| adeno-associated virus-mediated gene delivery. | several gene delivery vehicles are being developed for somatic gene therapy and each of these vectors has unique properties which makes them appropriate for different human disease applications. recombinant adeno-associated viral (raav) vectors are proving themselves to be safe and efficacious for the long-term expression of proteins and correction of genetic diseases following a single administration. the increasing number of tissues and diseases being targeted with raav vectors demonstrates th ... | 1999 | 10738565 |
| maxillary sinusitis as a surrogate model for cf gene therapy clinical trials in patients with antrostomies. | assessing the biological activity and clinical efficacy of gene therapy is critically important in cystic fibrosis (cf). it is widely accepted that clinical testing using surrogate markers including pulmonary function will be useful in assessing clinical efficacy. one problem with pulmonary surrogate markers of cf disease is the large number of patients and length of time required to demonstrate clinical efficacy. an alternative to pulmonary testing of new cf treatments is use of the maxillary s ... | 1999 | 10738581 |
| increased motoneuron survival and improved neuromuscular function in transgenic als mice after intraspinal injection of an adeno-associated virus encoding bcl-2. | mutations in the gene encoding cu/zn superoxide dismutase (sod1) underlie some familial cases of amyotrophic lateral sclerosis (als), a neurodegenerative disorder characterized by loss of cortical, brainstem and spinal motoneurons. transgenic mice over- expressing a mutated form of human sod1 containing a gly-->ala substitution at position 93 (sod1(g93a)) develop a severe, progressive motoneuron disease. we investigated the potential of recombinant adeno-associated virus (raav) to transfer neuro ... | 2000 | 10749988 |
| roles of adeno-associated virus rep protein and human chromosome 19 in site-specific recombination. | adeno-associated virus type 2 (aav) is the only known eucaryotic virus capable of targeted integration in human cells. aav integrates preferentially into human chromosome (ch) 19q13.3qter. the nonstructural proteins of aav-2, rep78 and rep68, are essential for targeted integration. rep78 and rep68 are multifunctional proteins with diverse biochemical activities, including site-specific binding to aav and ch-19 target sequences, helicase activity, and strand-specific, site-specific endonuclease a ... | 2000 | 10756007 |
| transfer of the feline erythropoietin gene to cats using a recombinant adeno-associated virus vector. | chronic renal failure and the associated erythropoietin-responsive anemia afflicts over 2 million domestic cats in the united states, resulting in morbidity that can affect the owner-pet relationship. although treatment of cats with recombinant human erythropoietin (epo) protein can be effective, response to the drug often dissipates over time, probably due to the development of antibodies reactive with the human protein. as an alternate approach to the treatment of this disease, we have develop ... | 2000 | 10757028 |
| adeno-associated virus site-specifically integrates into a muscle-specific dna region. | the nonpathogenic human virus adeno-associated virus type 2 (aav) has evolved the potentially unique strategy to establish latency by site-specifically integrating its genome into human chromosome 19 (19q13.3-qter) at a locus designated aavs1. this nonhomologous, site-specific recombination of viral dna with the human genome provides a basis for developing targeted gene therapy vectors. to assess whether the region surrounding aavs1 might have contributed to the selection of the specific integra ... | 2000 | 10758163 |
| construction of a recombinant adeno-associated virus (raav) vector expressing murine interleukin-12 (il-12). | il-12 is a heterodimeric cytokine that is known to induce tumor regression and long-term antitumor immunity. recombinant adeno-associated virus (raav) vectors are advantageous for gene therapy in that they lack pathogenicity in humans, infect dividing as well as nondividing cells, and show a broad range of infectivity. we constructed an raav vector expressing interleukin-12 (il-12) for cancer immunotherapy studies in a mouse model by inserting murine il-12 (mil-12) p35 and p40 cdnas into the pla ... | 2000 | 10770641 |
| chromosomal latency and expression at map unit 96 of a wild-type plus adeno-associated virus (aav)/neo vector and identification of p81, a new aav transcriptional promoter. | human adeno-associated virus (aav) is ubiquitous and known to establish latency by chromosomal integration. we have constructed a wild-type plus aav vector, ins96-0.9neo, containing the neomycin resistance gene open reading frame (neo orf) of 960 bases in length at map unit 96 of the virus. ins96-0.9neo was constructed in an unconventional manner in that the neo orf lacked a dedicated heterologous promoter. in this study, this wild-type plus aav vector was to used to test aav's packaging capacit ... | 1999 | 10774553 |
| design and packaging of adeno-associated virus gene targeting vectors. | adeno-associated virus (aav) vectors can transduce cells by several mechanisms, including (i) gene addition by chromosomal integration or episomal transgene expression or (ii) gene targeting by modification of homologous chromosomal sequences. the latter process can be used to correct a variety of mutations in chromosomal genes with high fidelity and specificity. in this study, we used retroviral vectors to introduce mutant alkaline phosphatase reporter genes into normal human cells and subseque ... | 2000 | 10775597 |
| improvement of erythropoiesis in beta-thalassemic mice by continuous erythropoietin delivery from muscle. | beta-thalassemias are highly prevalent genetic disorders that can cause severe hemolytic anemia. the main pathophysiologic feature of beta-thalassemia is the accumulation of unpaired alpha-globin chains in erythrocyte precursors and red blood cells (rbcs). this accumulation alters cell membrane function and results in early cell destruction and ineffective erythropoiesis. correction of globin chain imbalance through the induction of fetal hemoglobin (hbf) synthesis is a tentative therapeutic app ... | 2000 | 10779423 |
| adeno-associated virus mediated gene transfer into primary rat brain neuronal and glial cultures: enhancement with the ph-sensitive surfactant dodecyl 2-(1'-imidazolyl) propionate. | this study evaluated the effects of a novel, ph-sensitive surfactant, dodecyl 2-(1'-imidazolyl) propionate (dip), on cationic lipid mediated transfection in primary rat brain neuronal and glial cultures. the cationic lipid complex dotap/dope (1, 2-dioleoyl-3-trimethylammonium propionate and dioleoyl phosphatidylethanolamine, respectively) was added over a range of concentrations (0-120 microg/ml) with dna concentration kept constant (1.6 microg/ml). the neuron-specific enolase (nse) and cytomega ... | 2000 | 10781840 |
| gene delivery to human chondrocytes by an adeno associated virus vector. | to investigate the efficiency of gene transduction to human chondrocytes using an adeno associated virus (aav) vector. | 2000 | 10782826 |
