Exploring the Regulatory Mechanism of the Notch Ligand Receptor Jagged1 Via the Aryl Hydrocarbon Receptor in Breast Cancer Sean Alan Piwarski
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Marshall University Marshall Digital Scholar Theses, Dissertations and Capstones 2018 Exploring the Regulatory Mechanism of the Notch Ligand Receptor Jagged1 Via the Aryl Hydrocarbon Receptor in Breast Cancer Sean Alan Piwarski Follow this and additional works at: https://mds.marshall.edu/etd Part of the Biological Phenomena, Cell Phenomena, and Immunity Commons, Medical Molecular Biology Commons, Medical Toxicology Commons, and the Oncology Commons EXPLORING THE REGULATORY MECHANISM OF THE NOTCH LIGAND RECEPTOR JAGGED1 VIA THE ARYL HYDROCARBON RECEPTOR IN BREAST CANCER A dissertation submitted to the Graduate College of Marshall University In partial fulfillment of the requirements for the degree of Doctor of Philosophy In Biomedical Sciences by Sean Alan Piwarski Approved by Dr. Travis Salisbury, Committee Chairperson Dr. Gary Rankin Dr. Monica Valentovic Dr. Richard Egleton Dr. Todd Green Marshall University July 2018 APPROVAL OF DISSERTATION We, the faculty supervising the work of Sean Alan Piwarski, affirm that the dissertation, Exploring the Regulatory Mechanism of the Notch Ligand Receptor JAGGED1 via the Aryl Hydrocarbon Receptor in Breast Cancer, meets the high academic standards for original scholarship and creative work established by the Biomedical Sciences program and the Graduate College of Marshall University. This work also conforms to the editorial standards of our discipline and the Graduate College of Marshall University. With our signatures, we approve the manuscript for publication. ii © 2018 SEAN ALAN PIWARSKI ALL RIGHTS RESERVED iii DEDICATION To my mom and dad, This dissertation is a product of your hard work and sacrifice as amazing parents. I could never have accomplished something of this magnitude without your love, support, and constant encouragement. Thank you for teaching me that anything worth having must be earned and that what I am willing to accomplish is determined by what I am willing to sacrifice. Having you two in my life gave me the courage to acknowledge my self-worth and the tenacity to never quit, even in the most improbable situations. I love you and thank you for everything, Sean iv ACKNOWLEDGEMENTS The first and most important acknowledgement is for my advisor, Dr. Travis Salisbury. I’ll start off by saying that I would not have survived the program, nor would I have become the scientist I am today and want to continue to be, if it was not for him. He allowed me to pursue my original research that was outside the realm of his research, and he helped me turn an original idea into a complete story that I could make a reality. He has been an incredible mentor, an absolutely amazing role model, and scientist. He has been and continues to be extremely patient for putting up with my often-overzealous passion and tenacity. He has also helped me apply my creativity into research. His office door was always open to me, and he was always there in the lab with me to help me complete my experiments. He always challenged me to do my best work through encouraging me to think completely through my ideas. He helped me realize my strengths and weaknesses, and how to improve upon as well as work with my ideas. It is because of Dr. Salisbury that I learned what it truly means to be a scientist. I am forever grateful. I would also like to thank my amazing committee members Dr. Gary Rankin and Dr. Monica Valentovic for their professional advice and many letters of recommendation. They are two of the greatest scientists and people I know. Thank you so much for all that you have done for me, and for introducing me to the BMS program when I was studying forensic science. I would not have come to this program if it was not for you both. I also would like to extend my thanks to Dr. Richard Egleton for being an awesome teacher. I always enjoyed going to your lectures, and your door was always open for random conversations and professional advice, as well as introducing me to peanut butter powder. I also want to thank Dr. Green for serving on my committee, for keeping me on track to graduate, and for generously introducing me to the Chancellor’s Scholar Program. I would not have been given the opportunity to pursue this v research without you. Thank you all for being supportive of me, and for affording me the opportunities to learn and grow. For all of this, I am incredibly humbled, and will never take any of this for granted. Thank you all! To Katie and Carl, thank you for letting me into your family, becoming my home away from home and for everything you have done for me over these past seven years. You both are truly amazing, and I am truly blessed to have you guys in my life. Rick and Peggy, thank you for all the dinners you