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W2314806967 abstract "Table of Contents 1. Introduction 2. Methods of Transfer 2.1. Naked DNA transfer 2.2. Viral gene transfer 2.2.1. Adenoviruses 2.2.2. Retroviruses 2.2.3. Lentiviruses 2.2.4. Adeno-associated viruses 2.2.5. Herpes simplex viruses 2.2.6. Other viruses 2.3. Nonviral gene transfer 2.3.1. Chemical and physical methods 2.3.2. Liposomal gene transfer 2.3.3. Protein/peptide gene transfer 3. Disease Targets for Gene Therapy 3.1. Genetically determined protein deficiencies 3.1.1. Gaucher disease 3.1.2. Adenosine deaminase deficiency 3.1.3. γc Cytokine receptor deficiency 3.1.4. Cystic fibrosis 3.2. Gene therapy for acquired diseases 3.2.1. Immune stimulation in the treatment of cancer 3.2.2. Activation of ganciclovir by herpes thymidine kinase 3.2.3. Tumor suppressor genes 3.2.4. Acquired immunodeficiency syndrome (AIDS) 3.2.5. Rheumatoid arthritis 3.2.6. Cardiovascular disease 4. Conclusions 1. Introduction The treatment of genetic diseases has seemed to many a daunting challenge. What, after all, can be done if the immutable basic blueprints of the body have a serious imperfection? In reality, even in the middle of the last century it was possible to improve greatly the quality of life and, indeed, to save the lives of patients with some such genetic diseases. The successful approaches included dietary manipulation as in phenylketonuria or galactosemia, surgery to correct various deformities, and sometimes avoidance of inciting factors in the environment, as in acute intermittent porphyria. However, until the latter part of the century just past, the possibility of actually changing the faulty genetic blueprints was beyond the imagination of the realistic medical scientist. In 1972 Friedmann and Roblin (68) argued persuasively that defective genes could be replaced by those with the correct sequence. In the following 30 years a great deal of effort has been expended to bring such a therapeutic endeavor to fruition, thus far with limited success. 2. Methods of Transfer Although it has been possible to achieve expression of genes delivered to the cytoplasm (28,37,100,119) the most common and useful strategy is to deliver the gene of interest to the nucleus. There are 4 extracellular barriers to such delivery: 1) opsonins, 2) phagocytes, 3) extracellular matrices, and 4) degradative enzymes (59). An opsonin is a factor that attaches to foreign material and renders it more susceptible to ingestion by phagocytes. Thus, opsonins can inactivate a gene delivery system by attachment, leading to the inactivation of the gene and/or its carrier. Phagocytes are cells that can inactivate a gene delivery system by engulfment or digestion. The extracellular matrix represents a physical barrier of polymerized protein and carbohydrates present between cells protecting target cells from relatively large DNA carrier systems. Finally, the extracellular fluid contains DNases that can rapidly digest unprotected DNA. Once all these barriers are overcome, the gene delivery system attaches to the plasma membrane where it faces cellular barriers. The first of these is the plasma membrane. Then the gene must be protected from nucleases in the cytoplasm and overcome the possibility of endosomal entrapment. The nucleic acid must enter the nucleus where the gene of interest can proceed to be transcribed and translated, and finally the protein traffics to the cellular location where it has a function. The ideal gene delivery vector is nontoxic, nonimmunogenic, easy to produce in large quantities, and efficient in protecting and delivering DNA into cells, preferably with a specificity for a particular cell type. This ideal vector remains to be discovered (112,185). Properties of existing transfection agents are compared in Table 1.TABLE 1: Properties of transfection agents*2.1. Naked DNA transfer The principal obstacle to cellular DNA uptake is charge (56). In an aqueous solution, such as the milieu that bathes cells in the body, DNA has a net negative charge. DNA tends to be repelled from cell membranes, because they, too, are negatively charged. There are a few exceptions where cells appear to be able to assimilate naked DNA; this includes the successful target protein expression after direct muscular injections in mice (22,38,56,181). While the mechanism of this type of gene transfer is unclear, a small amount of tissue damage or increased pressure at the injection site may play a role (56). A few other types of cells and tissues can be transfected by the direct injection of naked DNA (70); these include the thyroid gland (154), certain tumor types (172), liver cells (84), skin (82), and myocardial cells (153). Other types of cells are quite resistant to transfection unless a carrier is used. Naked DNA, larger in size than oligonucleotides, is not readily endocytosed and must therefore be packaged into a delivery vehicle, or vector (171), capable of efficient entry into cells (23,56). Naked DNA does not provoke specific immune responses; however, DNA containing dinucleotide sequences comprising cytosine followed by guanine (CpG sequences) are unmethylated when produced in bacteria and, when flanked by 2 purines on 1 side and 2 pyrimidines on the other, they provoke the innate immune response (82,104). Naked DNA can be delivered without the use of needles with a gene gun or the Jet gun (Intraject, Weston Medical, Cambridge, UK, http://www.weston-medical.com). The gene gun uses a high pressure helium stream to deliver DNA coated onto gold particles into the cytoplasm, while the Jet gun uses liquid under pressure for delivery into interstitial spaces (82). 