| recent advances in liver-directed gene therapy: implications for the treatment of dyslipidemia. | somatic gene therapy for the treatment of dyslipidemia is an area of active investigation. a substantial body of data indicates that the transfer of various lipid-lowering genes to the liver is an effective method of restoring normal plasma lipids in animal models of dyslipidemia. most studies have used adenoviral vectors because of their excellent gene-transfer efficiency. however, the first and second-generation adenoviral vectors used in these experiments are highly toxic and are associated w ... | 2000 | 10787180 |
| gene vectors for cytokine expression in vivo. | the understanding of cytokine networks and the exploitation of these networks for the treatment of immune and inflammatory diseases as well as cancer depend on in vivo delivery of cytokines. due to instability of recombinant cytokine proteins, investigators have employed cytokine-encoding gene therapy vectors to induce high levels of cytokine expression in vivo. numerous gene therapy vectors have been developed recently which are suitable for this purpose. recent advances in the design of adenov ... | 2000 | 10788600 |
| dose response to a single intramuscular injection of recombinant adeno-associated virus-erythropoietin in monkeys. | anemia is a significant problem in many disease states. erythropoietin (epo) has been used in the treatment of anemia associated with numerous chronic diseases. this study investigates the dose-response profiles of a single intramuscular (im) injection of a recombinant adeno-associated virus vector (raav) containing the epo gene with the goal of achieving a sustained elevation of hematocrit (hct). | 2000 | 10792948 |
| production and purification of recombinant adeno-associated virus. | | 2000 | 10800712 |
| cross-species comparison of in vivo reporter gene expression after recombinant adeno-associated virus-mediated retinal transduction. | | 2000 | 10800714 |
| increasing the size of raav-mediated expression cassettes in vivo by intermolecular joining of two complementary vectors. | a major shortcoming to the use of adeno-associated virus (raav) vectors is their limited packaging size. to overcome this hurdle, we split an expression cassette and cloned it into two separate vectors. the vectors contained either a nuclear localizing escherichia coli lacz transgene (nlslacz) with a splice acceptor, or the human elongation factor 1alpha ( ef1alpha) gene enhancer/promoter(s) (ef1alphaep) with a splice donor. we co-injected a promoter-less nlslacz vector with a vector containing ... | 2000 | 10802620 |
| a new dual-vector approach to enhance recombinant adeno-associated virus-mediated gene expression through intermolecular cis activation. | | 2000 | 10802719 |
| overcoming adeno-associated virus vector size limitation through viral dna heterodimerization. | | 2000 | 10802720 |
| adeno-associated virus and other potential vectors for angiostatin and endostatin gene therapy. | | 2000 | 10810649 |
| differential expression of a recombinant adeno-associated virus 2 vector in human cd34+ cells and breast cancer cells. | the use of autologous hematopoietic stem cell (hsc) grafts after high-dose chemotherapy protocols may be hampered by contamination of the grafts with tumor cells. because epithelial cells seem to be the natural hosts of adeno-associated virus 2 (aav-2), we speculated that epithelial tumor cells in hsc grafts might be selective targets for aav-2-based vectors. to test this hypothesis, the breast cancer cell lines t47d and mcf-7 were infected with a recombinant aav-2 vector expressing the green fl ... | 2000 | 10811478 |
| detection of adeno-associated virus type 2 in patients with viral infection. | | 1999 | 10816616 |
| cation-exchange high-performance liquid chromatography of recombinant adeno-associated virus type 2. | there has been much interest recently in the development of recombinant viruses as vectors for gene therapy applications. we have constructed a recombinant adeno-associated viral (aav) vector containing the gene encoding cftr (cystic fibrosis transmembrane chloride regulator). this vector is currently being used in clinical trials as a treatment for cystic fibrosis. in the course of scale-up and process optimization efforts, a variety of analyses have been developed to characterize yield and qua ... | 2000 | 10821405 |
| somatic gene therapy for hypertension. | gene therapy for hypertension is needed for the next generation of antihypertensive drugs. current drugs, although effective, have poor compliance, are expensive and short-lasting (hours or one day). gene therapy offers a way to produce long-lasting antihypertensive effects (weeks, months or years). we are currently using two strategies: a) antisense oligodeoxynucleotides (as-odn) and b) antisense dna delivered in viral vectors to inhibit genes associated with vasoconstrictive properties. it is ... | 2000 | 10829100 |
| preclinical study on gene therapy of cervical carcinoma using adeno-associated virus vectors. | approximately 90% of cervical carcinomas are causally linked to infections with high-risk human papillomaviruses (hpvs), whose oncogenicity has been assigned to the continued expression of two early genes, e6 and e7. reversal of the transformed phenotype by inhibiting e6/e7 gene expression therefore provides a suitable goal for future tumor therapy. using recombinant adeno-associated virus type 2 (aav-2) vectors, two types of therapeutic genes were expressed in cervical carcinoma cells with the ... | 2000 | 10830724 |
| ribozyme gene therapy for autosomal dominant retinal disease. | gene delivery to cells of the retina, particularly to photoreceptor cells, has broad potential both for answering basic questions of retinal biology and for more applied therapeutic purposes. the use of ribozymes as therapy for autosomal dominant retinal diseases is a promising technique, and the theoretical and practical basis for their use is discussed. the process involves designing and testing ribozymes first in vitro and then in animal models of retinal disease. viral vectors based on the n ... | 2000 | 10834402 |
| adeno-associated virus as a gene delivery system. | adeno-associated virus (aav) has several characteristics which make it extremely attractive as a gene transfer vector: (1) no known pathogenicity; (2) high efficiency and the ability to remain latent; (3) a minimal number of antigens ensuring minimal immunogenicity; (4) the ability to transduce post-mitotic cells; (5) possible advantages of site-specific integration; and (6) a broad host and cell range. the human isolate, aav-2, is the best studied and has been the focus for gene delivery experi ... | 1997 | 10837553 |
| transplacental drug delivery: gene and virus delivery to the trophoblast. | the development of successful strategies for delivering genes to the placenta may provide new opportunities for modifying trophoblast function in order to learn more about trophoblast physiology and to offer novel therapeutic options for complications of pregnancy that result from placental dysfunction. replication-deficient recombinant viral vectors are useful vehicles for introducing genes into cells in vitro and in vivo. recombinant adenovirus and herpes simplex virus vectors are unable to ef ... | 1999 | 10837747 |
