Authors: Michelle McCulley, Pamela Greenwell
ISBN-13: 9780470019160, ISBN-10: 0470019166
Format: Hardcover
Publisher: Wiley, John & Sons, Incorporated
Date Published: January 2008
Edition: (Non-applicable)
Pamela Greenwell is Principal Lecturer at The University of Westminster. She leads a research team in glycobiology, molecular biology and bioinformatics and is actively involved with enabling research in clinical trials between academics, industry and Primary Care Trusts.
Michelle McCulley has a background in human genetics and experience teaching a broad range of students and health professionals, she is currently a Senior Teaching fellow at the Peninsula Medical School.
Book Synopsis
Molecular therapeutics incorporates genetics, recombinant DNA technology, biochemistry, protein production and purification, microbiology, molecular biology, immunology, pathobiology, and biotechnology. It addresses the treatment of human beings with "new drugs" and poses a range of ethical issues, particularly with respect to clinical trials, animal models, financial considerations and availability of treatment.
Recent advances in molecular therapeutics means that many of these cutting edge methods will soon be ready to be considered for clinical trial. It is essential that scientists, doctors, nurses and allied health professionals involved in these novel therapies understand how these techniques have evolved, what their benefits and drawbacks might be, the ethical issues associated with them and what their potential is for the future.
Targeted at postgraduate and undergraduate biomedical, allied health, medical and life science students, 'Molecular Therapeutics' provides an invaluable introduction to this rapidly evolving field.
Table of Contents
Prologue xi
Introduction 1
Microbial diseases 2
Cancer and heart disease 3
Cancer 4
Heart disease 5
Genetic diseases 5
Dominant diseases 6
Recessive diseases 6
Role of molecular biology in therapeutics 7
Prenatal diagnosis and pre-implantation 11
Should we treat inherited diseases? 11
Genetic screening 12
Pre-implementation genetic diagnosis 13
Counselling 15
Simple protein replacement therapy 17
Preventing transfusion-transmissible infectious diseases in the UK 18
Ensuring the safety of organ transplants 19
Preventing transfusion-transmissible infectious diseases worldwide 20
HIV 20
Recombinant protein production 23
Choice of organism 23
Somatostatin: an example of protein produced in E. coli 27
Insulin: an example of a recombinant protein 27
Alternatives to E. coli for the production of recombinant proteins 29
Insect cells 29
Whole insects 30
Mammalian cells 30
Plants 30
Transgenic animals 31
Problems with recombinant protein production 31
Problems with glycosylation 32
Effect of glycosylation 33
Erythropoietin: an example of protein produced in mammalian cells 35
Production method 36
Preparation of Factor VIII 36
Transgenic pigs and Factor VIII 37
All recombinants must be tested before they are given to humans 38
Why make recombinant proteins? 39
Recombinant products 40
Generics 40
Recombinant vaccines 43
Vaccine history 43
Vaccines 45
Vaccine methods 46
Types of vaccine 47
The limitations of vaccine programmes 48
The role of the WHO 50
Problems specific to developing countries 51
Economics and logistics of vaccinology 52
Recombinant vaccines 54
Simple recombinant protein vaccines 55
Gene vaccines: the vaccinia virus approach 57
DNA vaccines 57
Edible vaccines from transgenic plants 58
Rational design: bioinformatics and proteomics 59
Other interesting area for vaccine development 60
Conclusion 60
Therapeutic antibodies and immunotherapy 63
Monoclonal antibodies 63
Monoclonal production 64
Therapeutic monoclonal antibodies 66
Human monoclonals 66
Humanised antibodies 67
Transgenic monoclonals 69
The uses of monoclonal antibodies in therapy 69
Specific examples of therapeutic strategies 70
Unconjugated antibodies - treatment of kidney rejection 70
HIV treatment 71
Cancer therapy and monoclonal antibodies 71
Herceptin in breast cancer therapy 71
Treatment of multi-drug-resistant cancer cells 72
Anti-endotoxin antibodies 73
Conjugated antibodies 73
Delivery of radionuclides to leukaemia and lymphoma patients 74
Drug delivery 74
Toxin delivery 75
Bispecific antibodies 75
Other recombinant proteins used in immunotherapy 76
Cytokines 76
Colony-stimulating factors and growth factors 79
Transgenic animals 83
Why do we want to engineer the genomes of animals? 83
Experimental procedure 85
Method 1: Germline manipulation 85
Method 2: Using embryonic stem cells 87
DNA constructs, insertional mutagenesis and homologous recombination 90
