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A License to Kill: Understanding Natural Killer Cell Licensing to Fight Cancer

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A License to Kill: Understanding Natural Killer Cell Licensing to Fight Cancer The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 12-2017 A LICENSE TO KILL: UNDERSTANDING NATURAL KILLER CELL LICENSING TO FIGHT CANCER Jolie Schafer Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Medicine and Health Sciences Commons, and the Other Immunology and Infectious Disease Commons Recommended Citation Schafer, Jolie, "A LICENSE TO KILL: UNDERSTANDING NATURAL KILLER CELL LICENSING TO FIGHT CANCER" (2017). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 812. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/812 This Dissertation (PhD) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. A LICENSE TO KILL: UNDERSTANDING NATURAL KILLER CELL LICENSING TO FIGHT CANCER By Jolie Rae Schafer, B.S. APPROVED: _________________ Shulin Li, Ph.D. Advisory Professor ____________________ Dean Lee, M.D., Ph.D. Co-Mentor ____________________ Michael Curran, Ph.D. ____________________ Gregory Lizee, Ph.D. ____________________ Annemieke Kavelaars, Ph.D. _____________________ Silke Paust, Ph.D. APPROVED: _____________________ Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences I A LICENSE TO KILL: UNDERSTANDING NATURAL KILLER CELL LICENSING TO FIGHT CANCER A DISSERTATION Presented to the Faculty of The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY by Jolie Schafer, B.S. Houston, Texas December 2017 II Acknowledgements I would like to acknowledge all of the many people who helped me and encouraged me along the way. My love for science began in 7th grade, when I learned about genetics from Mr. Flori. Thank you Mr. Flori for instilling in me a facisination for science. My motivation for pursuing scientific research stemmed from my childhood friend, Chase McGowen, who has Cystic Fibrosis. Chase, you are a miracle, your fighting spirit keeps me fighting to learn more. Thank you to all of my undergraduate professors, from Houston Baptist University, who encouraged me to apply to graduate school, Drs. Hannah Wingate, Susan Cook, Jackie Horn, Brenda Whaley, Curtis Henderson, Rachel Hopp, and Saul Trevino. Thank you all for believing in me and seeing my potential. Thank you, Dr. Khandan Keyomarsi for my first laboratory position as a Cancer Prevention Research Institute of Texas undergraduate student, which turned into a yearlong research assistant position preceding my graduate work. Thank you to The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences for accepting me into this program. To Deans Barton and Blackburn, thank you for allowing me to have this opportunity. To my amazing mentor, Dr. Dean Lee for mentoring me, teaching me, growing me, generously allowing me to keep my project upon your departure. Thank you, Dr. Lee for your support over the last four years. You are an incredible mentor and friend and I am grateful to have trained under you. Thank you, Dr. Shulin Li for graciously adopting me into your lab and letting me continue my dissertation project. Thank you to the amazing members of both Dr. Lee and Dr. Shulin’s laboratories, your help and collaboration has been incredible. Thank you to Dr. Stefan Ciurea for allowing me to collaborate on the NK cell clinical trial, I am excited to see whats in store for the Phase II study. Thank you to my great friend Dr. Ariany Aquino-Lopez for being III an amazing collegue and friend. Our journey through graduate school together is one that I will always cherish. Thank you to the Pediatics Research department for a great training environment throughout graduate school. Thank you to my examining committee Drs. Steve Ullrich, Michael Curran, Neal Waxham, Cao, Lizee, for challenging me and preparing me for finishing graduate school. Thank you to my wonderful advisory committee, Drs. Dean Lee, Shulin Li, Silke Paust, Michael Curran, Gregory Lizee, and Annemieke Kavelaars. All of you have been a tremendous help and resource throughout graduate school. Thank you to the former Immunology Program Director, Dr. Ben Zhu for your service to the Immunology Program and for giving me a great family of researchers to be a part of. Thank you to our new Immunology Director, Dr. Jagan Sastry, and co-director, Dr. Kimberly Schluns, for your support and dedication to the Immunology students. Thank you, Dr. Melinda Yates, for your support and encouragement in the First-Generation Student Association. The First-Gen group provided me with a great support system throughout graduate school. Thank you to Dr. Andrew Bean, Dr. Marenda Wilson-Pham, my mentors, deans, program, and graduate school for giving me the opportunity to participate in the summer graduate program in public policy with the Archer Center and to intern with the US Department of Health and Human Services in the Office of the Assistant Secretary for Preparedness and Response. This was a once in a lifetime opportunity. Thank you to the Texas National Security Network for providing me with a scholarship. Thank you to Cancer Answers for awarding me a scholarship in cancer research. Thank you to my best friend, my husband, Dalton Schafer. Your support, encouragement, prayers, and undying love for me is what kept me from never giving up. You believed in me IV even when I didn’t. Thank you for putting up with crazy experiments, with all of my faults and failures, and for allowing me to chase my dreams. Thank you to my incredible family, my mom, dad, step-mom, brother, sister in-laws, grandparents, mother and father in-law. Thank you for being my biggest cheerleaders and my encouragement, and for always believing in me. Thank you to my incredible friends both in graduate school and outside of graduate school. I have the best support system and I would not be where I am without all of you. Thank you to God, for allowing me to be a part of discovering your Creation. My studies in science have only affirmed my faith. V Abstract A License to Kill: Understanding Natural Killer Cell Licensing to Fight Cancer Jolie Rae Schafer, B.S. (Advisor: Dean A. Lee, MD, PhD; Shulin Li, On-Site Advisor) Natural killer (NK) cell education is an essential developmental process for NK cell effector function, that renders some NK cells “licensed” and others “unlicensed” (with heightened or lowered effector function potential, respectively) against tumor and targets lacking self- molecules. However, the underlying mechanisms responsible for the heightened effector responses of licensed cells remain unknown. Using NK cells derived from humans and expanded ex vivo we performed high-throughput protein expression analysis, and identified multiple proteins that are differentially regulated in licensed and unlicensed human NK cells before and after inhibition by killer-cell immunoglobulin-like receptors (KIRs) and activation by the NKp46 natural cytotoxicity receptor, including several related to cellular metabolic pathways. We explored cellular metabolism in the two subsets and found that licensed NK cells are highly glycolytic, and use glycolysis and mitochondrial respiration for cytolysis of leukemia targets, whereas unlicensed NK cells are dependent on mitochondrial respiration. We determined the metabolic pathways that are necessary for licensed and unlicensed NK cells to elicit a cytolytic response using metabolic inhibitors to inhibit glycolysis or mitochondrial respiration metabolic pathways in the NK cells during a cytotoxicity assay. We observed that licensed NK cells utilize both glycolysis and mitochondrial respiration to perform cytolysis whereas unlicensed NK cells only use mitochondrial respiration for their cytolytic response against leukemia targets. To our knowledge, this is the first description of the underlying mechanisms that explain the cytolytic differences between licensed and unlicensed NK cells. Our findings provide a groundbreaking platform to further explore and manipulate metabolism in licensed and unlicensed NK cells to improve NK cell immunotherapy. VI Table of Contents Acknowledgements ................................................................................................................... III Abstract ....................................................................................................................................... VI List of Illustrations ...................................................................................................................... X Figure Twenty-Four: Comparison of intracellular metabolites found in expanded licensed and unlicensed single KIR positive NK cells. ........................................................................................... XI List of Tables ...........................................................................................................................
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