Altered Storage and Function of von Willebrand Factor in Human Cardiac Microvascular Endothelial Cells Isolated from Recipient Transplant Hearts

Author:

Meli Athinoula1,McCormack Ann1,Conte Ianina2,Chen Qu3,Streetley James4ORCID,Rose Marlene L.1,Bierings Ruben5ORCID,Hannah Matthew J.6ORCID,Molloy Justin E.7,Rosenthal Peter B.4,Carter Tom2ORCID

Affiliation:

1. Transplant Immunology, Heart Science Centre, Harefield Hospital, Hill End Road, Harefield UB9 6JH, UK

2. Molecular and Clinical Sciences Research Institute, St Georges University of London, London SW17 0RE, UK

3. Structural Biology Science Technology Platform, The Francis Crick Institute, London NW1 1AT, UK

4. Structural Biology of Cells and Viruses Laboratory, The Francis Crick Institute, London NW1 1AT, UK

5. Hematology, Erasmus University Medical Center, P.O. Box 2040, 3000 CA Rotterdam, The Netherlands

6. High Containment Microbiology, UK Health Security Agency, London NW9 5EQ, UK

7. Single Molecule Enzymology Laboratory, The Francis Crick Institute, London NW1 1AT, UK

Abstract

The assembly of von Willebrand factor (VWF) into ordered helical tubules within endothelial Weibel–Palade bodies (WPBs) is required for the efficient deployment of the protein at sites of vascular injury. VWF trafficking and storage are sensitive to cellular and environmental stresses that are associated with heart disease and heart failure. Altered storage of VWF manifests as a change in WPB morphology from a rod shape to a rounded shape and is associated with impaired VWF deployment during secretion. In this study, we examined the morphology, ultrastructure, molecular composition and kinetics of exocytosis of WPBs in cardiac microvascular endothelial cells isolated from explanted hearts of patients with a common form of heart failure, dilated cardiomyopathy (DCM; HCMECD), or from nominally healthy donors (controls; HCMECC). Using fluorescence microscopy, WPBs in HCMECC (n = 3 donors) showed the typical rod-shaped morphology containing VWF, P-selectin and tPA. In contrast, WPBs in primary cultures of HCMECD (n = 6 donors) were predominantly rounded in shape and lacked tissue plasminogen activator (t-PA). Ultrastructural analysis of HCMECD revealed a disordered arrangement of VWF tubules in nascent WPBs emerging from the trans-Golgi network. HCMECD WPBs still recruited Rab27A, Rab3B, Myosin-Rab Interacting Protein (MyRIP) and Synaptotagmin-like protein 4a (Slp4-a) and underwent regulated exocytosis with kinetics similar to that seen in HCMECc. However, secreted extracellular VWF strings from HCMECD were significantly shorter than for endothelial cells with rod-shaped WPBs, although VWF platelet binding was similar. Our observations suggest that VWF trafficking, storage and haemostatic potential are perturbed in HCMEC from DCM hearts.

Funder

UK Medical Research Council

Cancer Research UK

the Wellcome Trust

British Heart Foundation

Publisher

MDPI AG

Subject

Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis

Reference56 articles.

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