| [post-translational ligation and function of dual-vector transferred split cftr gene]. | the mutation of cystic fibrosis transmembrane conductance regulator (cftr) gene leads to an autosomal recessive genetic disorder cystic fibrosis (cf). the gene therapy for cf using adeno-associated virus (aav) vectors delivering cftr gene is restricted by the contents limitation of aav vectors. in this study the split cftr genes severed at its regulatory domain were delivered by a dual-vector system with an intein-mediated protein trans-splicing as a technique to investigate the post-translation ... | 2010 | 21351451 |
| [cellular immunotoxicity of raav gene medicine and possible solutions]. | gene medicine based on recombinant adeno-associated virus (raav) vector has rapidly become the prior-choose reagent for gene therapy, since it had been shown that the raav was able to stably express many genes in vivo without detectable side-effect. however, recent findings of ctl immune responses to aav capsid in a clinical trial highlighted a new issue regarding safety that previously was not identified in animal studies. obviously it is so important to understand the interaction of raav with ... | 2010 | 21351561 |
| in vitro evaluation of a double-stranded self-complementary adeno-associated virus type2 vector in bone marrow stromal cells for bone healing. | | 2011 | 21352585 |
| orthopaedic gene therapy using recombinant adeno-associated virus vectors. | use of recombinant adeno-associated virus (raav) vectors is increasingly gaining popularity in gene therapy because of their desirable properties, including lack of pathogenicity, efficient transduction of dividing and non-dividing cells, and sustained maintenance of the viral genome. it is these features of raav vectors that made them the focus for gene-based therapy of skeletal tissue regeneration. this review outlines the biological characteristics of adeno-associated virus (aav), states the ... | 2011 | 21354554 |
| comparison of il-10 and mcp-1-7nd gene transfer with aav9 vectors for protection from murine autoimmune myocarditis. | overexpression of therapeutic genes with potential disease-limiting effects, specifically at the site of inflammation, remains a major clinical challenge. in this study, we investigate the potential of adeno-associated virus (aav)-9-mediated cardiac expression of the anti-inflammatory mediators interleukin (il)-10 and a dominant-negative inhibitor of monocyte chemoattractant protein-1 (mcp1-7nd) on prevention of autoimmune myocarditis. | 2011 | 21354997 |
| gene transfer into cystic fibrosis airway epithelial cells. | gene transfer into airway epithelial cells becomes a particularly motivating goal as far as cystic fibrosis (cf) is concerned. as mentioned in chapter 15 , approx 90% of deaths caused by this devastating disease are the result of infections of the respiratory tract owing to dysfunction of the cl(-) transport in airway epithelial cells. efficient transfer of the cystic fibrosis transmembrane conductance regulator (cftr) gene into the airway epithelium of cf patients in vivo is one of the current ... | 1996 | 21359743 |
| engineering liver-detargeted aav9 vectors for cardiac and musculoskeletal gene transfer. | we report the generation of a new class of adeno-associated virus serotype 9 (aav9)-derived vectors displaying selective loss of liver tropism and demonstrating potential for cardiac and musculoskeletal gene transfer applications. random mutagenesis of residues within a surface-exposed region of the major aav9 capsid protein yielded a capsid library with mutations clustered at the icosahedral threefold symmetry axis. using a combination of sequence analysis, structural models, and in vivo screen ... | 2011 | 21364538 |
| development of a liver-specific tet-on inducible system for aav vectors and its application in the treatment of liver cancer. | recombinant adeno-associated virus (raav) are effective gene delivery vehicles that can mediate long-lasting transgene expression. however, tight regulation and tissue-specific transgene expression is required for certain therapeutic applications. for regulatable expression from the liver we designed a hepatospecific bidirectional and autoregulatory tetracycline (tet)-on system (tet(bidir)alb) flanked by aav inverted terminal repeats (itrs). we characterized the inducible hepatospecific system i ... | 2011 | 21364542 |
| a potential role of distinctively delayed blood clearance of recombinant adeno-associated virus serotype 9 in robust cardiac transduction. | recombinant adeno-associated virus serotype 9 (raav9) vectors show robust in vivo transduction by a systemic approach. it has been proposed that raav9 has enhanced ability to cross the vascular endothelial barriers. however, the scientific basis of systemic administration of raav9 and its transduction mechanisms have not been fully established. here, we show indirect evidence suggesting that capillary walls still remain as a significant barrier to raav9 in cardiac transduction but not so in hepa ... | 2011 | 21364543 |
