Dynamic Changes in Macrophage Polarization Correlate with Progression and Response in CNS Lymphoma: Insights Into the Transcriptome of Lymphoma-Associated M2 Macrophages,

Abstract Abstract 3219 Background: Macrophages are an important component of the tumor microenvironment and the immune response to malignancy. Classically activated (M1-polarized) macrophages exhibit anti-tumor effect, while alternatively activated (M2-polarized) macrophages promote tissue repair, angiogenesis, immunosuppression and tumor progression. To date, the vast majority of studies on macrophage phenotype and polarization have been based upon in vitro studies or murine model systems. We tested the hypothesis that distinct macrophage subsets can be identified within the microenvironment of lymphoid tumors and that the relative proportion of M1:M2 macrophages correlates with therapeutic response and/or resistance. In addition, we postulated that transcriptional analysis of distinct macrophage subpopulations would provide insights into mechanisms by which M2 macrophages promote tumor progression and reveal novel mechanisms which direct the interconversion of activated macrophages between classical and alternative activation states. Our focus has been on the evaluation of the tumor microenvironment in CNS lymphomas, the cerebrospinal fluid (CSF), which is enriched in inflammatory cells including activated CD14+ macrophages that we hypothesized would exhibit phenotypic features consistent with M1 and M2 polarization. Methods: We developed a novel 9-parameter flow-cytometric method to isolate distinct subpopulations of activated macrophages from CSF and peripheral blood. The method identifies macrophages with M1 features based upon expression of nitric oxide synthase (iNOS) and M2 macrophages on the basis of low-expression of iNOS and high expression of scavenger receptors CD206, CD209 and CD36. Subpopulations of macrophages were quantified and analyzed by FACS and transcriptional profiling (Affymetrix Gene Chip 1.0 ST). Serial analysis of CSF was performed in CNS lymphoma patients to correlate macrophage polarization states with response and/or progression. Parallel analyses were performed in blood in patients with non-Hodgkin lymphoma (NHL) to control conditions including infections and/or sarcoid. Results: At least four distinct subpopulations of activated macrophages were identified in CSF in association with CNS lymphoma (N=30) and controls (N=28). A greater than six-fold increase in the proportion of M2 macrophages was detected in CSF of CNS lymphoma patients compared to control subjects (p<0.001). By contrast, the proportion of CSF macrophages with M1 features was similar between NHL and controls. Moreover, peripheral blood macrophages from CNS lymphoma patients did not demonstrate significant M2-polarization. Serial analysis demonstrated that M2 polarization of CSF macrophages correlated with tumor progression and/or response. Transcriptional profiling analysis was conducted on macrophage subpopulations isolated from CSF of CNS lymphoma patients and controls. While few significant transcriptional differences were identified between M1 and unpolarized (Mnaive) macrophages, the transcriptional comparison of M2 vs. M1 as well as M2 vs. Mnaive macrophage subpopulations revealed several hundred differentially expressed genes with significant adjusted p-values. Pathway Analysis (Ingenuity) suggested mechanisms strongly associated with the M2 phenotype including activation of Notch signaling. Transcripts associated with short survival were most strongly expressed by M2 macrophages and included Ephrin A4 and lymphoid chemokines such as CXCL-13. The prognostic significance of high CXCL-13 and Ephrin A4 expression were confirmed by ELISA in an independent validation set of cases. Conclusions: We believe this to be the first application of flow-cytometry to define the phenotypes and dynamic interconversion of intratumoral macrophages within the lymphoma microenvironment and the first correlation of M2 macrophages with the evolution of resistance to therapy. In addition, this dataset provides the first in-depth transcriptional profiling analysis of in vivo human macrophage subpopulations and suggests novel mechanisms by which tumor-associated macrophages may facilitate lymphoma progression via the regulation of the metabolic microenvironment, angiogenesis, tumor invasion and immunosuppression. Based upon our additional preliminary data, we hypothesize that these results may be relevant to a variety of lymphoid tumors. Disclosures: Off Label Use: We will discuss the use of rituximab within the leptomeningeal compartment to treat recurrent/refractory CNS lymphomas.

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Macrophage Polarization and Acquired Resistance to Rituximab in CNS Lymphoma.

