Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution
Abstract Bone marrow ablation is routinely performed before hematopoietic stem cell transplantation (HSCT). Hematopoietic stem and progenitor cells (HSPCs) require a stable bone marrow microenvironment to expand and refill the peripheral blood cell pool after ablation. Roundabout guidance receptor 4...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Publishing Group
2024-02-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-024-06546-4 |
| _version_ | 1797273022244061184 |
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| author | Seyram Yao Adzraku Can Cao Qi Zhou Ke Yuan Xiaowen Hao Yue Li Shengnan Yuan Yujin Huang Kailin Xu Jianlin Qiao Wen Ju Lingyu Zeng |
| author_facet | Seyram Yao Adzraku Can Cao Qi Zhou Ke Yuan Xiaowen Hao Yue Li Shengnan Yuan Yujin Huang Kailin Xu Jianlin Qiao Wen Ju Lingyu Zeng |
| author_sort | Seyram Yao Adzraku |
| collection | DOAJ |
| description | Abstract Bone marrow ablation is routinely performed before hematopoietic stem cell transplantation (HSCT). Hematopoietic stem and progenitor cells (HSPCs) require a stable bone marrow microenvironment to expand and refill the peripheral blood cell pool after ablation. Roundabout guidance receptor 4 (Robo4) is a transmembrane protein exclusive to endothelial cells and is vital in preserving vascular integrity. Hence, the hypothesis is that Robo4 maintains the integrity of bone marrow endothelial cells following radiotherapy. We created an endothelial cell injury model with γ-radiation before Robo4 gene manipulation using lentiviral-mediated RNAi and gene overexpression techniques. We demonstrate that Robo4 and specific mesenchymal proteins (Fibronectin, Vimentin, αSma, and S100A4) are upregulated in endothelial cells exposed to irradiation (IR). We found that Robo4 depletion increases the expression of endoglin (CD105), an auxiliary receptor for the transforming growth factor (TGF-β) family of proteins, and promotes endothelial-to-mesenchymal transition (End-MT) through activation of both the canonical (Smad) and non-canonical (AKT/NF-κB) signaling pathways to facilitate Snail1 activation and its nuclear translocation. Endothelial Robo4 overexpression stimulates the expression of immunoglobulin-like adhesion molecules (ICAM-1 and VCAM-1) and alleviates irradiation-induced End-MT. Our coculture model showed that transcriptional downregulation of endothelial Robo4 reduces HSPC proliferation and increases HSC quiescence and apoptosis. However, Robo4 overexpression mitigated the damaged endothelium’s suppressive effects on HSC proliferation and differentiation. These findings indicate that by controlling End-MT, Robo4 preserves microvascular integrity after radiation preconditioning, protects endothelial function, and lessens the inhibitory effect of damaged endothelium on hematopoietic reconstitution. |
| first_indexed | 2024-03-07T14:37:36Z |
| format | Article |
| id | doaj.art-24d1a43ab8b849df850fb1dc90a8c5ad |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-03-07T14:37:36Z |
| publishDate | 2024-02-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-24d1a43ab8b849df850fb1dc90a8c5ad2024-03-05T20:31:12ZengNature Publishing GroupCell Death and Disease2041-48892024-02-0115211610.1038/s41419-024-06546-4Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitutionSeyram Yao Adzraku0Can Cao1Qi Zhou2Ke Yuan3Xiaowen Hao4Yue Li5Shengnan Yuan6Yujin Huang7Kailin Xu8Jianlin Qiao9Wen Ju10Lingyu Zeng11Blood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityBlood Diseases Institute, Xuzhou Medical UniversityAbstract Bone marrow ablation is routinely performed before hematopoietic stem cell transplantation (HSCT). Hematopoietic stem and progenitor cells (HSPCs) require a stable bone marrow microenvironment to expand and refill the peripheral blood cell pool after ablation. Roundabout guidance receptor 4 (Robo4) is a transmembrane protein exclusive to endothelial cells and is vital in preserving vascular integrity. Hence, the hypothesis is that Robo4 maintains the integrity of bone marrow endothelial cells following radiotherapy. We created an endothelial cell injury model with γ-radiation before Robo4 gene manipulation using lentiviral-mediated RNAi and gene overexpression techniques. We demonstrate that Robo4 and specific mesenchymal proteins (Fibronectin, Vimentin, αSma, and S100A4) are upregulated in endothelial cells exposed to irradiation (IR). We found that Robo4 depletion increases the expression of endoglin (CD105), an auxiliary receptor for the transforming growth factor (TGF-β) family of proteins, and promotes endothelial-to-mesenchymal transition (End-MT) through activation of both the canonical (Smad) and non-canonical (AKT/NF-κB) signaling pathways to facilitate Snail1 activation and its nuclear translocation. Endothelial Robo4 overexpression stimulates the expression of immunoglobulin-like adhesion molecules (ICAM-1 and VCAM-1) and alleviates irradiation-induced End-MT. Our coculture model showed that transcriptional downregulation of endothelial Robo4 reduces HSPC proliferation and increases HSC quiescence and apoptosis. However, Robo4 overexpression mitigated the damaged endothelium’s suppressive effects on HSC proliferation and differentiation. These findings indicate that by controlling End-MT, Robo4 preserves microvascular integrity after radiation preconditioning, protects endothelial function, and lessens the inhibitory effect of damaged endothelium on hematopoietic reconstitution.https://doi.org/10.1038/s41419-024-06546-4 |
| spellingShingle | Seyram Yao Adzraku Can Cao Qi Zhou Ke Yuan Xiaowen Hao Yue Li Shengnan Yuan Yujin Huang Kailin Xu Jianlin Qiao Wen Ju Lingyu Zeng Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution Cell Death and Disease |
| title | Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| title_full | Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| title_fullStr | Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| title_full_unstemmed | Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| title_short | Endothelial Robo4 suppresses endothelial-to-mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| title_sort | endothelial robo4 suppresses endothelial to mesenchymal transition induced by irradiation and improves hematopoietic reconstitution |
| url | https://doi.org/10.1038/s41419-024-06546-4 |
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