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The Effects of GSPT1 Degradation on Serum Calcium, Parathyroid Hormone, and Fibroblast Growth Factor 23 Concentrations in Human Cereblon Knock-In Mice

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The Effects of GSPT1 Degradation on Serum Calcium, Parathyroid Hormone, and Fibroblast Growth Factor 23 Concentrations in Human Cereblon Knock-In Mice Virginia Commonwealth University VCU Scholars Compass Theses and Dissertations Graduate School 2020 The Effects of GSPT1 Degradation on Serum Calcium, Parathyroid Hormone, and Fibroblast Growth Factor 23 Concentrations in Human Cereblon Knock-in Mice Kamran Ghoreishi Virginia Commonwealth University Follow this and additional works at: https://scholarscompass.vcu.edu/etd Part of the Other Medicine and Health Sciences Commons © The Author Downloaded from https://scholarscompass.vcu.edu/etd/6490 This Dissertation is brought to you for free and open access by the Graduate School at VCU Scholars Compass. It has been accepted for inclusion in Theses and Dissertations by an authorized administrator of VCU Scholars Compass. For more information, please contact [email protected]. The Effects of GSPT1 Degradation on Serum Calcium, Parathyroid Hormone, and Fibroblast Growth Factor 23 Concentrations in Human Cereblon Knock-in Mice by Kamran Ghoreishi Bachelor of Science in Microbiology and Clinical Laboratory Sciences, San Diego State University, 1995 Master of Science in Public Health (Toxicology), San Diego State University, 2003 Master of Science in Regulatory Affairs, San Diego State University, 2013 A dissertation submitted in the partial fulfillment of the requirements for the degree of Doctor of Philosophy at Virginia Commonwealth University. Virginia Commonwealth University, 2020 Advisor: William J. Korzun, Ph.D. Associate Professor Department of Clinical Laboratory Sciences Virginia Commonwealth University Richmond, Virginia November 19, 2020 ACKNOWLEDGEMENT Firstly, I would like to express my sincere gratitude to my advisor Dr. William Korzun for the continuous support and guidance of my research, which incented me to widen my research from various perspectives. My sincere thanks also go to Dr. Joseph Piccotti who provided me an opportunity to conduct this research, without his endless support it would not be possible to conduct this research. I am also thankful for his patience, thoughtful support, and immense knowledge of the field. His guidance helped me in all aspects of my research and writing of this thesis. I could not have imagined having a better counselor and mentor for my PhD study. Besides my advisor, I would like to thank the rest of my thesis committee: Dr. Teresa Nadder and Dr. Melissa Jamerson for their insightful comments and encouragement. I thank my colleagues for their support and contribution to this research, their skills, expertise, knowledge, and most importantly, their willingness to push the boundaries in the laboratory techniques, produced valuable data for this thesis. I like to thank Mr. Luis Alvarado, Mr. Ayan Alavi, Ms. Yan Ren, Ms. Tatiana Galvez, Ms. Kelsie Smith, Ms. Yang Tang, and Dr. Roberto Guzman for their contributions. Finally, but most importantly, I would like to thank my family: my parents, my wife Sahar, my daughter Parisa, and my son Parsa for their emotional support and encouragement, which made this journey so much more pleasant. I am forever grateful for their patience and understanding, they have earned this degree right along with me. TABLE OF CONTENTS TABLE OF CONTENTS ................................................................................................................. I LIST OF FIGURES ........................................................................................................................ V LIST OF TABLES ....................................................................................................................... VII GLOSSARY OF ABBREVIATIONS AND DEFINITIONS OF TERMS .................................. IX Abstract ......................................................................................................................................1 CHAPTER 1: INTRODUCTION ...................................................................................................3 Purpose of this Study and Research Questions ..................................................................10 Hypotheses .........................................................................................................................11 Summary ............................................................................................................................13 CHAPTER 2: LITERATURE REVIEW ......................................................................................14 Calcium ..............................................................................................................................14 Phosphate ...........................................................................................................................15 Magnesium .........................................................................................................................16 Calcium Homeostasis.........................................................................................................20 Hypocalcemia ....................................................................................................................21 Drug-Induced Hypocalcemia .............................................................................................24 Vitamin D...........................................................................................................................26 The Fibroblast Growth Factor 23 .......................................................................................28 The Parathyroid Glands .....................................................................................................30 Rodent Parathyroid ............................................................................................................31 Parathyroid Hormone .........................................................................................................31 Calcium Sensing Receptor .................................................................................................34 Hypoparathyroidism ..........................................................................................................41 Allosteric Modulators of The Calcium Sensing Receptor .................................................45 Positive allosteric modulator of CaSR NPS R-568............................................................45 Negative Allosteric Modulator of CaSR NPS 2143 ..........................................................47 Translation Termination Factor G1 to S phase Transition 1 (GSPT1) Degradation and ...............................................................................................................................48 Hypocalcemia ....................................................................................................................49 Parathyroid Hormone mRNA ............................................................................................50 Cereblon Modulators and Hypocalcemia...........................................................................51 i Human Cereblon Knock-in Mice Use in Research ............................................................52 Summary ............................................................................................................................52 Significance of This Research ...........................................................................................53 CHAPTER THREE: METHODS ..................................................................................................54 Hypotheses .........................................................................................................................55 Pilot Study Design .............................................................................................................58 Main Study Design ............................................................................................................59 Experimental Animals .......................................................................................................59 Test Substances 1 ...............................................................................................................59 Test Substances 2 ...............................................................................................................60 Dose Selection Criteria ......................................................................................................61 Dose Preparation and Administration ................................................................................62 CC-325 ...............................................................................................................................62 NPS 2143 ...........................................................................................................................64 Formulation Analysis .........................................................................................................65 Experimental Groups .........................................................................................................66 Treatment Descriptions ......................................................................................................68 Measurement Procedures ...................................................................................................69 Clinical observations ..........................................................................................................69 Body weights .....................................................................................................................69 Clinical laboratory
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