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Development and Characterization of an Immunologically Humanized And Iowa State University Capstones, Theses and Graduate Theses and Dissertations Dissertations 2019 Development and characterization of an immunologically humanized and cancer xenograft model in pigs with severe combined immunodeficiency (SCID) Adeline Nicole Boettcher Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/etd Part of the Cell Biology Commons, Developmental Biology Commons, and the Molecular Biology Commons Recommended Citation Boettcher, Adeline Nicole, "Development and characterization of an immunologically humanized and cancer xenograft model in pigs with severe combined immunodeficiency (SCID)" (2019). Graduate Theses and Dissertations. 16975. https://lib.dr.iastate.edu/etd/16975 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Graduate Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Development and characterization of an immunologically humanized and cancer xenograft model in pigs with severe combined immunodeficiency (SCID) by Adeline N. Boettcher A dissertation submitted to the graduate faculty in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Major: Molecular, Cellular, and Developmental Biology Program of Study Committee: Christopher K. Tuggle; Major Professor Joan E. Cunnick Crystal L. Loving Jason W. Ross Randy E. Sacco The student author, whose presentation of the scholarship herein was approved by the program of study committee, is solely responsible for the content of this dissertation. The Graduate College will ensure this dissertation is globally accessible and will not permit alterations after a degree is conferred. Iowa State University Ames, Iowa 2019 Copyright © Adeline Boettcher, 2019. All rights reserved. ii TABLE OF CONTENTS Page LIST OF FIGURES ........................................................................................................ v LIST OF TABLES .......................................................................................................... viii NOMENCLATURE ....................................................................................................... ix ACKNOWLEDGMENTS ............................................................................................... xii ABSTRACT………………………………. ................................................................................ xiv CHAPTER 1 INTRODUCTION AND DISSERTATON STRUCTURE ......................... 1 CHAPTER 2 LITERATURE REVIEW ...................................................................... 3 Abstract ......................................................................................................... 3 Introduction ........................................................................................................ 5 Existing SCID pig models ..................................................................................... 7 Leaky T cell development in SCID pigs and Artemis function ............................. 11 SCID pig cancer xenotransplantation studies ..................................................... 13 Porcine immunological similarities to human .................................................... 15 Routes for humanization and applications ......................................................... 16 Future outlooks on the utilization of SCID pigs for cancer therapies and research ........................................................................................................ 24 Concluding remarks ............................................................................................ 27 References ......................................................................................................... 28 Figures ......................................................................................................... 41 Tables ......................................................................................................... 49 Supplemental Tables ........................................................................................... 51 CHAPTER 3 PORCINE SIRPA BINDS TO HUMAN CD47 TO INHIBIT PHAGOCYTOSIS: IMPLICATIONS FOR HUMAN HEMATOPOIETIC STEM CELL TRANSPLANTATION INTO SCID PIGS .......................... 52 Abstract ......................................................................................................... 52 Introduction ........................................................................................................ 54 Results ......................................................................................................... 56 Discussion ......................................................................................................... 61 Materials and Methods ....................................................................................... 64 References ......................................................................................................... 67 Figures ......................................................................................................... 73 Supplemental Figures.......................................................................................... 77 iii CHAPTER 4 A COMPREHENSIVE PROTOCOL FOR LAPAROTOMY IN SWINE TO FACILITATE ULTRASOUND-GUIDED INJECTION INTO THE FETAL INTRAPERITONEAL SPACE .............................................................. 79 Abstract ......................................................................................................... 79 Introduction ........................................................................................................ 80 Results ......................................................................................................... 82 Discussion ......................................................................................................... 85 Materials and Methods ....................................................................................... 88 References ......................................................................................................... 92 Figures ……………………………………………………………………………………………………..... .. 96 Tables …………………………………………………………………............................................ 99 CHAPTER 5 ENGRAFTMENT OF HUMAN IMMUNE CELLS IN CRISPR/CAS9 Art-/- IL2RG-/Y SCID PIGS AFTER IN UTERO INJECTION OF HUMAN CD34+ CELLS .................................................................................................. 103 Abstract ......................................................................................................... 104 Introduction ........................................................................................................ 104 Results ......................................................................................................... 107 Discussion ......................................................................................................... 113 Materials and Methods ....................................................................................... 118 References ......................................................................................................... 127 Figures ......................................................................................................... 133 Tables ......................................................................................................... 139 Supplemental Figures.......................................................................................... 140 Supplemental Tables ........................................................................................... 146 CHAPTER 6 HUMAN OVARIAN CANCER TUMOR FORMATION IN SEVERE COMBINED IMMUNODEFICIENT (SCID) PIGS ............................... 153 Abstract ......................................................................................................... 153 Introduction ........................................................................................................ 154 Results ......................................................................................................... 157 Discussion ......................................................................................................... 159 Materials and Methods ....................................................................................... 161 References ......................................................................................................... 164 Figures ......................................................................................................... 168 Tables ......................................................................................................... 170 CHAPTER 7 MUTATIONS IN ARTEMIS CAN LEAD TO LEAKY CD3ε+ CELL DEVELOPMENT IN SWINE WITH SEVERE COMBINED IMMUNODEFICIENCY (SCID) .......................................................... 171 Abstract ......................................................................................................... 171 Introduction ........................................................................................................ 172 Results ......................................................................................................... 173 Discussion ........................................................................................................
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