2.2. Viral gene transfer Viruses have evolved specialized mechanisms for cellular binding and intracellular delivery, and capitalizing on these mechanisms has been a favored approach in attempting to achieve highly efficient transfer of genes to mammalian cells. Viral gene delivery has been analyzed using a number of vectors including adenoviral, retroviral, lentiviral, adenoassociated viral, and herpes simplex virus vectors. 2.2.1. Adenoviruses: Adenoviruses are highly effective vectors for transient gene transfer to different cell types (95). These are double-stranded DNA viruses whose capsid is composed of hexons and pentons, namely the penton base and fiber monomers (15). These viruses are relatively easy to produce in large quantities (67), and offer a large capacity for foreign genes, up to 38 kb (67,91). First-generation adenoviral vectors retain all essential adenoviral genes except E1A and E1B, the early functions required for the activation of most other viral genes during a productive infection (91). Second-generation adenoviral vectors are characterized by the additional inactivation of the E2 functions of DNA-binding protein or viral DNA polymerase, or by the inactivation of the E4 gene that has oncogenic potential and key regulatory functions (91). Vectors with deletions of both the E1 and E3 genes can accommodate transgenes of up to 8 kb in size; E3 is dispensable for virus propagation in cell culture (91). Adenoviral gene delivery leads to the episomal persistence of the target gene of interest within the viral vector, and it triggers a potent host immune response that makes this type of virus-mediated gene delivery unsuitable for repeated administration (67). The immune response to viral proteins expressed from adenovectors and transgene products leads to clearance of transfected cells (91). This immune response is humoral with neutralizing antibody to viral capsid proteins (7), and cellular with a cytotoxic T lymphocyte response against transfected cells (146). The host immune response consists of inflammation, production of neutralizing antibodies, and a virusspecific cytotoxic T lymphocyte response (67). The recent death of a volunteer in an adenoviral gene therapy experiment has prompted a review of the safety and efficacy of adenoviral vectors (110,111). The patient was a young man with an inherited defect of ammonia metabolism, namely ornithine-transcarbamylase (OTC) deficiency. He died in September 1999, 4 days after a genetically altered adenovirus had been infused into his liver (110,112). Over the past few years, several groups have created fully “gutless” or third-generation adenoviral vectors by removing all viral genes and replacing them with substitutes produced by helper viruses (91,112). In some reports virtually no toxicity was observed when such vectors were given to mice at high doses (112); nonhuman primate trials with gutless viruses have not yet been reported (112). In addition, encapsulated adenoviral minichromosomes with genomes of approximately 13 kb have been developed for gene transfer; as in the case of “gutless” adenoviral vectors, adenoviral minichromosomes require helper viruses and several serial passages for production (164). It is currently difficult to produce high-titer batches of gutless adenovirus and to eliminate contamination by live helper virus (91,112). There is also some suggestion that empty capsids of adenovirus are immunogenic; this possibility requires further exploration before future clinical trials (112). 2.2.2. Retroviruses: Retroviruses represent a group of viruses whose RNA genome is reverse transcribed into genomic DNA in the infected cell. The basic retroviral genome contains 3 genes known as gag, pol, and env; these genes are flanked by elements known as long terminal repeats (LTRs) (171). LTRs define the beginning and the end of the viral genome and are required for the integration of the host genome; they also serve as enhancer-promoter sequences. Viral LTRs can control the transcription of a transgene, or specific enhancer-promoter elements can be engineered in with the transgene. The retroviral genome also contains a packaging sequence, Ψ, that enables the viral RNA to be distinguished from other RNAs in the cell (171). Retroviruses offer the potential advantage of integrating genes into host chromosomes for