Blood ◽

10.1182/blood.v114.22.3749.3749 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3749-3749

Author(s):

Cigall Kadoch ◽

Valerie D. Wong ◽

Tara Rambaldo ◽

William Hyun ◽

Clifford Lowell ◽

...

Keyword(s):

Flow Cytometry ◽

Macrophage Polarization ◽

Acquired Resistance ◽

Fold Increase ◽

Cns Lymphoma ◽

M2 Macrophage ◽

M2 Macrophages ◽

Non Hodgkin Lymphoma ◽

Macrophage Subpopulations ◽

M2 Phenotype

Abstract Abstract 3749 Poster Board III-685 Macrophages are a critical component of anti-tumor immunity but may be subverted from the classically-activated, or M1 phenotype, which mediates tumor elimination, to an alternatively-activated, M2 phenotype, which promotes tumor progression. Interleukin-4 (IL-4) signaling is a pivotal regulator of macrophage polarization to the M2 phenotype. The efficacy of rituximab appears to be mediated predominantly by antibody-dependent cellular cytotoxicity (ADCC) with tumor-associated macrophages being the dominant effector cell. We hypothesize that the phenotype of tumor-associated macrophages (M1 vs. M2) may be a potential determinant of rituximab efficacy or resistance. Relevant to this hypothesis is our demonstration of the intratumoral expression of IL-4 by CNS lymphoma (Blood, 2006; Clin Cancer Res., 2009) as well as our observation that the M2 polarization of macrophages by IL-4 treatment in vitro (18 h) results in an 8-fold decrease in rituximab-mediated ADCC of Raji lymphoma cells compared to ADCC-mediated by naïve or M1 polarized macrophages (p<0.001). There is an extreme paucity of molecular information regarding the role and phenotype of tumor macrophages in non-Hodgkin lymphoma in general, including CNS lymphoma. While macrophages from the cerebrospinal fluid (CSF) are routinely quantified in cytospin analyses, to date there is no established information regarding the phenotype and state of differentiation of macrophages within the leptomeningeal compartment. We have developed a novel flow-cytometry-based protocol for the isolation and phenotypic characterization of macrophages from the CSF of patients with CNS lymphoma. We have used candidate markers of M2 differentiation based upon our gene expression studies of CD14+ activated CSF macrophages, including CD206 and Factor XIII (each of which have previously been shown to be induced in macrophages upon IL-4 stimulation). We have identified M1 differentiation of CSF CD14+ macrophages by high fluorescence when incubated with DAF-FM diacetate, a cell-permeable marker of nitric oxide synthase (iNOS). Using flow-cytometry to evaluate the relative expression of DAF-FM and CD206, we demonstrate the presence of at least four subpopulations of activated macrophages in the CSF of CNS lymphoma patients. M1 macrophages, which highly express iNOS are denoted as DAF-FM(+)/CD206(-). M2 macrophages, which weakly express iNOS but which highly express CD206 are denoted as DAF-FM (-)/CD206(+). In addition, we have detected a mixed phenotype with features of both M1 and M2 macrophages, DAF-FM(+)/CD206(+), which we have termed dual-activated. A fourth subpopulation of activated macrophages within the CSF do not express iNOS or CD206. Thus far, CSF macrophage subpopulations have been characterized and sorted from thirty-five subjects: 18 patients with CNS lymphoma and 17 control subjects with non-neoplastic conditions. We demonstrate for the first time the association of M2 macrophages within the CSF with the pathogenesis of CNS lymphoma: there was a greater than six-fold increase in the proportion of macrophages with M2 features in immunocompetent subjects with CNS lymphoma compared to controls (p<0.001). By contrast, the proportion of macrophages with M1 features was similar between lymphoma and controls. In addition, we reproducibly detected an increase in the ratio of M1:M2 macrophages which correlated with therapeutic response to intrathecal methotrexate or cytarabine (within 94 h) in CNS lymphoma patients. By contrast, the intraventricular administration of rituximab was reproducibly associated with a greater than three-fold increase in the relative ratio of M2:M1 macrophages which was sustained compared to macrophages analyzed pre-intrathecal rituximab (p<0.001). In each of four cases, increases in M2 macrophage polarization anticipated the onset of intrathecal rituximab resistance and tumor progression. We believe this to be the first application of flow-cytometry to define the polarization states of intratumoral macrophages in non-Hodgkin lymphoma as well as the first description of dynamic changes in macrophage phenotypes during the evolution of resistance to rituximab therapy. The elucidation of distinct macrophage subpopulations based upon the expression of candidate markers of M1 vs. M2 phenotype may provide insight into tumor pathogenesis and prognosis. Disclosures: Off Label Use: While we describe the use of intrathecal administration of rituximab in patients with recurrent CNS and intraocular lymphomaa, the focus of our study is on the relationship between M2 macrophage polarization and the pathogenesis of CNS lymphoma as well as the potential relationship of macrophage polarization to acquired resistance to rituximab in CNS lymphoma patients.

