Development and Use of Recombinant Oncolytic Measles Viruses for the Treatment of Medulloblastoma Dissertation Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Brian John Hutzen, B.A. Graduate Program in Molecular, Cellular and Developmental Biology The Ohio State University 2012 Dissertation Committee: Corey Raffel, Advisor Balveen Kaur Stefan Niewiesk Christopher Pierson Copyright by Brian John Hutzen 2012 Abstract Medulloblastoma is the most common malignant brain tumor of early childhood. Our understanding of this disease, its etiology, and treatment has improved considerably over the past several years and is reflected in 5-year survival rates that now exceed 70%. Despite these advancements, numerous challenges in the effective treatment of medulloblastoma remain. Conventional therapy, which consists of surgical resection and craniospinal irradiation with or without chemotherapy, is associated with significant neurocognitive morbidity. In addition, a sizable subset of medulloblastoma patients will effectively remain incurable because of medulloblastoma’s propensity to disseminate along the spinal canal within the cerebrospinal fluid. Alternative treatment modalities for medulloblastoma are thus clearly needed. One promising approach is the development and use of recombinant oncolytic measles viruses. These viruses, which are based on the attenuated Edmonston vaccine strain virus, display a natural tropism for malignant cells and induce their fusion and subsequent death via apoptosis following infection. We have demonstrated that medulloblastoma tumor cells are highly susceptible to measles virus-induced oncolysis, and measles virus treatment can significantly prolong survival in mouse xenograft models of localized and disseminated disease. We have also utilized recombinant viruses expressing transgenes for the thyroidal sodium iodide symporter and the angiogenesis inhibitors endostatin and angiostatin to further augment the efficacy of measles virotherapy. Our initial results are encouraging and suggest that measles virus-based therapies may be of clinical utility in the treatment of medulloblastoma. ii Dedicated to my loving parents, Christina, and all the friends I met along the way. iii Acknowledgments I would first and foremost like to thank my advisor, Corey Raffel, for his guidance, friendship and all the opportunities he provided me over the last few years. It was a pleasure serving in your lab. I would also like to extend my sincerest gratitude to Adam Studebaker, who took me under his wing and helped me become a better scientist. Your advice and your friendship were truly appreciated. I would also like thank the members of my graduate advisory committee, Balveen Kaur, Stefan Niewiesk and Chris Pierson for their insights and guidance in developing my research career. There are also many friends I need to thank, in particular my fellow classmates Tom Bebee and Kevin Bosse. You both helped me in ways you are probably not even aware of, and your friendship meant the world to me. I would also like to thank my former lab mates in the Lin lab, whose companionship got me through some crazy and hectic times. I wish you all nothing but success and happiness down the road. Finally, I must thank my two wonderful parents, Harold and Diane, my sister, Kristin, and my beautiful fiancée Christina. Your love and support carried me through the darkest of days, and when my confidence in myself wavered, yours stood all the more firm. No amount of words on my part can ever truly express my gratitude and love for you all. Thank you so much. iv Vita November 19, 1980 ……………………………………………….. Born in Youngstown, Ohio 1999 ……………………………………………………………………….. Graduated from Poland Seminary High School 2003 ………………………………………………………………………. Bachelor of Arts, Biology, Washington and Jefferson College 2005 – present ………………………………………………………. Graduate Research Associate, Molecular, Cellular and Developmental Biology 2008 ………………………………………………………………………. Graduate Teaching Associate, Center for Life Sciences Education, Biology 113 2011 – present ………………………………………………………. Pelotonia Graduate Fellow Publications 1. Studebaker AW, Hutzen B, Pierson CR, Russell SJ, Galanis E, et al. (2012) Oncolytic measles virus prolongs survival in a murine model of cerebral spinal fluid-disseminated medulloblastoma. Neuro Oncol 14(4): 459-470. 2. Lin L, Hutzen B, Li PK, Ball S, Zuo M, et al. (2010) A novel small molecule, LLL12, inhibits STAT3 phosphorylation and activities and exhibits potent growth-suppressive activity in human cancer cells. Neoplasia 12: 39-50. v 3. Lin L, Hutzen B, Zuo M, Ball S, Deangelis S, et al. (2010) Novel STAT3 phosphorylation inhibitors exhibit potent growth-suppressive activity in pancreatic and breast cancer cells. Cancer Res 70: 2445-2454. 4. Canner JA, Sobo M, Ball S, Hutzen B, DeAngelis S, et al. (2009) MI-63: a novel small-molecule inhibitor targets MDM2 and induces apoptosis in embryonal and alveolar rhabdomyosarcoma cells with wild-type p53. Br J Cancer 101: 774-781. 5. Cen L, Hutzen B, Ball S, DeAngelis S, Chen CL, et al. (2009) New structural analogues of curcumin exhibit potent growth suppressive activity in human colorectal carcinoma cells. BMC Cancer 9: 99. 6. Fuh B, Sobo M, Cen L, Josiah D, Hutzen B, et al. (2009) LLL-3 inhibits STAT3 activity, suppresses glioblastoma cell growth and prolongs survival in a mouse glioblastoma model. Br J Cancer 100: 106-112. 7. Hutzen B, Friedman L, Sobo M, Lin L, Cen L, et al. (2009) Curcumin analogue GO-Y030 inhibits STAT3 activity and cell growth in breast and pancreatic carcinomas. Int J Oncol 35: 867-872. 8. Lin L, Hutzen B, Ball S, Foust E, Sobo M, et al. (2009) New curcumin analogues exhibit enhanced growth-suppressive activity and inhibit AKT and signal transducer and activator of transcription 3 phosphorylation in breast and prostate cancer cells. Cancer Sci 100: 1719-1727. 9. Hutzen B, Willis W, Jones S, Cen L, Deangelis S, et al. (2009) Dietary agent, benzyl isothiocyanate inhibits signal transducer and activator of transcription 3 phosphorylation and collaborates with sulforaphane in the growth suppression of PANC-1 cancer cells. Cancer Cell Int 9: 24. ii 10. Chen CL, Cen L, Kohout J, Hutzen B, Chan C, et al. (2008) Signal transducer and activator of transcription 3 activation is associated with bladder cancer cell growth and survival. Mol Cancer 7: 78. 11. Lieblein JC, Ball S, Hutzen B, Sasser AK, Lin HJ, et al. (2008) STAT3 can be activated through paracrine signaling in breast epithelial cells. BMC Cancer 8: 302. Fields of Study Major Field: Molecular, Cellular and Developmental Biology iii Table of Contents Abstract ............................................................................................................................................ ii Acknowledgments........................................................................................................................... iv Vita ................................................................................................................................................... v Publications ...................................................................................................................................... v Fields of Study ................................................................................................................................. iii List of Tables .................................................................................................................................... ii List of Figures .................................................................................................................................. iii List of Abbreviations ....................................................................................................................... iv Chapter 1: Background .................................................................................................................... 1 Medulloblastoma ......................................................................................................................... 1 Diagnosis and treatment ......................................................................................................... 4 Sequelae ................................................................................................................................... 6 The Measles Virus ...................................................................................................................... 10 Basic Biology .......................................................................................................................... 12 Oncolytic virotherapy and rationale for measles virus-based therapies................................ 19 Oncolytic measles viruses....................................................................................................... 22 Considerations for measles virotherapy................................................................................. 28 Treatment of medulloblastoma with oncolytic MV ................................................................... 30 Chapter 2: Oncolytic measles virus prolongs survival in a murine model of cerebral spinal fluid- disseminated medulloblastoma ..................................................................................................... 35 Introduction ..............................................................................................................................