long-term stability in dividing cells. One potential disadvantage to this form of gene transfer is the random retroviral insertion that occurs into the host genome, leading to the potential activation or inactivation of genes critical to the normal functioning of the host (67,117). In addition, human complement rapidly inactivates retroviruses; this has lead to the design of modified retroviral vectors that prevent complement activation and complement-mediated elimination (146). Retroviruses can recombine with cellular or viral DNA or RNA to produce new oncogenic viruses or replicationcompetent retroviruses (146). Indeed, the reverse transcriptase-polymerase chain reaction analyses of T-cell lymphomas that developed 6 months after the engraftment of autologous stem cells in 3 of 10 rhesus macaques during a retroviral gene transfer trial revealed the presence of several different recombinant murine leukemia viruses (MuLV) (142). Specifically, the env gene of the helper packaging virus recombined with the LTR of the Moloney MuLVderived vector to form a retrovirus that does not exist in nature (142). The retroviral vector used in this study has been replaced by a new generation of vectors possessing shorter regions of homology with the helper virus; regions of homology as short as 8–10 nucleotides have been associated with breakouts of replication-competent retroviruses (142). Another important problem that is usually encountered when retroviruses are used as vector is their inactivation by the host. In transgenic animals microinjected DNA constructs give rise to more predictable expression than infection with recombinant retroviruses; these findings suggest that host cells have the ability to recognize and inactivate retroviral DNA (17). Eukaryotic genomes are protected by elaborate defense systems that prevent the expression of alien or abnormal transcription units. The insertion of retroviral DNA into a mammalian host can trigger transcriptional silencing of the inserted sequences via mechanisms that usually involve DNA methylation within regulatory regions (16). To enhance the applicability of retroviral gene transfer, efforts have been made to devise strategies to modify the host range of the retroviral envelope glycoproteins to achieve retroviral vectors capable of delivering their genes to target human cells with high specificity (151), so-called pseudotyping of the vector. These strategies can be characterized based on the nature of the molecular target, the targeting element, and the host range modification. The molecular target is usually either a receptor or a protease. It can be approached using soluble adaptor molecules, for example, streptavidin, that have 1 binding site that interacts with the retrovirus and a second that interacts with the targeted cell (151). Alternatively, the retroviral envelope glycoproteins can tolerate a variety of genetically encoded modifications by mutations that enhance or hinder specific structures, for example, receptor binding domains, making them more susceptible to retroviral infection (151). Another approach is to block virus infectivity by grafting a protease-cleavable blocking domain onto the viral coat protein. For example, vectors containing a factor Xa-cleavable EGF domain poorly infected EGF receptor-positive target cells until they were treated with factor Xa protease (151). Additional advances in retroviral targeting should become possible with the further elucidation of crystal structures of retroviral envelope glycoproteins and the development of strategies to select optimally targeted vectors from vector display libraries (151). The transduction of hematopoietic stem cells is enhanced by the presence of fibronectin or of peptides derived from fibronectin, such as that designated CH-296 (76,183). This effect is thought to be due to colocalization of fibronectin on retrovirus on the cell surface. 2.2.3. Lentiviruses: Lentiviral vectors are human immunodeficiency virus (HIV)-based retroviral vectors that have the advantage of integrating into the genome of both proliferating and nonproliferating cells (126). They can be modified to widen their host cell range to infect most dormant cells, such as neurons (155). However, as the most commonly known lentiviral vectors are HIV derived, the safety of these vectors must be tested extensively. Before human studies can be undertaken, these vectors must be shown to be unable to recombine or interact with another virus to produce an active HIV strain. The packaging cells used to produce high-titer virus currently contain HIV proteins, which presumably will be eliminated in the near future (155). HIV contains the gag, pol, and env genes. In addition, it carries genes for 6 accessory proteins termed tat, rev, vpr, vpu, nef, and vif(3,171). These accessory proteins have the potential to cause cell damage or disease; in fact, nef can produce an acquired immunodeficiency syndrome (AIDS)-like disease if expressed in transgenic mice (77,155). The HIV env gene