| endosomal processing limits gene transfer to polarized airway epithelia by adeno-associated virus. | the restriction of viral receptors and coreceptors to the basolateral surface of airway epithelial cells has been blamed for the inefficient transfer of viral vectors to the apical surface of this tissue. we now report, however, that differentiated human airway epithelia internalize raav type-2 virus efficiently from their apical surfaces, despite the absence of known adeno-associated virus-2 (aav-2) receptors or coreceptors at these sites. the dramatically lower transduction efficiency of raav ... | 2000 | 10841516 |
| trans-splicing vectors expand the utility of adeno-associated virus for gene therapy. | adeno-associated viral (aav) vectors have demonstrated considerable promise for gene therapy of inherited diseases. however, with a packaging size of <5 kb, applications have been limited to relatively small disease genes. based on the finding that aav genomes undergo intermolecular circular concatamerization after transduction in muscle, we have developed a paradigm to increase the size of delivered transgenes with this vector through trans-splicing between two independent vectors coadministere ... | 2000 | 10841568 |
| gene delivery to in situ veins: differential effects of adenovirus and adeno-associated viral vectors. | gene transfer offers the potential to modify vein graft biology at the time of surgical implantation. efficiency of gene delivery, stability of expression, and host responses are critical parameters for candidate vectors. we compared the effects of intraluminal exposure with adenovirus (ad) and adeno-associated virus (aav) vectors on transgene expression and monocyte adhesion (ma) in treated vein segments. | 2000 | 10842152 |
| gene therapy in the inner ear. mechanisms and clinical implications. | the application of gene therapy to the inner ear is an emerging field of study. most studies report the expression of marker genes (e.g., galactosidase) within the tissues of the cochlea. the first biologic response of an inner ear tissue (i.e., auditory neurons) to transduction by a gene therapy vector expressing a therapeutic gene (a herpes amplicon vector containing a bdnf gene) was observed in spiral explants obtained from early postnatal rat cochleae. this study was important because it dem ... | 1999 | 10842605 |
| expression of human herpesvirus 6b rep within infected cells and binding of its gene product to the tata-binding protein in vitro and in vivo. | we have characterized the human herpesvirus 6b (hhv-6b) rep gene, which is a homologue of the adeno-associated virus type 2 rep and is unique in the herpesvirus family. three transcripts, 9.0, 5.0, and 2. 7 kb (the major transcript), were detected by northern blotting using an hhv-6b rep probe under late conditions. we investigated the expression kinetics of the rep gene using cycloheximide (chx) and phosphonoformic acid (pfa), which are inhibitors of protein synthesis and viral dna synthesis, r ... | 2000 | 10846093 |
| dna sequence motifs which direct adeno-associated virus site-specific integration in a model system. | the dna sequence motifs which direct adeno-associated virus type 2 site-specific integration are being investigated using a shuttle vector, propagated as a stable episome in cultured cell lines, as the target for integration. previously, we reported that the minimum episomal targeting elements comprise a 16-bp binding motif (rep binding site [rbs]) for a viral regulatory protein (rep) separated by a short dna spacer from a sequence (terminal resolution site [trs]) that can serve as a substrate f ... | 2000 | 10846109 |
| adeno-associated viral vectors as gene delivery vehicles. | adeno-associated virus (aav), a non-pathogenic human parvovirus, is gaining attention for its potential use as a human gene therapy vector. one of the most attractive features of recombinant aav vectors is the ability to be stably maintained in host cells as integrated proviruses. this property is particularly desireable for therapies requiring long-term correction of a genetic defect. this review highlights recent advances made in the aav field and will discuss some limitations of raav vector i ... | 2000 | 10851261 |
| method to decrease the titers of contaminating helper adenovirus during the production of recombinant adeno-associated virus. | | 2000 | 10868274 |
| site-specific integration of a transgene mediated by a hybrid adenovirus/adeno-associated virus vector using the cre/loxp-expression-switching system. | as vectors, adenoviruses (ads) have many attractive advantages for in vivo gene therapy. however, ads do not usually integrate into the host genome and gene expression is, thus, transient. adeno-associated virus (aav) integrates into a specific locus (aavs1) on the human host's chromosome 19, while conventional recombinant aav (raav) vectors do not possess this property because such vectors lack the rep gene. aav vectors carrying the rep gene do not have enough space for insertion of a transgene ... | 2000 | 10873630 |
| ubiquitous human adeno-associated virus type 2 autonomously replicates in differentiating keratinocytes of a normal skin model. | since its discovery in 1966, adeno-associated virus type 2 (aav) has been described as a helper-dependent parvovirus. however, in this study we demonstrate that aav undergoes its complete life cycle, devoid of helper viruses or genotoxic agents, in the organotypic epithelial raft tissue culture system, a model of normal skin. aav progeny production directly correlated with epithelial differentiation, as nondifferentiating keratinocytes were defective for this activity. large nuclear virus arrays ... | 2000 | 10873777 |
| gene transfer into rat renal cells using adeno-associated virus vectors. | adeno-associated virus (aav) vectors have a number of attractive features, including lack of cytotoxicity, ability to transduce nondividing cells, and long-term transgene expression. we investigated whether rat renal cells could be efficiently transduced with aav vectors. rat glomerular mesangial cells were transduced with aav-lacz vector containing beta-galactosidase gene in vitro, and the expression of beta-galactosidase was evaluated by x-gal staining and elisa. for ex vivo experiments, secti ... | 2000 | 10878409 |
| adeno-associated virus vectors: activity and applications in the cns. | transgenic strategies are useful for functional studies and they may also lead to novel therapies. controlling transgene expression in defined cell populations over time is increasingly important for both functional and gene therapy experiments. the adeno-associated virus (aav) vector may provide sufficient spatio-temporal control of gene expression for these purposes. this paper reviews in vivo somatic gene transfer methodology using aav. advantageous features of this system include neuronal ge ... | 2000 | 10880823 |
