I am investigating the function of endothelial CD63, a long-established component of Weibel Palade Bodies (WPB). This universally expressed member of the tetraspanin family is best known as a marker of the intra-lumenal vesicles within multi-vesicular bodies or endosomes, but has no established endothelial-specific function in inflammation or haemostasis. In addition, I am using high resolution scanning electron microscopy to analyse the morphology of WPB exocytosis at the plasma membrane and the behaviour of the released von Willebrand factor strings under flow.
During my first post-doctoral experience, I worked on neuroendocrine cells developing an interest in secretory granules, the compartment used by cells to store bioactive peptides and hormones.
For this reason I moved to London to work on the regulated secretory pathway of endothelial cells with Dan Cutler at the MRC Laboratory for Molecular and Cell Biology. Dan’s is one of the few laboratories interested in the biogenesis of the Weibel-Palade Body (WPB), the endothelial-specific secretory granule.
My main focus here has been on the isolation/purification of WPBs in order to analyze its protein composition by mass spectrometry. We believe this will allow to identify, on one side, the molecular machinery responsible for the biogenesis and exocytosis of WPBs; on the other, new cargo with potential function in haemostasis, inflammation and angiogenesis.
I am also working to probe the relationships between the trans-Golgi network and Weibel-Palade Bodies, with the aim of identifying structural requirements for their biogenesis.
I manage the lab, train new staff and students, maintain reagents and equipment and supply technical advice, protocols and constructs. I also do research, which currently involves the use of electron and light microscopy to image the formation of Weibel-Palade bodies at the TGN.
I am interested in the molecular basis for regulation of thrombus formation and inflammation. I study the formation of Weibel-Palade bodies at the trans-Golgi network and exocytosis at the plasma membrane, particularly the role of SNARE and associated proteins in these processes. I am also using zebrafish to examine the role of Weibel-Palade body proteins in thrombosis.
I am interested in identifying novel regulators of Weibel Palade Body biogenesis and function and, more specifically, the role of kinases, which have been under-studied in the context of this secretory organelle. Whilst a few kinases have been identified as involved in regulated secretion of von Willebrand’s Factor, no roles for kinases have been reported during the earlier stages of Weibel-Palade Body formation, trafficking and regulation. I am therefore using high-throughput microscopy to carry out a screen of all human kinases, looking for candidate regulators of this organelle’s structure and behaviour.
I research the role of Rabs and the cytoskeleton in the exocytosis of Weibel-Palade Bodies. I have recently been working on the role of Rab27a and its effectors in inhibiting their fusion, preventing premature release of incompletely multimerised VWF that would be functionally impaired. I use live cell light microscopy in this work, and the long hours spent alone in a darkened room producing great movies also inspires my other role as chief supplier of (bad) puns to the Cutler lab.