DIAGNOSIS AND THERAPIES FOR MUCOPOLYSACCHARIDOSES by Francyne Kubaski A dissertation submitted to the Faculty of the University of Delaware in partial fulfillment of the requirements for the degree of Doctor of Philosophy in Biological Sciences Spring 2017 © 2017 Francyne Kubaski All Rights Reserved DIAGNOSIS AND THERAPIES FOR MUCOPOLYSACCHARIDOSES by Francyne Kubaski Approved: __________________________________________________________ Robin W. Morgan, Ph.D. Chair of the Department of Biological Sciences Approved: __________________________________________________________ George H. Watson, Ph.D. Dean of the College of Arts and Sciences Approved: __________________________________________________________ Ann L. Ardis, Ph.D. Senior Vice Provost for Graduate and Professional Education I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Erica M. Selva, Ph.D. Professor in charge of dissertation I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Shunji Tomatsu, Ph.D. Member of dissertation committee I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Robert W. Mason, Ph.D. Member of dissertation committee I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Anja Nohe, Ph.D. Member of dissertation committee I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Colin Thorpe, Ph.D. Member of dissertation committee I certify that I have read this dissertation and that in my opinion it meets the academic and professional standard required by the University as a dissertation for the degree of Doctor of Philosophy. Signed: __________________________________________________________ Donna S. Woulfe, Ph.D. Member of dissertation committee ACKNOWLEDGMENTS I would like to express my deepest acknowledgements to my family and dedicate this dissertation to them. I could not have achieved this dream without the support of my parents, grandparents, aunts, uncles and cousins. I have to thank my mom for her never-ending encouragement, love, and support from the very beginning when all of this was just a dream. I have to thank my dad for his support, love, care, and for always encouraging me to be the best that I could be. Thank you both for inspiring me to be who I am, for believing in me and making me believe in myself. I love you both so much, and I could not have done any of this without you both by my side. I also would like to dedicate this achievement to the brightest star in the sky, my grandma Carmen. I know how hesitant you were when I left and I know this was not easy for you. Unfortunately, you are not physically here to see me walking across the stage, but I know that wherever you are you will be sending me your love and you will be looking out for me. I would like to acknowledge my advisor, Dr. Shunji Tomatsu, for all of your mentoring, teaching, thorough guidance, encouragement, trust, and care, and for this, I am tremendously thankful. You have inspired me to be a better scientist, as well as instilled in me the traits I wish to possess as a mentor for my future students. I am extremely lucky to have had the opportunity to not only work with you, but also learn from you. I would also like to acknowledge my Co-advisor, Dr. Robert W. Mason. You were more of an advisor and true mentor to me rather than a Co-advisor. You have v taught me how to critically analyze data. You have taught me everything I know about mass spectrometry, in which I would have otherwise not been able to obtain any data without your thorough guidance. You have fixed my grammar innumerable times, giving my work better quality. You have helped me countless days in all scientific matters, in developing methods and analyses, in writing manuscripts and scientific proposals, and with this dissertation. I am assured that I have become a better scientist under your guidance, and I cannot thank you enough for everything that you have done for me. I would like to thank my advisor of record, Dr. Erica M. Selva. You are an amazing person and I am so grateful for all of the help, support, and guidance you have provided me throughout this Ph.D. I would also like to thank all of my committee members, Dr. Anja Nohe, Dr. Colin Thorpe, and Dr. Donna S. Woulfe. I am extremely thankful to all of you for the critical analysis provided during our discussions, in which excellent suggestions have highly improved the quality of my work. I would like to express my gratitude to Dr. Roberto Giugliani for believing in my potential and for awarding me with a Ph.D. scholarship, which was essential to make this dream come true. I will be forever grateful for your trust and countless help. I would like to acknowledge all of the support and inspiration from my Master`s advisor, Dr. Sandra Leistner-Segal. Profe, you are more than a mentor to me. You have encouraged me from the beginning and have stood there by my side. You have taught me so much, and I am so appreciative of your trust and support. I would like to thank my best friend, Victória Bernardes Guimarães (que lindooo), you know how important you are to me. None of this would have been vi possible without your support. You have been there through some really rough times, and you have never given up. You always found a way to make me smile, even if that meant eating corn with me on Skype. Thanks for being always there for me, and know that I will always be by your side. I would also like to thank the three best friends that Delaware gave me: Effie Halakos, Kimberly Eagle, and Miho Maeda. You were the greatest gifts I received from Udel. You have done so much for me, and mostly you have always believed in me. Thank you so much from the bottom of my heart. You can always count on me. Throughout this Ph.D. journey, I have met some amazing people that I will take with me wherever I go. Zainab, Vratasha, Aparna, Soma, Mike, Sambee, Christian, Cinsley, Katie, Sam, Karen, Shiva, Pravin, Molly, and Hemanth. You guys made this journey so much easier, and I cannot thank you enough for all of the care and the good moments we have shared. Soma, you are the best! You are such a good friend to me, you have helped me so many times with all sorts of things, and I am so grateful for everything. I know you are going very far and I am really proud of the scientist you are. I also need to express my deepest gratitude to Anthony J. Quagliano (human), you have done so much for me, and you have never left my side besides all the ups and downs. Thank you for your true support and care. You were absolutely always there for me. I also need to acknowledge all the girls from the “SheWolfpack”! You were all true gifts, and you made me part of the family. I also need to express my gratitude for special friends from home. Estela, Michelle, Renata, Nathalie, Elisa, Ana, Fe Bender, Gabi, Aline, Gui, Isabel, and Kris. You are all very special to me, and I really appreciate all the care, love and true support. vii I would like to acknowledge all my current and previous lab members for their support, guidance, and help. Eriko, Tsutomu, Kazuki, Molly, Shaukat, Caitlin, Melodie, Hira, and Jess. You guys make this group truly special. I would like to thank Betty Cowgill for all of her help. You are truly the best. Thank you for everything you have done and for always helping without any hesitation. I would like to thank Virginia Redmond for her amazing help with the dissertation formatting and Dianna McKellar for her tremendous support with Refworks. I would also like to acknowledge Dr. Thomas H. Shaffer, Dr. Theodore Harcke and Heidi Kecskemethy for their help and knowledge. You have taught me so much about lung function and bone mineral density. I would also like to acknowledge all the MPS patients and families that were involved in this study. All the work we do is to improve your life and all of this would be meaningless without your support and hope. Lastly, I would also like to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and the Instituto Nacional de Genética Médica Populacional (INAGEMP) for all of the funding and support to make this Ph.D. possible. As well as National Institutes of Health, International Morquio Organization (Carol Ann Foundation) and Austrian MPS Society for research funds. viii “The most important thing… is not to win but to take part, just as the most important thing in life is not the triumph but the struggle. The essential thing is not to have conquered but to have fought well.” -Pierre de Coubertin ix TABLE OF CONTENTS LIST OF TABLES .......................................................................................................