Inhibition of High Affinity Basic Fibroblast Growth Factor Binding by Oligonucleotides

Abstract

Oligonucleotides can be used to inhibit the binding of basic fibroblast growth factor to cells. Though standard phosphodiester oligonucleotides show a slight inhibition of binding, the oligonucleotides with phosphorothioate internucleoside linkages have inhibition levels equivalent to that of the polyanion heparin. Variations in sequence of the oligonucleotides does lead to differences in the inhibitory action of the oligonucleotides. This inhibition of basic fibroblast growth factor by phosphorothioate oligonucleotides may account for much of the published data on inhibition of various genes by proposed antisense oligonucleotides and needs to be taken into account when considering the mechanism of action of oligonucleotides in biological systems.

Phosphorothioate-Modified TLR9 Ligands Protect Cancer Cells against TRAIL-Induced Apoptosis

“…In agreement with this finding, it has been previously shown that the phosphorothioate backbone of ODN sequences strongly increased nonspecific binding to various proteins. In particular, phosphorothioate sequences inhibit basic fibroblast growth factor binding to its receptor (31,32). This stickiness of phosphorothioate sequences is also observed in the binding to TLR9, as a phosphorothioate-modified sequence, regardless of the bases, has a higher TLR9 affinity than does the same phosphodiester sequence (29).…”

Suppression of fibroblast growth factor 2 expression by antisense oligonucleotides inhibits embryonic chick neural retina cell differentiation and survival in vivo

“…Because 3Ј exonucleases are thought to be more active than 5Ј exonucleases, an amine group was added at the 3Ј end, a modification that has been shown previously to enhance both the stability of classical phosphodiester antisense ODNs to levels comparable to phosphorothioate ODNs and the stability of the ODN-target complex (Orson et al, 1991;Svinarchuk et al, 1996). These phosphodiester ODNs do not exhibit toxicity or heparin-like activity by themselves, in contrast to phosphorothioate ODNs (Benimetskaya et al, 1995;Fennewald and Rando, 1995;Guvakova et al, 1995). We used Lipofectin as a cationic lipid to deliver the ODNs, because this method results in high uptake and stability of phosphodiester ODNs in the intracellular compartment (Bennet et al, 1992;Cappacioli et al, 1993) without changing their final nuclear location (Loke et al, 1989;Léonetti et al, 1991;Clarenc et al, 1993;Lezoualc'h et al, 1995;Svinarchuk et al, 1996) after endocytosis and escape from the endocytic compartment.…”

Applications of CpG Motifs from Bacterial DNA in Cancer Immunotherapy

“…The phosphorothioate ODN backbone dramatically increases the non-specific ODN binding to a wide variety of proteins [69,70] Ç Phosphorothioate ODN bind much more avidly to cell membranes, and generally have a much higher degree of cell uptake [63,71,72] Ç The phosphorothioate backbone results in sequence-independent activities including the activation of SP1 transcription factor activity [73], inhibition of smooth muscle cell proliferation and migration [74,75] Ë inhibition of basic fibroblast growth factor binding to its receptor [76,77] and angiogenic activity [78], reduction of the sequence specific binding of transcription factors to their binding sites [79], inhibition of cellular adhesion to extracellular matrix [80], enhancement of LPS-induced TNF production [81], and some degree of non-sequence specific immune stimulation [82]. The immune stimulatory effects of PS ODN are reduced by further modification with 2' methoxyethoxy modifications [83].…”