Cell Fusion in Health and Disease (eBook)

Preface 6
Contents 9
Contributors 10
1 Introduction 12
2 Heterochronic Control of AFF-1-Mediated Cell-to-Cell Fusion in C. elegans 15
2.1 Introduction 15
2.2 Heterochronic Genes Regulate the Timing of Developmental Events 15
2.3 The Heterochronic Gene lin-29 Determines the Final Fate of the Seam Cells 16
2.4 LIN-29 Controls the Terminal Differentiation of the Epidermal Seam Cells 17
2.5 AFF-1 Protein Mediates the Terminal Fusion of the Hypodermal Seam Cells 18
References 19
3 Role of SNAREs in Membrane Fusion 22
3.1 Introduction 22
3.2 Materials and Methods 24
3.2.1 Preparation of Lipid Bilayer 24
3.2.2 Lipid Membrane on Mica Surface 25
3.2.3 Atomic Force Microscopy 25
3.2.4 EPC9 Electrophysiological Lipid Bilayer Setup 25
3.2.5 Preparation of Lipid Vesicles and SNARE Protein Reconstitutions 25
3.2.6 Circular Dichroism Spectroscopy 26
3.2.7 Wide-Angle X-Ray Diffraction 26
3.3 Discussion 26
3.3.1 V-SNARE and t-SNAREs Need to Reside in Opposing Membrane to Appropriately Interact and Establish Continuity Between Those Membranes 26
3.3.2 Membrane Curvature Dictate the Size of the SNARE Ring Complex 29
3.3.3 Disassembly of the SNARE Complex 31
3.3.4 CD Spectroscopy Confirm the Requirement of Membrane for Appropriate t-/v-SNARE Complex Assembly, and that NSF-ATP Alone Can Mediated SNARE Disassembly 35
3.3.5 SNAREs Bring Opposing Bilayers Closer, Enabling Calcium Bridging and Membrane Fusion 38
References 40
4 Molecular and Cellular Mechanisms of Mammalian Cell Fusion 42
4.1 Introduction 42
4.2 Programming Cellular Competence for Fusion 44
4.2.1 Cytokines 44
4.2.2 DNAX Activating Protein 12 45
4.2.3 Phosphatidylserine 45
4.2.4 Calcium 46
4.2.5 Proteases 47
4.2.6 Glis3 47
4.3 ChemoattractantReceptor Interactions and Cell Migration 48
4.3.1 Secretion and Function of Chemoattractants During Myogenesis 48
4.3.1.1 Hepatocyte Growth Factor 48
4.3.1.2 SDF-1/CXCR4 Axis 48
4.3.1.3 IL-4 48
4.3.2 Mouse Odorant Receptor 23 49
4.3.3 Monocyte Chemoattractant Protein-1 49
4.3.4 Progesterone 49
4.3.5 Integrins 50
4.3.6 The d2 Isoform of Vacuolar ATPase V0 Domain 50
4.3.7 Actin Cytoskeleton Regulators 51
4.3.8 Mannose Receptor 51
4.3.9 Matrix Metalloproteinases 51
4.4 Membrane Recognition and Adhesion 52
4.4.1 Immunoglobulin Super Family 52
4.4.1.1 Orthologs of Drosophila Ig Super Family Proteins 52
4.4.1.2 Other Mammalian Ig Proteins Involved in Myoblast Fusion 52
4.4.1.3 Izumo 53
4.4.1.4 CD47/SIRP-aInteraction 53
4.4.2 Cadherins 54
4.4.2.1 N-Cadherin 54
4.4.2.2 M-Cadherin 54
4.4.2.3 E-Cadherin and Cadherin-11 54
4.4.3 Tetraspanins 55
4.4.4 Integrins 55
4.4.5 Glycosyl-Phosphatidylinositol (GPI)-Anchored proteins 55
4.4.6 A Disintegrin And Metalloproteinase (ADAM) 56
4.4.7 Dendritic Cell-Specific Transmembrane Protein (DC-STAMP) 56
4.5 Fusion Pore Formation and Expansion 57
4.5.1 Actin Cytoskeleton 57
4.5.2 Lipid Rafts 59
4.5.3 A Novel Model of Plasma Membrane Fusion 59
4.5.4 Syncytin-1 and -2 60
4.6 Post-fusion Resetting and Cell Survival 61
4.6.1 Myoferlin 61
4.6.2 Bcl-2 and c-Flip 61
4.7 Conclusions 62
References 63
5 Membrane Fusions During Mammalian Fertilization 74
5.1 Introduction 74
5.2 Surface Remodeling of Gametes Prior to Zona Binding 76
5.2.1 The Cumulus-Oocyte Complex in the Oviduct 76
