Novel Approaches to Improving Opioid Analgesic Therapy Item Type text; Electronic Dissertation Authors Duron, David I. Publisher The University of Arizona. Rights Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction, presentation (such as public display or performance) of protected items is prohibited except with permission of the author. Download date 29/09/2021 11:09:47 Link to Item http://hdl.handle.net/10150/641705 1 NOVEL APPROACHES TO IMPROVING OPIOID ANALGESIC THERAPY by David Isaiah Duron Copyright © David Isaiah Duron 2020 A Dissertation Submitted to the Faculty of the DEPARTMENT OF PHARMACOLOGY In Partial Fulfillment of the Requirements For the Degree of DOCTOR OF PHILOSOPHY In the Graduate College THE UNIVERSITY OF ARIZONA 2020 3 ACKNOWLEDGEMENTS I first want to acknowledge my mentor John Streicher, PhD for giving me the opportunity to work within his lab and promoting free expression of thoughts and ideas within his lab. I would like to acknowledge my committee members as well for providing me with direction and furthering my academic development. I would also like to thank Wei Lei, PhD for providing the scientific basis for my initial project and also for his training and troubleshooting assistance of various procedures within the lab. I would next like to thank the other graduate students within my lab Justin LaVigne and Seph Palomino for their intellectual contributions and also for their friendship. I would like to also thank the rest of my current/former lab members namely Atilla Keresztes, PhD, Carrie Stine, and Gaby Molnar for their assistance and support within the lab. Finally, I would like to thank my friends and family for their emotional support. 4 DEDICATION I dedicate this to my parents who despite their differences have always supported me throughout my life in all my endeavors. 5 TABLE OF CONTENTS • LIST OF FIGURES ................................................................................................... 6 • LIST OF TABLES ...................................................................................................... 8 • ABSTRACT ............................................................................................................... 9 • CHAPTER 1: INTRODUCTION TO PAIN ................................................................ 11 • CHAPTER 2: TREATMENT OF PAIN: OPIOIDS ..................................................... 19 • CHAPTER 3: IMPROVING OPIOID TREATMENT .................................................. 24 o OPIOID SIGNALING MODULATION: HEAT SHOCK PROTEIN 90 ............. 24 o NON-PHARMACOLOGICAL OPIOID MODULATION: DIET AND INTERMITTENT FASTING .......................................................................... 26 • CHAPTER 4: HSP90 MODULATION OF OPIOID SIGNALING IN THE SPINAL CORD ...................................................................................................................... 29 o INTRODUCTION ......................................................................................... 29 o METHODS AND MATERIALS ...................................................................... 31 o RESULTS .................................................................................................... 39 o DISCUSSION ............................................................................................... 56 • CHAPTER 5: IMPROVING THE THERAPUTIC INDEX OF SYSTEMIC MORPHINE THROUGH SPINAL CORD HSP90 INHIBITION ..................................................... 64 o INTRODUCTION ......................................................................................... 64 o METHODS AND MATERIALS ...................................................................... 66 o RESULTS .................................................................................................... 70 o DISCUSSION ............................................................................................... 78 • CHAPTER 6: NON-PHARMACOLOGICAL IMPROVEMENT OF THE THERAPUTIC INDEX OF MORPHINE: INTERMITTENT FASTING ............................................... 84 o INTRODUCTION ......................................................................................... 84 o METHODS AND MATERIALS ...................................................................... 87 o RESULTS .................................................................................................... 93 o DISCUSSION ............................................................................................. 106 • CHAPTER 7: GENERAL DISCUSSION AND CONCLUSIONS ............................. 111 o SPINAL CORD HSP90 .............................................................................. 