product restricts the infection of HIV-based vectors to CD4+ cells. This gene has been replaced with env sequences from other RNA viruses that have a broader infection spectrum; for example, glycoprotein from vesicular stomatitis virus (VSV-G) binds to many cells, including neurons (155,171). Self-inactivating (SIN) viruses have a deletion inactivating the U3 region, which is duplicated and flanks the virus on both sides; this region contains active promoter elements that can drive the synthesis of viral RNA and downstream cellular RNA (155). The injection of lentiviral vectors has been shown to result in sustained expression for over 6 months in rodent brain, liver, muscle, eye, and pancreatic islet cells, without the inactivation of expression as is seen in prototypical retroviral vectors (171). 2.2.4. Adeno-associated viruses: Adeno-associated viruses (AAV) are single-stranded DNA parvoviruses that integrate their genes into human chromosomes of dividing and nondividing cells (44,171). They are not known to cause disease in the human population. Nonetheless, 80% of adults have circulating antibodies to AAV (171). AAV have 2 genes, cap and rep, that are located between inverted terminal repeats defining the beginning and the end of the virus, and containing the packaging sequence (171). The AAV inverted terminal repeats mediate the chromosomal integration of the host genome (105). The cap gene encodes the viral capsid or coat proteins, while the rep gene product is involved in viral integration and regulation (171). In the presence of the rep gene product, AAV vectors integrate selectively into a region of chromosome 19, near a breakpoint commonly observed in chronic lymphocytic leukemias; this site specificity seems to be lost when the viral genome of vectors has been replaced with the gene to be transferred (44,171). Helper viruses provide the additional genes that AAV require to replicate. Their gene therapy applications are limited because they can transfer only small foreign genes (93), and AAV vector preparation is laborious due to the toxic nature of the rep gene product and some of the adenoviral helper proteins (171). Helper viruses are no longer necessary to produce AAV vectors; this function can be performed with plasmids that essentially eliminate contaminating wildtype virus (30). AAV have been used successfully for long-term correction of Hemophilia B in a dog model with portal vein (159) or muscular (83) injections of an AAV vector expressing clotting factor IX, the factor deficient in Hemophilia B (106). A phase 1 trial of AAV CFTR in sinusitis has also been described. The maxillary sinuses were used as a model of cystic fibrosis because they have ion transport systems and microbiology similar to those of the lower respiratory tract. The results of this randomized, nonblinded dose escalation protocol revealed that successful gene transfer in this setting is dose-dependent, with DNA transfer detected in 2 of 10 patients for at least 40 days (177). 2.2.5. Herpes simplex viruses: Herpes simplex virus type 1 (HSV-1) is another vector that has been studied for its capacity to transfer genes to mammalian cells. It is a relatively large virus that contains more than 80 genes (171). Approximately half of the genes of HSV encode accessory functions implicated in the host virus life cycle, genes that can be deleted without consequence to viral replication in cell culture (61). HSV infects the cells of the nervous system, specifically neurons of sensory ganglia where it can establish a lifelong presence as an extrachromosomal element in a quiescent phase referred to as latency (61). Reactivation from latency of unmodified HSV results in viral replication and clinical signs of infection (61). In the latency phase of HSV, even in the case of virus mutants defective for growth and reactivation, lytic genes are repressed and intranuclear RNA latency-associated transcripts are expressed (61). HSV replication-defective mutants are nonetheless toxic for primary neurons in culture, and many viral and nonviral vector promoters are rapidly silenced after injection into the brain (61). Some progress has been made in the development of defective viral mutants with reduced toxicity (61). The use of HSV as a vector for gene transfer is limited because many people have already been infected by this virus and develop immunity to components of HSV (171). Gutless vectors, those from which essentially all viral proteins are deleted, have relatively low yields of 106 virus particles per mL, whereas vectors containing a unique deletion of the IE3 gene produce 108-109 viral particles per mL (171). 