Cargando…
Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing
Bromodomain (BRD) proteins are histone code interpreters that recognize acetylated lysines and link the dynamic state of chromatin with the transcriptional machinery. Here, we demonstrate that ablation of the Brd4 gene in primary mouse bone marrow–derived mesenchymal stem cells via a conditional Brd...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley & Sons, Inc.
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8520065/ https://www.ncbi.nlm.nih.gov/pubmed/34693189 http://dx.doi.org/10.1002/jbm4.10520 |
_version_ | 1784584586604314624 |
---|---|
author | Paradise, Christopher R De La Vega, Rodolfo E Galvan, M Lizeth Carrasco, Margarita E Thaler, Roman van Wijnen, Andre J Dudakovic, Amel |
author_facet | Paradise, Christopher R De La Vega, Rodolfo E Galvan, M Lizeth Carrasco, Margarita E Thaler, Roman van Wijnen, Andre J Dudakovic, Amel |
author_sort | Paradise, Christopher R |
collection | PubMed |
description | Bromodomain (BRD) proteins are histone code interpreters that recognize acetylated lysines and link the dynamic state of chromatin with the transcriptional machinery. Here, we demonstrate that ablation of the Brd4 gene in primary mouse bone marrow–derived mesenchymal stem cells via a conditional Brd4(fl/fl) allele suppresses osteogenic lineage commitment. Remarkably, loss of Brd4 function also enhances expression of genes in engineered adenoviral vectors, including Cre recombinase and green fluorescent protein (GFP). Similarly, vector‐based expression of BMP2 mRNA and protein levels are enhanced upon Brd4 depletion in cells transduced with an adenoviral vector that expresses BMP2 (Ad‐BMP2). Importantly, Brd4 depletion in MC3T3‐E1 and human adipose‐derived mesenchymal stem cells (AMSCs) transduced with Ad‐BMP2 enhances osteogenic differentiation of naïve MC3T3‐E1 cells via paracrine mechanisms based on transwell and conditioned medium studies. Our studies indicate that Brd4 depletion enhances adenoviral transgene expression in mammalian cells, which can be leveraged as a therapeutic strategy to improve viral vector‐based gene therapies. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. |
format | Online Article Text |
id | pubmed-8520065 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | John Wiley & Sons, Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-85200652021-10-22 Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing Paradise, Christopher R De La Vega, Rodolfo E Galvan, M Lizeth Carrasco, Margarita E Thaler, Roman van Wijnen, Andre J Dudakovic, Amel JBMR Plus Original Article Bromodomain (BRD) proteins are histone code interpreters that recognize acetylated lysines and link the dynamic state of chromatin with the transcriptional machinery. Here, we demonstrate that ablation of the Brd4 gene in primary mouse bone marrow–derived mesenchymal stem cells via a conditional Brd4(fl/fl) allele suppresses osteogenic lineage commitment. Remarkably, loss of Brd4 function also enhances expression of genes in engineered adenoviral vectors, including Cre recombinase and green fluorescent protein (GFP). Similarly, vector‐based expression of BMP2 mRNA and protein levels are enhanced upon Brd4 depletion in cells transduced with an adenoviral vector that expresses BMP2 (Ad‐BMP2). Importantly, Brd4 depletion in MC3T3‐E1 and human adipose‐derived mesenchymal stem cells (AMSCs) transduced with Ad‐BMP2 enhances osteogenic differentiation of naïve MC3T3‐E1 cells via paracrine mechanisms based on transwell and conditioned medium studies. Our studies indicate that Brd4 depletion enhances adenoviral transgene expression in mammalian cells, which can be leveraged as a therapeutic strategy to improve viral vector‐based gene therapies. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. John Wiley & Sons, Inc. 2021-06-23 /pmc/articles/PMC8520065/ /pubmed/34693189 http://dx.doi.org/10.1002/jbm4.10520 Text en © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research. https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Paradise, Christopher R De La Vega, Rodolfo E Galvan, M Lizeth Carrasco, Margarita E Thaler, Roman van Wijnen, Andre J Dudakovic, Amel Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title | Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title_full | Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title_fullStr | Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title_full_unstemmed | Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title_short | Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing |
title_sort | brd4 inactivation increases adenoviral delivery of bmp2 for paracrine stimulation of osteogenic differentiation as a gene therapeutic concept to enhance bone healing |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8520065/ https://www.ncbi.nlm.nih.gov/pubmed/34693189 http://dx.doi.org/10.1002/jbm4.10520 |
work_keys_str_mv | AT paradisechristopherr brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT delavegarodolfoe brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT galvanmlizeth brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT carrascomargaritae brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT thalerroman brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT vanwijnenandrej brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing AT dudakovicamel brd4inactivationincreasesadenoviraldeliveryofbmp2forparacrinestimulationofosteogenicdifferentiationasagenetherapeuticconcepttoenhancebonehealing |