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Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA
Autologous macrophage transfer is an emerging platform for cell therapy. It is anticipated that conventional macrophage reprogramming based on ex vivo polarization using cytokines and ligands of TLRs may enhance the therapeutic effect. We describe an alternative approach based on small interfering R...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406732/ https://www.ncbi.nlm.nih.gov/pubmed/36010574 http://dx.doi.org/10.3390/cells11162498 |
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author | Halimani, Noreen Nesterchuk, Mikhail Andreichenko, Irina N. Tsitrina, Alexandra A. Elchaninov, Andrey Lokhonina, Anastasia Fatkhudinov, Timur Dashenkova, Nataliya O. Brezgina, Vera Zatsepin, Timofei S. Mikaelyan, Arsen S. Kotelevtsev, Yuri V. |
author_facet | Halimani, Noreen Nesterchuk, Mikhail Andreichenko, Irina N. Tsitrina, Alexandra A. Elchaninov, Andrey Lokhonina, Anastasia Fatkhudinov, Timur Dashenkova, Nataliya O. Brezgina, Vera Zatsepin, Timofei S. Mikaelyan, Arsen S. Kotelevtsev, Yuri V. |
author_sort | Halimani, Noreen |
collection | PubMed |
description | Autologous macrophage transfer is an emerging platform for cell therapy. It is anticipated that conventional macrophage reprogramming based on ex vivo polarization using cytokines and ligands of TLRs may enhance the therapeutic effect. We describe an alternative approach based on small interfering RNA (siRNA) knockdown of selected molecular cues of macrophage polarization, namely EGR2, IRF3, IRF5, and TLR4 in Raw264.7 monocyte/macrophage cell line and mouse-bone-marrow-derived macrophages (BMDMs). The impact of IRF5 knockdown was most pronounced, curtailing the expression of other inflammatory mediators such as IL-6 and NOS2, especially in M1-polarized macrophages. Contrary to IRF5, EGR2 knockdown potentiated M1-associated markers while altogether abolishing M2 marker expression, which is indicative of the principal role of EGR2 in the maintenance of alternative phenotypes. IRF3 knockdown suppressed M1 polarization but upregulated Arg 1, a canonical marker of alternative polarization in M1 macrophages. As anticipated, the knockdown of TLR4 also attenuated the M1 phenotype but, akin to IRF3, significantly induced Arginase 1 in M0 and M1, driving the phenotype towards M2. This study validates RNAi as a viable option for the alteration and maintenance of macrophage phenotypes. |
format | Online Article Text |
id | pubmed-9406732 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-94067322022-08-26 Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA Halimani, Noreen Nesterchuk, Mikhail Andreichenko, Irina N. Tsitrina, Alexandra A. Elchaninov, Andrey Lokhonina, Anastasia Fatkhudinov, Timur Dashenkova, Nataliya O. Brezgina, Vera Zatsepin, Timofei S. Mikaelyan, Arsen S. Kotelevtsev, Yuri V. Cells Article Autologous macrophage transfer is an emerging platform for cell therapy. It is anticipated that conventional macrophage reprogramming based on ex vivo polarization using cytokines and ligands of TLRs may enhance the therapeutic effect. We describe an alternative approach based on small interfering RNA (siRNA) knockdown of selected molecular cues of macrophage polarization, namely EGR2, IRF3, IRF5, and TLR4 in Raw264.7 monocyte/macrophage cell line and mouse-bone-marrow-derived macrophages (BMDMs). The impact of IRF5 knockdown was most pronounced, curtailing the expression of other inflammatory mediators such as IL-6 and NOS2, especially in M1-polarized macrophages. Contrary to IRF5, EGR2 knockdown potentiated M1-associated markers while altogether abolishing M2 marker expression, which is indicative of the principal role of EGR2 in the maintenance of alternative phenotypes. IRF3 knockdown suppressed M1 polarization but upregulated Arg 1, a canonical marker of alternative polarization in M1 macrophages. As anticipated, the knockdown of TLR4 also attenuated the M1 phenotype but, akin to IRF3, significantly induced Arginase 1 in M0 and M1, driving the phenotype towards M2. This study validates RNAi as a viable option for the alteration and maintenance of macrophage phenotypes. MDPI 2022-08-11 /pmc/articles/PMC9406732/ /pubmed/36010574 http://dx.doi.org/10.3390/cells11162498 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 Halimani, Noreen Nesterchuk, Mikhail Andreichenko, Irina N. Tsitrina, Alexandra A. Elchaninov, Andrey Lokhonina, Anastasia Fatkhudinov, Timur Dashenkova, Nataliya O. Brezgina, Vera Zatsepin, Timofei S. Mikaelyan, Arsen S. Kotelevtsev, Yuri V. Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title | Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title_full | Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title_fullStr | Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title_full_unstemmed | Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title_short | Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA |
title_sort | phenotypic alteration of bmdm in vitro using small interfering rna |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9406732/ https://www.ncbi.nlm.nih.gov/pubmed/36010574 http://dx.doi.org/10.3390/cells11162498 |
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