2.2.6. Other viruses: The Semliki Forest and Sindbis viruses are alpha viruses that are under investigation as possible cytoplasmic vectors (87). The replication-incompetent vectors derived from these viruses deliver their RNA genomes to the cytoplasm of target cells where they potentially can produce large amounts of protein from high levels of vector-generated message encoding the gene of interest (87). They do not integrate into the genome, and thus produce foreign protein only transiently (171). Vaccinia virus-, or poxvirus-, based DNA vectors are under consideration primarily for generating vaccines (171). Their genome consists of 180 kb of linear double-stranded DNA connected at the ends (87). The viral system is attractive for use in gene therapy because of its extensive use as a smallpox vaccine, its large insert capacity, and the increasingly detailed understanding of its molecular virology (87). The virus core contains a DNA-dependent RNA polymerase, a transcription factor, capping and methylating enzymes, and a poly(A) polymerase, enabling the expression of genes within the cytoplasm of mammalian cells (121). Simian virus-40 (SV40) is a double-stranded DNA papova virus under investigation as a vector for gene transfer. It has a circular genome lacking the terminal repeats that characterize many other viral vectors, and it can transfect both quiescent and dividing cells (168). It has been tested in immunocompetent animals, and it does not elicit neutralizing antibody or cytotoxic immune responses against infected cells (168). Live, tagged, wildtype SV40 was an unrecognized contaminant of early batches of the Salk polio vaccine, with anecdotal reports of some human tumors carrying SV40 taglike sequences (168). Nonetheless, SV40 is oncogenic in sucking hamsters and it integrates randomly into cellular genomes (168). Consequently, its safety must be monitored carefully before it can be used as a vector for human gene transfer trials. 2.3. Nonviral gene transfer Nonviral gene transfer with synthetic vectors (79) provides an alternative method of efficient gene delivery intended to result in lower levels of toxicity. The goal of nonviral gene therapy is to mimic the successful viral mechanisms for overcoming cellular barriers that block efficient expression of the target gene while minimizing the toxicities associated with gene delivery. The capabilities of a synthetic nonviral vector could include specific binding to the cell surface, entry, endosomal escape, translocation to the nucleus, and, in some cases, stable integration into the target cell genome. The rate limiting step of current nonviral gene delivery techniques is the transfer of encapsulated plasmids from the endosomes to the nucleus (56). In this setting, plasmids are endocytosed by cells into the endosomal compartment. The acidity of this compartment, together with its nuclease activity, would be expected to degrade plasmids rapidly (56). Chloroquine is known to retard acidification of endosomes, and has been used in some gene therapy protocols to promote endosomal release of intact DNA (69). 2.3.1. Chemical and physical methods: Techniques have been developed to combine DNA with cationic polymers, such as DEAE-dextran (114,146,150), polybrene (127,146), and the mineral calcium phosphate (35,108), to neutralize DNA electrostatically and allow it to be more readily taken up by cells. Chemical techniques are considered too inefficient for clinical gene therapy. Physical methods such as microinjection (187) and electroporation (128,132) are also relatively inefficient means for gene delivery. Electroporation involves the application of an electric field that transiently breaks down the cell membrane, allowing DNA to enter the cytoplasm (82). More recently, nonlinear cationic polymer, for example, dendrimers, and the polycation polyethylenimine (PEI) have been synthesized possibly to overcome some of the barriers to efficient gene transfer of the earlier generation of cationic polymers (146). In particular, PEI has an intrinsic endosome buffering capacity that is thought to be a key element in its efficient gene transfer (104). In addition, transfection by immunoporation has recently been described. In this technique, Immunofect beads (Immunoporation, Colchester, UK) are coated with antibodies that bind to specific cell surface transmembrane proteins; they can create holes in the cells when the beads are dislodged by centrifugation, allowing transfection of cells with DNA or other macromolecules (18). The beads are removed using a magnetic separator, and the cells are returned to tissue culture medium for 48 hours. This method has the advantage of targeting specific cell types, and it allows targeted cells to take up a variety of different molecules. The inventors claim that this technique transfects 40%-80% of cells as determined by flow cytometry, and that it results in less than 20% nonviable cells (18). 