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Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model

Control of the expression of oncogenic small non-coding RNAs, notably microRNAs (miRNAs), is an attractive therapeutic approach. We report a design platform for catalytic knockdown of miRNA targets with artificial, sequence-specific ribonucleases. miRNases comprise a peptide [(LeuArg)(2)Gly](2) capa...

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Autores principales: Patutina, Olga A., Miroshnichenko, Svetlana K., Mironova, Nadezhda L., Sen’kova, Aleksandra V., Bichenkova, Elena V., Clarke, David J., Vlassov, Valentin V., Zenkova, Marina A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698794/
https://www.ncbi.nlm.nih.gov/pubmed/31456683
http://dx.doi.org/10.3389/fphar.2019.00879
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author Patutina, Olga A.
Miroshnichenko, Svetlana K.
Mironova, Nadezhda L.
Sen’kova, Aleksandra V.
Bichenkova, Elena V.
Clarke, David J.
Vlassov, Valentin V.
Zenkova, Marina A.
author_facet Patutina, Olga A.
Miroshnichenko, Svetlana K.
Mironova, Nadezhda L.
Sen’kova, Aleksandra V.
Bichenkova, Elena V.
Clarke, David J.
Vlassov, Valentin V.
Zenkova, Marina A.
author_sort Patutina, Olga A.
collection PubMed
description Control of the expression of oncogenic small non-coding RNAs, notably microRNAs (miRNAs), is an attractive therapeutic approach. We report a design platform for catalytic knockdown of miRNA targets with artificial, sequence-specific ribonucleases. miRNases comprise a peptide [(LeuArg)(2)Gly](2) capable of RNA cleavage conjugated to the miRNA-targeted oligodeoxyribonucleotide, which becomes nuclease-resistant within the conjugate design, without resort to chemically modified nucleotides. Our data presented here showed for the first time a truly catalytic character of our miR-21-miRNase and its ability to cleave miR-21 in a multiple catalytic turnover mode. We demonstrate that miRNase targeted to miR-21 (miR-21-miRNase) knocked down malignant behavior of tumor cells, including induction of apoptosis, inhibition of cell invasiveness, and retardation of tumor growth, which persisted on transplantation into mice of tumor cells treated once with miR-21-miRNase. Crucially, we discover that the high biological activity of miR-21-miRNase can be directly related not only to its truly catalytic sequence-specific cleavage of miRNA but also to its ability to recruit the non-sequence specific RNase H found in most cells to elevate catalytic turnover further. miR-21-miRNase worked synergistically even with low levels of RNase H. Estimated degradation in the presence of RNase H exceeded 10(3) miRNA target molecules per hour for each miR-21-miRNase molecule, which provides the potency to minimize delivery requirements to a few molecules per cell. In contrast to the comparatively high doses required for the simple steric block of antisense oligonucleotides, truly catalytic inactivation of miRNA offers more effective, irreversible, and persistent suppression of many copy target sequences. miRNase design can be readily adapted to target other pathogenic microRNAs overexpressed in many disease states.
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spelling pubmed-66987942019-08-27 Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model Patutina, Olga A. Miroshnichenko, Svetlana K. Mironova, Nadezhda L. Sen’kova, Aleksandra V. Bichenkova, Elena V. Clarke, David J. Vlassov, Valentin V. Zenkova, Marina A. Front Pharmacol Pharmacology Control of the expression of oncogenic small non-coding RNAs, notably microRNAs (miRNAs), is an attractive therapeutic approach. We report a design platform for catalytic knockdown of miRNA targets with artificial, sequence-specific ribonucleases. miRNases comprise a peptide [(LeuArg)(2)Gly](2) capable of RNA cleavage conjugated to the miRNA-targeted oligodeoxyribonucleotide, which becomes nuclease-resistant within the conjugate design, without resort to chemically modified nucleotides. Our data presented here showed for the first time a truly catalytic character of our miR-21-miRNase and its ability to cleave miR-21 in a multiple catalytic turnover mode. We demonstrate that miRNase targeted to miR-21 (miR-21-miRNase) knocked down malignant behavior of tumor cells, including induction of apoptosis, inhibition of cell invasiveness, and retardation of tumor growth, which persisted on transplantation into mice of tumor cells treated once with miR-21-miRNase. Crucially, we discover that the high biological activity of miR-21-miRNase can be directly related not only to its truly catalytic sequence-specific cleavage of miRNA but also to its ability to recruit the non-sequence specific RNase H found in most cells to elevate catalytic turnover further. miR-21-miRNase worked synergistically even with low levels of RNase H. Estimated degradation in the presence of RNase H exceeded 10(3) miRNA target molecules per hour for each miR-21-miRNase molecule, which provides the potency to minimize delivery requirements to a few molecules per cell. In contrast to the comparatively high doses required for the simple steric block of antisense oligonucleotides, truly catalytic inactivation of miRNA offers more effective, irreversible, and persistent suppression of many copy target sequences. miRNase design can be readily adapted to target other pathogenic microRNAs overexpressed in many disease states. Frontiers Media S.A. 2019-08-08 /pmc/articles/PMC6698794/ /pubmed/31456683 http://dx.doi.org/10.3389/fphar.2019.00879 Text en Copyright © 2019 Patutina, Miroshnichenko, Mironova, Sen’kova, Bichenkova, Clarke, Vlassov and Zenkova http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Pharmacology
Patutina, Olga A.
Miroshnichenko, Svetlana K.
Mironova, Nadezhda L.
Sen’kova, Aleksandra V.
Bichenkova, Elena V.
Clarke, David J.
Vlassov, Valentin V.
Zenkova, Marina A.
Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title_full Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title_fullStr Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title_full_unstemmed Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title_short Catalytic Knockdown of miR-21 by Artificial Ribonuclease: Biological Performance in Tumor Model
title_sort catalytic knockdown of mir-21 by artificial ribonuclease: biological performance in tumor model
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6698794/
https://www.ncbi.nlm.nih.gov/pubmed/31456683
http://dx.doi.org/10.3389/fphar.2019.00879
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