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AMPK and Mtor Impose Dualistic Regulation of Mitophagy and Autophagy in in Vitro Models of Prostate Cancer

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AMPK and Mtor Impose Dualistic Regulation of Mitophagy and Autophagy in in Vitro Models of Prostate Cancer Philadelphia College of Osteopathic Medicine DigitalCommons@PCOM PCOM Biomedical Studies Student Scholarship Student Dissertations, Theses and Papers 7-2020 AMPK and mTOR Impose Dualistic Regulation of Mitophagy and Autophagy in In Vitro Models of Prostate Cancer Brianne Richardson Philadelphia College of Osteopathic Medicine Follow this and additional works at: https://digitalcommons.pcom.edu/biomed Part of the Medicine and Health Sciences Commons Recommended Citation Richardson, Brianne, "AMPK and mTOR Impose Dualistic Regulation of Mitophagy and Autophagy in In Vitro Models of Prostate Cancer" (2020). PCOM Biomedical Studies Student Scholarship. 200. https://digitalcommons.pcom.edu/biomed/200 This Thesis is brought to you for free and open access by the Student Dissertations, Theses and Papers at DigitalCommons@PCOM. It has been accepted for inclusion in PCOM Biomedical Studies Student Scholarship by an authorized administrator of DigitalCommons@PCOM. For more information, please contact [email protected]. Philadelphia College of Osteopathic Medicine Graduate Program in Biomedical Sciences Department of Bio-Medical Sciences AMPK and mTOR Impose Dualistic Regulation of Mitophagy and Autophagy in In Vitro Models of Prostate Cancer A Thesis in Biomedical Sciences by Brianne Richardson Copyright 2020 Brianne Richardson Submitted in Partial Fulfillment of the Requirements for the Degree of Masters in Biomedical Sciences July 2020 This thesis has been presented to and accepted by the Associate Dean for Curriculum and Research Office of Philadelphia College of Osteopathic Medicine in partial fulfillment of the requirements for the degree of Master of Science in Biomedical Sciences. We, the undersigned, duly appointed committee have read and examined this manuscript and certify it is adequate in scope and quality as a thesis for this master’s degree. We approve the content of the thesis to be submitted for processing and acceptance. ______________________________________ Dr. Dennis Peffley, Thesis Advisor Professor of Physiology and Pharmacology Department of Bio-Medical Sciences _______________________________________ Dr. Harold Komiskey, Thesis Committee Member Professor of Neuroscience, Physiology and Pharmacology Department of Bio-Medical Sciences ________________________________________ Dr. Xinyu (Eric) Wang, Thesis Committee Member Associate Professor of Pharmaceutical Sciences Department of Pharmaceutical Sciences _________________________________________ Dr. Richard White, Program Director, Research Concentration Professor Department of Bio-Medical Sciences 5 SUMMARY Prostate cancer is the abnormal growth and proliferation of previously normal cells of the prostate and has the second highest incidence in men worldwide. Physiologic manipulation of AMP-activated protein kinase (AMPK), a highly conserved enzyme responsible for regulation of energy homeostasis during metabolic stress, is a potential treatment, especially for metastatic, castration-resistant prostate cancers. AMPK is a known inhibitor of the enzyme mTOR, the final enzyme in the PKT/AKT/mTOR pathway responsible for cell growth and proliferation signaling. The hypothesis of this project is that activation of AMPK leads to increased mTOR-dependent mitophagy and subsequent autophagy in prostate cancer cells. The overall aim of this project is to establish a role between activated AMPK and mTOR dependent mitophagy and autophagy. Although preliminary data remains inconclusive, future research is promising. 6 TABLE OF CONTENTS SUMMARY ...................................................................................................................................... 6 ACKNOWLEDGEMENTS ................................................................................................................ 10 INTRODUCTION ............................................................................................................................ 11 A Brief Introduction to Cancer ................................................................................................. 11 The cell cycle ......................................................................................................................... 13 Mitosis .................................................................................................................................. 13 Mitigation of DNA mutations .............................................................................................. 15 The Warburg Effect .............................................................................................................. 18 Prostate Cancer ........................................................................................................................ 22 Prostate development ......................................................................................................... 22 The prostate ......................................................................................................................... 23 Prostatitis, benign prostatic hyperplasia, and prostate cancer ......................................... 26 Prostate cancer ..................................................................................................................... 26 Genomic variability between prostate cancer genomes .................................................... 28 In vitro models of prostate cancer....................................................................................... 30 AMP-Activated Protein Kinase ................................................................................................ 31 The alpha subunit ................................................................................................................. 32 The beta subunit .................................................................................................................. 32 7 The gamma subunit .............................................................................................................. 33 Activation of AMPK .............................................................................................................. 33 Inhibition of AMPK ............................................................................................................... 34 Downstream targets of AMPK ............................................................................................. 34 The mTOR Pathway .................................................................................................................. 35 Mitophagy and Autophagy ...................................................................................................... 39 Autophagy ............................................................................................................................ 39 Mitophagy ............................................................................................................................ 40 AIMS AND GOALS ......................................................................................................................... 41 Previous Research .................................................................................................................... 41 Peffley et al. .......................................................................................................................... 41 Rae et al. ............................................................................................................................... 41 Hypothesis ................................................................................................................................ 42 Specific Aim 1 ........................................................................................................................... 42 Specific Aim 2 ........................................................................................................................... 43 Specific Aim 3 ........................................................................................................................... 44 RESULTS ........................................................................................................................................ 51 Future Research ........................................................................................................................ 58 REFERENCES .................................................................................................................................. 60 8 FIGURE LEGEND ............................................................................................................................ 81 9 ACKNOWLEDGEMENTS To the faculty of my committee, the Biomedical Sciences Department, and PCOM Georgia’s Department of Research, thank you for your invaluable guidance during this chapter of my academic and personal endeavors. To my family, thank you for your support in chasing my dreams. To Dr. Kasia Dinkeloo, this thesis could not have been finished without the inspiration you always seem to provide and your daily reminders to drink coffee. Most importantly, I am forever indebted to my dog, Bocephus, whose unconditional love has held me together through every whirlwind in my life’s journey. I would like to note that he will never understand this because, in the six years we have gone to school together, I have never taught him to read. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. This research received no
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