Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells
Endothelial cells (ECs) respond to concurrent stimulation by biochemical factors and wall shear stress (SS) exerted by blood flow. Disruptions in flow-induced responses can result in remodeling issues and cardiovascular diseases, but the detailed mechanisms linking flow-mechanical cues and biochemic...
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MDPI AG
2024-02-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/13/3/285 |
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| author | Ya-Wen Cheng Anthony R. Anzell Stefanie A. Morosky Tristin A. Schwartze Cynthia S. Hinck Andrew P. Hinck Beth L. Roman Lance A. Davidson |
| author_facet | Ya-Wen Cheng Anthony R. Anzell Stefanie A. Morosky Tristin A. Schwartze Cynthia S. Hinck Andrew P. Hinck Beth L. Roman Lance A. Davidson |
| author_sort | Ya-Wen Cheng |
| collection | DOAJ |
| description | Endothelial cells (ECs) respond to concurrent stimulation by biochemical factors and wall shear stress (SS) exerted by blood flow. Disruptions in flow-induced responses can result in remodeling issues and cardiovascular diseases, but the detailed mechanisms linking flow-mechanical cues and biochemical signaling remain unclear. Activin receptor-like kinase 1 (ALK1) integrates SS and ALK1-ligand cues in ECs; <i>ALK1</i> mutations cause hereditary hemorrhagic telangiectasia (HHT), marked by arteriovenous malformation (AVM) development. However, the mechanistic underpinnings of ALK1 signaling modulation by fluid flow and the link to AVMs remain uncertain. We recorded EC responses under varying SS magnitudes and ALK1 ligand concentrations by assaying pSMAD1/5/9 nuclear localization using a custom multi-SS microfluidic device and a custom image analysis pipeline. We extended the previously reported synergy between SS and BMP9 to include BMP10 and BMP9/10. Moreover, we demonstrated that this synergy is effective even at extremely low SS magnitudes (0.4 dyn/cm<sup>2</sup>) and ALK1 ligand range (femtogram/mL). The synergistic response to ALK1 ligands and SS requires the kinase activity of ALK1. Moreover, ALK1’s basal activity and response to minimal ligand levels depend on endocytosis, distinct from cell–cell junctions, cytoskeleton-mediated mechanosensing, or cholesterol-enriched microdomains. However, an in-depth analysis of ALK1 receptor trafficking’s molecular mechanisms requires further investigation. |
| first_indexed | 2024-03-08T03:59:41Z |
| format | Article |
| id | doaj.art-7c0733122f2b41919d12f958533a995e |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-08T03:59:41Z |
| publishDate | 2024-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-7c0733122f2b41919d12f958533a995e2024-02-09T15:09:53ZengMDPI AGCells2073-44092024-02-0113328510.3390/cells13030285Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial CellsYa-Wen Cheng0Anthony R. Anzell1Stefanie A. Morosky2Tristin A. Schwartze3Cynthia S. Hinck4Andrew P. Hinck5Beth L. Roman6Lance A. Davidson7Department of Bioengineering, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Structural Biology, School of Medicine, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Human Genetics, School of Public Health, University of Pittsburgh, Pittsburgh, PA 15261, USADepartment of Bioengineering, Swanson School of Engineering, University of Pittsburgh, Pittsburgh, PA 15261, USAEndothelial cells (ECs) respond to concurrent stimulation by biochemical factors and wall shear stress (SS) exerted by blood flow. Disruptions in flow-induced responses can result in remodeling issues and cardiovascular diseases, but the detailed mechanisms linking flow-mechanical cues and biochemical signaling remain unclear. Activin receptor-like kinase 1 (ALK1) integrates SS and ALK1-ligand cues in ECs; <i>ALK1</i> mutations cause hereditary hemorrhagic telangiectasia (HHT), marked by arteriovenous malformation (AVM) development. However, the mechanistic underpinnings of ALK1 signaling modulation by fluid flow and the link to AVMs remain uncertain. We recorded EC responses under varying SS magnitudes and ALK1 ligand concentrations by assaying pSMAD1/5/9 nuclear localization using a custom multi-SS microfluidic device and a custom image analysis pipeline. We extended the previously reported synergy between SS and BMP9 to include BMP10 and BMP9/10. Moreover, we demonstrated that this synergy is effective even at extremely low SS magnitudes (0.4 dyn/cm<sup>2</sup>) and ALK1 ligand range (femtogram/mL). The synergistic response to ALK1 ligands and SS requires the kinase activity of ALK1. Moreover, ALK1’s basal activity and response to minimal ligand levels depend on endocytosis, distinct from cell–cell junctions, cytoskeleton-mediated mechanosensing, or cholesterol-enriched microdomains. However, an in-depth analysis of ALK1 receptor trafficking’s molecular mechanisms requires further investigation.https://www.mdpi.com/2073-4409/13/3/285endothelial cell biologymechanotransductionmechanobiologymechanosensingpSMAD1/5/9ALK1 receptor trafficking |
| spellingShingle | Ya-Wen Cheng Anthony R. Anzell Stefanie A. Morosky Tristin A. Schwartze Cynthia S. Hinck Andrew P. Hinck Beth L. Roman Lance A. Davidson Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells Cells endothelial cell biology mechanotransduction mechanobiology mechanosensing pSMAD1/5/9 ALK1 receptor trafficking |
| title | Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells |
| title_full | Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells |
| title_fullStr | Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells |
| title_full_unstemmed | Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells |
| title_short | Shear Stress and Sub-Femtomolar Levels of Ligand Synergize to Activate ALK1 Signaling in Endothelial Cells |
| title_sort | shear stress and sub femtomolar levels of ligand synergize to activate alk1 signaling in endothelial cells |
| topic | endothelial cell biology mechanotransduction mechanobiology mechanosensing pSMAD1/5/9 ALK1 receptor trafficking |
| url | https://www.mdpi.com/2073-4409/13/3/285 |
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