CRISPR-mediated deletion of the PECAM-1 cytoplasmic domain increases receptor lateral mobility and strengthens endothelial cell junctional integrity. Life Sci 2018 Jan 15;193:186-193
Date
11/11/2017Pubmed ID
29122551Pubmed Central ID
PMC5754039DOI
10.1016/j.lfs.2017.11.002Scopus ID
2-s2.0-85033432699 (requires institutional sign-in at Scopus site) 5 CitationsAbstract
AIMS: PECAM-1 is an abundant endothelial cell surface receptor that becomes highly enriched at endothelial cell-cell junctions, where it functions to mediate leukocyte transendothelial migration, sense changes in shear and flow, and maintain the vascular permeability barrier. Homophilic interactions mediated by the PECAM-1 extracellular domain are known to be required for PECAM-1 to perform these functions; however, much less is understood about the role of its cytoplasmic domain in these processes.
MAIN METHODS: CRISPR/Cas9 gene editing technology was employed to generate human endothelial cell lines that either lack PECAM-1 entirely, or express mutated PECAM-1 missing the majority of its cytoplasmic domain (∆CD-PECAM-1). The endothelial barrier function was evaluated by Electric Cell-substrate Impedance Sensing, and molecular mobility was assessed by fluorescence recovery after photobleaching.
KEY FINDINGS: We found that ∆CD-PECAM-1 concentrates normally at endothelial cell junctions, but has the unexpected property of conferring increased baseline barrier resistance, as well as a more rapid rate of recovery of vascular integrity following thrombin-induced disruption of the endothelial barrier. Fluorescence recovery after photobleaching analysis revealed that ∆CD-PECAM-1 exhibits increased mobility within the plane of the plasma membrane, thus allowing it to redistribute more rapidly back to endothelial cell-cell borders to reform the vascular permeability barrier.
SIGNIFICANCE: The PECAM-1 cytoplasmic domain plays a novel role in regulating the rate and extent of vascular permeability following thrombotic or inflammatory challenge.
Author List
Liao D, Mei H, Hu Y, Newman DK, Newman PJAuthors
Debra K. Newman PhD Professor in the Pharmacology and Toxicology department at Medical College of WisconsinDebra K. Newman PhD Investigator in the Blood Research Institute department at BloodCenter of Wisconsin
MESH terms used to index this publication - Major topics in bold
Capillary PermeabilityCell Adhesion
Cell Line
Cytoplasm
Endothelial Cells
Humans
Intercellular Junctions
Platelet Endothelial Cell Adhesion Molecule-1
Protein Binding
Protein Domains
Thrombin
