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A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells

Synthetic double-stranded small interfering RNAs (siRNAs) mimic interference RNAs (RNAi) and can bind target mRNAs with a high degree of specificity, leading to selective knockdown of the proteins they encode. However, siRNAs are very labile and must be both protected and transported by nanoparticle...

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Autores principales: de la Torre, Cristina, Játiva, Pablo, Posadas, Inmaculada, Manzanares, Darío, Blanco, José L. Jiménez, Mellet, Carmen Ortiz, Fernández, José Manuel García, Ceña, Valentín
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692777/
https://www.ncbi.nlm.nih.gov/pubmed/36365241
http://dx.doi.org/10.3390/pharmaceutics14112424
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author de la Torre, Cristina
Játiva, Pablo
Posadas, Inmaculada
Manzanares, Darío
Blanco, José L. Jiménez
Mellet, Carmen Ortiz
Fernández, José Manuel García
Ceña, Valentín
author_facet de la Torre, Cristina
Játiva, Pablo
Posadas, Inmaculada
Manzanares, Darío
Blanco, José L. Jiménez
Mellet, Carmen Ortiz
Fernández, José Manuel García
Ceña, Valentín
author_sort de la Torre, Cristina
collection PubMed
description Synthetic double-stranded small interfering RNAs (siRNAs) mimic interference RNAs (RNAi) and can bind target mRNAs with a high degree of specificity, leading to selective knockdown of the proteins they encode. However, siRNAs are very labile and must be both protected and transported by nanoparticles to be efficiently delivered into cells. In this work, we used a Janus-type polycationic amphiphilic β-cyclodextrin derivative to efficiently transfect siRNAs targeting mRNAs encoding mitogen-activated protein kinase (p42-MAPK) or Ras homolog enriched in brain (Rheb) into different cancer cell lines as well as astrocytes. We took advantage of this high transfection efficiency to simultaneously knock down p42-MAPK and Rheb to boost docetaxel (DTX)-mediated toxicity in two human prostate cancer cell lines (LNCaP and PC3). We found that double knockdown of p42-MAPK and Rheb increased DTX-toxicity in LNCaP but not in PC3 cells. However, we also observed the same effect when scramble siRNA was used, therefore pointing to an off-target effect. Indeed, we found that the siRNA we used in this work induced toll-like receptor 3 activation, leading to β-interferon production and caspase activation. We believe that this mechanism could be very useful as a general strategy to elicit an immune response against prostate cancer cells.
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spelling pubmed-96927772022-11-26 A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells de la Torre, Cristina Játiva, Pablo Posadas, Inmaculada Manzanares, Darío Blanco, José L. Jiménez Mellet, Carmen Ortiz Fernández, José Manuel García Ceña, Valentín Pharmaceutics Article Synthetic double-stranded small interfering RNAs (siRNAs) mimic interference RNAs (RNAi) and can bind target mRNAs with a high degree of specificity, leading to selective knockdown of the proteins they encode. However, siRNAs are very labile and must be both protected and transported by nanoparticles to be efficiently delivered into cells. In this work, we used a Janus-type polycationic amphiphilic β-cyclodextrin derivative to efficiently transfect siRNAs targeting mRNAs encoding mitogen-activated protein kinase (p42-MAPK) or Ras homolog enriched in brain (Rheb) into different cancer cell lines as well as astrocytes. We took advantage of this high transfection efficiency to simultaneously knock down p42-MAPK and Rheb to boost docetaxel (DTX)-mediated toxicity in two human prostate cancer cell lines (LNCaP and PC3). We found that double knockdown of p42-MAPK and Rheb increased DTX-toxicity in LNCaP but not in PC3 cells. However, we also observed the same effect when scramble siRNA was used, therefore pointing to an off-target effect. Indeed, we found that the siRNA we used in this work induced toll-like receptor 3 activation, leading to β-interferon production and caspase activation. We believe that this mechanism could be very useful as a general strategy to elicit an immune response against prostate cancer cells. MDPI 2022-11-09 /pmc/articles/PMC9692777/ /pubmed/36365241 http://dx.doi.org/10.3390/pharmaceutics14112424 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
de la Torre, Cristina
Játiva, Pablo
Posadas, Inmaculada
Manzanares, Darío
Blanco, José L. Jiménez
Mellet, Carmen Ortiz
Fernández, José Manuel García
Ceña, Valentín
A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title_full A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title_fullStr A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title_full_unstemmed A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title_short A β-Cyclodextrin-Based Nanoparticle with Very High Transfection Efficiency Unveils siRNA-Activated TLR3 Responses in Human Prostate Cancer Cells
title_sort β-cyclodextrin-based nanoparticle with very high transfection efficiency unveils sirna-activated tlr3 responses in human prostate cancer cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9692777/
https://www.ncbi.nlm.nih.gov/pubmed/36365241
http://dx.doi.org/10.3390/pharmaceutics14112424
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