Gene Therapy - technologies, markets and companies |
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0. Executive Summary ............... 18 1. Introduction . 20 Definitions ............. 20 Historical evolution of gene therapy .................. 20 Relation of gene therapy to other biotechnologies................ 22 Molecular biological basics for gene therapy...... 22 Genome..............22 DNA .23 RNA.23 Alternative RNA splicing .......24 Genes.................25 Gene regulation ....25 Gene expression ...27 Chromosomes.......27 Telomeres ............28 Mitochondrial DNA.28 Proteins ...............29 2. Gene Therapy Technologies ... 30 Classification of gene therapy techniques.......... 30 Ex vivo and in vivo gene therapy..... 31 Ex vivo gene therapy................31 In vivo gene therapy ................32 Physical methods of gene transfer.. 32 Electroporation .....32 Applications of electroporation ...33 Clinical applications of electroporation ............34 Advantages of electroporation.....34 Limitations of electroporation35 Hydrodynamic ......35 Microinjection .......35 Particle bombardment...............36 Ultrasound-mediated transfection.....38 Molecular vibration 39 Application of pulsed magnetic field and superparamagnetic nanoparticles ...........39 Gene transfection using laser irradiation ..............39 Photochemical transfection........40 Chemical methods of gene transfer. 40 Gene repair and replacement.... 40 Gene repair by single-stranded oligonucleotides....41 History and current status of chimeraplasty..........41 Spliceosome-mediated RNA trans-splicing......... 41 mRNA gene therapy .................. 42 Vectors for gene therapy........... 42 Basic considerations .................42 Use of genes as pharmaceuticals43 The ideal vector for gene therapy.....43 Viral vectors .........44 Retroviral vectors................44 Oncognic potential of retroviral vectors ...........46 Adenovirus vectors..............47 Adeno-associated virus vectors ...49 Herpes simplex virus vectors 51 Lentiviral vectors.................53 Baculovirus vectors .............54 Alphavirus vectors...............54 Multicistronic retroviral vectors ...55 Companies using viral vectors.....55 Non-viral vectors for gene therapy ...57 Liposomes for gene therapy..58 Cationic lipid-DNA complexes59 Anionic lipid-DNA complexes.60 HVJ-liposome method ..........60 Polycation-DNA complexes (polyplexes) ..........61 Synthetic peptide complexes 61 Polymer molecules ..............62 - 4 - Nanobiotechnology for gene transfer 62 Nanoparticles as non-viral vectors for gene therapy.............62 Dendrimers .....63 Cochleates ......63 Calcium phosphate nanoparticles as non-viral vectors..........64 Lipid nanoparticles for nucleic acid delivery .....64 Silica nanoparticles as a nonviral vector for gene delivery ....65 Gelatin nanoparticles for gene delivery ...........65 Nonionic polymeric micelles for oral gene delivery ...............65 Biological nanoparticle technology ..................66 Receptor-mediated endocytosis .66 Artificial viral vectors ................67 Directed evolution of AAV to create efficient gene delivery vectors ...68 Bacterial ghosts as DNA delivery systems.............68 Bacteria plus nanoparticles for gene delivery into cells..............68 Chromosome-based vectors for gene therapy .......69 Companies using non-viral vectors ...71 Concluding remarks about vectors....72 Cell-mediated gene therapy ...... 73 Fibroblasts ...........74 Skeletal muscle cells.................74 Vascular smooth muscle cells ....75 Keratinocytes .......75 Hepatocytes .........75 Lymphocytes ........76 Regulating protein delivery by genetically encoded lymphocytes .76 Implantation of microencapulated genetically modified cells ......76 Stem cell gene therapy .............77 Therapeutic applications for hematopoietic stem cell gene transfer .................77 Improving delivery of genes to stem cells........78 Lentiviral vectors for gene transfer to marrow stem cells......78 Use of mesenchymal stem cells for gene therapy ................78 In utero gene therapy using stem cells ...........78 Gene delivery to stem cells by artificial chromosome expression .79 Linker based sperm-mediated gene transfer technology.......79 Combination of gene therapy with therapeutic cloning .........79 Expansion of transduced HSCs in vivo.............80 The future of hematopoietic stem cell gene therapy.............80 Routes of administration for gene therapy......... 80 Direct injection of naked DNA ....81 Intramuscular injection .............81 Intravenous DNA injection.........81 Intraarterial delivery.................81 Companies with gene delivery devices/ technologies ................82 Targeted gene therapy.............. 