University of Massachusetts Medical School eScholarship@UMMS GSBS Dissertations and Theses Graduate School of Biomedical Sciences 2017-09-12 Different Journeys, Same Destination: Exploring the Role of a PYHIN Protein and Involvement of Caspase-8 in the Regulation and Activation of Inflammasomes Sreya Ghosh University of Massachusetts Medical School Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/gsbs_diss Part of the Immunity Commons, Immunology of Infectious Disease Commons, and the Microbiology Commons Repository Citation Ghosh S. (2017). Different Journeys, Same Destination: Exploring the Role of a PYHIN Protein and Involvement of Caspase-8 in the Regulation and Activation of Inflammasomes. GSBS Dissertations and Theses. https://doi.org/10.13028/M2CD6Z. Retrieved from https://escholarship.umassmed.edu/ gsbs_diss/928 Creative Commons License This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in GSBS Dissertations and Theses by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. Different Journeys, Same Destination: Exploring the Role of a PYHIN Protein and Involvement of Caspase-8 in the Regulation and Activation of Inflammasomes A Dissertation Presented By Sreya Ghosh Submitted to the Faculty of the University of Massachusetts Graduate School of Biomedical Sciences, Worcester in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY September 12, 2017 Immunology and Microbiology Program Different Journeys, Same Destination: Exploring the Role of a PYHIN Protein and Involvement of Caspase-8 in the Regulation and Activation of Inflammasomes A Dissertation Presented By Sreya Ghosh The signatures of the Dissertation Defense Committee signify Completion and approval as to style and content of the Dissertation ____________________________________________ Katherine A. Fitzgerald, Ph.D., Thesis Advisor ____________________________________________ Egil Lien, Ph.D., Member of Committee ____________________________________________ Evelyn Kurt-Jones, Ph.D., Member of Committee ____________________________________________ Jeremy Luban, M.D., Member of Committee ____________________________________________ Alexander Poltorak, Ph.D. Member of Committee The signature of the Chair of the Committee signifies that the written dissertation meets the requirements of the Dissertation Committee ____________________________________________ Neal Silverman, Ph.D., Chair of Committee The signature of the Dean of the Graduate School of Biomedical Sciences signifies that the student has met all graduation requirements of the school. ____________________________________________ Anthony Carruthers, Ph.D., Dean of the Graduate School of Biomedical Sciences Immunology and Microbiology Program September 12, 2017 In the loving memory of my grandmother, Anjana Mitra, my staunchest ally 3 Acknowledgements “There are those that look at things the way they are, and ask why? I dream of things that never were, and ask why not?” – R. F. Kennedy My journey as a Ph.D. student has been the most rewarding, exciting and challenging phase of my life. Away from home and every thing familiar, I wouldn’t have been able to live my dream without the friendship, support and guidance of the wonderful people I had the privilege of knowing. I thank every one of you for the encouragement. First and foremost, I would like to thank my mentor, Prof. Kate Fitzgerald. Her unequivocal support, candor, continuous encouragement and sense of pragmatism have motivated and inspired me over the years. While I surely haven’t been the most easy-going protégée, her faith in my scientific abilities, thinking and aspirations never wavered. Sometimes, when things were too trying, it was only her conviction in me, and her accompanying support, that kept me focused and going. And I believe, being a part of her team, made me enjoy and appreciate science more. I would like to thank Prof. Neal Silverman for his guidance over the years- from my mock Qualifiers to my Thesis Defense, he has motivated me to challenge my own boundaries every time. I would like to thank Prof. Egil Lien who has also been a part of my journey from the beginning, pointing out the strengths and weaknesses of my science in the kindest manner. I would like to thank Profs. Evelyn Kurt-Jones and Jeremy Luban who have also been instrumental in my development as a scientist; their insights and advice have helped hone my scientific applications. The Fitzgerald lab has a wonderful environment for doing science. I would like to thank the past and current post-docs Vijay, Shruti, Sivapriya, Maninjay, Roland, Ragunath, Yongzhi, Juliet for their enthusiasm and encouragement; Zhaozhao and Gaia deserve special mention, ZZ for being a rock and Gaia- her friendship and supply of chocolates eased me through the final phase of my graduate studies. A big thank-you goes to Susan Carpenter, my post-doc mentor in the lab- I have learned so much from her, and it was grand to have had the opportunity to work with her. I am very grateful for the friendship and support of my co-mentees in the lab – Mikayla, Stefan, John, Pontus, Ankit, especially Jennie, Mona, Sandhya and Shiuli for all the laughs and brainstorming; and for the help from other lab members, prominently Jennifer, Jason and Lisa; and Kelly and Pallavi. Another special thank-you goes to Natália Ketelut-Carneiro, with whom I had the opportunity to work on a part of both our theses- learning, teaching and discussing science with her always motivated me. I would like to 4 thank all my colleagues in the Golenbock lab, Silverman lab, Lien lab, Levitz lab and the Department of Medicine who make it an exciting atmosphere for science. In my time here at UMass and in the United States, I have had the honor of meeting many interesting and wonderful people. I wouldn’t have made it without the constancy of my best friends, Sonal and Anubhab; my fellow researchers, Swapnil, Divya, Siva, Arvind, Mihir, Ami, Meetu, Nisha, Dwijit, Sanjoy, Navonil, Sanchaita, Paurav, Pallavi, Amena, Aditya, Mayuri, Ankita, Sneha, Naveen, Sri Devi, Samyabrata, Seemin; and my dear roommate for six years, Vahbiz. Outside of UMass, I cherish the friendship, support and the good times with Sutapa-Di/Samik-Da, Gargi-Di/Debasish-Da, Kamna/Tathagat, Debjani, Nitin, Debarpita, and especially the unconditional love that I get from Rashi, Ruhi, Leo and Tanni. I would like to thank my teachers at my alma mater, Gokhale Memorial Girls’ School (Kolkata, India), and University of Calcutta, India as well as my mentors at Indian Institute at Chemical Biology (Kolkata, India) – their advice, teachings and encouragement were unparalleled. Finally, nothing would have been possible without the support of my family and all the long-distance phone and video calls - my cousins, my childhood friends have always rooted for me; especially my family away from home: Anita/Deepak, Shatarupa-mashi/Arijit-da and my nephews, Dhruv and Simba, Doyel-didi, my aunt (Bhaloma), and Neha. I wouldn’t have made it so far without Tirtha and his steadfast presence. My grandmother, who would have been proud to see me earn this degree, is my beacon. She taught me to aspire and persevere, and this thesis is dedicated to her. My sister, Sreeta, is my shining star, who never lets me lose hope or faith in myself. And lastly, my parents, Pew and Subhajit - whatever I am, whatever good I have achieved today, is all because of them. The journey has just begun. 5 Table of Contents Acknowledgements......................................................................................................4 Abstract..........................................................................................................................8 List of Publications.......................................................................................................9 List of Figures…..........................................................................................................10 List of Abbreviations .................................................................................................12 CHAPTER I: Introduction ...........................................................................................15 1.1 Introduction to Innate Immunity..........................................................................16 1.2 Pattern Recognition Receptors and their ligands...............................................19 Toll-like receptors..........................................................................................19 RIG-I like receptors.......................................................................................20 NOD-like receptors........................................................................................21 C-type lectin receptors...................................................................................22 AIM2-like receptors & cGAS..........................................................................23 1.3 Innate Immune Signaling Pathways...................................................................24 Complement System.....................................................................................24 Type-I & Type-II Interferons..........................................................................26 NF-κB and MAPK pathways..........................................................................28