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Universidad Autónoma De Madrid UNIVERSIDAD AUTÓNOMA DE MADRID DEPARTAMENTO DE BIOLOGÍA MOLECULAR Identification and functional characterization of epigenetic determinants of pancreatic CSCs Sladjana Zagorac Madrid, 2015 DEPARTAMENTO DE BIOLOGÍA MOLECULAR FACULTAD DE CIENCIAS UNIVERSIDAD AUTÓNOMA DE MADRID Identification and functional characterization of epigenetic determinants of pancreatic CSCs Sladjana Zagorac Licenciatura en Biología Molecular y Fisiología Director de Tesis: Prof. Christopher Heeschen, M.D. PhD Centro de células madre en el cáncer y el envejecimiento Barts Instituto de Cancer (BCI) This thesis, submitted for the degree of Doctor of Philosophy at the Universidad Autónoma de Madrid, has been completed in the Stem Cells & Cancer Group at the Spanish National Cancer Research Centre (CNIO) and Centre for Stem Cells in Cancer and Ageing, Barts Cancer Instutute (BCI) under the supervision of Prof. Dr. Christopher Heeschen. Dedicated to my parents, my sister Ivana and my friends who always supported me. ACKNOWLEDGEMENTS It is strange to look back in time and see myself entering true the door of CNIO my first day in Christopher’s laboratory back in October 2010. For some maybe little and for other’s so much. Thank you Christopher for replaying to my email and opening me the door of your lab! It was just a start of an incredible journey that led me where I am now, and that will mark my next trails to run. So many people I need to mention and say thanks for their unconditional help and support. Enza thank you for accepting me as your student and for being patient with me. I will never forget how you would always tell me to think why I am doing things. Your school was taught, but set the stage for me to become independent young scientist. Hundred thanks to Iker, Jorge, Susana, Merxe, Jenifer, Javi, Michele. Thank you Patrick for chasing the single cells with me, and for always having a nice joke to share with me. Irene special thanks to you too. Thank you for sharing your autofluorescent secrets with me, and for being with me when I needed a hand. I need to express my deep gratitude to Sonia. Always armed with patients, and willingness to jump and help or answer any question I had. Thank you for being my friend and lab mate that anyone can just wish for. Yolanda, I think that time that I spent with you was the funniest part of my PhD. Thank you for support, and always positive attitude. Thank you for taking me on the roller TET-coaster, the ride I enjoyed from the first day, and the journey that made me love science and discoveries even more! This PhD was not regular one, I would say. Took me from my home town, Belgrade, to Madrid, where I have spent three wonderful years. And then this last year, giving me opportunity to come and live in London. Gave me opportunity to meet new great scientist and appreciate differences even more. Special thanks to my London’s co-workers, though still with touch of Madrid’s one. Thank you Emma, Sholah, Andy, David, Ahmet, Deepak, Michaela, Minghsin, Marina. Tony I will never forget your words. Thank you for reminding me that life is constant process of learning and for being my London’s supervisor. Matt, can’t say your name without the smile on my face. Thank you for making this year cheerful, always having a nice word for ‘’Good morning’’. Thank you for all help and support, time and patients that you had for me and for setting up my first step in the world of ‘’grown up scientists’’. Mara, you always had time for me with just words of encouragement and support. Thank you for all that times when you just was there for me to help me with animals, histology or any other thing I would bother you about. Shan, how can I thank you more for all nice words and help you gave me during also writing up this thesis. I am so happy I had opportunity to share my time in the lab with you, I will surely miss you a lot. Petra, always breaking my fear. We have more than time in the lab, talk about successful and/or unsuccessful experiments. This year in London wouldn’t be the same without you. I admire you as a young scientist and as a person. I hope to celebrate with you your big publication because I believe in your talent and your brightness. Sara and Patricia, you are just more than co-workers or flatmates. How can I thank you for all past and present support. Patricia with her strong but always positive opinions. I can say that I admire you deeply as a person, scientist and as a friend. I am grateful that I had opportunity to work, live and learn from you. Sara, when I hear your laugh that just makes things so easier. Thank you for sharing next-bench with me, for taking the ’’grey clouds away’’, for listening me and for supporting me. Alexandra, present from the beginning! Sometimes directly involved and many times in the shadow, but always prepared to give me a hand and refresh with ideas. Many thanks for all unconditional help. Hours and weekends that we shared, reminds me that hard work pays back sooner or later. Thank you very much. I give my thanks, as well to people from my PhD committee, Manuel Serrano, Maria Jose Barrero and Mario Fraga. Thank you for all support during these years and for the meaningful and productive meetings. I have to mix a bit now and say word of thanks to other group of people in my life before I move on, and give my thanks two persons very important in my scientific life. I need to thank to my friends, Ksenija and Dragana, many years of our friendship is yet to come, but thank you for support, encouragement and patients in keeping our long distance friendship live and strong! Miguel, your support was more than important. You were there for every lab-meeting to listen to me, and ask questions even not having clue what I was talking about. Spending weekends with me in CNIO. Thank you for holding my hand during this journey and for always having a smile and joke under your sleeves. I give my thanks to my parents that were always there for me. Without your support I wouldn’t be able to achieve what I did. Special thanks to my family members. I know that you are happy to see me building my one castle. Ivana, if you haven’t opened the door…this PhD is like a marathon and running it next to you, as a colleague, friend, sister…was the best part of it. Hope that our scientific paths will continue to overlap, because I won’t tell how much I admire you as a sister, you already know that, but I admire you very much as a scientist as well. Keep on running toward your thesis, and I will be there for you just like you were there for me!! Bruno, I was thinking when I come to the part to thank you that words will just start to flow. Instead I am stunned how small words are. When you took me under your ‘’scientific shelter’’, I knew that doesn’t matter how hard science is, and how difficult is to keep up being motivated, that having someone brilliant next to, is sufficient to overcome all obstacles. Hours you spent with me, lessons you thought me, times when you had a handkerchief for me…all these things you have done for me, since the begging and at the end. Was an honour growing up as a scientist next to you. Hope that one day I would be able to pay you back. Thank you for everything!!! Lastly, as I started I would like to finish. Christopher, it’s a ‘’cliché’’ to say in Doctoral thesis, but I need to say it. Thank you for giving me opportunity to do my PhD in your lab. It was a hard rock to climb, but I managed to do it! I am more than grateful for every time you were there to say words of encouragement and support. For being tough and demanding, because only in that way I managed to mature as a scientist and as a person! You are an admirable tutor! At the end I think I will keep on pipetting and thanks to people I mentioned I hope I have set up good grounds to one day make a difference. TABLE OF CONTENTS TABLE OF CONTENTS SUMMARY .............................................................................................................................. 9 RESUMEN.............................................................................................................................. 13 ABBREVIATIONS ................................................................................................................ 17 INTRODUCTION.................................................................................................................. 21 1. PANCERATIC DUCTAL ADENOCARCINOMA ............................................................ 23 1.1. Risk factors for PDAC. ................................................................................................. 23 1.2. Molecular biology of PDAC. ........................................................................................ 24 Figure I1. ....................................................................................................................... 25 1.3. Treatment options for PDAC patients. .......................................................................... 25 Table I1. ........................................................................................................................ 28 2. CANCER STEM CELLS IN CANCER ............................................................................... 28 2.1.The cancer stem cell concept in cancer ......................................................................... 28 Figure I2. ...................................................................................................................... 31 2.2. Identification of cancer stem cells..................................................................................31
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