Table of content
RÉSUMÉ 7
ABBREVIATIONS 9
CHAPTER I 13
1. INTRODUCTION 13
1.1. INTERSTITIAL CELLS OF CAJAL 14
1.1.1. Organization of ICC networks in the gastrointestinal tract 15 1.1.2. Physiological role of ICC in the gastrointestinal tract 16
1.1.3. ICC markers 17
1.2. RECEPTOR TYROSINE KINASE KIT 18
1.2.1. RTK KIT structure 19
1.2.2. Stem cell factor/KIT signal transduction 21
1.2.3. Regulation of KIT signaling 27
1.3. GASTROINTESTINAL STROMAL TUMORS (GIST) 29
1.3.1. GIST epidemiology 29
1.3.2. GIST immunohistochemical markers 30
1.3.3. Oncogenic mutations in GIST 31
1.3.4. Therapeutic targets in GIST 32
1.3.5. In vitro and in vivo GIST models to study GIST 33 1.3.6. Transcriptome study of KitK641E/K641E mouse 36
CHAPTER II 42
2. AIMS AND FRAMEWORK OF THE STUDY 42
2.1. AIMS OF THE STUDY 42
2.2.1. Selection of putative genes of interest 42 2.2.2. Expression and regulation of putative targets using in vitro cellular GIST model 43 2.2.3. Localization of selected putative targets by immunohistochemistry in human GIST
pathological material 43
2.2.4. Expression and regulation of selected putative targets using in vivo mouse models 44
CHAPTER III 45
3. MATERIAL AND METHODS 45
3.1. CELL CULTURE AND CELL GROWTH ASSAY 45
3.2. ANIMALS 45
3.3. TOTAL GASTROINTESTINAL TRANSIT TIME IN ADULT MICE 46
3.4. REAL TIME QUANTITATIVE PCR(QPCR) 47
3.5. WESTERN BLOT 48
3.6. RADIOLABELING OF NT-XIX 48
3.7. BINDING ASSAYS 49
3.8. IMMUNOFLUORESCENCE 50
3.9. QUANTIFICATION OF ICC USING IMMUNOREACTIVITY ON CIRCUMFERENTIAL SECTIONS. 52
3.10. CONFOCAL MICROSCOPY 52
3.11. IMMUNOHISTOCHEMISTRY ON HUMAN TISSUE MICROARRAY 53
CHAPTER IV 55
4. RADIO-LABELED NEUROTENSIN RECEPTOR 1 LIGAND AS A NEW IN VIVO IMAGING TOOL FOR
CHAPTER V 59
5. ENDOGLIN AS A PUTATIVE THERAPEUTIC TARGET FOR GASTROINTESTINAL STROMAL
TUMORS? 59
CHAPTER VI 63
6. GLYPICAN 6 EXPRESSION IN HUMAN GASTROINTESTINAL STROMAL TUMORS? 63
CHAPTER VII 65
7. ERK ACTIVITY AND SPROUTY HOMOLOG PROTEINS REGULATION IN A HUMAN K642EGIST
CELL LINE MODEL 65
CHAPTER VIII 71
8. HYPERPLASIA OF INTERSTITIAL CELLS OF CAJAL IN SPROUTY HOMOLOG 4 DEFICIENT MICE
–THYS ET AL,PLOSONE,2015– 71
CHAPTER VIII 113
9. HYPERPLASIA OF INTERSTITIAL CELLS OF CAJAL IN SPROUTY HOMOLOG 4 DEFICIENT MICE
–SUPPLEMENTARY DATA – 113
CHAPTER X 118
10. DISCUSSION AND PERSPECTIVES 118
10.1. INCREASED ICC DENSITY IN SPRY4KO ANIMALS 118
10.1.1. PDE3A as a novel robust ICC marker 118
10.1.2. Quantification of ICC area in mouse gut 119
10.2. ROLE OF SPRY IN CELL SIGNALING 121
10.3. SPRY4 AND EMBRYONIC ICC DEVELOPMENT 126
10.4. CONCLUDING WORDS 126
ACKNOWLEDGEMENTS 127
REFERENCES 129
ANNEX I 149
NEUROTENSIN RECEPTOR 1 IS EXPRESSED IN GASTROINTESTINAL STROMAL TUMORS BUT NOT
INTERSTITIAL CELLS OF CAJAL 149
ANNEX II 163
ENDOGLIN/CD105 IS EXPRESSED IN KIT POSITIVE CELLS IN THE GUT AND IN GASTROINTESTINAL
STROMAL TUMORS 163
ANNEX III 185
HIGH EXPRESSION OF THE RNA-BINDING PROTEIN RBPMS2 IN GASTROINTESTINAL STROMAL
TUMORS 185
ANNEX V 195
