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Interrogating DOT1L Recruitment by MLL-Fusion Proteins MLL-AF9 and MLL-ENL Towards the Development of Novel Targeted Therapy

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Interrogating DOT1L Recruitment by MLL-Fusion Proteins MLL-AF9 and MLL-ENL Towards the Development of Novel Targeted Therapy Interrogating DOT1L Recruitment by MLL-Fusion Proteins MLL-AF9 and MLL-ENL Towards the Development of Novel Targeted Therapy by Sierrah Marie Grigsby A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Molecular and Cellular Pathology) in the University of Michigan 2019 Doctoral Committee: Associate Professor Zaneta Nikolovska-Coleska, Chair Professor Yali Dou Associate Professor Ivan Maillard Assistant Professor Andrew Muntean Sierrah Marie Grigsby [email protected] ORCID iD: 0000-0002-5410-8792 © Sierrah Marie Grigsby 2019 DEDICATION This work is dedicated to my Guardian Angels: My dearest Mother Bettye Lois Albert Grigsby February 1, 1966 – February 7, 2016 My dearest maternal Grandmother Cloteal Albert August 31, 1936 – November 12, 2014 My dearest paternal Grandfather Tommie Lee Grigsby October 10, 1932 - August 8, 2014 ii ACKNOWLEDGEMENTS My journey to my Ph.D. has been riddled with many peaks and valleys, but through it all, I have been supported by my faith, family, friends, and colleagues. First and foremost, I would like to give thanks and honor to my Lord and Savior Jesus Christ. Isaiah 40:31 states, “but those who hope in the LORD will renew their strength. They will soar on wings like eagles; they will run and not grow weary, they will walk and not be faint.” My faith provided me with the strength needed to finish this project with grace and courage. I want to honor my parents Archie Edwin Grigsby and Dr. Bettye Lois Albert Grigsby for their undying love and support throughout my educational career. Unfortunately, my mother is not physically here to support me further, but I will carry her legacy with me as I pursue my dreams. My half-siblings Legend, Leah, and Camille who inspire me to be the best role model that I can be. To my maternal grandparents J.W. and Cloteal Albert, to whom I am deeply indebted for their love and support me during my early years, and I am truly grateful. To my paternal grandparents Murd and Tommie Lee Grigsby and Jimmie Nell Headrick for their words of encouragement. I would also like to acknowledge my God-parents Rev. Marion Phillips III, Dr. Rhonda Osborne, Harold Hankins and Cornelia Hankins who have been instrumental in my upbringing as surrogate parents when I have needed them. To my aunts Dr. DeEadra Albert- Green and Sheila Headrick-Sisk and extended family for their prayers and support throughout my journey in graduate school. My Aunt, DeEadra, and mother, Bettye, have always been my role models and friends. They were the first in my immediate family to receive their Ph.D., and I am exceedingly blessed to have my aunt’s continued support. iii Since I chose to pursue both my undergraduate and graduate degrees from institutions far away from my home state, Texas, it was vital for me to establish a new family network. I was truly blessed to have met my biggest supporter Prince Diaby, who has selflessly put up with my changing moods and missed texts due to lack of sleep and many hours in the lab. He and his family treat me as their own, and I look forward to us continuing to push each other towards our dreams. I would also like to thank my lovely sorority sisters of Alpha Kappa Alpha Sorority, Inc. for their many cards, hugs, support, and meals throughout my undergraduate and graduate career. They have always been there for me in my times of need and invaluable additions to my family. During undergrad and graduate school, I was able to make amazing friends. My undergraduate roommates Beverly Wong and Katrina Manzi who always remind me to eat correctly and take breaks when needed. Thank you to My recent roommates Lindsey Harris and Paloma Garcia for putting up with all of the many faces of graduate student Sierrah that changed on a regular basis. My undergraduate friends Thanh Ngo, Soyoun Kim, and Edward Chung have been supportive throughout all of my studies. To my friends back home, Megan Sumpton, Harold Booker, Grace O’Neal, and Sharon Ogidan, thank you so much for keeping in touch with me even when I ignore your text messages and phone calls. You all have always been there for me and reminding me of where I come from and who I am. Next, I would also like to thank all of the Professors who were instrumental in my training as a scientist — first, Dr. Gregory Whitlock for introducing me to academic research and clinical laboratory sciences. Working