Thesis Anna Daniel

On 5 February 2016 Anna Daniel defended her thesis ‘“In sickness and in health” Regulation of junctional integrity in Inflamed and healthy endothelial cells’ at the University of Amsterdam.

Promotor: Prof PL Hordijk PhD
Co-promotor: JD van Buul PhD

The research for this thesis was conducted at the Dept of Molecular Cell Biology of Sanquin Research and was supported by a grant of the Landsteiner Foundation for Blood Transfusion Research (#1701).

Summary

Endothelial cells form a single layer of cells on the luminal side of all blood vessels. This layer of cells represents a dynamic physical barrier that restricts and regulates the movement of blood cells, solutes and fluids from the blood to the tissues surrounding blood vessels. Within this thesis we address several important aspects of endothelial junction regulation. 
We then investigated plasminogen activator inhibitor 1 (PAI-1) as a novel regulator of endothelial junction stability. We show that PAI-1, which is expressed by endothelial cells, protects endothelial junctions in the steady state by ensuring efficient vascular endothelial cadherin (VE-cadherin) trafficking to and from the plasma membrane. Moreover, we demonstrated the importance of the phosphorylation of two adjacent serine residues in the RhoGEF Trio for the reformation of endothelial cell-cell junctions after stimulation with thrombin. Further to previous work from our lab we demonstrated here that serine phosphorylation primes Trio for rapid reformation of endothelial junctions. Finally, we returned to endothelial junction regulation in inflammation by demonstrating a role for the ubiquitin ligase cellular inhibitor of apoptosis 1 (c-IAP-1), the ubiquitin-editing enzyme tumor necrosis factor α (TNFα) -induced protein 3 (TNFAIP3) and the deubiquitinase ubiquitin-specific peptidase 8 (USP8) in limiting the extent of TNFα-induced changes to endothelial morphology, permeability and adhesion receptor expression.
Overall we have show that the serine protease inhibitor PAI-1, serine phosphorylation of the RhoGEF Trio and ubiquitination of proteins involved in TNFα-mediated signaling are important regulators of endothelial junction permeability.

Chapters

Download PhD thesis (university repository)

Chapter 1

General introduction and scope of the thesis.

Chapter 2

Endothelial junction regulation: a prerequisite for leukocytes crossing the vessel wall.
J Innate Immun 2013;(4):324-35.

Chapter 3

Leukocytes crossing the endothelium: A matter of communication.

Chapter 4

Real-time imaging of endothelial cell-cel l junctions during neutrophil transmigration under physiological flow.
J Vis Exp 2014; (90):e51766.

Chapter 5

Plasminogen activator inhibitor-1 controls vascular integrity by regulating VE-cadherin trafficking.

Chapter 6

Serine phosphorylation of the RhoGEF Trio stabilizes endothelial cell-cell junctions.

Chapter 7

Silencing of the ubiquitin ligase c-IAP-1, the ubiquitin-editing enzyme TNFAIP3 or the deubiquitinase USP8 leads to the exacerbation of TNFT-stimulated responses.

Chapter 8

Summary and concluding remarks

Download

Download the PhD thesis (university repository)