| vascular endothelial growth factor-b gene transfer exacerbates retinal and choroidal neovascularization and vasopermeability without promoting inflammation. | the role of vascular endothelial growth factor (vegf)-b in the eye is poorly understood. the present study was conducted to evaluate the effect of overexpression of vegf-b via adeno-associated virus (aav) gene transfer on ocular angiogenesis, inflammation, and the blood-retinal barrier (brb). | 2011 | 21364963 |
| simultaneous pomc gene transfer to hypothalamus and brainstem increases physical activity, lipolysis and reduces adult-onset obesity. | pro-opiomelanocortin (pomc) neurons are identified in two brain sites, the arcuate nucleus of the hypothalamus and nucleus of the solitary tract (nts) in brainstem. earlier pharmacological and pomc gene transfer studies demonstrate that melanocortin activation in either site alone improves insulin sensitivity and reduces obesity. the present study, for the first time, investigated the long-term efficacy of pomc gene transfer concurrently into both sites in the regulation of energy metabolism in ... | 2011 | 21366729 |
| enhanced bone regeneration with sequential delivery of basic fibroblast growth factor and sonic hedgehog. | bone regeneration approaches that mimic the natural processes of bone repair have generated significant attention. we hypothesized that early delivery of an angiogenic factor combined with sustained exposure to an osteogenic factor would recapitulate the critical aspects of natural bone repair. | 2011 | 21367413 |
| recombinant-adeno-associated viral vector-mediated gene therapy for cardiovascular diseases. | adeno-associated virus is a kind of dna defective parvovirus which is non-pathogenic. recombinant-adeno-associated virus vector comes from wild-type non-pathogenic adeno-associated virus and is highly secure, and it also has the advantages of broad host range. recombinant-adeno-associated virus vector has become a hot spot for gene therapy and is widely used in gene therapy for cardiovascular diseases, especially for hypertension, heart failure, arteriosclerosis, and myocardial infarction. | 2011 | 21368431 |
| lethal toxicity caused by expression of shrna in the mouse striatum: implications for therapeutic design. | therapeutic rna interference (rnai) has emerged as a promising approach for the treatment of many incurable diseases, including cancer, infectious disease or neurodegenerative disorders. demonstration of efficacy and safety in animal models is necessary before planning human application. our group and others have previously shown the potential of this approach for the dominantly inherited neurological disease dyt1 dystonia by achieving potent short-hairpin rna (shrna)-mediated silencing of the d ... | 2011 | 21368900 |
| construction of recombinant adeno-associated virus (aav). | efficient and stable transfer of foreign dna into cells both in vitro and in vivo has become a powerful tool in the study of the pathogeneses of various diseases, such as cancer and infectious diseases. the study of hepatitis c virus (hcv) pathogenesis has been hampered by the lack of an easily available animal model or in vitro replication system. a new approach to the establishment of an hcv model are cell-culture systems stably expressing viral genes. | 1999 | 21374394 |
| reproducible high yields of recombinant adeno-associated virus produced using invertebrate cells in 0.02- to 200-liter cultures. | abstract the large amounts of recombinant adeno-associated virus (raav) vector needed for clinical trials and eventual commercialization require robust, economical, reproducible, and scalable production processes compatible with current good manufacturing practice. raav produced using baculovirus and insect cells satisfies these conditions; however, recovering raav particles from 200-liter bioreactors is more complicated than bench-scale vector preparations. using a variety of processing media, ... | 2011 | 21381980 |
| correction of neurological disease of mucopolysaccharidosis iiib in adult mice by raav9 trans-blood-brain barrier gene delivery. | the greatest challenge in developing therapies for mucopolysaccharidosis (mps) iiib is to achieve efficient central nervous system (cns) delivery across the blood-brain barrier (bbb). in this study, we used the novel ability of adeno-associated virus serotype 9 (aav9) to cross the bbb from the vasculature to achieve long-term global cns, and widespread somatic restoration of +¦-n-acetylglucosaminidase (naglu) activity. a single intravenous (iv) injection of raav9-cmv-hnaglu, without extraneous t ... | 2011 | 21386820 |
