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Effect of small molecule signaling in PepFect14 transfection

Cell-penetrating peptides can be used to deliver oligonucleotide-based cargoes into cells. Previous studies have shown that the use of small molecule drugs could be an efficient method to increase the efficacy of delivery of oligonucleotides by cell-penetrating peptides either as targeting agents th...

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Autores principales: Gestin, Maxime, Helmfors, Henrik, Falato, Luca, Lorenzon, Nicola, Michalakis, Filip Ilias, Langel, Ülo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992163/
https://www.ncbi.nlm.nih.gov/pubmed/31999754
http://dx.doi.org/10.1371/journal.pone.0228189
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author Gestin, Maxime
Helmfors, Henrik
Falato, Luca
Lorenzon, Nicola
Michalakis, Filip Ilias
Langel, Ülo
author_facet Gestin, Maxime
Helmfors, Henrik
Falato, Luca
Lorenzon, Nicola
Michalakis, Filip Ilias
Langel, Ülo
author_sort Gestin, Maxime
collection PubMed
description Cell-penetrating peptides can be used to deliver oligonucleotide-based cargoes into cells. Previous studies have shown that the use of small molecule drugs could be an efficient method to increase the efficacy of delivery of oligonucleotides by cell-penetrating peptides either as targeting agents that can be used in formulation with the cell-penetrating peptide and its cargo or as cell signaling modulators that facilitates the cellular uptake of the treatment. This study presents two aims. The first aim is the identification of small molecule drugs that would induce a synergic effect on the transfection of splice correcting oligonucleotides assisted by PepFect14. The second aim is to identify the mechanisms behind the effect of small molecule drugs modulation of cell-penetrating peptide assisted transfection of oligonucleotides. Through an optimized, high-throughput luciferase assay for short oligonucleotide delivery using cell-penetrating peptides, and the simultaneous addition of a small molecule drug library, we show that three small molecule drugs (MPEP, VU0357121 and Ciproxifan) induced an increase in the transfection efficacy of PepFect14 in complex with a short single-stranded oligonucleotide in HeLa pLuc705 cells. These three drugs are described in the literature to be highly specific for their respective target receptors. However, none of those receptors are expressed in our cell line, indicating a yet non-described pathway of action for these small molecules. We show that the indicated small molecules, without interfering with the particles formed by PepFect14 and the oligonucleotide, interfere via still unidentified interactions in cell signaling, leading to an up-regulation of endocytosis and a higher efficacy in the delivery of short splice correcting oligonucleotides in complex with PepFect14.
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spelling pubmed-69921632020-02-20 Effect of small molecule signaling in PepFect14 transfection Gestin, Maxime Helmfors, Henrik Falato, Luca Lorenzon, Nicola Michalakis, Filip Ilias Langel, Ülo PLoS One Research Article Cell-penetrating peptides can be used to deliver oligonucleotide-based cargoes into cells. Previous studies have shown that the use of small molecule drugs could be an efficient method to increase the efficacy of delivery of oligonucleotides by cell-penetrating peptides either as targeting agents that can be used in formulation with the cell-penetrating peptide and its cargo or as cell signaling modulators that facilitates the cellular uptake of the treatment. This study presents two aims. The first aim is the identification of small molecule drugs that would induce a synergic effect on the transfection of splice correcting oligonucleotides assisted by PepFect14. The second aim is to identify the mechanisms behind the effect of small molecule drugs modulation of cell-penetrating peptide assisted transfection of oligonucleotides. Through an optimized, high-throughput luciferase assay for short oligonucleotide delivery using cell-penetrating peptides, and the simultaneous addition of a small molecule drug library, we show that three small molecule drugs (MPEP, VU0357121 and Ciproxifan) induced an increase in the transfection efficacy of PepFect14 in complex with a short single-stranded oligonucleotide in HeLa pLuc705 cells. These three drugs are described in the literature to be highly specific for their respective target receptors. However, none of those receptors are expressed in our cell line, indicating a yet non-described pathway of action for these small molecules. We show that the indicated small molecules, without interfering with the particles formed by PepFect14 and the oligonucleotide, interfere via still unidentified interactions in cell signaling, leading to an up-regulation of endocytosis and a higher efficacy in the delivery of short splice correcting oligonucleotides in complex with PepFect14. Public Library of Science 2020-01-30 /pmc/articles/PMC6992163/ /pubmed/31999754 http://dx.doi.org/10.1371/journal.pone.0228189 Text en © 2020 Gestin et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Gestin, Maxime
Helmfors, Henrik
Falato, Luca
Lorenzon, Nicola
Michalakis, Filip Ilias
Langel, Ülo
Effect of small molecule signaling in PepFect14 transfection
title Effect of small molecule signaling in PepFect14 transfection
title_full Effect of small molecule signaling in PepFect14 transfection
title_fullStr Effect of small molecule signaling in PepFect14 transfection
title_full_unstemmed Effect of small molecule signaling in PepFect14 transfection
title_short Effect of small molecule signaling in PepFect14 transfection
title_sort effect of small molecule signaling in pepfect14 transfection
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6992163/
https://www.ncbi.nlm.nih.gov/pubmed/31999754
http://dx.doi.org/10.1371/journal.pone.0228189
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