Inflammatory Breast Cancer: the Immune Perspective
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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 5-2013 Inflammatory Breast Cancer: The Immune Perspective Evan N. Cohen Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Immunity Commons, and the Medical Sciences Commons Recommended Citation Cohen, Evan N., "Inflammatory Breast Cancer: The Immune Perspective" (2013). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 369. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/369 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]. Inflammatory Breast Cancer: The Immune Perspective by Evan Nathaniel Cohen, B.S., B.A. APPROVED:!!!! ______________________________! James M. Reuben, Ph.D., M.B.A. Supervisory Professor ______________________________! Bradley McIntyre, Ph.D. ______________________________! Naoto T. Ueno, M.D., Ph.D. ______________________________! Stephen E. Ullrich, Ph.D. ______________________________! Wendy A. Woodward, M.D., Ph.D. APPROVED:!!!!____________________________ Dean, The University of Texas Graduate School of Biomedical Sciences at Houston Inflammatory Breast Cancer: The Immune Perspective Presented to the Faculty of The University of Texas Health Science Center at Houston and The University of Texas MD Anderson Cancer Center Graduate School of Biomedical Sciences in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY by Evan Nathaniel Cohen, B.S., B.A. Houston, Texas May, 2013 DEDICATION This dissertation is dedicated to my parents, Stephen and Marin Cohen, and the patients of the Morgan Welch Inflammatory Breast Cancer Research Program and Clinic at the University of Texas MD Anderson Cancer Center. In memory of Marilyn Kretzer ACKNOWLEDGEMENTS This study was funded in part by the following grants: Assessment of Circulating Breast Cancer Stem Cells To Predict Recurrent Disease, W81XWH-09-1-0031 01, DOD (Evan N. Cohen), Cancer Center Support Grant, NIH/NCI; State of Texas Rare and Aggressive Breast Cancer Research Program (Naoto T. Ueno); Human Breast Cancer Stem Cell Surrogates, CA138239-02, NIH/NCI (James Reuben and Wendy Woodward). I would like to thank my parents who have supported me at every step. My parents and sister have given me their unequivocal support and inspiration throughout, as always, for which my mere expression of thanks does not suffice. It would not have been possible to write this doctoral thesis without the help and support of the kind people around me, to only some of whom it is possible to give particular mention here. First and foremost, I would like to thank my advisor, James Reuben. I could not have asked for a more supportive advisor. I appreciate all his contributions of time, ideas, and funding to make my Ph.D. experience productive and stimulating. The joy and enthusiasm he has for research was contagious and motivational for me, even during tough times in the Ph.D. pursuit. I am also thankful for the excellent example he has provided as a genuinely good person and professor. Above all, he made me welcomed as a friend, which I appreciate from my heart. The members of the Dr. Reuben’s lab have contributed immensely to my personal and professional time at MD Anderson. The group has been a source of friendships as well as good advice and collaboration. I cannot imagine how I would have survived these last years without the help and friendship of Hui Gao. Hui has taught me most of iv what I learned in the lab through grad school and certainly everything about flow cytometry. Most importantly, she has been a true friend and I cannot possibly thank her properly. This thesis is a compilation of massive amounts of data that has been amassed by a wonderful team of dedicated people. Sanda Tin was responsible for coordinating the LAB08-199 protocol within the lab that is the basis for many of the patient data presented here. Summer Jackson, Juanita Lara, and Charla Parker were responsible for recruiting the patients. Qiong Wu and YingDong Li diligently processed the samples. Raul “Josh” Garza and Matt Galland analyzed many of the serum samples. Antonio Giordano has lent his clinical expertise to put our blood samples into context. Simone Anfossi and Hyun-Jun “Robert” Park, my fellow travelers through this education, helped me learn RT-PCR and countless other assays. Gail Hammond helped with all the little and not so little things I seem incapable of doing. As mentioned above, Hui Gao’s careful eye was always present at every step. Under Dr. Reuben, we have been more than co-workers. I think of these people as my extended family. Additionally over the years, I have