Enhanced biological activity of antisense oligonucleotides complexed with glycosylated poly-L-lysine.
Identifieur interne : 000408 ( France/Analysis ); précédent : 000407; suivant : 000409Enhanced biological activity of antisense oligonucleotides complexed with glycosylated poly-L-lysine.
Auteurs : A. Stewart ; Chantal Pichon [France] ; L. Meunier ; P. Midoux [France] ; M. Monsigny [France] ; A. Roche [France]Source :
- Molecular Pharmacology [ 0026-895X ] ; 1996-12.
Abstract
We sought to exploit glycosylated poly-L-lysine (pLK) to increase the uptake and biological antisense activity of a phosphorothioate oligonucleotide (pt-odn) [pt-odn complementary to the 3' noncoding region of intercellular adhesion molecule-1 (ICAM-1) (odn(ICAM-1))] complementary to the 3'-noncoding region of ICAM-1 in A549 cells. Dose-dependent inhibition of ICAM-1 expression was obtained (IC50 = 500 nM) through treatment of cells with odn(ICAM-1) complexed with pLK carrying fucose residues in the presence of 100 microM chloroquine. Alteration in the charge ratio between fucosylated pLK and pt-odn had a significant effect on the efficacy of inhibition (optimal conditions, charge ratio = 1.1). This effect was also dependent on the number of fucose moieties per pLK. Free pt-odn or pt-odn complexed with nonglycosylated pLK gave no inhibition at concentrations of < or = 2 microM. Two control pt-odn (one was targeted against an unrelated gene not present in these cells, gag(HIV), and the other had a randomized sequence) gave no inhibition of ICAM-1 expression in the presence or absence of pLK carrying fucose residues at concentrations of < or = 2 microM. When complexed with pLK carrying 100 fucose residues, the amount of cell-associated pt-odn was increased by 15-fold compared with the free pt-odn. Nongycosylated pLK also increased the amount of cell-associated pt-odn by >10 fold but did not alter the biological activity. These results demonstrate clearly the potential of glycosylated pLK as a pt-odn transporter.
Url:
Affiliations:
- France
- Centre-Val de Loire, Région Centre
- Orléans
- Centre Val de Loire Université, Université d'Orléans
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Links to Exploration step
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<orgName>Université d'Orléans</orgName>
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<desc> <address> <addrLine>Château de la Source - Avenue du Parc Floral - BP 6749 - 45067 Orléans cedex 2</addrLine>
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<ref type="url">http://www.univ-orleans.fr/</ref>
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<orgName>Institut National de la Santé et de la Recherche Médicale</orgName>
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<desc> <address> <addrLine>101, rue de Tolbiac, 75013 Paris </addrLine>
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<orgName>Centre National de la Recherche Scientifique</orgName>
<orgName type="acronym">CNRS</orgName>
<date type="start">1939-10-19</date>
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<country>France</country>
<placeName><settlement type="city">Orléans</settlement>
<region type="old region" nuts="2">Région Centre</region>
<region type="region" nuts="2">Centre-Val de Loire</region>
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<orgName type="university">Université d'Orléans</orgName>
<orgName type="institution" wicri:auto="newGroup">Centre Val de Loire Université</orgName>
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<author><name sortKey="Roche, A" sort="Roche, A" uniqKey="Roche A" first="A." last="Roche">A. Roche</name>
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<orgName>Centre de biophysique moléculaire</orgName>
<orgName type="acronym">CBM</orgName>
<date type="start">1967-01-01</date>
<desc> <address> <addrLine>Rue Charles Sadron 45071 ORLEANS CEDEX 2</addrLine>
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</address>
<ref type="url">http://cbm.cnrs-orleans.fr/</ref>
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<orgName>Université d'Orléans</orgName>
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<desc> <address> <addrLine>Château de la Source - Avenue du Parc Floral - BP 6749 - 45067 Orléans cedex 2</addrLine>
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<orgName>Institut National de la Santé et de la Recherche Médicale</orgName>
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<orgName>Centre National de la Recherche Scientifique</orgName>
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<date type="start">1939-10-19</date>
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</hal:affiliation>
<country>France</country>
<placeName><settlement type="city">Orléans</settlement>
<region type="old region" nuts="2">Région Centre</region>
<region type="region" nuts="2">Centre-Val de Loire</region>
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<orgName type="university">Université d'Orléans</orgName>
<orgName type="institution" wicri:auto="newGroup">Centre Val de Loire Université</orgName>
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<series><title level="j">Molecular Pharmacology</title>
<idno type="ISSN">0026-895X</idno>
<imprint><date type="datePub">1996-12</date>
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<front><div type="abstract" xml:lang="en"> <p>We sought to exploit glycosylated poly-L-lysine (pLK) to increase the uptake and biological antisense activity of a phosphorothioate oligonucleotide (pt-odn) [pt-odn complementary to the 3' noncoding region of intercellular adhesion molecule-1 (ICAM-1) (odn(ICAM-1))] complementary to the 3'-noncoding region of ICAM-1 in A549 cells. Dose-dependent inhibition of ICAM-1 expression was obtained (IC50 = 500 nM) through treatment of cells with odn(ICAM-1) complexed with pLK carrying fucose residues in the presence of 100 microM chloroquine. Alteration in the charge ratio between fucosylated pLK and pt-odn had a significant effect on the efficacy of inhibition (optimal conditions, charge ratio = 1.1). This effect was also dependent on the number of fucose moieties per pLK. Free pt-odn or pt-odn complexed with nonglycosylated pLK gave no inhibition at concentrations of < or = 2 microM. Two control pt-odn (one was targeted against an unrelated gene not present in these cells, gag(HIV), and the other had a randomized sequence) gave no inhibition of ICAM-1 expression in the presence or absence of pLK carrying fucose residues at concentrations of < or = 2 microM. When complexed with pLK carrying 100 fucose residues, the amount of cell-associated pt-odn was increased by 15-fold compared with the free pt-odn. Nongycosylated pLK also increased the amount of cell-associated pt-odn by >10 fold but did not alter the biological activity. These results demonstrate clearly the potential of glycosylated pLK as a pt-odn transporter.</p>
</div>
</front>
</TEI>
<affiliations><list><country><li>France</li>
</country>
<region><li>Centre-Val de Loire</li>
<li>Région Centre</li>
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<settlement><li>Orléans</li>
</settlement>
<orgName><li>Centre Val de Loire Université</li>
<li>Université d'Orléans</li>
</orgName>
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<tree><noCountry><name sortKey="Meunier, L" sort="Meunier, L" uniqKey="Meunier L" first="L" last="Meunier">L. Meunier</name>
<name sortKey="Stewart, A" sort="Stewart, A" uniqKey="Stewart A" first="A." last="Stewart">A. Stewart</name>
</noCountry>
<country name="France"><region name="Région Centre"><name sortKey="Pichon, Chantal" sort="Pichon, Chantal" uniqKey="Pichon C" first="Chantal" last="Pichon">Chantal Pichon</name>
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<name sortKey="Midoux, P" sort="Midoux, P" uniqKey="Midoux P" first="P" last="Midoux">P. Midoux</name>
<name sortKey="Monsigny, M" sort="Monsigny, M" uniqKey="Monsigny M" first="M." last="Monsigny">M. Monsigny</name>
<name sortKey="Roche, A" sort="Roche, A" uniqKey="Roche A" first="A." last="Roche">A. Roche</name>
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