2.3.2. Liposomal gene transfer: Liposomes are minute hollow spheres composed of a lipid membrane surrounding an aqueous sphere. The net charge of liposomes depends on the type and ratio of the constituent lipids. Cationic liposomes form complexes with DNA called lipoplexes (57), at least in part via an electrostatic interaction with the negatively charge phosphate backbone of DNA (39,57,166). The hydrophobic cations in liposomes can condense DNA (59). Liposomal gene transfer has several advantages including lack of immunogenicity, ease of preparation, and the ability to package large DNA molecules (58,166). While this method of gene transfer has potentially useful applications, the ratio of liposome to DNA must be carefully controlled to circumvent the development of toxic aggregates (58,166); the mechanism of liposome-based nuclear DNA delivery and DNA release remains to be fully elucidated (39,55,98,166,184,189). In addition, liposomes have a limited efficiency of delivery and gene expression, and they have potentially adverse interactions with negatively charged macromolecules. Immunoliposomes target specific cells using ligands such as monoclonal antibodies attached to the surface of lipid vesicles; they can be prepared as pHsensitive and non-pH-sensitive liposomes (146). Proteoliposomes integrate reconstituted viral envelope glycoproteins into the lipid bilayer to mediate the cellular fusion and entry of liposomes at the cell surface or within the endosomal compartment (146). 2.3.3. Protein/peptide gene transfer: Complex formation with DNA in protein and peptide gene transfer, that is, polyplex formation (57), is mediated through electrostatic interactions between the positively charged lysine and arginine residues and the negatively charged phosphates in the DNA backbone (166). The hydrophilic polycations of cationic polymers and polypeptides have been utilized to condense DNA through ionic interactions and to facilitate in vitro cellular uptake of plasmids through nonspecific adsorptive mechanisms (146). Complexes of DNA and polycationic polymers are designated polyplexes, while DNA complexes with mixtures of lipids and polymers are called lipopolyplexes (59). DNA binding elements that are effective in gene transfer include protamines (103,161); histones H1, H2A, H3, and H4 (8,9,11,36,39,42,69,73,75,94,157,188); the nonhistone nuclear protein HMG1 (188); poly-L- lysine (175); and cationic amphiphilic-helical oligopeptides with repeated sequences (129). Polylysine chain length appears to be critical to particle size and in vitro gene transfer efficiency. Peptides with 8 or fewer lysine residues, as well as those with 19 or more lysine residues, bind DNA weakly using a dye exclusion assay and produce large particles as determined by light scattering at 90, that is, 0.7–3 m; peptides with lysine repeats of 13–18 bind DNA tightly and produced condensates that decreased in mean diameter from 231 to 53 nm as the lysine chain increased (173). The addition of an alkylated cysteine, as a potential ligand attachment site, followed by a tryptophan to the N-terminus of polylysine18 resulted in a 40-fold reduction in particle size and 1000-fold amplification of transfection efficiency as determined by luciferase production in Hep G2 cells as compared with polylysine19(173). There is active, ongoing research to understand and control DNA condensation and packaging with these agents, as well as the determination of the structure of such complexes (59). Once DNA has been condensed successfully, the efficiency of gene delivery can be enhanced by the introduction of ligands into the complexes. These ligands may target specific cells via a cell surface receptor. Receptor-mediated gene delivery constructs contain a receptor-binding ligand and a DNA-binding moiety, usually poly-L-lysine (98). Cells have been targeted using a number of different ligands including transferrin (156,176,182), asialoglycoprotein (174), immunoglobulins (60), insulin (149), EGF (64), and an integrin-binding peptide (78,80,152). Some examples of peptide gene transfer exploit the physiologic cellular process of receptor-mediated endocytosis for internalization (51,81,139). Other strategies to enhance gene delivery include ligands with the following properties: 1) nuclear localization signals to enhance nuclear delivery; 2) pH-sensitive ligands to encourage endosomal release; and 3) steric stabilizing agents to avoid interaction with biologic factors that would destabilize the complexes after introduction into the biologic milieu (59). Some investigators have chosen random peptide-presenting phage libraries to select cell-binding peptides with potential gene therapy applications (12,123). Others have synthesized 2 component peptide systems that emulate viral functions, for example, a DNA-condensing agent and a pH-dependent endosomal releasing agent (72). We have developed a method of gene transfer mediated by histone H2A (8,9). An N-terminal 37-mer of histone H2A mediates DNA transfection via DNA binding and nuclear localization. Histone H2A was also tested in vivo in a murine neuroblastoma model (11). The transfection of plasmids secreting interleukin-2 (IL-2) and single chain interleukin-12 (sc IL-12) resulted in superior antitumor immunity with histone H2A, com" @default.
W2314806967 created "2016-06-24" @default.
W2314806967 creator A5064609126 @default.
W2314806967 creator A5066463616 @default.