Uses of inducible and tissue-specific promoters 91
Introduction of the DNA into the cells 92
Uses of transgenics 93
Recombinant protein production 93
Animal models of human diseases 94
Transplantation: a form of gene therapy 99
Introduction 99
Bone marrow 100
Logistical problems with BMT 102
Solid organ transplantation 102
Heart transplantation 103
Lung 104
Kidney 105
Liver 105
Ovarian tissue 106
Other cells and tissues 106
Summary of the problems associated with transplantation 107
Transplantation statistics 108
Legislation 108
Religious beliefs and transplantation 110
Xenotransplantation 113
Introduction 113
Rationale for the use of non-human donors 114
Organs from non-human primates 114
Pigs 115
Can we pretreat the recipient to prevent rejection? 116
Problems with pigs 117
Will pig hearts function in humans? 117
Xenozoonoses 117
Religious objections 118
Animal activists 119
Government legislation 119
When will xenotransplantation start? 120
Patient attitudes 120
Ethics 121
Alternatives to xenotransplants 121
Reproductive cloning 125
History 125
Problems 127
Why was there so much interest in Dolly? 128
Was Dolly a lone example? 129
Why is cloning useful? 129
Is human cloning a reality? 130
Why can we not produce human clones that are identical? 131
So why clone humans? 132
What are the ethical and moral problems? 132
Stem cell therapy 137
The potency of cells 137
Cloning 137
Potency of stem cells 138
Potential sources of stem cells 138
Stem cells and therapeutic cloning 138
Legislation and therapeutic cloning 140
Other sources of stem cells 142
What can be done? 142
Experiments on embryonic cells 143
Experiments on fetal tissue and cord blood 143
Stem cells from adult tissues 143
Safety and technical problems 144
Perceived scope of therapy 145
Clinical trials of stem cell therapy 145
What are the future prospects for stem cell research? 146
Gene augmentation therapy 149
Introduction 149
Strategy 150
DNA delivery 151
Viral vectors and gene therapy 152
Artificial viruses 155
Non-viral delivery in gene therapy 155
What tissues can we currently target? 157
Targeting gene expression 159
Problems associated with augmentation therapy 160
Gene augmentation therapy vs. recombinant DNA therapy or transplantation 161
Current criteria for the use of gene therapy 162
The bystander effect 163
Candidates for gene therapy 163
Gene therapy trials for inherited diseases 165
Introduction 165
Examples of disease treated with retroviral gene therapy 166
Severe combined immunodeficiency 166
Hypercholesterolaemia 169
Cystic fibrosis 170
Rationale for adenoviral vectors 171
Early animal trials 171
Is CF gene therapy 'safe but not useful'? 172
Problems also found in in vivo delivery 172
Animal trials with Factor IX 173
Adenoviruses have also been used to introduce genes into brain 174
Duchenne's muscular dystrophy 175
Problems with adenoviruses 175
The uses of adeno-associated viruses 176
Haemophilia B treatment with Factor IX gene augmentation 176
AAV therapy for DMD 177
Liposome vector trials 178
Trials with polymer mareix delivery 178
Gene silencing technologies 181
Antisense therapy 181
Modification of antisense molecules 183
Replacement of oxygens in the phosphate bridge 184
Modifications can be made to the bases themselves 184
Other types of modifications 185
The ideal oligonucleotide 185
Uptake 185
Uses of antisense oligonucleotides 187
Examples 187
Catalytic antisense molecules 191
Triple helix (triplex) technology 191
Problems 193
Advantages over antisense strategies 193
Experimental data 194
Ribozymes 196
Examples of ribozyme therapies 197
Small interfering RNAs (siRNAs) 198
Clinical trials and siRNA 202
Is RNAi better than antisense? 202
Gene therapy for cancer 205
What causes cancer? 205
Cancer: a multifactorial disease 206
Cancer statistics 207
Best treatment currently available 208
Avoidance 208
Screening 208
Surgery 209
Chemotherapy 209
Radiotherapy 210
Do chemo- and radiotherapy cause problems? 210
New cancer therapies 210
Cancer models in animals 211
What kinds of gene therapy can we use to treat cancer? 212
Perceived problems in cancer gene augmentation therapy 213
Killing cells with ganciclovir or suicide therapy 213
Prodrug activation therapy 214
Enhancing the immune system with gene therapy 215
Gene silencing technologies and cancer 217
Conclusion 219
Single-nucleotide polymorphisms (SNPs) and therapy 223
Legislation, clinical trials and ethical issues 231
Legislative bodies 231
Clinical trials 233
The problems of placebo controlled trials 236
The need for informed consent 238
Trials in developing countries 239
Recent trial issues 241
Conclusion 242
Epilogue 245
Sourcing references 245
Index 247
Subjects