| induction and prevention of severe hyperammonemia in the spfash mouse model of ornithine transcarbamylase deficiency using shrna and raav-mediated gene delivery. | urea cycle defects presenting early in life with hyperammonemia remain difficult to treat and commonly necessitate liver transplantation. gene therapy has the potential to prevent hyperammonemic episodes while awaiting liver transplantation, and possibly also to avert the need for transplantation altogether. ornithine transcarbamylase (otc) deficiency, the most prevalent urea cycle disorder, provides an ideal model for the development of liver-targeted gene therapy. while we and others have succ ... | 2011 | 21386824 |
| [post-translational ligation of split cftr severed before tmd2 and its chloride channel function]. | mutations of cystic fibrosis transmembrane conductance regulator (cftr) gene leads to cystic fibrosis, an autosomal recessive genetic disorder affecting a number of organs including the lung airways, pancreas and sweat glands. in order to investigate the post-translational ligation of cftr with reconstructed functional chloride ion channel and the split ssp dnab intein-mediated protein trans-splicing was explored to co-deliver cftr gene into eukaryotic cells with two vectors. the human cftr cdna ... | 2010 | 21387835 |
| rescue of severe infantile hypophosphatasia mice by aav-mediated sustained expression of soluble alkaline phosphatase. | abstract hypophosphatasia (hpp) is an inherited disease caused by a deficiency of tissue-nonspecific alkaline phosphatase (tnalp). the major symptom of human hpp is hypomineralization, rickets, or osteomalacia, although the clinical severity is highly variable. the phenotypes of tnalp knockout (akp2(-/-)) mice mimic those of the severe infantile form of hpp. akp2(-/-) mice appear normal at birth, but they develop growth failure, epileptic seizures, and hypomineralization and die by 20 days of ag ... | 2011 | 21388343 |
| chondromodulin 1 stabilizes the chondrocyte phenotype and inhibits endochondral ossification of porcine cartilage repair tissue. | to investigate the effect of chondromodulin 1 on the phenotype of osteochondral progenitor cells in cartilage repair tissue. | 2011 | 21391200 |
| generation of recombinant adeno-associated virus. | adeno-associated virus (aav) was discovered about 30 yr ago as a contaminant of adenovirus preparations. since its discovery, researchers have described many unique characteristics of aav biology that have made it attractive as a potential vector for gene therapy. for example, aav is not pathogenic, approx 80% of adults in the united states are seropositive, but in no case has the virus been implicated as the etiological agent for a human disease. aav is a defective parvovirus with a single-stra ... | 2001 | 21394584 |
| direct gene transfer to the cns prevents emergence of neurologic disease in a murine model of mucopolysaccharidosis type i. | the mucopolysaccharidoses (mpss) are a group of 11 storage diseases caused by disruptions in glycosaminoglycan (gag) catabolism, leading to their accumulation in lysosomes. resultant multisystemic disease is manifested by growth delay, hepatosplenomegaly, skeletal dysplasias, cardiopulmonary obstruction, and, in severe mps i, ii, iii, and vii, progressive neurocognitive decline. some mpss are treated by allogeneic hematopoietic stem cell transplantation (hsct) and/or recombinant enzyme replaceme ... | 2011 | 21397026 |
| aav5-mediated sflt01 gene therapy arrests retinal lesions in ccl2(-/-)/cx3cr1(-/-) mice. | to test the effects of adeno-associated virus encoding sflt01 (aav5.sflt01) on the retinal lesions in ccl2(-/-)/cx3cr1(-/-) mice, a model for age-related macular degeneration (amd), aav5.sflt01 was injected into the subretinal space of the right eyes and the left eyes served as controls. histology found no retinal toxicity due to the treatment after 3 months. the treated eyes showed lesion arrest compared with lesion progression in the left eyes by fundus monitoring monthly and histological eval ... | 2011 | 21397984 |
| microrna-221 regulates fas-induced fulminant liver failure. | death receptor-mediated apoptosis of hepatocytes contributes to hepatitis and fulminant liver failure. micrornas (mirnas), 19-25 nucleotide-long noncoding rnas, have been implicated in the posttranscriptional regulation of the various apoptotic pathways. here we report that global loss of mirnas in hepatic cells leads to increased cell death in a model of fas/cd95 receptor-induced apoptosis. mirna profiling of murine liver identified 11 conserved mirnas, which were up-regulated in response to fa ... | 2011 | 21400558 |
