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Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency

Despite research efforts, cell uptake processes determining siRNA silencing efficiency remain unclear. Here, we examine the relationship between in vitro cell culture models, cellular trafficking and siRNA silencing efficiency to provide a mechanistic insight on siRNA delivery system design. Model s...

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Detalles Bibliográficos
Autores principales: Capel, Victoria, Vllasaliu, Driton, Watts, Peter, Stolnik, Snow
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4948577/
https://www.ncbi.nlm.nih.gov/pubmed/27349867
http://dx.doi.org/10.1016/j.bbrc.2016.06.054
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author Capel, Victoria
Vllasaliu, Driton
Watts, Peter
Stolnik, Snow
author_facet Capel, Victoria
Vllasaliu, Driton
Watts, Peter
Stolnik, Snow
author_sort Capel, Victoria
collection PubMed
description Despite research efforts, cell uptake processes determining siRNA silencing efficiency remain unclear. Here, we examine the relationship between in vitro cell culture models, cellular trafficking and siRNA silencing efficiency to provide a mechanistic insight on siRNA delivery system design. Model siRNA-polyplexes, based on chitosan as a ‘classical’ condensing agent, were applied to a panel of lung epithelial cell lines, H1299, A549 and Calu-3 and cell internalization levels, trafficking pathways and gene silencing assessed on exposure to pharmacological inhibitors. The data reveal striking differences in the internalization behaviour and gene silencing efficiency in the tested cell lines, despite their common lung epithelial origins. The model system’s silencing was lower where clathrin internalization pathway predominated in Calu-3, relative to silencing in H1299 cells where a non-clathrin internalization appears dominant. Increased silencing on endosomal disruption was apparent in Calu-3 cells, but absent when cellular internalization was not predominantly clathrin-mediated in A549 cells. This highlights that identifying cell trafficking pathways before incorporation of functional components to siRNA delivery systems (e.g. endosomolytic compounds) is crucial. The study hence stresses the importance of selection of appropriate cell culture model, relevant to in vivo target, to assess the gene silencing efficiency and decide which functionalities the ‘stratified siRNA silencing vector’ requires.
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spelling pubmed-49485772016-08-19 Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency Capel, Victoria Vllasaliu, Driton Watts, Peter Stolnik, Snow Biochem Biophys Res Commun Article Despite research efforts, cell uptake processes determining siRNA silencing efficiency remain unclear. Here, we examine the relationship between in vitro cell culture models, cellular trafficking and siRNA silencing efficiency to provide a mechanistic insight on siRNA delivery system design. Model siRNA-polyplexes, based on chitosan as a ‘classical’ condensing agent, were applied to a panel of lung epithelial cell lines, H1299, A549 and Calu-3 and cell internalization levels, trafficking pathways and gene silencing assessed on exposure to pharmacological inhibitors. The data reveal striking differences in the internalization behaviour and gene silencing efficiency in the tested cell lines, despite their common lung epithelial origins. The model system’s silencing was lower where clathrin internalization pathway predominated in Calu-3, relative to silencing in H1299 cells where a non-clathrin internalization appears dominant. Increased silencing on endosomal disruption was apparent in Calu-3 cells, but absent when cellular internalization was not predominantly clathrin-mediated in A549 cells. This highlights that identifying cell trafficking pathways before incorporation of functional components to siRNA delivery systems (e.g. endosomolytic compounds) is crucial. The study hence stresses the importance of selection of appropriate cell culture model, relevant to in vivo target, to assess the gene silencing efficiency and decide which functionalities the ‘stratified siRNA silencing vector’ requires. Elsevier 2016-08-19 /pmc/articles/PMC4948577/ /pubmed/27349867 http://dx.doi.org/10.1016/j.bbrc.2016.06.054 Text en © 2016 The Authors. Published by Elsevier Inc. http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Capel, Victoria
Vllasaliu, Driton
Watts, Peter
Stolnik, Snow
Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title_full Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title_fullStr Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title_full_unstemmed Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title_short Insight into the relationship between the cell culture model, cell trafficking and siRNA silencing efficiency
title_sort insight into the relationship between the cell culture model, cell trafficking and sirna silencing efficiency
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4948577/
https://www.ncbi.nlm.nih.gov/pubmed/27349867
http://dx.doi.org/10.1016/j.bbrc.2016.06.054
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