5.2.2 Sperm Cell Surface Remodeling 76
5.3 Zona Binding and Initiation of the Acrosome Reaction 77
5.3.1 Zona Pellucida Contains Acrosome Exocytosis Inducing Binding Sites 77
5.3.2 Acrosome Exocytosis 77
5.4 Zona Penetration After the Acrosome Reaction 78
5.5 Gamete Membrane Fusion and the Oocyte-to-Embryo Transition 79
5.6 The Membrane (Oolemma) Block to Polyspermy 81
5.6.1 Redundant Sperm Around the Fertilized Oocyte 81
5.6.2 Prevention of Polyspermy at the Oolemma 81
5.7 Cortical Reaction and the Zona Pellucida Block to Polyspermic Fertilization 82
5.7.1 Cortical Granules Content Can Modify the Zona Pellucida Structure 82
5.7.2 Maturation Dependent Exocytotic Fusion Machinery of the Cortical Reaction 82
5.8 Conclusion 83
References 84
6 Trophoblast Fusion 90
6.1 Introduction 90
6.2 The Trophoblast 91
6.2.1 The Villous Trophoblast 91
6.3 Initiation of Syncytial Fusion in Villous Trophoblast 92
6.3.1 Fusogenic Proteins 92
6.3.2 Preparation of Syncytial Fusion in Villous Trophoblast 93
6.3.2.1 Growth Factors and Cytokines 93
6.3.2.2 Protein Kinases 96
6.3.2.3 Transcription Factor GCM1 96
6.3.2.4 Externalization of Phosphatidylserine 96
6.4 Caspases and Their Roles in Syncytial Fusion 98
6.4.1 Localization of Caspase 8 98
6.4.2 Contributions of Caspase 8 to Trophoblast Fusion 99
6.4.2.1 Caspase 8 and Externalization of Phosphatidylserine 99
6.4.2.2 Caspase 8 and Cytoskeletal Remodeling 99
6.4.3 Caspase 8 Regulation During and After Fusion 100
6.5 Location Where Fusion Is Initiated in the Trophoblast 101
6.5.1 Syncytiotrophoblast 101
6.5.2 Cytotrophoblast 101
6.6 Concluding Remarks 101
References 102
7 Macrophage Fusion and Multinucleated Giant Cells of Inflammation 105
7.1 Introduction 105
7.2 Morphological Types of Multinucleated Giant Cells 106
7.3 Differential Signaling Pathways for Macrophage Multinucleation 107
7.3.1 Interleukin-4 and Interleukin-13 107
7.3.2 Interferon-. 109
7.3.3 a-Tocopherol 109
7.4 Mechanisms of Adhesion that Support FBCG Formation 109
7.4.1 Adhesion Receptors (Integrins) 110
7.4.2 RGD and Vitronectin 110
7.4.3 Adhesion Kinases 111
7.4.4 Microfilaments and Microtubules 111
7.4.5 Podosomes 111
7.4.6 Adhesion Failure (Anoikis) 112
7.5 Molecular Mechanism of Macrophage Fusion 112
7.5.1 A Role for Mannose Receptors (MR) 112
7.5.2 A Phagocytic Mechanism for Fusion with Participation of the Endoplasmic Reticulum 113
7.5.3 Diacyl Glycerol (DG)-Dependent and -Independent PKCs 113
7.5.4 Diacylglycerol Kinase 113
7.5.5 Matrix Metalloproteinase (MMP)-13 114
7.6 Phenotypic and Functional Profiles of Multinucleated Giant Cells 114
7.6.1 FBGC Versus Osteoclasts 115
7.7 Summary 116
References 116
8 Molecular Mechanisms of Myoblast Fusion Across Species 120
8.1 Introduction 120
8.2 Current Model Systems for Studying Myoblast Fusion 120
8.3 General Aspects of Myoblast Fusion in Drosophila melanogaster 121
8.4 Molecules Regulating Myoblast Fusion in Drosophila melanogaster 122
8.4.1 Recognition and Adhesion 123
8.4.2 Cytoplasmic Adaptors and Actin Cytoskeletal Regulators 125
8.4.2.1 Signaling Pathways Activating DRac1 125
8.4.2.2 Signaling Pathways Directly Upstream of Arp2/3 126
8.5 General Aspects of Myoblast Fusion in Danio rerio 126
8.6 Molecules Regulating Myoblast Fusion in Danio rerio 127
8.6.1 Recognition and Adhesion 127