had the unique privilege to work with a number of post-docs and fellows that have come through the lab. First, I would like to thank Bang-Ning Lee who helped guide me through my early days in the lab. And Carolina Gutierrez, who I don’t think taught me anything about science, but so much about medicine and even more about life. We were all most saddened when Michal Mego, who developed the test we used for circulating tumor cells in EMT with Prof. Sendurai Mani, returned to his home in Slovakia. We miss him as much as he misses his hair. Perhaps one of the greatest experiences of my time at MD Anderson was the ability to work with and guide some truly outstanding high school and college students v including Nicole Kruijs, Grant Dougherty, Thomas Yang, Lilly Zhang, Albert Huang and Ken Tong. Meshaal Nadeem spent several of her summers and winters with us. If her work here is any indication, I have been very fortunate to meet her so early in her career. I am also grateful I had the opportunity to work with Xiaoyan “Sunny” Wang, Changping Li, Mario Giuliano, De-Yu Shen, “Jessica” Li Li, Gracie Rodriguez, Monica Martinez, Ugo de Giorgi, Paloma “Jimena” de Andres, Laura Camacho, Karin Görner, Zarixia Zavala and her student Amilcar Rivera, Tiffany LaFortune, Ujjwala Warwdekar, Lamees Ali Aziz Alsayed Atteya, Neslihan Duzkale, Tamer Fouad, Sriram Yennu and countless other fellows that Dr. Reuben has hosted. This institution has been a wonderful opportunity to meet people from a variety of fields. I am especially grateful to Prof. Gabriel Lopez-Berestein in Experimental Therapeutics and Prof. Val Johnson in Statistics who hosted me in first year rotations. These were valuable experience that broadened my horizons. Through Dr. Reuben, I also had the opportunity to collaborate with Prof. Charles Cleeland in Symptom Research and Prof. Sriram Yennu in Palliative Care and Rehabilitative Medicine. I would like to thank Neelima G Reddy from Prof. Raja Luthra’s lab for helping with the Fluidigm experiments and Bisrat Debeb from Dr. Woodward’s lab who helped with the mammosphere experiments. In addition, the members of Hematopatholgy, Breast Medical Oncology and the Morgan Welch Inflammatory Breast Cancer Research Program and Clinic have all been very supportive; providing know-how, reagents, sounding boards, guidance and advice. I particularly need to thank Dave Imperial, Nikhil Chari, Hatice Duzkale, and Archie Tamayo in HemPath and Lara C. Alvarez de Lacerda in BMO. I have been fortunate to have the guidance of a wonderful committee. I would like to thank Drs. Massimo Cristofanilli, Bradley McIntye, Fredika Robertson, Naoto T. vi Ueno, Stephen Ullrich, and Wendy Woodward for their time, interest, helpful comments and insightful questions. Their support has truly enriched my time and experience here. Finally, I wish to thank the patients of the MD Anderson Nellie B. Connally Breast Center and Morgan Welch Inflammatory Breast Cancer Research Program and Clinic who eagerly await answers. vii Inflammatory Breast Cancer: The Immune Perspective Publication No.________ Evan Nathaniel Cohen, PhD Supervisory Professor: James M. Reuben, Ph.D. ABSTRACT Inflammatory breast cancer (IBC) is the most insidious form of locally advanced disease. Although rare and less than 2% of all breast cancer, IBC is responsible for up to 10% of all breast cancer deaths. Despite the name, very little is known about the role of inflammation or immune mediators in IBC. Therefore, we analyzed blood samples from IBC patients and non-IBC patients, as well as healthy donor controls to establish an IBC- specific profile of peripheral blood leukocyte phenotype and function of T cells and dendritic cells and serum inflammatory cytokines. Emerging evidence suggests that host factors in the microenviromement may interact with underlying IBC genetics to promote the aggressive nature of the tumor. An integral part of the metastatic process involves epithelial to mesenchymal transition (EMT) where primary breast cancer cells gain motility and stem cell-like features that allow distant seeding. Interestingly, the IBC consortium microarray data found no clear evidence for EMT in IBC tumor tissues. It is becoming increasingly evident that inflammatory factors can induce EMT. However, it is unknown if EMT-inducing soluble factors secreted by activated immune cells in the IBC microenvironment can" account for the absence of viii EMT in studies of the tumor cells themselves. We hypothesized that soluble factors from immune cells are capable of inducing EMT in IBC. We tested the ability of immune conditioned media to induce EMT in IBC cells.