Washington University in St. Louis Washington University Open Scholarship All Theses and Dissertations (ETDs) Summer 9-1-2014 DAPK3 Suppresses Mammary Acini Morphogenesis and is Required for Mouse Development Brandon Kocher Washington University in St. Louis Follow this and additional works at: https://openscholarship.wustl.edu/etd Recommended Citation Kocher, Brandon, "DAPK3 Suppresses Mammary Acini Morphogenesis and is Required for Mouse Development" (2014). All Theses and Dissertations (ETDs). 1312. https://openscholarship.wustl.edu/etd/1312 This Dissertation is brought to you for free and open access by Washington University Open Scholarship. It has been accepted for inclusion in All Theses and Dissertations (ETDs) by an authorized administrator of Washington University Open Scholarship. For more information, please contact [email protected]. WASHINGTON UNIVERSITY IN ST. LOUIS Division of Biology and Biomedical Sciences Molecular Cell Biology Dissertation Examination Committee: David Piwnica-Worms, Chair Deborah Novack, Co-chair David Denardo Fanxin Long Sheila Stewart Zhongsheng You DAPK3 Suppresses Mammary Acini Morphogenesis and is Required for Mouse Development by Brandon Anthony Miller Kocher A dissertation presented to the Graduate School of Arts and Sciences of Washington University in partial fulfillment of the requirements for the degree of Doctor of Philosophy August 2014 St Louis, Missouri © 2014, Brandon Anthony Miller Kocher TABLE OF CONTENTS Acknowledgements ....................................................................................................................... vii Abstract of Dissertation ............................................................................................................... viii CHAPTER 1: INTRODUCTION ................................................................................................... 1 1.1 Death-associated protein kinase family (DAPK) .................................................................. 1 1.2 DAPK3 .................................................................................................................................. 2 1.3 DAPK3 & Cancer ................................................................................................................. 4 1.4 The mTOR Pathway ............................................................................................................. 7 1.5 mTOR & Cancer ................................................................................................................. 11 1.6 DAPK and mTOR pathway interactions ............................................................................. 12 1.7 Three-Dimensional Cell Culture as a Powerful Model of Breast Cancer for Elucidating Novel DAPK3 Functions .......................................................................................................... 13 1.8 Figures................................................................................................................................. 16 1.9 References ........................................................................................................................... 19 CHAPTER 2: DAPK3 INHIBITION OF mTOR SIGNALING SUPPRESSES MAMMARY ACINI MORPHOGENESIS ......................................................................................................... 28 2.1 Introduction ......................................................................................................................... 28 2.2 Materials & Methods .......................................................................................................... 30 2.3 Results ................................................................................................................................. 34 2.4 Discussion ........................................................................................................................... 37 2.5 Figures................................................................................................................................. 40 2.6 References ........................................................................................................................... 53 CHAPTER 3: DAPK3 IS REQUIRED FOR EARLY MOUSE DEVELOPMENT & DISPLAYS DISTINCT EXPRESSION PATTERNS IN EMBRYONIC AND ADULT TISSUES ............... 55 3.1 Introduction ......................................................................................................................... 55 3.2 Materials & Methods .......................................................................................................... 56 3.3 Results ................................................................................................................................. 60 3.4 Discussion ........................................................................................................................... 63 3.5 Figures................................................................................................................................. 65 3.6 Table ................................................................................................................................... 70 3.7 References ........................................................................................................................... 71 CHAPTER 4: CONCLUSIONS & FUTURE DIRECTIONS ...................................................... 73 4.1 Conclusions ......................................................................................................................... 73 4.2 Future Directions ................................................................................................................ 73 ii 4.3 References ........................................................................................................................... 79 APPENDIX A: ILLUMINATING CANCER SYSTEMS WITH GENETICALLY- ENGINEERED MOUSE MODELS AND COUPLED LUCIFERASE REPORTERS IN VIVO ....................................................................................................................................................... 80 A.1 Published Manuscript ......................................................................................................... 80 A.2 References ........................................................................................................................ 105 A.3 Figures .............................................................................................................................. 114 A.4 Tables ............................................................................................................................... 117 APPENDIX B: DEVELOPMENT OF TUMOR-MICROENVIRONMENT ACTIVATED ANTI-CANCER SALMONELLA ............................................................................................. 121 B.1 Contribution to authorship ............................................................................................... 121 B.2 Published manuscript ....................................................................................................... 121 B.3 References ........................................................................................................................ 150 B.4 Figures .............................................................................................................................. 154 B.5 Table ................................................................................................................................. 164 B.6 Supplementary Figure ...................................................................................................... 165 B.7 Supplementary Table ........................................................................................................ 166 iii LIST OF FIGURES CHAPTER 1: INTRODUCTION Figure 1.1 .................................................................................................................................. 16 Figure 1.2 .................................................................................................................................. 17 Figure 1.3 .................................................................................................................................. 18 CHAPTER 2: DAPK3 INHIBITION OF mTOR SIGNALING SUPPRESSES MAMMARY ACINI MORPHOGENESIS Figure 2.1 .................................................................................................................................. 40 Figure 2.2 .................................................................................................................................. 41 Figure 2.3 .................................................................................................................................. 42 Figure 2.4 .................................................................................................................................. 43 Figure 2.5 .................................................................................................................................. 44 Figure 2.6 .................................................................................................................................. 45 Figure 2.7 .................................................................................................................................. 46 Figure 2.8 .................................................................................................................................. 47 Figure