| binding of the human papillomavirus type 16 e7 oncoprotein and the adeno-associated virus rep78 major regulatory protein in vitro and in yeast and the potential for downstream effects. | both human papillomavirus (hpv) and adeno-associated virus (aav) are common anogenital viruses and likely co-infect the epithelium in vivo. however, whereas hpvs are positively associated with cervical cancer, aav appears to be negatively associated. in tissue culture, aav-encoded rep78--which is essential for aav--inhibits gene expression and oncogenic transformation by hpv-16/18 and bovine papillomavirus type 1. here we observed whether the hpv-16 e7 oncoprotein might recognize and bind rep78. ... | 2000 | 10881991 |
| restoration of photoreceptor ultrastructure and function in retinal degeneration slow mice by gene therapy. | the gene prph2 encodes a photoreceptor-specific membrane glycoprotein, peripherin-2 (also known as peripherin/rds), which is inserted into the rims of photoreceptor outer segment discs in a complex with rom-1 (ref. 2). the complex is necessary for the stabilization of the discs, which are renewed constantly throughout life, and which contain the visual pigments necessary for photon capture. mutations in prph2 have been shown to result in a variety of photoreceptor dystrophies, including autosoma ... | 2000 | 10888879 |
| protease-deleted adenovirus vectors and complementing cell lines: potential applications of single-round replication mutants for vaccination and gene therapy. | a new kind of versatile adenoviral vector (adv) has been constructed, one that is completely replication disabled in the absence of ad-e1 proteins but is capable of a single round of replication when ad-e1 is present. this was made possible by deletion of the ad protease gene (ps), which is essential for many steps of the ad life cycle. the ps-deleted virus can be propagated in 293-derived cell lines engineered to express ps. in these new complementing cells, the ps gene was expressed from a tet ... | 2000 | 10890743 |
| safety and biological efficacy of an adeno-associated virus vector-cystic fibrosis transmembrane regulator (aav-cftr) in the cystic fibrosis maxillary sinus. | the host immune response and low vector efficiency have been key impediments to effective cystic fibrosis transmembrane regulator (cftr) gene transfer for cystic fibrosis (cf). an adeno-associated virus vector (aav-cftr) was used in a phase i dose-escalation study to transfer cftr cdna into respiratory epithelial cells of the maxillary sinus of 10 cf patients. | 1999 | 10890777 |
| aspartoacylase gene transfer to the mammalian central nervous system with therapeutic implications for canavan disease. | with the ultimate goal of developing safe and effective in vivo gene therapy for the treatment of canavan disease and other neurological disorders, we developed a non-viral lipid-entrapped, polycation-condensed delivery system (lpd) for central nervous system gene transfer, in conjunction with adeno-associated virus (aav)-based plasmids containing recombinant aspartoacylase (aspa). the gene delivery system was tested in healthy rodents and primates, before proceeding to preliminary studies in 2 ... | 2000 | 10894213 |
| adeno-associated virus-based vectors in gene therapy. | adeno-associated virus (aav) vectors were shown capable of high efficiency transduction of both dividing and nondividing cells and tissues. aav-mediated transduction leads to stable, long-term transgene expression in the absence of apparent immune response. these properties and the broad host range of aav vectors indicate that they constitute a powerful tool for gene therapy purposes. an additional potential benefit of aav vectors is their ability to integrate site-specifically in the presence o ... | 2000 | 10895050 |
| transduction of hepatocellular carcinoma (hcc) using recombinant adeno-associated virus (raav): in vitro and in vivo effects of genotoxic agents. | adeno-associated virus (aav) is an attractive tool for gene therapy. here we investigated the in vitro and in vivo transduction of hepatocellular carcinoma (hcc) cells by an aav vector and the efficacy of different strategies to enhance the transduction of the tumor. | 2000 | 10898318 |
| adeno-associated virus (aav)-3-based vectors transduce haematopoietic cells not susceptible to transduction with aav-2-based vectors. | although adeno-associated virus (aav)-2 has a broad tissue-host range and can transduce a wide variety of tissue types, some cells, such as erythro-megakaryoblastoid cells, are non-permissive and appear to lack the aav-2 receptor. however, limited studies have been reported with the related dependovirus aav-3. we have previously cloned this virus, characterized its genome and produced an infectious clone. in this study, the gene for green fluorescent protein (gfp) was inserted into aav-2- and aa ... | 2000 | 10900047 |
| characteristics of the adeno-associated virus preintegration site in human chromosome 19: open chromatin conformation and transcription-competent environment. | adeno-associated virus (aav) establishes latency in infected cells by integrating into the cellular genome, with a high preference for a unique region, called aavs1, of the human chromosome 19. the aav proteins rep78 and -68 are postulated to initiate the site-specific integration process by binding to a rep binding site (rbs) in aavs1. we provide further evidence to corroborate this model by demonstrating that the aavs1 rbs in human cell lines is located near a dnase i hypersensitive "open" chr ... | 2000 | 10906224 |
| adeno-associated virus vector-mediated bcl-2 gene transfer into post-ischemic gerbil brain in vivo: prospects for gene therapy of ischemia-induced neuronal death. | the proto-oncogene bcl-2 is known as an anti-apoptotic gene that confers the ability to block neuronal cell death after transient ischemia. in order to examine whether the bcl-2 gene can be used for protection of ischemic brain injury, we generated adeno-associated virus (aav) vectors capable of expressing human bcl-2. replication-defective aav vectors were found effectively to transfer and express bcl-2 gene in the gerbil hippocampal neurons. transduction with aav bcl-2 5 days before forebrain ... | 2000 | 10918494 |
| mechanism of rep-mediated adeno-associated virus origin nicking. | the single-stranded adeno-associated virus type 2 (aav) genome is flanked by terminal repeats (trs) that fold back on themselves to form hairpinned structures. during aav dna replication, the trs are nicked by the virus-encoded rep proteins at the terminal resolution site (trs). this origin function apparently requires three sequence elements, the rep binding element (rbe), a small palindrome that comprises a single tip of an internal hairpin within the tr (rbe'), and the trs. previously, we det ... | 2000 | 10933682 |
| cd40 ligand-dependent activation of cytotoxic t lymphocytes by adeno-associated virus vectors in vivo: role of immature dendritic cells. | recombinant adeno-associated virus type 2 (raav) is being explored as a vector for gene therapy because of its broad host range, good safety profile, and persistent transgene expression in vivo. however, accumulating evidence indicates that administration of aav vector may initiate a detectable cellular and humoral immune response to its transduced neo-antigen in vivo. to elucidate the cellular basis of the aav-mediated immune response, c57bl/6 mouse bone marrow-derived immature and mature dendr ... | 2000 | 10933709 |