8.6.2 Cytoplasmic Adaptors and Actin Cytoskeletal Regulators 127
8.7 General Aspects of Myoblast Fusion in Mus musculus 128
8.8 Molecules Regulating Specific Processes During Myoblast Fusion in Mus musculus 129
8.8.1 Elongation and Membrane Alterations 129
8.8.2 Migration 129
8.8.3 Recognition and Adhesion 132
8.8.4 Actin Cytoskeletal Dynamics and Integrin Signaling 132
8.8.5 Cell Fusion with Nascent Myotubes 133
8.9 Other Molecules Regulating Myoblast Fusion in Mus musculus 133
8.10 Integrated Pathways Regulating Myoblast Fusion in Mus musculus 134
8.10.1 Molecules that Enhance Follistatin Expression 134
8.10.2 Molecules Upstream and Downstream of NFATc2 134
8.10.3 Upstream and Downstream Molecules Regulating M-Cadherin Signaling 136
8.11 Conclusions and Future Directions 137
References 137
9 Cell-Fusion-Mediated Reprogramming: Pluripotency or Transdifferentiation Implications for Regenerative Medicine 143
9.1 Cell-Cell Fusion Methodologies 143
9.1.1 PEG-Mediated Fusion 144
9.1.2 Electrofusion 144
9.2 Somatic Cell Reprogramming 144
9.2.1 Somatic Cell Nuclear Transfer 145
9.2.2 Nuclear Reprogramming of Somatic Cells by Cell--Cell Fusion 146
9.2.3 Direct Reprogramming of Somatic Cells 146
9.3 Induced Pluripotency Through Cell-Fusion-Mediated Reprogramming 147
9.3.1 Fusion of Somatic Cells with ECCs 147
9.3.2 Fusion of Somatic Cells with ESCs 149
9.3.3 Fusion of Somatic Cells with EGCs 150
9.3.4 Mechanisms Controlling Somatic Cell Reprogramming 150
9.3.4.1 Nanog 151
9.3.4.2 The Wnt/ß-Catenin Pathway 151
9.3.4.3 AKT Signaling 151
9.3.4.4 Sall4 152
9.3.4.5 Epigenetic Modulation: Roles of PRC2, AID and G9a 152
9.4 Lineage Reprogramming by Cell Fusion 154
9.4.1 Synkaryon Versus Heterokaryon Strategies to Investigate the Mechanisms of Cell-Fusion-Mediated Lineage Reprogramming 155
9.4.2 The Importance of Gene Dosage in the Direction of Lineage Reprogramming by Cell Fusion 156
9.4.3 Global Chromatin Changes in Cell-Fusion-Induced Lineage Reprogramming 156
9.5 Cell-Fusion-Mediated Reprogramming as a Regeneration Mechanism 157
9.5.1 Transdifferentiation Versus Cell--Cell Fusion Theories to Determine Cellular Plasticity 158
9.6 Concluding Remarks 160
References 160
10 Cell Fusion and Tissue Regeneration 166
10.1 Introduction 166
10.2 Cell Fusion as a Cell Plasticity Mechanism 166
10.3 Regenerative Potential of Cell Fusion 169
10.3.1 Liver Regeneration by Cell Fusion 169
10.3.2 Skeletal Muscle and Cell Fusion 170
10.3.3 Cell Fusion After Heart Infarct 171
10.3.4 From Blood to Brain 171
10.3.5 Breathing Cell Fusion 173
10.3.6 Other Tissues 173
10.4 The Challenges of Cell Fusion to Become a Therapy 173
10.4.1 Fusion Induction 174
10.4.2 Fusion Partners and Delivery 175
10.4.3 Safety 175
10.5 Conclusions 176
References 176
11 Dendritic Cell-Tumor Cell Fusion Vaccines 181
11.1 Introduction 181
11.2 Dendritic Cell Based Immunotherapy 181
11.3 Principle of Dendritic Cell-Tumor Fusion Hybrids 182
11.4 Generating DC-Tumor Hybrids 183
11.5 Verification of True DC-Tumor Fusion 183
11.6 Pre-clinical Studies 184
11.7 Additional 3rd Signal with DC-Tumor Hybrids 184
11.8 Allogeneic Fusion Partners 185
11.8.1 Allogeneic Tumor 185
11.8.2 Allogeneic DC 185
11.9 Delivery Method 186
11.10 Clinical Studies 186
11.11 Future Challenges and Directions 187
References 187
Index 191