83 Targeted integration .................83 Bacteriophage integrase system for site-specific gene delivery...84 Controlled-release delivery of DNA ...84 Controlled gene therapy............ 85 Controlled delivery of genetic material .................85 Controlled induction of gene expression ...............85 Drug-inducible systems for control of gene expression..............86 Timed activation of gene therapy by a circuit based on signaling network............86 Small molecules for post-transcriptional regulation of gene expression ................86 Light Activated Gene Therapy ....87 Engineered zinc finger DNA binding proteins for gene correction 87 Companies with regulated /targeted gene therapy.87 Gene marking......... 89 Germline gene therapy.............. 89 Potential applications of human germline genome modification ..89 Pros and cons of human germline genome modification ............90 Role of gene transfer in antibody therapy.......... 91 Genetically engineered vaccines ..... 91 DNA vaccines........91 DNA inoculation technology ..92 Methods for enhancing the potency of DNA vaccines............92 Advantages of DNA vaccines.93 Vaccine vectors.....93 Challenges and limitations of genetically engineered vaccines....94 Vaccines based on reverse genetics..95 Technologies for gene suppression . 95 - 5 - Antisense oligonucleotides.........95 Transcription factor decoys........96 Aptamers .............96 Ribozymes ...........97 Peptide nucleic acid ..................97 Intracellular delivery of PNAs 97 Locked nucleic acid98 Zorro-LNA .......98 Gene silencing ......98 Post-transcriptional gene silencing ...98 Definitions and terminology of RNAi..99 RNAi mechanisms .99 Inhibition of gene expression by antigene RNA.... 100 RNAi gene therapy ................. 100 Application of molecular diagnostic methods in gene therapy .. 101 Use of PCR to study biodistribution of gene therapy vector ...... 101 PCR for verification of the transcription of DNA ... 101 In situ PCR for direct quantification of gene transfer into cells.. 101 Detection of retroviruses by reverse transcriptase (RT)-PCR.... 102 Confirmation of viral vector integration .............. 102 Monitoring of gene expression . 102 Monitoring of gene expression by green fluorescent protein 102 Monitoring in vivo gene expression by molecular imaging... 103 Advantages of gene therapy compared with protein therapy ..... 103 3. Clinical Applications of Gene Therapy ..... 105 Introduction........ 105 Bone and joint disorders ......... 105 Bone fractures .... 105 Gene therapy for intervertebral disc degeneration .................. 106 Spinal fusion....... 106 Osteogenesis imperfecta ......... 107 Rheumatoid arthritis............... 107 Local or systemic treatment 108 In vivo or ex vivo gene therapy of RA ........... 108 Clinical trials.. 109 Gene therapy for osteoarthritis 110 Sports injuries .... 111 Repair of articular cartilage defects............... 111 Regeneration and replacement of bone by gene therapy ......... 112 Dentistry............. 113 Tissue engineering in dental implant defects....... 113 Endocrine disorders ................ 113 Introduction ....... 113 Diabetes mellitus 113 Methods of gene therapy of diabetes mellitus ..... 114 Viral vector-mediated gene transfer in diabetes ................ 115 Gene delivery with ultrasonic microbubble destruction technology ................ 115 Genetically engineered cells for diabetes mellitus .............. 116 Genetically altered liver cells .... 116 Genetically modified stem cells . 116 Genetically engineered dendritic cells ........... 117 Insertion of gene encoding for IL-4............... 117 Concluding remarks about cell and gene therapy of diabetes .... 117 Gene therapy of growth-hormone deficiency....... 118 Gene therapy of obesity.......... 119 Ad viral vector-mediated transfer of leptin gene................ 119 AAV vector-mediated delivery of GDNF for obesity ............ 119 Gastrointestinal disorders....... 120 Introduction ....... 120 Methods of gene transfer to the gastrointestinal tract ............. 120 Direct delivery of genes ..... 120 Naked plasmid DNA into the submucosa........ 120 Viral vectors .. 121 Receptor-mediated endocytosis. 121 Indications for gastrointestinal gene therapy ...... 121 Gene therapy for inflammatory disorders of the bowel ....... 122 Gene transfer to the salivary glands .................. 122 Potential clinical applications of salivary gene therapy........ 123 Hematology.......... 123 Hemophilias ....... 124 - 6 - Gene therapy of hemophilia 124 Hemophilia A . 125 Hemophilia B . 125 Concluding remarks about gene therapy of hemophilias ..... 126 Hemoglobinopathies ............... 126 Gene therapy for b-thalassemia 128Gene therapy of Fanconi's anemia ................ 