| long-term cardiac pro-b-type natriuretic peptide gene delivery prevents the development of hypertensive heart disease in spontaneously hypertensive rats. | diastolic dysfunction associated with high blood pressure (bp) leads to cardiac remodeling and fibrosis and progression to congestive heart failure. b-type natriuretic peptide (bnp) has bp-lowering, antifibrotic, and antihypertrophic properties, which makes bnp an attractive agent for attenuating the adverse cardiac remodeling associated with hypertension. in the current study, we tested the effects of sustained cardiac probnp gene delivery on bp, cardiac function, and remodeling in spontaneousl ... | 2011 | 21403100 |
| good manufacturing practice production of self-complementary serotype 8 adeno-associated viral vector for a hemophilia b clinical trial. | to generate sufficient clinical-grade vector to support a phase i/ii clinical trial of adeno-associated virus serotype 8 (aav8)-mediated factor ix (fix) gene transfer for hemophilia b, we have developed a large-scale, good manufacturing practice (gmp)-compatible method for vector production and purification. we used a 293t-based two-plasmid transient transfection system coupled with a three-column chromatography purification process to produce high-quality self-complementary aav2/8 fix clinical- ... | 2011 | 21410419 |
| specific cellular immune responses in mice immunized with dna, adeno-associated virus and adenoviral vaccines of epstein-barr virus-lmp2 alone or in combination. | cellular immune responses, particularly those associated with cd3(+)cd8(+) cytotoxic t lymphocytes (ctl), are critical factors in controlling viral infection. nasopharyngeal carcinoma (npc) is closely associated with persistent epstein-barr virus (ebv) infection. npc vaccine studies have focused on enhancing specific antiviral ctl responses. in this study, three vaccines capable of expressing the ebv-latent membrane protein 2 (lmp2) (a dna vector, an adeno-associated virus (aav) vector, and a re ... | 2011 | 21416326 |
| metabolic activities and chondrogenic differentiation of human mesenchymal stem cells following recombinant adeno-associated virus-mediated gene transfer and overexpression of fibroblast growth factor 2. | the genetic manipulation of bone marrow-derived mesenchymal stem cells (mscs) is an attractive approach to produce therapeutic platforms for settings that aim at restoring articular cartilage defects. here, we examined the effects of recombinant adeno-associated virus (raav)-mediated overexpression of human fibroblast growth factor 2 (hfgf-2), a mitogenic factor also known to influence msc differentiation, upon the proliferative and chondrogenic activities of human mscs (hmscs) in a three-dimens ... | 2011 | 21417714 |
| intracellular localization of adeno-associated viral proteins expressed in insect cells. | production of vectors derived from adeno-associated virus (aavv) in insect cells represents a feasible option for large-scale applications. however, transducing particles yields obtained in this system are low compared with total capsid yields, suggesting the presence of genome encapsidation bottlenecks. three components are required for aavv production: viral capsid proteins (vp), the recombinant aav genome, and rep proteins for aav genome replication and encapsidation. little is known about th ... | 2011 | 21425251 |
| adenosine kinase determines the degree of brain injury after ischemic stroke in mice. | adenosine kinase (adk) is the major negative metabolic regulator of the endogenous neuroprotectant and homeostatic bioenergetic network regulator adenosine. we used three independent experimental approaches to determine the role of adk as a molecular target for predicting the brain's susceptibility to ischemic stroke. first, when subjected to a middle cerebral artery occlusion model of focal cerebral ischemia, transgenic fb-adk-def mice, which have increased adk expression in striatum (164%) and ... | 2011 | 21427729 |
| [preliminary study on transdifferentiation of human amniotic epithelial cells and its intrasplenic transplantation]. | the human amniotic epithelial cells (haecs) are a recently identified new type of stem cells. it has previously been shown that haecs express hepatocyte-related gene and possess intracellular features and functional properties of hepatocytes. the haecs may be a candidate seed cell for liver regeneration. to research the survival and migration in vivo of haecs via adeno-associated virus-mediated the green fluorescent protein gene (aav-gfp) transfection, and to explore the expression of hepatocyte ... | 2011 | 21427840 |