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Interstitial flow promotes macrophage polarization toward an M2 phenotype

Molecular Biology of the Cell ◽

10.1091/mbc.e18-03-0164 ◽

2018 ◽

Vol 29 (16) ◽

pp. 1927-1940 ◽

Cited By ~ 22

Author(s):

Ran Li ◽

Jean Carlos Serrano ◽

Hao Xing ◽

Tara A. Lee ◽

Hesham Azizgolshani ◽

...

Keyword(s):

Fluid Flow ◽

Tumor Microenvironment ◽

Tumor Progression ◽

Cancer Cell ◽

Interstitial Fluid ◽

Macrophage Polarization ◽

Interstitial Flow ◽

Interstitial Fluid Flow ◽

M2 Polarization ◽

Tumor Tissues

Tumor tissues are characterized by an elevated interstitial fluid flow from the tumor to the surrounding stroma. Macrophages in the tumor microenvironment are key contributors to tumor progression. While it is well established that chemical stimuli within the tumor tissues can alter macrophage behaviors, the effects of mechanical stimuli, especially the flow of interstitial fluid in the tumor microenvironment, on macrophage phenotypes have not been explored. Here, we used three-dimensional biomimetic models to reveal that macrophages can sense and respond to pathophysiological levels of interstitial fluid flow reported in tumors (∼3 µm/s). Specifically, interstitial flow (IF) polarizes macrophages toward an M2-like phenotype via integrin/Src-mediated mechanotransduction pathways involving STAT3/6. Consistent with this flow-induced M2 polarization, macrophages treated with IF migrate faster and have an enhanced ability to promote cancer cell migration. Moreover, IF directs macrophages to migrate against the flow. Since IF emanates from the tumor to the surrounding stromal tissues, our results suggest that IF could not only induce M2 polarization of macrophages but also recruit these M2 macrophages toward the tumor masses, contributing to cancer cell invasion and tumor progression. Collectively, our study reveals that IF could be a critical regulator of tumor immune environment.

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Giant cell tumor stromal cells: osteoblast lineage-derived cells secrete IL-6 and IL-10 for M2 macrophages polarization

PeerJ ◽

10.7717/peerj.9748 ◽

2020 ◽

Vol 8 ◽

pp. e9748

Author(s):

Kuan Yang ◽

Lihui Bao ◽

Xiaoning He ◽

Wanmin Zhao ◽

Dongdong Fei ◽

...

Keyword(s):

Giant Cell Tumor ◽

Giant Cell ◽

Stromal Cells ◽

Macrophage Polarization ◽

Giant Cells ◽

Interleukin 10 ◽

Cell Tumor ◽

M2 Macrophages ◽

M2 Polarization ◽

Macrophages Polarization

Background The giant cell tumor (GCT) is a benign tumor which consists of three types cells: mononuclear histiocytic cells (MNHCs), multinuclear giant cells (MNGCs), and GCT stromal cells (GCTSCs). Numerous studies claim that GCTSCs have mesenchymal stem cells (MSCs) characters and play an important role in osteoclastogenesis; however, there are no research studies concerning macrophage polarization among GCT, which can be regarded as an ingredient for tumor aggression. Method We tested the effect of GCTSCs from three GCT samples which were collected from patients on proliferation, apoptosis and polarization of macrophage. Result In this article, we verified that GCTSCs expressed MSCs markers and had higher proliferation and relative lower differentiation abilities compared with BMMSCs. What’s more, we found a higher proportion of M2 macrophages among neoplasm. Co-culturing GCTSCs with macrophages resulted in prominent macrophage M2 polarization and increased the release of IL-6 (Interleukin-6) and IL-10 (Interleukin-10)from GCTSCs. In conclusion, GCTSCs, as originating from MSCs, can secret IL-6 and IL-10, which may play a significant role in macrophage M2 polarization.