112 o INTERMITTENT FASTING ........................................................................ 121 o CLOSING STATEMENTS .......................................................................... 128 • REFERENCES ...................................................................................................... 129 6 LIST OF FIGURES • CHAPTER 1: INTRODUCTION TO PAIN o Fig. 1 - Diagram of classical nociceptive circuitry ......................................... 13 • CHAPTER 4: HSP90 MODULATION OF OPIOID SIGNALING IN THE SPINAL CORD o Fig. 1 - Spinal cord Hsp90 inhibition enhances morphine anti-nociception ... 41 o Fig. 2 Brain Hsp90 inhibition overrides spinal cord Hsp90 inhibition with respect to opioid anti-nociception ................................................................. 44 o Fig. 3 - Spinal Hsp90 inhibition enables opioid activation of ERK MAPK, leading to enhanced anti-nociception ........................................................... 47 o Fig. 4 - Spinal Hsp90 inhibition evokes rapid protein translation that is necessary for enhanced morphine anti-nociception ...................................... 51 o Fig. 5 - Quantitative proteomic analysis reveals a protein network altered by spinal Hsp90 inhibition ................................................................................. 53 o Fig. 6 - Spinal Hsp90 inhibition activates RSK1/2 phosphorylation, which is necessary for enhanced morphine anti-nociception ...................................... 57 o Fig. 7 - Proposed model of Hsp90 regulation of opioid signaling in the spinal cord .............................................................................................................. 60 • CHAPTER 5: IMPROVING THE THERAPUTIC INDEX OF SYSTEMIC MORPHINE THROUGH SPINAL CORD HSP90 INHIBITION o Fig. 1 - Intrathecal KU-32 enhances morphine anti-nociception dose dependently in the tail flick model ................................................................. 72 o Fig. 2 - Intrathecal KU-32 enhances morphine anti-nociception dose dependently in the post-operative paw incision model .................................. 73 o Fig. 3 - Intrathecal KU-32 enhances morphine anti-nociception dose dependently in the HIV neuropathic pain model ........................................... 75 o Fig. 4 - Intrathecal KU-32 reduces morphine anti-nociceptive tolerance in the tail flick model .............................................................................................. 76 o Fig. 5 - Intrathecal KU-32 reverses morphine anti-nociceptive tolerance in the tail flick model .............................................................................................. 77 o Fig. 6 - Intrathecal KU-32 has no significant impact on morphine induced constipation .................................................................................................. 79 o Fig. 7 - Intrathecal KU-32 has no significant impact on morphine induced reward .......................................................................................................... 80 • CHAPTER 6: NON-PHARMACOLOGICAL IMPROVEMENT OF THE THERAPUTIC INDEX OF MORPHINE: INTERMITTENT FASTING o Fig. 1 - Daily intermittent fasting enhances morphine anti-nociception ......... 94 o Fig. 2 - Daily intermittent fasting blocks morphine-induced reward ............... 97 o Fig. 3 - Daily intermittent fasting reduces opioid tolerance and constipation . 99 o Fig. 4 - Altered pharmacokinetics do not explain benefits of intermittent fasting ........................................................................................................ 101 o Fig. 5 - Daily intermittent fasting enhances MOR efficacy and reduces tolerance in spinal cord and PAG ............................................................... 103 7 o Fig. 6 - Daily intermittent fasting does not change MOR protein expression in spinal cord and PAG .............................................................................................107 8 LIST OF TABLES • CHAPTER 6: NON-PHARMACOLOGICAL IMPROVEMENT OF THE THERAPUTIC INDEX OF MORPHINE: INTERMITTENT FASTING o Table 1 - Daily intermittent fasting enhances MOR efficacy and reduces tolerance in spinal cord and PAG ............................................................... 105 9 ABSTRACT Despite their extensive side effect profile and potential for abuse, clinically used opioids such as morphine are still the most effective analgesics for most chronic pain patients. A negative stigma has been cast over opioids due to the recent opioid epidemic, which has claimed hundreds of thousands of lives over the past few decades in the United States alone. Despite this, there is still great potential for the use of these drugs if negative side effect