Memory Potential Molecular Characterization and Translational A

The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences 12-2017 Memory Potential, Molecular Characterization, and Translational Applications of the Novel ThEO/TcEO T Cell Phenotype Todd Bartkowiak Todd Bartkowiak Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Immunopathology Commons, Medical Immunology Commons, Oncology Commons, and the Therapeutics Commons Recommended Citation Bartkowiak, Todd and Bartkowiak, Todd, "Memory Potential, Molecular Characterization, and Translational Applications of the Novel ThEO/TcEO T Cell Phenotype" (2017) The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) 816 https://digitalcommons.library.tmc.edu/utgsbs_dissertations/816 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 digitalcommons@library.tmc.edu MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF THE NOVEL ThEO/TcEO T CELL PHENOTYPE by Todd Jacob Bartkowiak, M.S APPROVED: Michael A Curran, Ph.D Advisory Professor Chantale Bernatchez, Ph.D Joya Chandra, Ph.D Roza Nurieva, Ph.D Kimberly Schluns, Ph.D APPROVED: Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF THE NOVEL ThEO/TcEO T CELL PHENOTYPE 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 Todd Jacob Bartkowiak, M.S Houston, Texas December 2017 COPYRIGHT The Proceedings of the National Academy of Sciences allows authors and their affiliated institution to retain extensive rights to the use of materials after publication, including, but not limited to “the right to include your article in your thesis or dissertation.” The full list of authorship rights can be found at http://www.pnas.org/site/aboutpnas/rightpermfaq.xhtml Frontiers in Oncology is an open access journal wherein articles are “published under the CC-BY license, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and the source are credited.” More information about the publishing policies of Frontiers journals can be found at http://home.frontiersin.org/about/open-access iii DEDICATION I would like to dedicate this dissertation to my family without whom I would not be here My father, James, and my mother, Renalda, have been instrumental in guiding my passion for science My brothers (Patrick and Matthew), my sister-in-law, Nikki, and my nephews Luke and Logan, and my niece, Lily, all of whom encouraged me to pursue my love for science iv ACKNOWLEDGEMENTS I would like to acknowledge all of the current and past members of the Curran lab and all of my friends and coworkers who helped me succeed in graduate school: Ashvin, Casey, Priya, Chao-Hsien, Renee, Pratha, Krishna, Midan, Matt, Courtney, Brittany, Rachel, Shail, Felix, Scott, Spencer, Stephen, Colm, Lucy, Nana-Ama, and all of the graduate students, faculty, and staff at MD Anderson I would also like to thank all of my advisors and collaborators at MD Anderson who have helped me reach this point in my life Dr Michael Curran, whose mentorship was vital to helping me succeed in graduate school and grow as a scientist To my advisory committee members especially Dr Kimber ly Schluns, who was always willing to take the time to help me with manage experiments and provide a helping hand I would also like to extend my gratitude to collaborators Dr Jagan Sastry who was always willing to listen and provide valuable advice to h elp me navigate graduate school and to Dr Amy Heimberger; all of whom provided guidance and helped me to secure my future in science I would like to especially thank Shailbala Singh and Guojun Yang for their assistance with the HPV immunotherapy experim ents and Ashvin Jaiswal and Casey Ager for their help with the liver toxicity studies Thank you all v MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF THE NOVEL ThEO/TcEO T CELL PHENOTYPE Todd Jacob Bartkowiak, M.S Advisory Professor: Michael A Curran, Ph.D T cells comprise a substantial arm of the immune system and are exquisitely adapted to combat pathogens and tumors The inflammatory environment largely dictates the nature of T cell response A hallmark of T cell-mediated immunity is formation of immunological memory; the ability to respond more potently to re encounter with pathogens The immune system is also capable of recognizing tumors as foreign, much like viral or bacterial pathogens Tumors have evolved, though, to generate an immunosuppressive environment to avoid destruction The field of immunotherapy seeks to overcome immune suppression, in part by targeting T cell coreceptors on the cell surface with either agonist or antagonist antibodies Targeting the T cell co-stimulatory receptor 4-1BB with agonist antibodies engenders strong antitumor responses in multiple murine tumor models, in part by expanding the proliferative capacity, survival, and cytotoxicity of T cells within the tumor microenvironment We have previously shown that systemic administration of α4 -1BB antibodies induces a novel T cell program typified by expression of the T-box transcription factor Eomesodermin (Eomes) and the co-inhibitory receptor Killer Cell Lectin-like Receptor G1 (KLRG1) which we collectively term ThEO Herein, we demonstrate that the ThEO phenotype