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Macrophage polarization in periodontal ligament stem cells enhanced periodontal regeneration

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1409-4 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 9

Author(s):

Jiaying Liu ◽

Bin Chen ◽

Jun Bao ◽

Yangheng Zhang ◽

Lang Lei ◽

...

Keyword(s):

Stem Cell ◽

Stem Cell Transplantation ◽

Periodontal Ligament ◽

Early Stage ◽

Macrophage Polarization ◽

Periodontal Regeneration ◽

Treated Group ◽

M2 Macrophages ◽

M2 Polarization ◽

Tnf Α

Abstract Background The inflammation and regeneration process may be accompanied by the shift in the M1/M2 polarization of macrophages to adapt to extracellular signals. How the macrophages responded to the altered immunological environment in the periodontal niche after stem cell transplantation has never been explored. The purpose of present study is to investigate whether M1/M2 polarization of macrophages participated in the tissue homeostasis and wound healing during periodontal ligament stem cell (PDLSC)-based periodontal regeneration. Methods A rat periodontal defect model was utilized to observe the regeneration process in the PDLSC transplantation-enhanced periodontal repair. Dynamic changes in the markers of M1/M2 macrophages were observed on days 3, 7, and 21 post surgery. In addition, the outcome of regeneration was analyzed on day 21 after surgery. To further investigate the effect of PDLSCs on macrophage polarization, the conditioned medium of PDLSCs was utilized to treat M0, M1, and M2 macrophages for 24 h; markers of M1/M2 polarization were evaluated in macrophages. Results Elevated bone volume and average thickness of bone trabecular was observed in the PDLSC-treated group by micro-computed tomography on day 21. In addition, enhanced periodontal regeneration was observed in the PDLSC-treated group with cementum-like structure regeneration and collagen fiber formation, which inserted into the newly formed cementum. On day 3, PDLSC transplantation increased IL-10 level in the periodontal tissue, while decreased TNF-α in the early stage of periodontal regeneration. On day 7, enhanced CD163+ cell infiltration and heightened expression of markers of M2 macrophages were observed. Furthermore, conditioned medium from PDLSC culture induced macrophage polarization towards the anti-inflammatory phenotype by downregulating TNF-α and upregulating IL-10, Arg-1, and CD163 in vitro. Conclusions PDLSCs could induce macrophage polarization towards the M2 phenotype, and the shift in the polarization towards M2 macrophages in the early stage of tissue repair contributed to the enhanced periodontal regeneration after stem cell transplantation. Therefore, signals from the transplanted PDLSCs might alter the immune microenvironment to enhance periodontal regeneration.

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Mesenchymal stromal cells release CXCL1/2/8 and induce chemoresistance and macrophage polarization

10.1101/482513 ◽

2018 ◽

Author(s):

Bettina Couderc ◽

Augustin Le Naour ◽

Mélissa Prat ◽

Renaud Mevel ◽

Benoit Thibault ◽

...

Keyword(s):

Tumor Progression ◽

Mesenchymal Stromal Cells ◽

Stromal Cells ◽

Therapeutic Strategy ◽

Ex Vivo ◽

Macrophage Polarization ◽

Activated Macrophages ◽

High Concentrations ◽

Ovarian Tumor Cells

Factors released by surrounding cells such as cancer-associated mesenchymal stromal cells (CA-MSCs) are involved in tumor progression and chemoresistance. We determine the mechanisms by which a naïve MSC could become a CA-MSC and characterize CA-MSCs. Ovarian tumor cells (OTC) trigger the transformation of MSCs to CA-MSCs expressing different pro-tumoral, genes and secreting high amounts of CXCR1/2 ligands (CXCL1, CXCL2 and IL-8) implicated in the chemoresistance of cancer cells. CXCR1/2 ligands can also inhibit the immune response against OTC. Indeed, through their released factors, CA-MSCs can trigger the differentiation of monocytes to pro-tumoral M2 phenotype macrophages known to promote the tumor progression. When CXCR1/2 receptors are inhibited, these CA-MSC-activated macrophages lose their M2 functions and acquire an anti-tumoral phenotype. Both ex vivo and in vivo a CXCR1/2 inhibitor can sensitize OTC to carboplatin even in the presence of a pro-tumoral microenvironment. This inhibitor can circumvent the pro-tumoral effects of CA-MSCs. As high concentrations of CXCR1/2 ligands in blood from patients can be associated with chemoresistance, CXCR1/2 inhibition could be a potential therapeutic strategy to revert chemoresistance.

