AKAP12 deficiency impairs VEGF‐induced endothelial cell migration and sprouting

• DOI: 10.1111/apha.13325
• Publication Date: 2019-06-04T17:16:13Z
• Authors: Peter M. Benz; Yindi Ding; Heike Stingl; Annemarieke E. Loot; Joana Zink; Ilka Wittig; Rüdiger Popp; Ingrid Fleming
• Keywords: Mice, Knockout; Vascular Endothelial Growth Factor A; 0301 basic medicine; Microfilament Proteins; A Kinase Anchor Proteins; Endothelial Cells; Cell Cycle Proteins; Cardiovascular Physiology; Phosphoproteins; Cyclic AMP-Dependent Protein Kinases; Mice; 03 medical and health sciences; Gene Expression Regulation; Cell Movement; Animals; Phosphorylation; Cell Adhesion Molecules

AbstractAimProtein kinase (PK) A anchoring protein (AKAP) 12 is a scaffolding protein that anchors PKA to compartmentalize cyclic AMP signalling. This study assessed the consequences of the downregulation or deletion of AKAP12 on endothelial cell migration and angiogenesis.MethodsThe consequences of siRNA‐mediated downregulation AKAP12 were studied in primary cultures of human endothelial cells as well as in endothelial cells and retinas from wild‐type versus AKAP12−/− mice. Molecular interactions were investigated using a combination of immunoprecipitation and mass spectrometry.ResultsAKAP12 was expressed at low levels in confluent endothelial cells but its expression was increased in actively migrating cells, where it localized to lamellipodia. In the postnatal retina, AKAP12 was expressed by actively migrating tip cells at the angiogenic front, and its deletion resulted in defective extension of the vascular plexus. In migrating endothelial cells, AKAP12 was co‐localized with the PKA type II‐α regulatory subunit as well as multiple key regulators of actin dynamics and actin filament‐based movement; including components of the Arp2/3 complex and the vasodilator‐stimulated phosphoprotein (VASP). Fitting with the evidence of a physical VASP/AKAP12/PKA complex, it was possible to demonstrate that the VEGF‐stimulated and PKA‐dependent phosphorylation of VASP was dependent on AKAP12. Indeed, AKAP12 colocalized with phospho‐Ser157 VASP at the leading edge of migrating endothelial cells.ConclusionThe results suggest that compartmentalized AKAP12/PKA signalling mediates VASP phosphorylation at the leading edge of migrating endothelial cells to translate angiogenic stimuli into altered actin dynamics and cell movement.