PMID- 36010574 OWN - NLM STAT- MEDLINE DCOM- 20220829 LR - 20221126 IS - 2073-4409 (Electronic) IS - 2073-4409 (Linking) VI - 11 IP - 16 DP - 2022 Aug 11 TI - Phenotypic Alteration of BMDM In Vitro Using Small Interfering RNA. LID - 10.3390/cells11162498 [doi] LID - 2498 AB - 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. FAU - Halimani, Noreen AU - Halimani N AUID- ORCID: 0000-0001-6004-226X AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. FAU - Nesterchuk, Mikhail AU - Nesterchuk M AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. FAU - Andreichenko, Irina N AU - Andreichenko IN AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. FAU - Tsitrina, Alexandra A AU - Tsitrina AA AD - Koltzov Institute of Developmental Biology of Russian Academy of Sciences, 26 Vavilov Street, Moscow 119334, Russia. FAU - Elchaninov, Andrey AU - Elchaninov A AD - National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov, Russian Federation, 4 Oparina Street, Moscow 117997, Russia. AD - Department of Histology, Pirogov Russian National Research Medical University, Ministry of Healthcare of The Russian Federation, 1 Ostrovitianov Street, Moscow 117997, Russia. FAU - Lokhonina, Anastasia AU - Lokhonina A AD - National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov, Russian Federation, 4 Oparina Street, Moscow 117997, Russia. AD - Department of Histology, Cytology and Embryology, Peoples' Friendship University of Russia, 6 Miklukho-Maklaya Street, Moscow 117198, Russia. FAU - Fatkhudinov, Timur AU - Fatkhudinov T AD - Department of Histology, Pirogov Russian National Research Medical University, Ministry of Healthcare of The Russian Federation, 1 Ostrovitianov Street, Moscow 117997, Russia. AD - Scientific Research Institute of Human Morphology, 3 Tsurupa Street, Moscow 117418, Russia. FAU - Dashenkova, Nataliya O AU - Dashenkova NO AD - Koltzov Institute of Developmental Biology of Russian Academy of Sciences, 26 Vavilov Street, Moscow 119334, Russia. FAU - Brezgina, Vera AU - Brezgina V AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. FAU - Zatsepin, Timofei S AU - Zatsepin TS AUID- ORCID: 0000-0003-0030-9174 AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. FAU - Mikaelyan, Arsen S AU - Mikaelyan AS AD - Koltzov Institute of Developmental Biology of Russian Academy of Sciences, 26 Vavilov Street, Moscow 119334, Russia. FAU - Kotelevtsev, Yuri V AU - Kotelevtsev YV AUID- ORCID: 0000-0002-7255-7794 AD - Vladimir Zelman Center for Neurobiology and Brain Rehabilitation and Center of Life Sciences, Skolkovo Institute of Science and Technology, Moscow 143025, Russia. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20220811 PL - Switzerland TA - Cells JT - Cells JID - 101600052 RN - 0 (Biomarkers) RN - 0 (Interferon Regulatory Factors) RN - 0 (Irf5 protein, mouse) RN - 0 (RNA, Small Interfering) RN - 0 (Toll-Like Receptor 4) SB - IM MH - Animals MH - Biomarkers/metabolism MH - Interferon Regulatory Factors/genetics/metabolism MH - *Macrophage Activation MH - Mice MH - Phenotype MH - RNA, Small Interfering/genetics/metabolism MH - *Toll-Like Receptor 4/genetics/metabolism PMC - PMC9406732 OTO - NOTNLM OT - EGR2 OT - IRF3 OT - IRF5 OT - TLR4 OT - macrophages OT - polarization OT - siRNA COIS- The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results. EDAT- 2022/08/27 06:00 MHDA- 2022/08/30 06:00 PMCR- 2022/08/11 CRDT- 2022/08/26 01:12 PHST- 2022/07/14 00:00 [received] PHST- 2022/08/05 00:00 [revised] PHST- 2022/08/09 00:00 [accepted] PHST- 2022/08/26 01:12 [entrez] PHST- 2022/08/27 06:00 [pubmed] PHST- 2022/08/30 06:00 [medline] PHST- 2022/08/11 00:00 [pmc-release] AID - cells11162498 [pii] AID - cells-11-02498 [pii] AID - 10.3390/cells11162498 [doi] PST - epublish SO - Cells. 2022 Aug 11;11(16):2498. doi: 10.3390/cells11162498.