Processing of Chimeric Antisense Oligonucleotides by Human Vascular Smooth Muscle Cells and Human Atherosclerotic Plaque

Abstract

S-chimeric oligonucleotides are stable and can specifically inhibit gene expression in human VSMCs. Nuclear transport is a feature of oligonucleotide processing by human VSMCs, indicating a potential influence at the nuclear level rather than with cytoplasmic mRNA. Cationic liposomes increased oligonucleotide uptake but not intracellular bioavailability, and S-chimeric oligonucleotides can be incorporated into cells within human atherosclerotic plaque, despite the presence of a dense extracellular matrix.

Central Role of the MAPK Pathway in Ang II–Mediated DNA Synthesis and Migration in Rat Vascular Smooth Muscle Cells

“…More recently, Lu et al 22 have used these ODNs to deplete ERK1 and ERK2 in rat VSMCs. Although transfection of VSMCs with plasmid DNA occurs with low efficiency, ODNs complexed with liposomes transfect VSMCs at high efficiency; a recent study by Pickering et al 23 showed that virtually all VSMCs treated with as little as 0.2 mol/L for 1 hour incorporated the ODN into both the nuclear and the cytoplasmic compartments. A similar high efficiency of transfection can be inferred from our results, which showed that MAPK protein levels were depleted by 65% in antisense ODN-treated VSMCs compared with sense and scrambled ODNs ( Figure 5).…”

Endoproteolytic Activation of α v Integrin by Proprotein Convertase PC5 Is Required for Vascular Smooth Muscle Cell Adhesion to Vitronectin and Integrin-Dependent Signaling

“…A 19-mer asODNs targeting ␣ 1 -collagen was used as additional control. 21 Transfection was done with the help of OligofectAMINE (GIBCO), used according to the manufacturer. Briefly, subconfluent VSMCs were growth arrested overnight.…”

Antisense oligodeoxynucleotides of sulfonylurea receptors inhibit ATP-sensitive K + channels in cultured neonatal rat ventricular cells

“…They were designed as phosphorothioate-modified ODNs (18-meric) complementary to the sequence adjacent to the initiation codon of SUR1 or SUR2 (A and B) mRNA (+22 to about +39), and designated as AS-SUR1 and AS-SUR2, respectively. The phosphorothioate modification was made at the first four and last four internucleotide linkages as recommended previously [28,32,34,35]. AS prepared against SUR2A and 2B was termed AS-SUR2 because cDNA sequences of SUR2A and 2B are identical at this region (see below).…”

“…The cells were treated with antisense oligodeoxynucleotides (AS-ODNs) complementary to the mRNA for each type of SUR. Phosphorothioate modification of these AS-ODNs (18-meric) was restricted to four internucleotide linkages at both 5′-and 3′-terminals to diminish possible non-specific effects [9,14,28,34,39]. This chimeric modification by phosphorothioate has been reported to retain stability against nucleases [28,32,34,35].…”