| t2 imaging in monitoring of intraparenchymal real-time convection-enhanced delivery. | real-time convection-enhanced delivery (rcd) of adeno-associated viral vectors by co-infusion of gadoteridol allows t1 magnetic resonance imaging (t1 mri) prediction of areas of subsequent gene expression. the use of t2 mri in rcd is less developed. in addition, the effect of flushing a dead-space volume on subsequent distribution of a therapeutic agent is not known. | 2011 | 21430597 |
| targeting neurons of rat nucleus tractus solitarii with the gene transfer vector adeno-associated virus type 2 to up-regulate neuronal nitric oxide synthase. | adeno-associated virus (aav) has distinct advantages over other viral vectors in delivering genes of interest to the brain. aav mainly transfects neurons, produces no toxicity or inflammatory responses, and yields long-term transgene expression. in this study, we first tested the hypothesis that aav serotype 2 (aav2) selectively transfects neurons but not glial cells in the nucleus tractus solitarii (nts) by examining expression of the reporter gene, enhanced green fluorescent protein (egfp), in ... | 2011 | 21431420 |
| adeno-associated virus serotype 2 mediated transduction and coexpression of the human apoai and sr-bi gene in hepg2 cells. | cholesterol efflux is the first step in the reverse cholesterol transport (rct) pathway, removing excess cholesterol from tissues, including the arterial wall, thus preventing the development of atherosclerosis. adeno-associated virus (raav) has demonstrated significant promise as a dna-delivery vector to treat serious human diseases. in this study, we constructed recombinant adeno-associated viruses coexpressing apoai and sr-bi successfully, the double gene mrna and protein were both strongly e ... | 2011 | 21431865 |
| retroviral-mediated transduction and clonal integration analysis of human hematopoietic stem and progenitor cells. | this chapter provides information on the methods used to introduce genes into human hematopoietic stem and progenitor cells, using moloney murine leukemia (momulv)-based retroviral vectors. momulv-based vectors have the ability to efficiently transfer genes into mammalian cells, leading to permanent integration of a single copy of the gene of interest into the cellular chromosomes. the technique of single-colony inverse [polymerase chain reaction (pcr) can be used to track individual descendants ... | 2002 | 21437813 |
| current status and future directions of gene and cell therapy for cystic fibrosis. | although the development of gene therapy for cystic fibrosis (cf) was high priority for many groups in academia and industry in the first 10-15 years after cloning the cystic fibrosis transmembrane conductance regulator (cftr) gene, more recently active research into cf gene therapy is only being performed by a small number of committed groups. however, despite the waning enthusiasm, which is largely due to the realization that gene transfer into lungs is more difficult than originally thought, ... | 2011 | 21443272 |
| preclinical evaluation of gene delivery methods for the treatment of loco-regional disease in breast cancer. | preclinical results with various gene therapy strategies indicate significant potential for new cancer treatments. however, many therapeutics fail at clinical trial, often due to differences in tissue physiology between animal models and humans, and tumor phenotype variation. clinical data relevant to treatment strategies may be generated prior to clinical trial through experimentation using intact patient tissue ex vivo. we developed a novel tumor slice model culture system that is universally ... | 2011 | 21444371 |
| high-efficiency transduction of liver cancer cells by recombinant adeno-associated virus serotype 3 vectors. | recombinant vectors based on a non-pathogenic human parvovirus, the adeno-associated virus 2 (aav2) have been developed, and are currently in use in a number of gene therapy clinical trials. more recently, a number of additional aav serotypes have also been isolated, which have been shown to exhibit selective tissue-tropism in various small and large animal models. of the 10 most commonly used aav serotypes, aav3 is by far the least efficient in transducing cells and tissues in vitro as well as ... | 2011 | 21445055 |
| release of bioactive adeno-associated virus from fibrin scaffolds: effects of fibrin glue concentrations. | fibrin glue (fg) is used in a variety of clinical applications and in the laboratory for localized and sustained release of factors potentially important for tissue engineering. however, the effect of different fibrinogen concentrations on fg scaffold delivery of bioactive adeno-associated viruses (aavs) has not been established. this study was performed to test the hypothesis that fg concentration alters aav release profiles, which affect aav bioavailability. gene transfer efficiency of aav-gfp ... | 2011 | 21449684 |
