Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line
The pathogenesis of atherosclerosis is complex, evolves, and involves many cell types. Macrophages and vascular smooth muscle cells (VSMCs) are critically involved in atherosclerosis development and progression. Several studies have shown that WNT5A protein is abundantly expressed in human atheroscl...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2020-11-01
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| Series: | Frontiers in Cardiovascular Medicine |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcvm.2020.567837/full |
| _version_ | 1819225156641357824 |
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| author | Ian Ackers Candice Szymanski Mitchell J. Silver Ramiro Malgor Ramiro Malgor Ramiro Malgor |
| author_facet | Ian Ackers Candice Szymanski Mitchell J. Silver Ramiro Malgor Ramiro Malgor Ramiro Malgor |
| author_sort | Ian Ackers |
| collection | DOAJ |
| description | The pathogenesis of atherosclerosis is complex, evolves, and involves many cell types. Macrophages and vascular smooth muscle cells (VSMCs) are critically involved in atherosclerosis development and progression. Several studies have shown that WNT5A protein is abundantly expressed in human atherosclerotic lesions; however, the mechanism and role of WNT signaling pathway activation is not clearly known. Using THP-1 derived macrophages, and human aortic VSMC cells, we evaluated in vitro how oxidized low-density lipoprotein (oxLDL) and WNT5A signaling interact in these two cell lines. We used western blot, scratch assay, metabolic proliferation assay, as well as immunostaining to analyze the effect of Wnt signaling activation. The results demonstrated that oxLDL, as well as WNT5A (control), induced Disheveled-2 (DVL2) activation and Kif26b degradation, indicating activation of non-canonical Wnt signaling. We found that oxLDL and WNT5A induced FZD5-ROR2 co-localization at the cellular membrane in vitro in THP-1 derived macrophages. Box5 (FZD5 receptor antagonist) inhibited oxLDL-induced DVL2/JNK activation secondary to newly secreted WNT protein from THP-1 derived macrophages. We found that WNT3A (canonical Wnt) and WNT5A showed different roles in this VSMC cell line. These findings indicate that WNT5A is upregulated by oxLDL, promotes foam cell formation, and affects VSMC phenotype and migration in these two cell lines. Also, in these cell lines FZD5 signaling seems to be necessary for lipid accumulation and, through this mechanism, WNT5A could modulate foam cell formation. Thus, our results suggest that WNT5A may contribute to the pathogenesis of vascular disease through modulating macrophage and VSMC behavior. |
| first_indexed | 2024-12-23T10:05:07Z |
| format | Article |
| id | doaj.art-8a42b9f887bf432d8c1d95dffe745306 |
| institution | Directory Open Access Journal |
| issn | 2297-055X |
| language | English |
| last_indexed | 2024-12-23T10:05:07Z |
| publishDate | 2020-11-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cardiovascular Medicine |
| spelling | doaj.art-8a42b9f887bf432d8c1d95dffe7453062022-12-21T17:51:07ZengFrontiers Media S.A.Frontiers in Cardiovascular Medicine2297-055X2020-11-01710.3389/fcvm.2020.567837567837Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell LineIan Ackers0Candice Szymanski1Mitchell J. Silver2Ramiro Malgor3Ramiro Malgor4Ramiro Malgor5Osteopathic Heritage Foundation, Translational Biomedical Sciences Program, Ohio University, Athens, OH, United StatesDepartment of Biomedical Sciences, Ohio University, Athens, OH, United StatesMidwest Cardiology Research Foundation, Columbus, OH, United StatesDepartment of Biomedical Sciences, Ohio University, Athens, OH, United StatesThe Diabetes Institute, Ohio University, Athens, OH, United StatesMolecular and Cellular Biology Graduate Program, Ohio University, Athens, OH, United StatesThe pathogenesis of atherosclerosis is complex, evolves, and involves many cell types. Macrophages and vascular smooth muscle cells (VSMCs) are critically involved in atherosclerosis development and progression. Several studies have shown that WNT5A protein is abundantly expressed in human atherosclerotic lesions; however, the mechanism and role of WNT signaling pathway activation is not clearly known. Using THP-1 derived macrophages, and human aortic VSMC cells, we evaluated in vitro how oxidized low-density lipoprotein (oxLDL) and WNT5A signaling interact in these two cell lines. We used western blot, scratch assay, metabolic proliferation assay, as well as immunostaining to analyze the effect of Wnt signaling activation. The results demonstrated that oxLDL, as well as WNT5A (control), induced Disheveled-2 (DVL2) activation and Kif26b degradation, indicating activation of non-canonical Wnt signaling. We found that oxLDL and WNT5A induced FZD5-ROR2 co-localization at the cellular membrane in vitro in THP-1 derived macrophages. Box5 (FZD5 receptor antagonist) inhibited oxLDL-induced DVL2/JNK activation secondary to newly secreted WNT protein from THP-1 derived macrophages. We found that WNT3A (canonical Wnt) and WNT5A showed different roles in this VSMC cell line. These findings indicate that WNT5A is upregulated by oxLDL, promotes foam cell formation, and affects VSMC phenotype and migration in these two cell lines. Also, in these cell lines FZD5 signaling seems to be necessary for lipid accumulation and, through this mechanism, WNT5A could modulate foam cell formation. Thus, our results suggest that WNT5A may contribute to the pathogenesis of vascular disease through modulating macrophage and VSMC behavior.https://www.frontiersin.org/articles/10.3389/fcvm.2020.567837/fullmacrophagesvascular smooth muscle cellsfoam cellsWnt signalingvascular biology |
| spellingShingle | Ian Ackers Candice Szymanski Mitchell J. Silver Ramiro Malgor Ramiro Malgor Ramiro Malgor Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line Frontiers in Cardiovascular Medicine macrophages vascular smooth muscle cells foam cells Wnt signaling vascular biology |
| title | Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line |
| title_full | Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line |
| title_fullStr | Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line |
| title_full_unstemmed | Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line |
| title_short | Oxidized Low-Density Lipoprotein Induces WNT5A Signaling Activation in THP-1 Derived Macrophages and a Human Aortic Vascular Smooth Muscle Cell Line |
| title_sort | oxidized low density lipoprotein induces wnt5a signaling activation in thp 1 derived macrophages and a human aortic vascular smooth muscle cell line |
| topic | macrophages vascular smooth muscle cells foam cells Wnt signaling vascular biology |
| url | https://www.frontiersin.org/articles/10.3389/fcvm.2020.567837/full |
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