Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format
Oncolytic adenoviruses are being developed as new anti-cancer agents. Their efficacy can be improved by incorporating RNA interference (RNAi) molecules. RNAi molecules can be expressed in various precursor formats. The aim of this study was to determine the most effective format. To this end, we con...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Elsevier
2020-12-01
|
Series: | Molecular Therapy: Oncolytics |
Subjects: | |
Online Access: | http://www.sciencedirect.com/science/article/pii/S2372770520301637 |
id |
doaj-1aaae3dbac784368bbd487059385b756 |
---|---|
record_format |
Article |
spelling |
doaj-1aaae3dbac784368bbd487059385b7562020-12-19T05:09:13ZengElsevierMolecular Therapy: Oncolytics2372-77052020-12-0119332343Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor FormatTereza Brachtlova0Jan-Willem van Ginkel1Mark J. Luinenburg2Renée X. de Menezes3Danijela Koppers-Lalic4D. Michiel Pegtel5Wenliang Dong6Tanja D. de Gruijl7Victor W. van Beusechem8Department of Medical Oncology, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, Amsterdam Infection & Immunity Institute, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands; ORCA Therapeutics B.V., 1081 HV Amsterdam, the NetherlandsORCA Therapeutics B.V., 1081 HV Amsterdam, the NetherlandsDepartment of Neurosurgery, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the NetherlandsDepartment of Epidemiology and Biostatistics, Amsterdam UMC, Vrije Universiteit Amsterdam, Netherlands Bioinformatics Center, De Boelelaan 1117, 1081 HV Amsterdam, the NetherlandsDepartment of Neurosurgery, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the NetherlandsDepartment of Pathology, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, De Boelelaan 1117, 1081 HV Amsterdam, the NetherlandsORCA Therapeutics B.V., 1081 HV Amsterdam, the NetherlandsDepartment of Medical Oncology, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, Amsterdam Infection & Immunity Institute, De Boelelaan 1117, 1081 HV Amsterdam, the NetherlandsDepartment of Medical Oncology, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, Amsterdam Infection & Immunity Institute, De Boelelaan 1117, 1081 HV Amsterdam, the Netherlands; Corresponding author: Victor W. van Beusechem, Department of Medical Oncology, Amsterdam UMC, Vrije Universiteit Amsterdam, Cancer Center Amsterdam, Amsterdam Infection & Immunity Institute, De Boelelaan 1117, Room CCA 3.50, P.O. Box 7057, 1007 MB Amsterdam, the Netherlands.Oncolytic adenoviruses are being developed as new anti-cancer agents. Their efficacy can be improved by incorporating RNA interference (RNAi) molecules. RNAi molecules can be expressed in various precursor formats. The aim of this study was to determine the most effective format. To this end, we constructed three Δ24-type oncolytic adenoviruses, with human microRNA-1 (miR-1) expression cassettes in short hairpin RNA (shRNA), precursor microRNA (pre-miRNA), and primary miRNA (pri-miRNA) format, respectively. The viruses were compared for virus replication, mature miR-1 expression, and target gene silencing in cancer cells. Incorporation of the cassettes had only minor effects on virus replication. Mature miR-1 expression from the pri-miRNA format reached on average 100-fold higher levels than from the other two formats. This expression remained stable upon long-term virus propagation. Infection with the pri-miR-1-expressing virus silenced the validated miR-1 targets FOXP1 and MET. Drosha knockout almost completely abrogated mature miR-1 expression, confirming that processing of adenovirus-encoded pri-miR-1 was dependent on the host cell miRNA machinery. Using simple in vitro recombination cloning, a similar virus expressing miR-26b was made and shown to silence the validated miR-26b target PTGS2. We thus provide a platform for construction of oncolytic adenoviruses with high expression of RNAi molecules of choice.http://www.sciencedirect.com/science/article/pii/S2372770520301637oncolytic virusoncolytic immunotherapygene silencinggene knockdown |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Tereza Brachtlova Jan-Willem van Ginkel Mark J. Luinenburg Renée X. de Menezes Danijela Koppers-Lalic D. Michiel Pegtel Wenliang Dong Tanja D. de Gruijl Victor W. van Beusechem |
spellingShingle |
Tereza Brachtlova Jan-Willem van Ginkel Mark J. Luinenburg Renée X. de Menezes Danijela Koppers-Lalic D. Michiel Pegtel Wenliang Dong Tanja D. de Gruijl Victor W. van Beusechem Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format Molecular Therapy: Oncolytics oncolytic virus oncolytic immunotherapy gene silencing gene knockdown |
author_facet |
Tereza Brachtlova Jan-Willem van Ginkel Mark J. Luinenburg Renée X. de Menezes Danijela Koppers-Lalic D. Michiel Pegtel Wenliang Dong Tanja D. de Gruijl Victor W. van Beusechem |
author_sort |
Tereza Brachtlova |
title |
Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format |
title_short |
Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format |
title_full |
Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format |
title_fullStr |
Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format |
title_full_unstemmed |
Expression of Oncolytic Adenovirus-Encoded RNAi Molecules Is Most Effective in a pri-miRNA Precursor Format |
title_sort |
expression of oncolytic adenovirus-encoded rnai molecules is most effective in a pri-mirna precursor format |
publisher |
Elsevier |
series |
Molecular Therapy: Oncolytics |
issn |
2372-7705 |
publishDate |
2020-12-01 |
description |
Oncolytic adenoviruses are being developed as new anti-cancer agents. Their efficacy can be improved by incorporating RNA interference (RNAi) molecules. RNAi molecules can be expressed in various precursor formats. The aim of this study was to determine the most effective format. To this end, we constructed three Δ24-type oncolytic adenoviruses, with human microRNA-1 (miR-1) expression cassettes in short hairpin RNA (shRNA), precursor microRNA (pre-miRNA), and primary miRNA (pri-miRNA) format, respectively. The viruses were compared for virus replication, mature miR-1 expression, and target gene silencing in cancer cells. Incorporation of the cassettes had only minor effects on virus replication. Mature miR-1 expression from the pri-miRNA format reached on average 100-fold higher levels than from the other two formats. This expression remained stable upon long-term virus propagation. Infection with the pri-miR-1-expressing virus silenced the validated miR-1 targets FOXP1 and MET. Drosha knockout almost completely abrogated mature miR-1 expression, confirming that processing of adenovirus-encoded pri-miR-1 was dependent on the host cell miRNA machinery. Using simple in vitro recombination cloning, a similar virus expressing miR-26b was made and shown to silence the validated miR-26b target PTGS2. We thus provide a platform for construction of oncolytic adenoviruses with high expression of RNAi molecules of choice. |
topic |
oncolytic virus oncolytic immunotherapy gene silencing gene knockdown |
url |
http://www.sciencedirect.com/science/article/pii/S2372770520301637 |
work_keys_str_mv |
AT terezabrachtlova expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT janwillemvanginkel expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT markjluinenburg expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT reneexdemenezes expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT danijelakopperslalic expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT dmichielpegtel expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT wenliangdong expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT tanjaddegruijl expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat AT victorwvanbeusechem expressionofoncolyticadenovirusencodedrnaimoleculesismosteffectiveinaprimirnaprecursorformat |
_version_ |
1724377616440360960 |