 99%

“…16 These oligonucleotides have increased RNA binding activity, superior nuclease resistance, greater potency and selective inhibition of mRNA/ protein expression but less toxicity than the phosphorothioate-only structures. 16 ISIS 13650 is an example of a 2 0 -MOE ASO targeted against Raf-1 and has the same base sequence as that of ISIS 5132. This ASO has been shown to have higher RNA-binding affinity than ISIS 5132 and inhibits Raf-1 at a lower IC 50 .…”

mentioning

confidence: 99%

“…This ASO has been shown to have higher RNA-binding affinity than ISIS 5132 and inhibits Raf-1 at a lower IC 50 . 16 An alternative strategy of mRNA knockdown, termed RNA interference (RNAi), has been developed recently, following the observation that double-stranded RNAs (dsRNAs) elicit potent degradation of complementary RNA sequences. 17 The active component of this pathway (small interfering RNAs or siRNAs) can be chemically synthesized and designed to target specific mRNA sequences.…”

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confidence: 99%

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Comparison of strategies targeting Raf-1 mRNA in ovarian cancer

et al. 2005

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In this study, we characterize the uptake and specificity of a firstgeneration Raf-1 antisense oligonucleotide (ASO) (ISIS 5132) and compare it with a second-generation ASO (ISIS 13650) and an RNA interference approach. All three approaches resulted in inhibition of both Raf-1 expression and cellular growth. Specificity of the Raf-1 ASOs was confirmed by comparison with ASOs targeted against another Raf isoform (B-Raf) as well as mismatch sequences. Cellular uptake studies with FAM-labelled ISIS 5132 revealed that whilst the majority of cells treated at a low-intermediate plating density were labelled within 3 hr, cells treated at high density demonstrated neither Raf-1 protein knockout nor significant growth inhibition, following similar treatment. This lack of response at high cell densities was associated with reduced pERK and Raf-1 inhibition. Cell cycle analysis revealed that whilst SKOV-3 cells both accumulated in the S-phase of the cell cycle and showed enhanced annexin V levels, following Raf-1 ASO treatment; these effects were also demonstrated with first-generation but not second-generation mismatch oligonucleotides. Bromodeoxyuridine incorporation analysis suggested that these effects may indeed be partly attributable to sequence nonspecific effects. Finally, the combination of ISIS 5132 with either carboplatin or taxol showed enhanced growth inhibition, supporting the view that such ASOs may have a more effective clinical role when used in combination with cytotoxic agents. ' 2005 Wiley-Liss, Inc.

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Second-generation antisense oligonucleotides: structure—activity relationships and the design of improved signal-transduction inhibitors

Cited by 104 publications

References 8 publications

Inhibition of Bcl-xL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli

Inhibition of Bcl-xL expression sensitizes normal human keratinocytes and epithelial cells to apoptotic stimuli

Synthetic Oligonucleotides: The Development of Antisense Therapeutics

Comparison of strategies targeting Raf-1 mRNA in ovarian cancer

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