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Targeting M2 Macrophages Alleviates Airway Inflammation and Remodeling in Asthmatic Mice via miR-378a-3p/GRB2 Pathway

Frontiers in Molecular Biosciences ◽

10.3389/fmolb.2021.717969 ◽

2021 ◽

Vol 8 ◽

Author(s):

Qiujie Wang ◽

Luna Hong ◽

Ming Chen ◽

Jiangting Shi ◽

Xiaoling Lin ◽

...

Keyword(s):

Airway Inflammation ◽

Alveolar Macrophages ◽

Target Gene ◽

Macrophage Polarization ◽

Growth Factor Receptor ◽

Luciferase Reporter ◽

M2 Macrophages ◽

Mirna Profiling ◽

M2 Polarization

Background: Asthma is a complex respiratory disease characterized by airway inflammation and remodeling. MicroRNAs (miRNAs) mediate various cellular processes including macrophage polarization and play an important role in the pathogenesis of asthma. In present study, we aimed to screen miRNA profiling involved in macrophage polarization and investigate its possible functions and mechanisms.Methods: An OVA-sensitized mouse model was established and 2-chloroadenosine (2-CA) was used to interfere with macrophages. The airway inflammation and remodeling were assessed. The identification and function of M2 alveolar macrophages were assessed by flow cytometry, RT-qPCR, arginase activity and co-culture experiment. Microarray screening was used to select miRNAs which were related to macrophage polarization and RNA interference (RNAi) technique was performed to confirm the function of the selected miRNA and its target gene.Results: Alveolar macrophages of asthmatic mice showed significant M2 polarization. 2-CA alleviated airway inflammation and remodeling as well as M2 polarization. In vitro, IL-4-induced M2 macrophages promoted the proliferation of α-SMA-positive cells. And miRNA profiling showed a remarkable increased expression of miR-378a-3p in IL-4 induced M2 macrophages. Dual luciferase reporter assay confirmed growth factor receptor binding protein 2 (GRB2) was a target gene of miR-378a-3p. A miR-378a-3p inhibitor and knockdown of GRB2 repolarized alveolar macrophages from M1 to M2 phenotype.Conclusion: Our findings suggest that miR-378a-3p/GRB2 pathway regulates the polarization of alveolar macrophages which acts as a potential therapeutic target for airway inflammation and remodeling in asthma.

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Version up-date: The CDKN2B-AS1/ MIR497/TXNIP axis regulated macrophages

10.21203/rs.3.rs-806497/v1 ◽

2021 ◽

Author(s):

JianZhong Xu

Keyword(s):

Rheumatoid Arthritis ◽

Macrophage Polarization ◽

Critical Role ◽

M2 Macrophages ◽

Macrophage Cell Line ◽

Macrophage Cell ◽

Surface Polarization ◽

M2 Polarization ◽

M1 Polarization ◽

Healthy Donors

Abstract The polarization of macrophages plays a critical role in the pathophysiology of rheumatoid arthritis. The macrophages can have pro-inflammatory M1 polarization and various types of alternative anti-inflammatory M2 polarization. Our preliminary results showed that the CDKN2B-AS1/MIR497/TXNIP axis might regulate macrophages of rheumatoid arthritis patients. Therefore, we hypothesized that this axis regulated the polarization of rheumatoid macrophages. Flow cytometry was used to determine the surface polarization markers in M1 or M2 macrophages from healthy donors and rheumatoid arthritis patients. The QPCR and Western Blotting were used to compare the expression of the CDKN2B-AS1/MIR497/TXNIP axis in these macrophages. We Knocked down and overexpressed the axis in the macrophage cell line MD to test its roles in macrophage polarization. Compared to cells from healthy donors, cells from rheumatoid arthritis patients expressed higher levels of CD40 and CD80 and lower levels of CD16, CD163, CD206, and CD200R after polarization, they also expressed higher CDKN2B-AS1, lower MIR497, and higher TXNIP. In macrophages from healthy donors, there was no correlation among CDKN2B-AS1, MIR497, and TXNIP. But in macrophages from patients, there were significant correlations. The CDKN2B-AS1 knockdown, MIR497 mimics suppressed the M1 polarization but promoted the M2 polarization in MD cells, while the MIR497 knockdown and the TXNIP overexpression did the opposite. This study demonstrated that elevated CDKN2B-AS1 in macrophages promotes the M1 polarization and inhibited the M2 polarization of macrophages by the CDKN2B-AS1/ MIR497/TXNIP axis.