| long-term and significant correction of brain lesions in adult mucopolysaccharidosis type vii mice using recombinant aav vectors. | most lysosomal storage diseases, including mucopolysaccharidosis, affect the central nervous system (cns). they often induce severe and progressive mental retardation. replacement therapy by purified enzyme infusions is a promising approach for the treatment of peripheral organs but without effect on cns pathology because the enzyme cannot cross the blood-brain barrier. intracranial injection of recombinant adeno-associated virus (aav) vectors offers an alternative for sustained local enzyme del ... | 2000 | 10933913 |
| improved adeno-associated virus vector production with transfection of a single helper adenovirus gene, e4orf6. | recent advances in adeno-associated virus (aav) vector production have eliminated the need for adenovirus infection by transfection of plasmids encoding the adenovirus e2a, e4orf6, and va rna transcription units. we report here the generation of significantly higher aav vector titers with transfection of the single adenovirus gene, e4orf6, when used in conjunction with the split aav packaging plasmids mtrep and cmvcap. transduction in a murine lung model with these higher titer vector stocks was ... | 2000 | 10933916 |
| rescue of skeletal muscles of gamma-sarcoglycan-deficient mice with adeno-associated virus-mediated gene transfer. | in humans, a subset of cases of limb-girdle muscular dystrophy (lgmd) arise from mutations in the genes encoding one of the sarcoglycan (alpha, beta, gamma, or delta) subunits of the dystrophin-glycoprotein complex. while adeno-associated virus (aav) is a potential gene therapy vector for these dystrophies, it is unclear if aav can be used if a diseased muscle is undergoing rapid degeneration and necrosis. the skeletal muscles of mice lacking gamma-sarcoglycan (gsg-/- mice) differ from the anima ... | 2000 | 10933922 |
| sustained expression of therapeutic level of factor ix in hemophilia b dogs by aav-mediated gene therapy in liver. | we demonstrate that a single intraportal vein injection of a recombinant adeno-associated virus (raav) vector encoding canine factor ix (cfix) cdna under the control of a liver-specific enhancer/promoter leads to a long-term correction of the bleeding disorder in hemophilia b dogs. stable expression of the therapeutic level of cfix (5% of normal level) was detected in the plasma of a dog injected with an aav vector at a dose of 4.6 x 10(12) particles/kg for over 7 months. both whole-blood clotti ... | 2000 | 10933925 |
| route of administration determines induction of t-cell-independent humoral responses to adeno-associated virus vectors. | vectors based on adeno-associated viruses (aav) type 2 show promise for treating chronic diseases because transgene expression appears to be stable. this study evaluated the impact of humoral immunity to the capsid proteins on vector uptake by hepatocytes following an intravascular approach. route of vector administration in mice had a qualitative effect on antivector b cell responses. administration of vector into the tail vein resulted in t-cell-dependent (td) b cell responses that were comple ... | 2000 | 10933950 |
| inhibition of s-phase progression by adeno-associated virus rep78 protein is mediated by hypophosphorylated prb. | adeno-associated virus (aav) has an antiproliferative action on cells. we investigated the effect of the aav replication proteins (rep) on the cell division cycle using retroviral vectors. rep78 and rep68 inhibited the growth of primary, immortalized and transformed cells, while rep52 and rep40 did not. rep68 induced cell cycle arrest in phases g(1) and g(2), with elevated cdk inhibitor p21 and reduced cyclin e-, a- and b1-associated kinase activity. rep78-expressing cells were also impaired in ... | 2000 | 10944118 |
| triple transduction with adeno-associated virus vectors expressing tyrosine hydroxylase, aromatic-l-amino-acid decarboxylase, and gtp cyclohydrolase i for gene therapy of parkinson's disease. | parkinson's disease (pd), a neurological disease suited to gene therapy, is biochemically characterized by a severe decrease in the dopamine content of the striatum. one current strategy for gene therapy of pd involves local production of dopamine in the striatum achieved by inducing the expression of enzymes involved in the biosynthetic pathway for dopamine. we previously showed that the coexpression of tyrosine hydroxylase (th) and aromatic-l-amino-acid decarboxylase (aadc), using two separate ... | 2000 | 10945765 |
| adeno-associated virus mediates long-term gene transfer and delivery of chondroprotective il-4 to murine synovium. | treatments for rheumatoid arthritis and other inflammatory arthropathies are often ineffective at preventing joint destruction. long-term genetic modification of the cells lining the joint space (synoviocytes) in vivo represents a potential method for the treatment of these chronic conditions. however, a vector capable of efficiently transducing synoviocytes in vivo for a persistent period has not been available. the present report describes the genetic modification of synoviocytes in vivo using ... | 2000 | 10947942 |
| combined effects of adeno-associated virus vector and a herpes simplex virus mutant as neoplastic therapy. | although surgical therapy for pancreatic cancer has not been successful, new gene therapies, such as adeno-associated virus (aav) vectors hold promise for treating cancer. however, expression of aav vectors alone is insufficient for adequate effects in vivo for cancer therapy. we describe a novel therapy using the combined herpes simplex virus-icp6 deletion mutant (icp6delta) and aav vector. | 2000 | 10951421 |
| efficient recombinant adeno-associated virus production by a stable rep-cap hela cell line correlates with adenovirus-induced amplification of the integrated rep-cap genome. | a possible procedure for the production of clinical grade recombinant adeno-associated virus type 2 (raav) would include the use of packaging cell lines, harboring the rep-cap genes and the vector, combined with a replication defective adenoviral plasmid to provide the helper activities. several studies have already shown that raav can be efficiently assembled by infecting the stable packaging cell line with adenovirus. however, the direct comparison with an adenoviral plasmid has never been rep ... | 2000 | 10953917 |
| mutational analysis of the adeno-associated virus type 2 (aav2) capsid gene and construction of aav2 vectors with altered tropism. | adeno-associated virus type 2 (aav2) has proven to be a valuable vector for gene therapy. characterization of the functional domains of the aav capsid proteins can facilitate our understanding of viral tissue tropism, immunoreactivity, viral entry, and dna packaging, all of which are important issues for generating improved vectors. to obtain a comprehensive genetic map of the aav capsid gene, we have constructed 93 mutants at 59 different positions in the aav capsid gene by site-directed mutage ... | 2000 | 10954565 |
