The University of Leeds Faculty of Medicine and Health Leeds Institute of Cardiovascular and Metabolic Medicine Functions and mechanisms of Rab46 in endothelial cells Lucia Pedicini October, 2019 Thesis submitted to The University of Leeds in accordance with the requirements for the Degree of Doctor of Philosophy II Intellectual Property and Publication Statements The candidate confirms that the work submitted is his own and that appropriate credit has been given where reference has been made to the work of others. This copy has been supplied on the understanding that it is copyright material and that no quotation from the thesis may be published without proper acknowledgement. The right of Lucia Pedicini to be identified as Author of this work has been asserted by her in accordance with the Copyright, Designs and Patents Act 1988. © 2019. The University of Leeds and Lucia Pedicini III Acknowledgments My PhD journey has been a great personal and professional challenge since the very beginning. I will never forget the first days when I had to face the Northern accent and quickly realise that my English knowledge was not enough in the North of England, a land far away from the posh Queen’s English that I learnt at school. Undertaking this PhD has been a truly life-changing experience for me and it would not have been possible without the people I met throughout this journey. First and foremost, I would like to thank the continued guidance and support of my supervisor, Dr Lynn McKeown. Thanks for your patience every time I asked you to slow down and ‘say it again’ during our lab meetings – how we have put up with each other accents I still do not know! I will be eternally grateful for believing in me and for always having an open door especially when things did not work out. I could not have imagined having a better advisor for my PhD study. I look forward to continuing working together. I gratefully acknowledge Professor David Beech for all his constant support and guidance throughout my PhD. All the invaluable advice helped me working at my full potential without being lost within my research. I feel privileged to have had the opportunity to carry out my PhD in his lab. I also would like to thank Dr Izzy Jayasinghe for being always very welcoming in his lab and for all the extremely useful advice on imaging analysis. My PhD has been a continuous learning curve and I am especially grateful to all the old and new members of the Beech lab for their help and support over the years. Hannah who has always been patient in teaching me all the techniques from scratch at the very beginning of my PhD. Beth, Nikki, Katy and Nele who have been part of these three years as good teammates. Beth with your posh accent, you have always made me laugh and smile with all the hilarious stories of your daily life. Thanks for always being a good friend and a good lab mate, especially with our fury friends. I’ll miss you so much but moving it’s part of the scientist’s life and I know you’ll be a brilliant and successful scientist. Nikki, I will always remember your bagels and your very precise western blots. We shared this last few months of writing and it was a good feeling not to being alone. Katy, thanks for your fluent Italian and for your mild Manchester accent, I am sorry for all the time you had to express yourself again. Also, you’re an artist, keep drawing you’re amazing! Nele, thanks for always being very supportive and for helping me since the beginning when I was struggling to adjust myself to live in a foreign country. Thanks for listening, for understanding my struggle about living abroad and for always being a very good friend. So many things have changed in these last years but definitely the best one is your little girl. IV Marj and Fiona thanks for being enormously helpful in this year. Fiona thanks for answering my very stupid PCR questions and Marj for sharing the pain of our western blot session together, we will never know which friends show up at our party! Thanks to both of you for sharing your Tuscany trip with me, you know how much I miss it. Anyway I’m very happy you’ll be part of whatever is coming next in my LICAMM experience. And please keep filming our daily life! Last but not least a huge thank you for someone who has been sharing this adventure for me since the very first day, Kat. It was hard at the beginning to find a balance between me and you as we were both experiencing hard time but now, I feel I couldn’t do it without you on my side. You have always been very helpful, you have never left me alone and most and foremost you have been the most supportive person in these last few months. You know better than anyone how hard it was for me, how much I sacrificed trying to work hard and finish this PhD on time. I am indebted to you for being a patient listener, I have always felt better after all our complain sessions! Kat you are really a good friend and the best lab mate ever, I’ll always be grateful for all the times you made me laugh during the experiments, to always keep a sense of humour when I had lost mine and for all the crimes that somehow always happen to us. Hope to return everything you did for me in the next year! I also like to thank the rest of the McKeown’s group. Ash and Sabina we couldn’t ask for better new entries than you. Thanks a lot for your help and proofreading and I really think we should do more Aberdeen trips together. It was a very good time. I also want to thank my new and old friends. My Italian friends who have always waited for my return maybe for just a dinner together and friends whose although the distance made hard to see each other, are still part of my life. Moving to Leeds was not easy and first and foremost I want to thank my friend Guada who is the first person I met here. We shared the house during my first year and since the beginning was like we knew each other from years. They say that Italians and Spanish have something in common, I have to say that maybe it’s true. You have been the closest friend here in the UK, you were always there to help me when I was alone and definitely think we built a long-lasting friendship. A very deep thank you must now go to my family, to my life-coaches: my granddad and my grandma and to the love of my life. Un grazie di cuore a voi mamma e papà che siete e sarete per sempre la mia “casa”, il mio rifugio sicuro. Grazie per avermi sempre sostenuto in tutte le mie scelte, so che è stato difficile lasciarmi andare. Ogni volta che parto lo vedo nei vostri occhi e lo sento in silenzio nel mio cuore…ma io vi porto sempre con me, porto con me i valori che mi avete insegnato e spero di potervi far sentire orgogliosi del futuro che sto cercando di costruire. Magari un giorno chissà a furia di venirmi a trovare vi verrà anche voglia di viaggiare! C’è un mondo da scoprire e anche se l’Italia sarà sempre il posto più bello del mondo ne vale la pena di scoprire il resto. V Francesco e Noemi, che dire non potevamo essere più diversi di così ma siete la parte diciamo frizzante della famiglia. Mi sono sempre sentita un po’ mamma e un po’ sorella per voi…chissà forse solo per tenere a bada i vostri animi ribelli…comunque vogliate io ci sarò sempre per voi anche perché cosa fareste senza di me? Chi sarà la parte saggia del trio? Grazie semplicemente di esserci. E poi è difficile ringraziare le persone che hanno fatto di me quella che sono oggi, trovare le parole per descrivere cosa siete per me è davvero difficile. Siete la mia più grande fortuna, ci siete sempre stati, mi avete regalato tutti i ricordi più belli e la gioia nel cuore di tornare a casa. Grazie a te nonno che sei sempre stato il mio più grande sostenitore, sei la mia motivazione e vedere nei tuoi occhi la tua soddisfazione quando parlo di scienza è il mio regalo più grande. Grazie per avermi trasmesso l’amore per scoprire cose nuove e soprattutto l’amore per viaggiare….sarete con me in ogni mio viaggio! Grazie a te a nonna per aspettare ogni volta le mie telefonate per sapere quando torno. Come dici tu non è semplice stare lontani (anche se non esattamente con queste parole). Ma non importa la distanza perché i momenti che passiamo insieme sono i più belli di sempre, abbiamo tante cose da fare e anche se ora forse ti sembra più difficile, niente è impossibile perché l’amore smuove le montagne e io ne ho abbastanza per smuoverti da quella poltrona. L’ultimo grazie è sempre per Te che hai sempre creduto in me, mi hai spinto a guardare in alto, ad essere una persona migliore prima di tutto per me stessa. Mi hai spesso, a volte con forza, posto davanti i miei limiti e spinto a superarli.