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Nucleobindin-2: a Novel Regulator of Immune-Metabolic Interactions" (2016)

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Nucleobindin-2: a Novel Regulator of Immune-Metabolic Interactions Louisiana State University LSU Digital Commons LSU Doctoral Dissertations Graduate School 2016 Nucleobindin-2: A Novel Regulator of Immune- Metabolic Interactions Anthony Ravussin Louisiana State University and Agricultural and Mechanical College, [email protected] Follow this and additional works at: https://digitalcommons.lsu.edu/gradschool_dissertations Recommended Citation Ravussin, Anthony, "Nucleobindin-2: A Novel Regulator of Immune-Metabolic Interactions" (2016). LSU Doctoral Dissertations. 3652. https://digitalcommons.lsu.edu/gradschool_dissertations/3652 This Dissertation is brought to you for free and open access by the Graduate School at LSU Digital Commons. It has been accepted for inclusion in LSU Doctoral Dissertations by an authorized graduate school editor of LSU Digital Commons. For more information, please [email protected]. NUCLEOBINDIN-2: A NOVEL REGULATOR OF IMMUNE-METABOLIC INTERACTIONS A Dissertation Submitted to the Graduate Faculty of the Louisiana State University and Agricultural and Mechanical College in partial fulfillment of the requirements for the degree of Doctor of Philosophy in The Department of Biological Sciences by Anthony Ravussin B.S., Louisiana State University, 2008 August 2016 Acknowledgements I would like to thank my thesis advisor, Dr. Vishwa D. Dixit, for having persuaded me in 2011 to pursue a Ph.D. and further my knowledge by conducting this doctoral work. His encouragement and mentorship has guided me during the course of my graduate career. Throughout my time in his lab, he has given me the necessary tools, both intellectually and physically, to conduct the experiments described here within my thesis. Furthermore, I cannot express the appreciation, veneration, and respect I have for him. I would also like to thank my thesis committee members, Drs. Jacqueline Stephens, Mark A. Batzer and Patrick J. DiMario, for their guidance during my PhD work. I have been inspired by each of them and they have been instrumental in the progression of my career. I would like to acknowledge that Dr. Stephens was highly involved in my decision to pursue a Ph.D., as well as knowledge and intellectual discussions about science and various projects. I would like to also thank all the members of Dr. Dixit’s lab, both past and present, which have helped me in innumerable ways. I would also like to express thanks to the students, postdocs and faculty of the Yale Immunobiology Department for accepting me in as one of their own student. I have had guidance, support, and gained immense knowledge during the past several years that have been instrumental to my work and intellect. I would also like to thank my family (Eric, Jacqueline, Yann and Jeremy Ravussin) for supporting me throughout my studies and always aiding during more difficult times of scientific research. I am grateful to Lene Hjelle, whose love and support have kept me grounded and sane during my thesis work. ii Table of Contents ACKNOWLEDGEMENTS ....................................................................................................................ii LIST OF FIGURES ............................................................................................................................... v LIST OF ABBREVIATIONS ................................................................................................................ viii ABSTRACT ........................................................................................................................................ ix CHAPTER 1: INTRODUCTION ........................................................................................................... 1 1.1 Background and Significance .................................................................................................... 1 1.2 Metabolism, Obesity and Type 2 Diabetes mellitus ................................................................. 2 1.3 Immunity ................................................................................................................................... 8 1.4 Immuno-metabolism .............................................................................................................. 12 1.5 Nucleobindin-2 ........................................................................................................................ 17 CHAPTER 2: EFFECTS OF NUCLEOBINDIN-2 ON GLUCOSE HOMEOSTASIS AND INSULIN RESISTANCE ................................................................................................................................... 20 2.1 Introduction ............................................................................................................................ 20 2.2 Materials and Methods ........................................................................................................... 21 2.3 Results ..................................................................................................................................... 23 2.4 Discussion ................................................................................................................................ 44 CHAPTER 3: EFFECTS OF NUCLEOBINDIN-2 ON MYELOID CELLS AND LYMPHOCYTES AND STERILE OBESITY-INDUCED INFLAMMATION ............................................................................................. 48 3.1 Introduction ............................................................................................................................ 48 3.2 Materials and Methods ........................................................................................................... 49 3.3 Results ..................................................................................................................................... 52 3.4 Discussion ................................................................................................................................ 74 CHAPTER 4: REGULATION OF LIPOPOLYSACCHARIDE (LPS)-INDUCED INFLAMMATION AND ENDOTOXEMIA BY NUCLEOBINDIN-2 ........................................................................................... 80 4.1 Introduction ............................................................................................................................ 80 4.2 Materials and Methods ........................................................................................................... 81 4.3 Results ..................................................................................................................................... 83 4.4 Discussion ................................................................................................................................ 89 iii CHAPTER 5: DISCUSSION AND FUTURE DIRECTIONS .................................................................... 91 REFERENCES .................................................................................................................................. 96 APPENDIX .................................................................................................................................... 112 VITA ............................................................................................................................................. 117 iv List of Figures Figure 1.1: Overview of typical energy metabolism processes in a macrophage .......................... 4 Figure 1.2: Overview of adipocyte insulin dependent glucose uptake via GLUT4 transporter ...... 8 Figure 1.3: Time course of a normal immune response with important cell types ..................... 10 Figure 1.4: Adipose tissue expand during obesity in parallel to inflammation and metabolic dysregulation ................................................................................................................................ 15 Figure 1.5: Graphical representation of Nucleobindin-2 gene, transcript and proteins .............. 19 Figure 1.6: Post translational peptide proteins and peptide motifs ............................................ 19 Figure 2.1: Age and diet modulates Nucb2 expression differently in different tissues ............... 25 Figure 2.2: High fat diet-induced obesity increases Nucb2 expression in visceral adipose tissue macrophages (ATMs) and T cells .................................................................................................. 25 Figure 2.3: Nucb2 -/- vector construct and embryonic stem cell PCR screens ............................. 27 Figure 2.4: Nucb2 floxed-neo/+ founder mouse and PCR genotyping screen ............................. 28 Figure 2.5: Nucb2-floxed/+ founder mouse and PCR genotyping screen .................................... 28 Figure 2.6: Nucb2 is ubiquitously expressed in many cell types and tissue ................................. 29 Figure 2.7: Nucb2 ablation does not alter detected circulating levels of nesfatin-1 or Nucb2 .... 31 Figure 2.8: Nucb2 ablation does not alter food intake ................................................................. 32 Figure 2.9: Nucb2 ablation has no effect on body weight and body composition ....................... 34 Figure 2.10: Nucb2 ablation does not impact adipocyte morphology and adipocyte size ......... 35 Figure 2.11: Nucb2 ablation does not affect circulating adipokines in HFD fed mice .................. 37 Figure 2.12: Nucb2 ablation does not affect circulating IL-18 in HFD fed mice ........................... 38 Figure 2.13: Nucb2 does not affect whole body glucose homeostasis in lean chow fed mice ... 39 Figure 2.14: Nucb2 does not affect plasma insulin
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