| novel transcriptional regulatory signals in the adeno-associated virus terminal repeat a/d junction element. | adeno-associated virus (aav) type 2 vectors transfer stable, long-term gene expression to diverse cell types in vivo. many gene therapy applications require the control of long-term transgene expression, and aav vectors, similar to other gene transfer systems, are being evaluated for delivery of regulated gene expression cassettes. previously, we (r. p. haberman, t. j. mccown, and r. j. samulski, gene ther. 5:1604-1611, 1998) demonstrated the use of the tetracycline-responsive system for long-te ... | 2000 | 10954575 |
| ngf gene transfer to intrinsic basal forebrain neurons increases cholinergic cell size and protects from age-related, spatial memory deficits in middle-aged rats. | administration of nerve growth factor (ngf) by intracerebroventricular infusion or transplantation of ngf-secreting cells to the basal forebrain improves spatial memory in aged animals. using the adeno-associated virus (aav) vector system, basal forebrain neurons were transduced to produce ngf ectopically for long intervals (at least 9 months). rats received intraseptal injections of either the control vector, ptr-uf4, or the ptr-ngfmyc at 3 months of age, prior to testing their performance in t ... | 2000 | 10967308 |
| transduction of murine cerebellar neurons with recombinant fiv and aav5 vectors. | our data demonstrate that vectors derived from recombinant feline immunodeficiency virus (rfiv) and adeno-associated virus type 5 (raav5) transduce cerebellar cells following direct injection into the cerebellar lobules of mice. both recombinant viruses mediated gene transfer predominantly to neurons, with up to 2500 and 1500 purkinje cells transduced for raav5 or rfiv-based vectors, respectively. the vectors also transduced stellate, basket and golgi neurons, with occasional transduction of gra ... | 2000 | 10976941 |
| hyaluronidase enhances recombinant adeno-associated virus (raav)-mediated gene transfer in the rat skeletal muscle. | skeletal muscle is a privileged target for long-term raav-mediated gene transfer in mouse, rat, dog and non-human primates. intramuscular injections of raav encoding human factor ix in hemophilia b patients have been initiated, based on promising results gathered in affected dogs. we found that intramuscular raav administration in rats resulted in restricted transduction essentially along the myofibers axis with poor lateral diffusion. this suggested that the transduction rate might be limited b ... | 2000 | 10981669 |
| selective cleavage of aavs1 substrates by the adeno-associated virus type 2 rep68 protein is dependent on topological and sequence constraints. | the adeno-associated virus type 2 (aav-2) rep78 and rep68 proteins are required for replication of the virus as well as its site-specific integration into a unique site, called aavs1, of human chromosome 19. rep78 and rep68 initiate replication by binding to a rep binding site (rbs) contained in the aav-2 inverted terminal repeats (itrs) and then specifically nicking at a nearby site called the terminal resolution site (trs). similarly, rep78 and rep68 are postulated to trigger the integration p ... | 2000 | 10982325 |
| adeno-associated virus type 2 rep protein inhibits human papillomavirus type 16 e2 recruitment of the transcriptional coactivator p300. | infection by human adeno-associated virus type 2 (aav2) is a possible protective factor in the development of cervical carcinomas associated with human papillomaviruses (hpv). the replicative proteins of aav2 (rep) have been implicated in the inhibition of papillomavirus replication and transforming activities, although the molecular events underlying these effects are poorly understood. we observed that each of the four forms of aav2 rep inhibited the e1- and e2-driven replication of oncogenic ... | 2000 | 10982355 |
| endocytosis and nuclear trafficking of adeno-associated virus type 2 are controlled by rac1 and phosphatidylinositol-3 kinase activation. | adeno-associated virus (aav) is a single-stranded dna parvovirus that causes no currently known pathology in humans. despite the fact that this virus is of increasing interest to molecular medicine as a vector for gene delivery, relatively little is known about the cellular mechanisms controlling infection. in this study, we have examined endocytic and intracellular trafficking of aav-2 using fluorescent (cy3)-conjugated viral particles and molecular techniques. our results demonstrate that inte ... | 2000 | 10982365 |
| monoclonal antibodies against the adeno-associated virus type 2 (aav-2) capsid: epitope mapping and identification of capsid domains involved in aav-2-cell interaction and neutralization of aav-2 infection. | the previously characterized monoclonal antibodies (mabs) a1, a69, b1, and a20 are directed against assembled or nonassembled adeno-associated virus type 2 (aav-2) capsid proteins (a. wistuba, a. kern, s. weger, d. grimm, and j. a. kleinschmidt, j. virol. 71:1341-1352, 1997). here we describe the linear epitopes of a1, a69, and b1 which reside in vp1, vp2, and vp3, respectively, using gene fragment phage display library, peptide scan, and peptide competition experiments. in addition, mabs a20, c ... | 2000 | 10982375 |
| use of the nadh-quinone oxidoreductase (ndi1) gene of saccharomyces cerevisiae as a possible cure for complex i defects in human cells. | the ndi1 enzyme of saccharomyces cerevisiae is a single subunit rotenone-insensitive nadh-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. we have shown previously that the ndi1 gene can be functionally expressed in chinese hamster cells (seo, b. b., kitajima-ihara, t., chan, e. k., scheffler, i. e., matsuno-yagi, a., and yagi, t. (1998) proc. natl. acad. sci. u. s. a. 95, 9167-9171) and human embryonal kidney 293 (hek 293) cells (seo, b. b., matsuno ... | 2000 | 10982813 |
| the potential role of antisense oligodeoxynucleotide therapy for cardiovascular disease. | current drugs used in the treatment of cardiovascular disease are effective but compliance is poor and they are short acting (hours or one day). gene therapy offers a way to produce long-lasting effects (weeks, months or years). antisense inhibition is being developed for the treatment of hypertension, myocardial ischaemia and improved allograft survival in human vascular bypass grafts. we are currently using 2 strategies: (i) antisense oligodeoxynucleotides (as-odns) which are delivered nonvira ... | 2000 | 10983731 |
| viral vectors for gene transfer: a review of their use in the treatment of human diseases. | the efficient delivery of therapeutic genes and appropriate gene expression are the crucial issues for clinically relevant gene therapy. viruses are naturally evolved vehicles which efficiently transfer their genes into host cells. this ability made them desirable for engineering virus vector systems for the delivery of therapeutic genes. the viral vectors recently in laboratory and clinical use are based on rna and dna viruses processing very different genomic structures and host ranges. partic ... | 2000 | 10983732 |
| evaluation of the possible protective role of adeno-associated virus type 2 infection in hpv-associated premalignant disease of the cervix. | our objective was to examine the prevalence of adeno-associated virus (aav) infection in women with normal cervical smears and those with hpv-associated cervical intraepithelial neoplasia (cin). | 2000 | 10985891 |
