mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor
Monomeric C-reactive protein (mCRP) has recently been implicated in the abnormal vascular activation associated with development of atherosclerosis, but it may act more specifically through mechanisms perpetuating damaged vessel inflammation and subsequent aggregation and internalization of resident...
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MDPI AG
2024-03-01
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| Online Access: | https://www.mdpi.com/1422-0067/25/6/3097 |
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| author | Ylenia Pastorello Doina Manu Xenia Sawkulycz Vittorio Caprio Claudia Banescu Minodora Dobreanu Lawrence Potempa Mario Di Napoli Mark Slevin |
| author_facet | Ylenia Pastorello Doina Manu Xenia Sawkulycz Vittorio Caprio Claudia Banescu Minodora Dobreanu Lawrence Potempa Mario Di Napoli Mark Slevin |
| author_sort | Ylenia Pastorello |
| collection | DOAJ |
| description | Monomeric C-reactive protein (mCRP) has recently been implicated in the abnormal vascular activation associated with development of atherosclerosis, but it may act more specifically through mechanisms perpetuating damaged vessel inflammation and subsequent aggregation and internalization of resident macrophages. Whilst the direct effects of mCRP on endothelial cells have been characterized, the interaction with blood monocytes has, to our knowledge, not been fully defined. Here we showed that mCRP caused a strong aggregation of both U937 cell line and primary peripheral blood monocytes (PBMs) obtained from healthy donors. Moreover, this increase in clustering was dependent on focal adhesion kinase (FAK) activation (blocked by a specific inhibitor), as was the concomitant adhesive attachment to the plate, which was suggestive of macrophage differentiation. Confocal microscopy confirmed the increased expression and nuclear localization of p-FAK, and cell surface marker expression associated with M1 macrophage polarization (CD11b, CD14, and CD80, as well as iNOS) in the presence of mCRP. Inclusion of a specific CRP dissociation/mCRP inhibitor (C10M) effectively inhibited PBMs clustering, as well as abrogating p-FAK expression, and partially reduced the expression of markers associated with M1 macrophage differentiation. mCRP also increased the secretion of pro-inflammatory cytokines Interleukin-8 (IL-8) and Interleukin-1β (IL-1β), without notably affecting MAP kinase signaling pathways; inclusion of C10M did not perturb or modify these effects. In conclusion, mCRP modulates PBMs through a mechanism that involves FAK and results in cell clustering and adhesion concomitant with changes consistent with M1 phenotypical polarization. C10M has potential therapeutic utility in blocking the primary interaction of mCRP with the cells—for example, by protecting against monocyte accumulation and residence at damaged vessels that may be predisposed to plaque development and atherosclerosis. |
| first_indexed | 2024-04-24T18:12:00Z |
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| language | English |
| last_indexed | 2024-04-24T18:12:00Z |
| publishDate | 2024-03-01 |
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| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-087617469567465bb3e47590146a4bdf2024-03-27T13:45:00ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672024-03-01256309710.3390/ijms25063097mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M InhibitorYlenia Pastorello0Doina Manu1Xenia Sawkulycz2Vittorio Caprio3Claudia Banescu4Minodora Dobreanu5Lawrence Potempa6Mario Di Napoli7Mark Slevin8Department of Anatomy and Embryology, George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, RomaniaCenter for Advanced Medical and Pharmaceutical Research (CCAMF), George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, RomaniaDepartment of Life Sciences, Manchester Metropolitan University, Manchester M15 6BH, UKDepartment of Life Sciences, Manchester Metropolitan University, Manchester M15 6BH, UKCenter for Advanced Medical and Pharmaceutical Research (CCAMF), George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, RomaniaCenter for Advanced Medical and Pharmaceutical Research (CCAMF), George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, RomaniaDepartment of Life Sciences, College of Science, Health and Pharmacy, Roosevelt University, Schaumburg, IL 60173, USANeurological Service, dell’ Annunziata Hospital, Sulmona, 67039 L’Aquila, ItalyCenter for Advanced Medical and Pharmaceutical Research (CCAMF), George Emil Palade University of Medicine, Pharmacy, Science and Technology of Targu Mures, 540142 Targu Mures, RomaniaMonomeric C-reactive protein (mCRP) has recently been implicated in the abnormal vascular activation associated with development of atherosclerosis, but it may act more specifically through mechanisms perpetuating damaged vessel inflammation and subsequent aggregation and internalization of resident macrophages. Whilst the direct effects of mCRP on endothelial cells have been characterized, the interaction with blood monocytes has, to our knowledge, not been fully defined. Here we showed that mCRP caused a strong aggregation of both U937 cell line and primary peripheral blood monocytes (PBMs) obtained from healthy donors. Moreover, this increase in clustering was dependent on focal adhesion kinase (FAK) activation (blocked by a specific inhibitor), as was the concomitant adhesive attachment to the plate, which was suggestive of macrophage differentiation. Confocal microscopy confirmed the increased expression and nuclear localization of p-FAK, and cell surface marker expression associated with M1 macrophage polarization (CD11b, CD14, and CD80, as well as iNOS) in the presence of mCRP. Inclusion of a specific CRP dissociation/mCRP inhibitor (C10M) effectively inhibited PBMs clustering, as well as abrogating p-FAK expression, and partially reduced the expression of markers associated with M1 macrophage differentiation. mCRP also increased the secretion of pro-inflammatory cytokines Interleukin-8 (IL-8) and Interleukin-1β (IL-1β), without notably affecting MAP kinase signaling pathways; inclusion of C10M did not perturb or modify these effects. In conclusion, mCRP modulates PBMs through a mechanism that involves FAK and results in cell clustering and adhesion concomitant with changes consistent with M1 phenotypical polarization. C10M has potential therapeutic utility in blocking the primary interaction of mCRP with the cells—for example, by protecting against monocyte accumulation and residence at damaged vessels that may be predisposed to plaque development and atherosclerosis.https://www.mdpi.com/1422-0067/25/6/3097macrophage M1 polarizationmCRPinflammationvascular activationsmall molecule inhibitors |
| spellingShingle | Ylenia Pastorello Doina Manu Xenia Sawkulycz Vittorio Caprio Claudia Banescu Minodora Dobreanu Lawrence Potempa Mario Di Napoli Mark Slevin mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor International Journal of Molecular Sciences macrophage M1 polarization mCRP inflammation vascular activation small molecule inhibitors |
| title | mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor |
| title_full | mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor |
| title_fullStr | mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor |
| title_full_unstemmed | mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor |
| title_short | mCRP-Induced Focal Adhesion Kinase-Dependent Monocyte Aggregation and M1 Polarization, Which Was Partially Blocked by the C10M Inhibitor |
| title_sort | mcrp induced focal adhesion kinase dependent monocyte aggregation and m1 polarization which was partially blocked by the c10m inhibitor |
| topic | macrophage M1 polarization mCRP inflammation vascular activation small molecule inhibitors |
| url | https://www.mdpi.com/1422-0067/25/6/3097 |
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