129 Acquired hematopoietic disorders... 129 Chronic acquired anemias .. 129 Neutropenia .. 130 Thrombocytopenia............. 131 Concluding remarks about gene therapy of hemoglobinopathies..... 131 Companies involved in gene thery of hematological disorders .. 132 In utero gene therapy............. 132 Fetal gene transfer techniques . 132 Animal models of fetal gene therapy.................. 133 Potential applications of fetal gene therapy......... 134 Fetal gene therapy for cystic fibrosis............. 134 Fetal intestinal gene therapy..... 134 Hearing disorders 135 Potential of gene therapy ........ 135 Vectors for gene therapy of hearing disorders..... 135 Auditory hair cell replacement and hearing improvement by gene therapy ......... 136 Kidney diseases ... 136 End-stage renal disease .......... 136 Methods of gene delivery to the kidney .............. 137 Gene transfer into kidney by adenoviral vectors................ 138 Non-viral gene transfer to the kidneys .......... 138 Gene transfer into the glomerulus by HVJ-liposome ........... 138 Bone marrow stem cells for renal disease...... 138 Mesangial cell therapy ....... 138 Liposome-mediated gene transfer into the tubules ............ 139 Gene transfer to tubules with cationic polymer polyethylenimine.................. 139 Gene therapy in animal experimental models of renal disease.. 140 Genetic manipulations of the embryonic kidney... 140 Antisense intervention in glomerulonephritis....... 140 Gene therapy for renal fibrosis. 141 Use of genetically engineered cells for uremia due to renal failure .. 141 Concluding remarks................ 141 Liver disorders ..... 142 Techniques of gene delivery to liver 142 Direct injection of DNA into liver .................. 143 Local gene delivery by isolated organ perfusion................. 143 Liposome-mediated direct gene transfer ....... 143 Retroviral vector for gene transfer to liver ..... 143 Adenoviral vectors for gene transfer to liver .. 144 Receptor-mediated approach .... 144 Cell therapy for liver disorders . 144 Transplantation of genetically modified hepatocytes .......... 145 Genetically modified hematopoietic stem cells 145 Gene therapy by ex vivo transduced liver progenitor cells .. 145 Gene therapy of genetic diseases affecting the liver................ 145 Crigler-Najjar syndrome..... 146 Hereditary tyrosinemia type I (HT1) ............. 146 Hereditary tyrosinemia type 3... 146 Gene therapy of acquired diseases affecting the liver.............. 146 Cirrhosis of liver................ 147 Ophthalmic disorders.............. 147 Introduction to gene therapy of ophthalmic disorders ............. 147 Degenerative retinal disorders . 148 Inherited retinal degenerations . 148 Leber's congenital amaurosis .... 148 X-linked juvenile retinoschisis ... 149 Retinitis pigmentosa .......... 149 Age-related macular degeneration................ 150 Proliferative retinopathies........ 151 Methods of gene transfer to retinal cells............. 151 DNA nanoparticles for nonviral gene transfer to the eye ..... 153 Prevention of complications associated with eye surgery ......... 153 Prevention of proliferative retinopathy by gene therapy ..... 153 DNA nanoparticles for gene therapy of retinal degenerative disorders ........... 153 - 7 - Posterior capsule opacification after cataract surgery......... 153 Autoimmune uveitis................ 154 Retinal ischemic injury............ 154 Corneal disorders 155 Glaucoma........... 155 Disorders of hearing ............... 156 Gene therapy for hearing loss.. 156 Organ transplantation............. 156 Introduction ....... 156 Gene therapy for prolonging allograft survival..... 157 Gene therapy in lung transplantation ................. 157 Role of gene therapy in liver transplantation....... 157 Gene therapy in kidney transplantation.............. 158 Pulmonary disorders............... 158 Techniques of gene delivery to the lungs............ 158 Adenoviral vectors............. 159 Non-viral vectors............... 160 Aerosolization as an aid to gene transfer to lungs.............. 160 Cystic fibrosis ..... 160 Genetics and clinical features.... 160 Gene therapy for CF .......... 161 CFTR gene transfer in CF.... 161 Concluding remarks about gene therapy of CF .................. 162 Miscellaneous pulmonary disorders. 163 Gene therapy for pulmonary arterial hypertension............. 163 Gene therapy for bleomycin-induced pulmonary fibrosis..... 164 Pulmonary complications of a1-antitrypsin deficiency......... 164 Gene therapy for asthma ... 165 Gene therapy for adult respiratory distress syndrome ........ 165 Gene therapy for lung injury..... 166 Gene therapy for bronchopulmonary |
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