Gene Therapy. The use of DNA as a drug. Edited by Gavin Brooks. BPharm, PhD, MRPharmS (PP) Pharmaceutical Press
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1 Gene Therapy The use of DNA as a drug Edited by Gavin Brooks BPharm, PhD, MRPharmS (PP) Pharmaceutical Press
2 Contents Preface xiii Acknowledgements xv About the editor xvi Contributors xvii An introduction to DNA and its use in gene therapy 1 DNA and RNA molecules 1 Chromosomes and chromatin 2 Base pairing 3 Transcription 4 Translation 6 Complementary DNA (cdna) 8 Regulation of gene expression 8 Analysis of DNA 9 Preparation and purification of DNA 10 Detection of specific DNA sequences 1 0 Analysis of RNA 11 Preparation and purification of mrna 1 1 Detection of specific RNA sequences 1 2 Analysis of proteins 1 3 Preparation and purification of proteins 1 3 Detection of specific proteins 15 Use of DNA as a drug - an introduction to gene therapy 17 Conclusions and summary 20 References 21 Further reading 21 Methods for delivering DNA to target tissues and cells 23 Viral vectors for DNA delivery 24 Retroviral delivery vectors 25 Adenoviral and adeno-associated viral (AAV) delivery vectors 29 Other viral methods for DNA delivery 34 Non-viral methods for DNA delivery 35 Naked DNA 36
3 viii Contents Cationic lipids and polymers 38 Other non-viral delivery methods 41 Targeting the delivery of therapeutic DNA 42 Summary and conclusions 43 References 44 The Human Genome Project 51 Human genome maps 52 The cytogenetic map 52 The genetic map 52 The transcript (radiation hybrid) map 55 The physical map 58 The human genome sequence 59 Identifying genes and their structures 63 Human genome sequence variation and SNPs 64 Conclusion 65 Note added in proof 65 Acknowledgements 66 References 66 Gene therapy for single gene disorders 71 Cystic fibrosis 71 Severe combined immunodeficiencies (SCIDs) 72 X-linked severe combined immunodeficiency 73 Adenosine deaminase and purine nucleoside phosphorylase deficiencies 73 Other autosomal recessive SCIDs 74 Somatic gene therapy for SCID 75 Treatment for ADA-SCID 75 Retroviral vectors for gene therapy of ADA-SCID 75 T lymphocyte gene therapy for ADA-SCID 77 Stem cell gene therapy for ADA-SCID 78 Limiting factors for stem cell gene therapy 79 Clinical gene therapy for X-SCID 79 Alternative vector systems 80 Lentiviral vectors 80 Adeno-associated viral vectors 8 1 Conclusions 82 References 82 Gene therapy approaches for cancer 87 Cancer gene therapy strategies 88 Immunotherapy 88 Introduction of tumour suppressor genes'and induction of apoptosis 94
4 Contents ix Virus-directed enzyme/prodrug therapy (VDEPT) 97 Inhibition of tumour angiogenesis Tumour targeting 114 Direct injection 1 14 Regional delivery 1 14 Tumour-specific vector binding Tumour- or tissue-specific promoters Tumour-specific virus replication 1 19 Conclusions 121 References 122 Gene therapy for the treatment of cardiovascular diseases 135 Gene therapy for the failing heart 136 Heart failure 1 36 Vectors for myocardial gene therapy 142 Molecular targets for myocardial gene therapy 143 Gene therapy for the vascular system 147 Coronary heart disease 149 Atherosclerosis 149 Vascular diseases associated with defects in lipid metabolism 151 Gene therapy for atherosclerosis 1 54 Ischaemic heart disease and angina 158 Classification of angina 158 Current medical treatment for angina 1 59 Angiogenesis 160 Surgical treatments for angina 164 Restenosis 164 In-stent stenosis 1 66 Gene therapy for restenosis and in-stent stenosis 167 Stroke: prevalence and mortality 170 Types of stroke 1 71 Diagnosis and current treatments for stroke 173 Gene therapy for stroke 175 Summary and conclusions 1 81 References 182 Gene therapy for AIDS and other infectious diseases 189 Gene therapy techniques 193 Gene therapy directed against the infecting organism 194 Gene therapy designed to manipulate the immune response 197 Gene therapy for HIV-1 infection 199 Antiviral gene therapy directed against HIV Immunomodulatory gene therapy directed against HIV-1 infection 202 Clinical trials for the study of gene therapy and HIV-1 infection 203
5 x Contents Gene therapy and other infectious diseases 204 Hepatitis viruses 204 Herpesviruses 205 Human papillomaviruses 205 Conclusions 206 Acknowledgement 206 References 206 Gene therapy approaches for rheumatoid arthritis 215 Clinical aspects of rheumatoid arthritis 218 Epidemiology and genetics of RA 218 The diagnosis of RA 219 The clinical course of RA 220 Pathophysiology and treatment of rheumatoid arthritis 221 The inflammatory process of RA 221 The limitations of current pharmacotherapy of RA 222 T cells, macrophages, interleukin 1 and tumour necrosis factor a as therapeutic targets in RA 225 Biological therapy of RA 228 Some important therapeutic target genes in rheumatoid arthritis 231 Inhibitors of proinflammatory cytokines 23 1 Anti-inflammatory cytokines 233 The transcription factor NF-KB as a therapeutic target 235 Gene delivery systems suitable for rheumatoid arthritis 237 Non-viral strategies 237 Viral strategies 239 Gene therapy in animal models of arthritis 243 Neutralisation of proinflammatory cytokines 243 Overexpression of anti-inflammatory cytokines 244 NF-KB as g,therapeutic target 246 Other approaches 247 What is the mechanism for the 'contralateral effect'? 248 Combination constructs and inducible promoters 249 Current clinical trials of gene therapy in rheumatoid arthritis 250 Discussion 251 References 253 The use of gene therapy in neurological diseases 267 The nervous system 267 Neurons, glia and myelin 268 Neuroimmunology 268 Activation of T cells 268 Cytokines 270 Mechanisms of damage and repair in the CNS 270
6 Contents xi Mechanisms of damage 270 Repair strategies 271 Gene transfer into the nervous system 271 Blood-brain barrier 271 Delivery systems 272 Gene therapy for neurological diseases 277 Multiple sclerosis 277 Guillain-Barre syndrome 281 Parkinson's disease 282 Motor neuron disease and amyotrophic lateral sclerosis 285 Huntington's chorea 286 Alzheimer's disease 287 Gene therapy in other neurological diseases 288 Epilepsy 288 Stroke/ischaemia 289 The future of gene therapy for neurological diseases 290 Acknowledgements 290 References Xenotransplantation: promise and problems 295 Meeting the demand for transplantation 296 The challenge to find suitable animal sources 297 Immunological aspects of xenotransplantation 299 Infection hazards of xenotransplantation 300 Porcine endogenous retroviruses (PERV) 302 Monitoring clinical infection by PERV 306 Transgenic pigs and human infection 307 Clinical applications of xenotransplantation 308 Conclusions and prospects 308 Acknowledgement 309 References 309 Glossary 313 Index 319
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