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| Status |
Public on Mar 29, 2021 |
| Title |
Transcriptome analysis of the role of SMAD6 in mediating endothelial cell response to laminar shear stress by high throughput RNA sequencing |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Laminar shear stress regulates blood vessel morphogenesis and subsequent quiescence, but how endothelial cells (EC) enact and maintain the vascular homeostasis required in most vessels for proper vessel function is poorly understood. SMAD6, a scaffold for several signaling pathways, is expressed in developing arteries and its expression is flow-regulated. We found that SMAD6 is essential for endothelial cell flow-mediated responses, and that it functions downstream of the mechanosensor Notch1. Endothelial cells with reduced SMAD6 levels failed to align under stable laminar shear flow that promotes vascular homeostasis, while forced SMAD6 expression rescued misalignment induced by reduced Notch1 signaling. SMAD6-dependent homeostatic laminar flow required the Notch ligand Dll4 and Notch transcriptional activity. Mechanistically, neither the N-terminal nor the C-terminal domain of SMAD6 alone rescued flow alignment upon loss of Notch signaling. Endothelial cells with reduced Smad6 levels has compromised barrier function, and RNA profiling revealed upregulation of proliferation-associated genes and down regulation of junction-associated genes. Among junction-related genes affected by SMAD6 levels, the proto-cadherin PCDH12 was upregulated by homeostatic flow and required for proper flow-mediated endothelial cell alignment. Thus, SMAD6 is a critical integrator of flow-mediated signaling inputs downstream of Notch1, as vessels transition from an angiogenic to a homeostatic phenotype.
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| Overall design |
To understand how SMAD6 mediate EC flow response, we performed bulk RNAseq of human umbilical vein endothelial cells (HUVEC) transfected with siNT non-targeting control siRNA or siSMAD6 (30 hrs of transfection) and then cultured under static condition or flowed under 15 dynes/cm2 laminar shear for 72 hrs.
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| Web link |
https://0-link-springer-com.brum.beds.ac.uk/article/10.1007/s10456-021-09777-7
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| |
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| Contributor(s) |
Liu Z, Ruter DL, Bautch VL |
| Citation(s) |
33779885 |
| Submission date |
Mar 16, 2020 |
| Last update date |
Jul 08, 2021 |
| Contact name |
Ziqing Liu |
| Organization name |
UNC-Chapel Hill
|
| Department |
Biology
|
| Lab |
Victoria Bautch
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| Street address |
416 Fordham Hall, University of North Carolina
|
| City |
Chapel Hill |
| State/province |
NC |
| ZIP/Postal code |
27514 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL20301 |
Illumina HiSeq 4000 (Homo sapiens) |
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| Samples (8)
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| Relations |
| BioProject |
PRJNA612830 |
| SRA |
SRP252947 |
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