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Biology of CNS lymphoma and the potential of novel agents

Hematology ◽

10.1182/asheducation-2017.1.556 ◽

2017 ◽

Vol 2017 (1) ◽

pp. 556-564 ◽

Cited By ~ 11

Author(s):

James L. Rubenstein

Keyword(s):

Tumor Microenvironment ◽

Clinical Investigation ◽

Transcriptional Profiling ◽

Large Fraction ◽

Cell Receptor ◽

Primary Response ◽

Comparative Genomic ◽

Cns Lymphoma ◽

Targeted Agents ◽

Cns Lymphomas

Abstract Primary and secondary CNS lymphomas are aggressive brain tumors that pose an immense challenge to define in terms of molecular pathogenesis, as well as to effectively treat. During the past 10 years improvements in survival have been achieved with the implementation of anti-CD20 immunotherapy and optimization of dose-intensive consolidation strategies. The applications of whole-exome sequencing, comparative genomic hybridization, transcriptional profiling, and examination of the tumor microenvironment, particularly in the context of clinical investigation, provide insights that create a roadmap for the development and implementation of novel targeted agents for this disease. A body of genetic evidence strongly suggested that primary CNS lymphomas (PCNSLs) are likely largely dependent on NF-κB prosurvival signals, with enrichment of mutations involving the B-cell receptor pathway, in particular myeloid differentiation primary response 88 and cluster of differentiation 79B. The first set of early-phase investigations that target NF-κB in PCNSL have now been completed and support the NF-κB hypothesis but at the same time reveal that much work needs to be done to translate these results into meaningful advances in survival for a large fraction of patients. Insights into secondary prosurvival pathways that mediate drug resistance is a priority for investigation. Similarly, further evaluation of the immune-suppressive mechanisms in the CNS lymphoma tumor microenvironment is requisite for progress. Combinatorial interventions that promote the antitumor immune response have significant potential. With increasing availability of targeted agents, there is also a need to develop more sensitive imaging tools, not only to detect this highly invasive brain neoplasm but also potentially to define an evolving molecular phenotype to facilitate precision medicine.

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Angelicin Alleviates Post-Trauma Osteoarthritis Progression by Regulating Macrophage Polarization via STAT3 Signaling Pathway

Frontiers in Pharmacology ◽

10.3389/fphar.2021.669213 ◽

2021 ◽

Vol 12 ◽

Author(s):

Zhansong Tian ◽

Fanchun Zeng ◽

Chunrong Zhao ◽

Shiwu Dong

Keyword(s):