made for me and for bringing me into your home for the holidays when I could not go to back to California. But most importantly, thank you for calling me your son (you will always be my other mom and dad). I will make it a point to come visit you all whenever I am back in West Virginia. To Naseeb, Quentin, Ateeq, Arnold, Will E., Will O., Jenna, Warren, Duran, Jason, Cindy and Delilah. You were all there with me from the very beginning when my research was nothing more than an idea and when I was going through my struggles of making it happen. Each of you have always kept me sane in your own unique way, whether it was watching football, chilling out and chatting, going on trips, or just calling and texting me to see how I was doing when you never heard from me for extended periods of time. Thank you for sticking around and looking out for me over the years; I would not have succeeded if I did not have you guys in my life. I am truly grateful to have such amazing friends. Last but not least, I’d like to acknowledge my dogs Bentley, Charlie, and Milo. Thank you for your unconditional love and loyalty. I could not ask for a better entourage to have in my life. vi TABLE OF CONTENTS Approval of Dissertation…………………………………………………………………………. ii Dedication………………………………………………………………………………………...iv Acknowledgements………………………………………………………………………………. v List of Tables……………………………………………………………………………………xiii List of Figures .............................................................................................................................. xiv Abstract ....................................................................................................................................... xvii Chapter 1: Overview of Current Knowledge in Breast Cancer ...................................................... 1 1.1. Survival Rates and Subtypes………………………………………………………….1 1.2. Breast Cancer Subtype Characteristics and Current Treatment Strategies…………...3 1.2.1. Luminal A and B Subtype..………………………………………………....3 1.2.2. HER2-enriched Subtype ............................................................................... 6 1.2.3. Triple Negative Subtype ............................................................................... 9 1.2.4. Breast Cancer Stem Cells………………………………………………….15 1.3. Drug Resistance in Breast Cancer .............................................................................. 24 Chapter 2: The Notch Signaling Pathway ..................................................................................... 29 2.1. Canonical Notch Signaling ........................................................................................ 29 2.2. Physiological Functions of Notch Signaling: Immunity, Inflammation, Hematopoiesis and Development ..................................................................................... 36 2.3. JAG1-dependent Notch Signaling in Cancer ............................................................. 42 2.3.1. Angiogenesis ............................................................................................... 44 2.3.2. Cancer Stem Cells ....................................................................................... 46 2.3.3. Epithelial-Mesenchymal Transition (EMT), Invasion, and Metastasis ...... 47 vii 2.3.4. Proliferation, Cell Cycle Regulation, Drug Resistance, and Survival ........ 49 2.3.5. T-cell Regulation ........................................................................................ 50 2.3.6. A Synopsis of JAG1-Notch Signaling in Breast Cancer ............................ 51 Chapter 3: The Aryl Hydrocarbon Receptor (AHR): A Multi-Faceted Transcription Factor ...... 53 3.1. A Brief History and Introduction to AHR Biology ................................................... 53 3.2. The Canonical AHR Signaling Pathway .................................................................... 55 3.3. Non-canonical AHR Signaling ................................................................................. 60 3.4. TCDD-induced Negative-feedback Loops that Limit AHR Signaling ...................... 64 3.5. Physiological Roles of AHR: Immunity, Development, and Reproduction .............. 67 3.6. AHR Signaling and Cancer ....................................................................................... 69 3.7. Current Problematic Observation and Overall Hypothesis ....................................... 75 Chapter 4: Methods for Studying AHR Signaling ........................................................................ 76 4.1. Materials .................................................................................................................... 76 4.2. RNA-sequencing ........................................................................................................ 76 4.3. Cell Culture and Treatments .....................................................................................