W2314806967 date "2002-01-01" @default.
W2314806967 modified "2023-10-10" @default.
W2314806967 title "Gene Therapy of Human Disease" @default.
W2314806967 cites W110839551 @default.
W2314806967 cites W134928066 @default.
W2314806967 cites W1483766007 @default.
W2314806967 cites W1484466993 @default.
W2314806967 cites W1487084354 @default.
W2314806967 cites W1490324405 @default.
W2314806967 cites W1497511116 @default.
W2314806967 cites W1513878643 @default.
W2314806967 cites W1530533037 @default.
W2314806967 cites W1565788467 @default.
W2314806967 cites W1576238436 @default.
W2314806967 cites W1579604458 @default.
W2314806967 cites W1585257520 @default.
W2314806967 cites W1587057560 @default.
W2314806967 cites W1596008808 @default.
W2314806967 cites W1599588446 @default.
W2314806967 cites W1616824889 @default.
W2314806967 cites W1663829637 @default.
W2314806967 cites W175604019 @default.
W2314806967 cites W1800005238 @default.
W2314806967 cites W1871369825 @default.
W2314806967 cites W1901002830 @default.
W2314806967 cites W1963861474 @default.
W2314806967 cites W1967187884 @default.
W2314806967 cites W1970812755 @default.
W2314806967 cites W1971290197 @default.
W2314806967 cites W1972832005 @default.
W2314806967 cites W1973919938 @default.
W2314806967 cites W1974698048 @default.
W2314806967 cites W1975121126 @default.
W2314806967 cites W1976215218 @default.
W2314806967 cites W1978952718 @default.
W2314806967 cites W1980869398 @default.
W2314806967 cites W1981059306 @default.
W2314806967 cites W1983863563 @default.
W2314806967 cites W1985527224 @default.
W2314806967 cites W1985584561 @default.
W2314806967 cites W1985678203 @default.
W2314806967 cites W1987599352 @default.
W2314806967 cites W1990187729 @default.
W2314806967 cites W1990668606 @default.
W2314806967 cites W1994617469 @default.
W2314806967 cites W1995240556 @default.
W2314806967 cites W1995315280 @default.
W2314806967 cites W1998165633 @default.
W2314806967 cites W1999598401 @default.
W2314806967 cites W2000615271 @default.
W2314806967 cites W2000695619 @default.
W2314806967 cites W2001332902 @default.
W2314806967 cites W2001514241 @default.
W2314806967 cites W2003017660 @default.
W2314806967 cites W2003021187 @default.
W2314806967 cites W200338635 @default.
W2314806967 cites W2006258756 @default.
W2314806967 cites W2007153121 @default.
W2314806967 cites W2009692654 @default.
W2314806967 cites W2009761723 @default.
W2314806967 cites W2010955557 @default.
W2314806967 cites W2013819225 @default.
W2314806967 cites W2014495927 @default.
W2314806967 cites W2015617337 @default.
W2314806967 cites W2015789556 @default.
W2314806967 cites W2016723478 @default.
W2314806967 cites W2018116983 @default.
W2314806967 cites W2019592706 @default.
W2314806967 cites W2019921823 @default.
W2314806967 cites W2020741949 @default.
W2314806967 cites W2021875656 @default.
W2314806967 cites W2022328339 @default.
W2314806967 cites W2026699829 @default.
W2314806967 cites W2027022352 @default.
W2314806967 cites W2027321452 @default.
W2314806967 cites W2028368276 @default.
W2314806967 cites W2029175314 @default.
W2314806967 cites W2029979670 @default.
W2314806967 cites W2031890808 @default.
W2314806967 cites W2032678451 @default.
W2314806967 cites W2037215798 @default.
W2314806967 cites W2038617586 @default.
W2314806967 cites W2038937549 @default.
W2314806967 cites W2039457108 @default.
W2314806967 cites W2039722728 @default.
W2314806967 cites W2040016439 @default.
W2314806967 cites W2041179764 @default.
W2314806967 cites W2041186543 @default.
W2314806967 cites W2043521465 @default.
W2314806967 cites W2043852940 @default.
W2314806967 cites W2044502352 @default.
W2314806967 cites W2045479594 @default.
W2314806967 cites W2046253878 @default.
W2314806967 cites W2047938450 @default.