Side Effects ◽

Macrophage Polarization ◽

Critical Factor ◽

Molecular Medicine ◽

M2 Macrophages ◽

Stat3 Pathway ◽

M2 Polarization ◽

Ifn Γ

Post-trauma osteoarthritis (PTOA) is the most common articular disease characterized by degeneration and destruction of articular cartilage (Bultink and Lems, Curr. Rheumatol Rep., 2013, 15, 328). Inflammatory response of local joint tissue induced by trauma is the most critical factor accelerating osteoarthritis (OA) progression (Sharma et al., 2019; Osteoarthritis. Cartilage, 28, 658–668). M1/M2 macrophages polarization and repolarization participates in local inflammation, which plays a major role in the progression of OA (Zhang et al., 2018; Ann. Rheum. Dis., 77, 1524–1534). The regulating effect of macrophage polarization has been reported as a potential therapy to alleviate OA progression. Synovitis induced by polarized macrophages could profoundly affect the chondrocyte and cartilage matrix (Zhang et al., 2018; Ann. Rheum. Dis., 77, 1524–1534). Generally, anti-inflammatory medications widely used in clinical practice have serious side effects. Therefore, we focus on exploring a new therapeutic strategy with fewer side effects to alleviate the synovitis. Angelicin (ANG) is traditional medicine used in various folk medicine. Previous studies have revealed that angelicin has an inhibitory effect on inflammation (Wei et al., 2016; Inflammation, 39, 1876–1882), tumor growth (Li et al., 2016; Oncology reports, 36, 3,504–3,512; Wang et al., 2017; Molecular Medicine Reports, 16, 5441–5449), DNA damage (Li et al., 2019; Exp. Ther. Med., 18, 1899–1906), and virus proliferation (Li et al., 2018; Front. Cell. Infect. Microbiol., 8, 178). But its specific effects on influencing the process of OA were rarely reported. In this study, the molecular mechanism of angelicin in vivo and in vitro was clearly investigated. Results showed that angelicin could regulate the M1/M2 ratio and function and alleviate the development of PTOA in the meanwhile. Bone marrow monocytes were isolated and induced by macrophage colony-stimulating factor (M-CSF), lipopolysaccharide (LPS) and interferon (IFN)-γ for M1 polarization and interleukin (IL)-4/IL-13 for M2 polarization. Subsequently, repolarization intervention was performed. The results indicate that angelicin can repolarize M1 toward M2 macrophages by upregulating the expression of CD9. Besides, angelicin can also protect and maintain M2 polarization in the presence of LPS/IFN-γ, and subsequently downregulate the expression of inflammatory mediators such as IL-1β and TNF-α. Mechanistically, angelicin can activate the p-STAT3/STAT3 pathway by conducting CD9/gp130 to repolarize toward M2 macrophages. These results suggest angelicin can alleviate the progression of OA by regulating M1/M2 polarization via the STAT3/p-STAT3 pathway. Therefore, angelicin may have a promising application and potential therapeutic value in OA clinical treatment.

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Cortisol Metabolism in Carp Macrophages: A Role for Macrophage-Derived Cortisol in M1/M2 Polarization

International Journal of Molecular Sciences ◽

10.3390/ijms21238954 ◽

2020 ◽

Vol 21 (23) ◽

pp. 8954

Author(s):

Magdalena Maciuszek ◽

Katarzyna Klak ◽

Leszek Rydz ◽

B. M. Lidy Verburg-van Kemenade ◽

Magdalena Chadzinska

Keyword(s):

Glucocorticoid Receptors ◽

Macrophage Polarization ◽

In Vitro Study ◽

Interleukin 10 ◽

Interferon Γ ◽

M2 Macrophages ◽

Cortisol Metabolism ◽

M2 Polarization ◽

Induced Changes

Macrophages are crucial not only for initiation of inflammation and pathogen eradication (classically polarized M1 macrophages), but also for inflammation inhibition and tissue regeneration (alternatively polarized M2 macrophages). Their polarization toward the M1 population occurs under the influence of interferon-γ + lipopolysaccharide (IFN-γ + LPS), while alternatively polarized M2 macrophages evolve upon, e.g., interlukin 4 (IL-4) or cortisol stimulation. This in vitro study focused on a possible role for macrophage-derived cortisol in M1/M2 polarization in common carp. We studied the expression of molecules involved in cortisol synthesis/conversion from and to cortisone like 11β-hydroxysteroid dehydrogenase type 2 and 3. (11β-HSD2 and 3) and 11β-hydroxylase (CYP11b), as well as the expression of glucocorticoid receptors (GRs) and proliferator-activated receptor gamma (PPARγ) in M1 and M2 macrophages. Lastly, we analyzed how inhibition of these molecules affect macrophage polarization. In M1 cells, upregulation of gene expression of GRs and 11β-HSD3 was found, while, in M2 macrophages, expression of 11β-hsd2 was upregulated. Moreover, blocking of cortisol synthesis/conversion and GRs or PPARγ induced changes in expression of anti-inflammatory interleukin 10 (IL-10). Consequently, our data show that carp monocytes/macrophages can convert cortisol. The results strongly suggest that cortisol, via intracrine interaction with GRs, is important for IL-10-dependent control of the activity of macrophages and for the regulation of M1/M2 polarization to finally determine the outcome of an infection.

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