in your lab in high school opened my eyes to a new world of discovery and clinical impact, and Dr. John Jaenike, who allowed me to work as a technician in his lab during my first year at the University of Rochester. Dr. Jacques Robert, who was a fantastic mentor and friend, and prepared me for graduate school. The time I spent in the lab iv taught me a lot about myself and managing a project. Outside of the lab, Dr. Robert and his wife Dewi were like my family in Rochester as they invited all of the lab members to their home to celebrate holidays and accomplishments. During my time in the lab I was also mentored by Dr. Hristina “Tina” Nedelkovska, who taught me many things about science, but also took the time to teach me about her Macedonian culture. I want to express my heartfelt gratitude to Dr. Zaneta-Nikolovska-Coleska for allowing me to join her lab and carry out my research without limits. I have learned so much about myself and how I learn from others based on her guidance. These lessons will be with me throughout my life as I develop as a researcher. I had no experience as a biochemist before joining the lab, and I was able to learn a variety of techniques from assay development to NMR spectroscopy and even dabbled in peptide and small molecule synthesis. Since I was the only woman in the lab for a while, she was also open to having girl talk which I appreciated. Zaneta has seen me at my absolute best and my absolute worst, and I am genuinely grateful for her support during my times of grief throughout my graduate studies. I would also like to thank my dissertation committee members Dr. Ivan Maillard, Dr. Andrew Muntean, and Dr. Yali Dou for their patience, guidance, and overall awesomeness. They worked tirelessly with me to make sure my paper drafts were ready for publication, my science was rigorously reviewed, and that I had access to the training and tools needed for my experiments. I would like to especially thank members in the Muntean lab, Justin Serio, James Ropa and Lilly Chen for their support while I was generating my cell lines. They took time out of their busy schedules to make sure I had all the resources I needed to succeed, and I am very appreciative. I would also like to thank Jennifer Chase and Ann Friedmann from Ivan Maillard’s v lab for their experimental contributions to Chapter 2 of my thesis. Their studies critically rounded out the second half of those studies, and words cannot express my gratitude. I had the opportunity to work with so many brilliant minds throughout my graduate studies, and I would like to thank the past lab members Dr. Chenxi Shen and Dr. Garrett Gibbons for their pioneering work on my project that helped lay the foundation for my studies and other projects that I worked on in the lab. Both Chenxi and Garrett trained me on biochemical, biophysical, and cell-based assays that I further developed throughout my time as a graduate student. I would also like to thank Lei Miao for his support while I tried to synthesize compounds and purify peptides for my project. I want to thank my current labmates, Karson Kump, Bing Zhou, Yuting Yang, Nurul Ansari, and Marcelo Murai for their fruitful lab discussions and for tolerating me as the “Lab Princess.” I want to give a special thanks to Ahmed Samir Ahmed Mady who has been with me throughout my entire graduate school career at U of M. To Ahmed, I am the little sister he never wanted, and he is my dearest friend and colleague. I will be so sad to miss his presence with me in the lab, but I am very excited for his future as a scientist and hope we will continue to be in touch as we both set out on our journeys. In the Department of Pathology, I would like to thank Laura Labut for all of her help and support throughout my graduate studies. Laura is not only a great administrator for the department but also a valued friend. On that same note, I would like to thank Tasha Thurman in the Procurement office for always helping me figure out ordering issues and being a listening ear when I need to talk through things that trouble me. I would also like to thank my many friends in the Pathology department. Those who graduated before me and those who will soon follow. You all are the absolute best! My graduate career would not be what it was without your love and support throughout all of these years. vi For those who I was unable to mention, I and want also to thank you. As you see, my life has been impacted in so many ways by a variety of people, and if I named everyone it would be longer than my actual dissertation work, but please know that I am grateful for the smallest gesture to the largest.
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