| adeno-associated virus expresses transgenes in hair follicles and epidermis. | adeno-associated virus (aav) vectors are nonpathogenic, integrating dna vectors capable of transducing dividing and nondividing cells with the potential of long-term expression. evaluating this interesting vector system in the skin for the first time, we found that an aav vector containing the lacz gene (aavlacz) led to the expression of beta-galactosidase for more than 6 weeks following in vivo injection. interestingly, expression was present not only in dividing and postmitotic epidermal kerat ... | 2000 | 10985948 |
| prolonged correction of hyperlipidemia in mice with familial hypercholesterolemia using an adeno-associated viral vector expressing very-low-density lipoprotein receptor. | adeno-associated viral vectors were used to deliver the gene for very-low-density lipoprotein (vldl) receptor (vldlr) to liver of a murine model of familial hypercholesterolemia (fh). infusion of adeno-associated virus-vldlr into the portal circulation of fh mice resulted in a 40% reduction in serum cholesterol and triglyceride that was stable for the duration of the study (30 weeks). fractionation of serum lipids revealed a reduction of both vldl and low-density lipoprotein. expression of trans ... | 2000 | 10985956 |
| induction of circular episomes during rescue and replication of adeno-associated virus in experimental models of virus latency. | the synthesis of linear duplex replicative structures (monomers, head-to-head, and tail-to-tail dimers) is an important hallmark of the productive phase of the adeno-associated virus (aav) life cycle. these structures are generated by a strand-displacement replication mechanism and believed to be a reservoir for single-stranded dna genomes. during the course of studies with recombinant versions of aav (raav), we discovered the assembly of circular duplex provirus derivatives in latently infected ... | 2000 | 10998340 |
| gene therapy vectors based on adeno-associated virus: characteristics and applications to acquired and inherited diseases (review). | adeno-associated virus (aav), a defective parvovirus, was discovered more than 30 years ago. interest in this virus for human gene therapy applications focuses on its non-pathogenicity, broad tropism and infectivity, site-specific integration and long-term persistence. the field of raav research has considerably advanced: titers of 1014 p/ml have been achieved, plasmid systems devised to produce helper-free viruses, chimaeric vectors combining properties of raav itrs and large sequence capacity ... | 2000 | 10998427 |
| adeno-associated virus type 2 rep78 induces apoptosis through caspase activation independently of p53. | adeno-associated virus (aav) type 2 rep78 is a multifunctional protein required for aav dna replication, integration, and gene regulation. the biochemical activities of rep78 have been described, but the effects of rep proteins on the cell have not been characterized. we have analyzed rep-mediated cytotoxicity. we demonstrated that rep78 expression is sufficient to induce cell death and disruption of the cell cycle. cell death was found to be mediated by apoptosis. rep78 expression resulted in t ... | 2000 | 11000213 |
| recruitment of single-stranded recombinant adeno-associated virus vector genomes and intermolecular recombination are responsible for stable transduction of liver in vivo. | recombinant adeno-associated virus (raav) vectors stably transduce hepatocytes in experimental animals. following portal-vein administration of raav vectors in vivo, single-stranded (ss) raav genomes become double stranded (ds), circularized, and/or concatemerized concomitant with a slow rise and, eventually, steady-state levels of transgene expression. over time, at least some of the stabilized genomes become integrated into mouse chromosomal dna. the mechanism(s) of formation of stable ds raav ... | 2000 | 11000214 |
| ribozyme rescue of photoreceptor cells in p23h transgenic rats: long-term survival and late-stage therapy. | ribozyme-directed cleavage of mutant mrnas appears to be a potentially effective therapeutic measure for dominantly inherited diseases. we previously demonstrated that two ribozymes targeted to the p23h mutation in rhodopsin slow photoreceptor degeneration in transgenic rats for up to 3 months of age when injected before significant degeneration at postnatal day (p) 15. we now have explored whether ribozyme rescue persists at older ages, and whether ribozymes are effective when injected later in ... | 2000 | 11005848 |
| retinal degeneration is slowed in transgenic rats by aav-mediated delivery of fgf-2. | we evaluated adeno-associated virus (aav)-mediated gene transfer of basic fibroblast growth factor (fgf-2) as a therapy for photoreceptor degeneration in a transgenic rat model of retinitis pigmentosa. | 2000 | 11006261 |
| adeno-associated virus vector transduction of vascular smooth muscle cells in vivo. | adeno-associated virus (aav) vectors might offer solutions for restenosis and angiogenesis by transducing nondividing cells and providing long-term gene expression. we investigated the feasibility of vascular cell transduction by aav vectors in an in vivo rabbit carotid artery model. time course of gene expression, inflammatory reaction to the vector, and effects of varying viral titer, exposure time, and intraluminal pressures on gene expression were examined. recombinant aav vectors with an ro ... | 2000 | 11015590 |
| safety of adeno-associated virus as cochlear gene transfer vector: analysis of distant spread beyond injected cochleae. | the adeno-associated virus (aav), inoculated into the perilymph, has been shown to be an effective vector for mediating intracochlear transgene expression. the unexpected finding of transgene expression in the contralateral cochlea in previous work raised concern about dissemination of the virus from the target tissue. the current study was undertaken to assess the extent of aav dissemination following its introduction into the inner ear. adult male guinea pigs were injected with recombinant aav ... | 2000 | 11020352 |
| selective rep-cap gene amplification as a mechanism for high-titer recombinant aav production from stable cell lines. | gene transfer vectors based on adeno-associated virus mediate high-level, stable gene expression in a variety of postmitotic tissues; thus, there is interest in developing improved production systems. we previously described the generation of raav producer cell lines that, upon infection with adenovirus, yielded biologically active raav particles. in these studies we show that the adenovirus multiplicity of infection (m.o.i.) is a critical variable for efficient production of cell line-derived r ... | 2000 | 11020356 |
| adeno-associated virus vector mediated gene transfer to pancreatic beta cells. | insulin-dependent diabetes mellitus (iddm) or type 1 diabetes is an autoimmune disease that results in destruction of the insulin-producing pancreatic islet beta cells. several factors induce the invasion of immune cells into islets and trigger inflammation. gene therapy approaches targeting the islet cells could be an effective treatment to prevent the onset or reverse type 1 diabetes. allogeneic islet transplantation provides short-term treatment. however, genetically modified islets, which re ... | 2000 | 11021593 |