constitutes a stable T cell polarity capable of recalling to subsequent antigen challenge Despite expression of terminal differentiation markers, ThEO cells phenotypically resemble discrete memory T cell subsets We also find that the activation of the Signal Transducer and Activator of Transcription (STAT) pathways, in particular STAT1 and STAT3, is critical to ThEO polarization ThEO cells possess clinical relevance Anti-4-1BB antibodies synergize with vi HPV peptide vaccination to eradicate HPV+ murine tumors, due to a honed tumorspecific ThEO response Further, ThEO phenotype cells infiltrate the livers of α4-1BB treated mice, which may play a role in 4-1BB mediated hepatotoxicity Finally, we show that melanoma patients enrolled in α4-1BB clinical trials upregulate key markers associated with the ThEO phenotype; hence formation of ThEO cells within patient blood may act as a biomarker for therapeutic outcome This body of work demonstrates that the ThEO phenotype constitutes a unique T cell polarity that may prove beneficial in cancer treatment vii TABLE OF CONTENTS Page Approval Signatures……………………………………………………………………………… i Title Page………………………………………………………………………………………… ii Copyright………………………………………………………………………………………… iii Dedication………………………………………………………………………………………… iv Acknowledgements……………………………………………………………………………… v Abstract…………………………………………………………………………………………… vi Table of Contents……………………………………………………………………………… viii List of Illustrations…………………………………………………………………………….xvi List of Tables……………………………………………………………………………… ….…xix Abbreviations………………………………………………………………………………….…xx Chapter 1: Background……………………………………………………………………….22 1.1 Tumor Immunology and Immune Evasion……………………………… …………22 1.2 Cancer Immunotherapy…………………………………….…………………………23 1.3 4-1BB…………………………….………………………………………………………25 1.4 Transcriptional Regulation of T cell subsets……………………………….………32 viii 1.5 T Cell Subsets…………………………………….…………………………… …….37 1.5.1: Th1……………………………………………………… …………… ……… 37 1.5.2: Th2…………………………………………………………… ……………… 37 1.5.3: Th17……………………………………………………………… ……… ……38 1.5.4: Treg……………………………………………………… …… ………………39 1.5.5: Tfh ………………………………………………………………… ……….……40 1.5.6: ThCTL…………………………………………………………… … ………….41 1.6 Adaptive Immunity and T cell memory………………………………………… …42 1.6.1: Central Memory………………………………… …………………………… 44 1.6.2: Effector Memory……………… …………………………………………… 45 1.6.3: Tissue Resident Memory………………………………………… ……….….45 1.6.4: Stem Cell Memory………………………………………… ………… ………46 Chapter 2: Specific Aims……………………………………….…………………………….48 2.1: Determine the phenotypic stability and memory potential of the novel TcEO/ThEO T cell polarity………………………………………………………….….48 2.2 Determine potential intrinsic and extrinsic factors that contribute to TcEO/ThEO cell development and/or persistence…………………………………48 2.3 Determine the clinical potential of the TcEO/ThEO phenotype in the context of therapeutic, pathologic, and prognostic potential………………………………48 Chapter 3: Materials and Methods………………………………… ……………………51 3.1 Animals……….……………………………………………………………………… ….51 3.2 Cell lines and reagents……………………………… …………………………… ……51 ix 347 Sun Y, Peng S, Qiu J, Miao J, Yang B, Jeang J, Hung CF, Wu TC 2015 Intravaginal HPV DNA vaccination with electroporation induces local CD8+ T-cell immune responses and antitumor effects against cervicovaginal tumors Gene Ther 22: 528-35 348 Selby MJ, Engelhardt JJ, Quigley M, Henning KA, Chen T, Srinivasan M, Korman AJ 2013 Anti-CTLA-4 antibodies of IgG2a isotype enhance antitumor activity through reduction of intratumoral regulatory T cells Cancer Immunol Res 1: 32-42 349 Simpson TR, Li F, Montalvo-Ortiz W, Sepulveda MA, Bergerhoff K, Arce F, Roddie C, Henry JY, Yagita H, Wolchok JD, Peggs KS, Ravetch 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September 19, 1986, the son of Renalda Marie Bartkowiak and James Patrick Bartkowiak After completing his work at Klein High School, Klein, Texas in 2005, he entered Sam Houston State University in Huntsville, Texas He received the degree of Bachelor of Science with a major in Biology from Sam Houston State University in May, 2008 For the next two years, he worked as a research technician in the Department of Diagnostic Sciences at the University of Texas Health Science Center In August of 2011, he then entered the University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences He received the degree of Master of Science with a major in Immunology in May, 2013 In August of 2013, he entered the University of Texas MD Anderson Cancer Center School of Biomedical Sciences Todd completed this dissertation under the mentorship of Dr Michael A Curran in the Immunology Department at the University of Texas MD Anderson Cancer Center Permanent address: 9711 Halkirk St Spring, Texas 77379 262 ... Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF.. .MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF THE NOVEL ThEO/TcEO T CELL PHENOTYPE by Todd Jacob Bartkowiak, M.S APPROVED: Michael A Curran,... ents and Ashvin Jaiswal and Casey Ager for their help with the liver toxicity studies Thank you all v MEMORY POTENTIAL, MOLECULAR CHARACTERIZATION, AND TRANSLATIONAL APPLICATIONS OF THE NOVEL