| adeno-associated virus rnas appear in a temporal order and their splicing is stimulated during coinfection with adenovirus. | we have used a quantitative rnase protection assay to characterize the relative accumulation and abundance of individual adeno-associated virus type 2 (aav) rnas throughout the course of aav-adenovirus coinfections and preinfections. we have demonstrated that there is a previously unrecognized temporal order to the appearance of aav rnas. first, unspliced p5-generated transcripts, which encode rep78, were detectable prior to the significant accumulation of other aav rnas. ultimately, as previous ... | 2000 | 11024114 |
| adeno-associated virus-mediated vascular endothelial growth factor gene transfer into cardiac myocytes. | vascular endothelial growth factor (vegf) is an angiogenic growth factor that stimulates endothelial cell proliferation, increases endothelial permeability, and promotes collateral vessel formation. we transferred human vegf gene into rat cardiac myocytes using adeno-associated virus (aav) vectors and investigated whether vegf secreted from the transduced cardiac myocytes promoted proliferation of endothelial cells. we produced vegf-expressing aav vectors (aav-vegf) by the adenovirus-free method ... | 2000 | 11026643 |
| self-amplification system for recombinant adeno-associated virus production. | a recently reported system for recombinant adeno-associated virus (raav) production does not require infection of a helper virus and depends on the transfection with a huge amount of three plasmids: aav-vector, aav-helper, and adenovirus-helper plasmids. toward simplifying raav production, as a first step, we tested the use of the raav itself instead of the aav-vector plasmid as a source of raav dna and determined the optimal timing of infection and dose of the input raav. when 293 cells were in ... | 2000 | 11027513 |
| the adeno-associated virus vector for orthopaedic gene therapy. | during the last decade researchers working with recombinant adeno-associated virus have shown the use of this vector for efficient and long-term gene transfer in various tissues including lung, muscle, brain, spinal cord, retina, and liver. in 1999 the first results documenting the use of this vector in transducing joint cells were published. additional advantages of recombinant adeno-associated virus for in vivo gene therapy are: (1) its ability to transduce nondividing cells; (2) site-specific ... | 2000 | 11039749 |
| gene therapy approaches for treating rheumatoid arthritis. | current gene therapy approaches for treating rheumatoid arthritis have made use of gene transfer technology as an improved delivery system for emerging proteins and other biologicals whose activities may have therapeutic value. preclinical research has focused on two primary directions, evaluation of methods of gene delivery and identification of gene products with antiarthritic potential. although there are reports involving systemic gene delivery, the bulk of effort has focused on local, intra ... | 2000 | 11039782 |
| adeno-associated virus (aav) rep protein enhances the generation of a recombinant mini-adenovirus (ad) utilizing an ad/aav hybrid virus. | mini-adenoviruses (mad) deleted of all viral coding regions represent an emerging approach for transgene expression. we have exploited the unique features of the adeno-associated virus (aav) terminal repeats within the context of an adenovirus-adeno-associated hybrid virus (ad/aav) as a strategy for rapid and efficient generation of mad. excision and generation of mad from the parental ad/aav hybrid vector was achieved in 293 cells through recombination but without selection for mad production. ... | 2000 | 11044082 |
| site-specific integration of an adeno-associated virus vector plasmid mediated by regulated expression of rep based on cre-loxp recombination. | recombinant adeno-associated virus (aav) type 2 has attracted attention because it appears to have the potential to serve as a vector for human gene therapy. an interesting feature of wild-type aav is its site-specific integration into aavs1, a defined locus on chromosome 19. this reaction requires the presence of two viral elements: inverted terminal repeats and rep78/68. accordingly, current aav vectors lacking the rep gene lack the capacity for site-specific integration. in this report, we de ... | 2000 | 11044107 |
| recombinant adeno-associated virus vector-based gene transfer for defects in oxidative metabolism. | defects in oxidative metabolism may be caused by mutations either in nuclear genes or in mitochondrial dna (mtdna). we tested the hypothesis that recombinant adeno-associated virus (raav) could be used to complement mtdna mutations. aav vector constructs were designed to express the reporter gene encoding green fluorescent protein (gfp), fused to a targeting presequence that directed gfp to be translocated into mitochondria. these vectors mediated expression of mitochondrial-localized gfp, as in ... | 2000 | 11044909 |
| purification of recombinant adeno-associated virus vectors by column chromatography and its performance in vivo. | recombinant adeno-associated virus (aav) holds much promise for human gene therapy. while evidence indicates that aav mediates long-term gene transfer in several different tissues, difficulty in preparing and purifying this viral vector in large quantities remains a major obstacle for evaluating aav vectors in clinical trials. the current method of purification, based on sedimentation through cesium chloride, is not scaleable and yields product of insufficient quality. in this article we report ... | 2000 | 11044910 |
| chronic ethanol increases adeno-associated viral transgene expression in rat liver via oxidant and nfkappab-dependent mechanisms. | recombinant adeno-associated virus (raav) transduction is limited in vivo, yet can be enhanced by hydroxyurea, ultraviolet-irradiation, or adenovirus coinfection, possibly via mechanisms involving stress in the host cell. because chronic ethanol induces oxidative stress, it was hypothesized that chronic ethanol would increase raav transduction in vivo. to test this hypothesis, raav encoding beta-galactosidase was given to wistar rats that later received either ethanol diet or high-fat control di ... | 2000 | 11050056 |
| adeno-associated virus vector-mediated gene transfer to somatic cells in the central nervous system. | | 2000 | 11050954 |
| production of recombinant adeno-associated virus. | currently, raav appears to be one of the most promising vectors for gene therapy applications. attractive features of the vector include nonpathogenicity, the ability to infect nondividing cells, escape from host immune responses, and integration into the host genome. tremendous progress has been made in the production of this vector, which makes it possible to start to examine the vector performance in large animals and to implement the transition to phase i human clinical trials with a variety ... | 2000 | 11050955 |