| a signaling cascade of nuclear calcium-creb-atf3 activated by synaptic nmda receptors defines a gene repression module that protects against extrasynaptic nmda receptor-induced neuronal cell death and ischemic brain damage. | synapse-to-nucleus signaling triggered by synaptic nmda receptors can lead to the buildup of a neuroprotective shield. nuclear calcium activating the camp response element binding protein (creb) plays a key role in neuroprotection acquired by synaptic activity. here we show that in mouse hippocampal neurons, the transcription factor atf3 (activating transcription factor 3) is a direct target of creb. induction of atf3 expression by creb in hippocampal neurons was initiated by calcium entry throu ... | 2011 | 21451036 |
| improvement of cardiac fibrosis in dystrophic mice by raav9-mediated microdystrophin transduction. | duchenne muscular dystrophy (dmd) is the most common form of the progressive muscular dystrophies characterized by defects of the dystrophin gene. although primarily characterized by degeneration of the limb muscles, cardiomyopathy is a major cause of death. therefore, the development of curative modalities such as gene therapy is imperative. we evaluated the cardiomyopathic features of mdx mice to observe improvements in response to intravenous administration of recombinant adeno-associated vir ... | 2011 | 21451578 |
| delivery of aav2/9-microdystrophin genes incorporating helix 1 of the coiled-coil motif in the c-terminal domain of dystrophin improves muscle pathology and restores the level of a1-syntrophin and a-dystrobrevin in skeletal muscles of mdx mice. | abstract duchenne muscular dystrophy is a severe x-linked inherited muscle wasting disorder caused by mutations in the dystrophin gene. adeno-associated virus (aav) vectors have been extensively used to deliver genes efficiently for dystrophin expression in skeletal muscles. to overcome limited packaging capacity of aav vectors (<5?kb), truncated recombinant microdystrophin genes with deletions of most of rod and carboxyl-terminal (ct) domains of dystrophin have been developed. we have previousl ... | 2011 | 21453126 |
| adeno-associated virus (aav) vectors in the cns. | adeno-associated virus (aav) vectors exhibit a number of properties that have made this vector system an excellent choice for both cns gene therapy and basic neurobiological investigations. in vivo, the preponderance of aav vector transduction occurs in neurons where it is possible to obtain long-term, stable gene expression with very little accompanying toxicity. promoter selection, however, significantly influences the pattern and longevity of neuronal transduction distinct from the tropism in ... | 2011 | 21453285 |
| aav-mediated gene targeting methods for human cells. | gene targeting with adeno-associated virus (aav) vectors has been demonstrated in multiple human cell types, with targeting frequencies ranging from 10(-5) to 10(-2) per infected cell. these targeting frequencies are 1-4 logs higher than those obtained by conventional transfection or electroporation approaches. a wide variety of different types of mutations can be introduced into chromosomal loci with high fidelity and without genotoxicity. here we provide a detailed protocol for gene targeting ... | 2011 | 21455185 |
| [characterization and application of adeno-associated virus serotype 8 as a gene transfer vector]. | | 2011 | 21462514 |
| conformational changes in adeno-associated virus type 1 induced by genome packaging. | adeno-associated virus (aav) is frequently used as a vector for gene therapy. the viral capsid consists of three structural proteins (vp1, vp2, and vp3) that have a common c-terminal core (vp3), with n-terminal extensions of increasing length in vp2 and vp1. the capsid encloses a single-stranded genome of up to 4.7 kb, which is packaged into empty capsids. the n-terminal extension of vp1 carries a phospholipase domain that becomes accessible during infection in the endosomal pathway. we have use ... | 2011 | 21463638 |
| secretory expression of par-4 sac-ha2tat following adeno-associated virus-mediated gene transfer induces apoptosis in hepg2 cells. | prostate apoptosis response-4 (par-4) is a tumor-suppressor protein that induces apoptosis in cancer cells, but not in normal cells. the cancer-specific pro-apoptotic action of par-4 is encoded in its centrally located sac domain. in this study, to further enhance the anti-cancer effect of par-4 in order to overcome the limitations of peptide therapy, a recombinant adeno-associated virus was constructed using the following strategies: the secretory expression of therapeutic peptide, a ha2tat-med ... | 2010 | 21472309 |
| enhanced real-time monitoring of adeno-associated virus trafficking by virus-quantum dot conjugates. | the unique spectral properties of semiconductor quantum dots (qds) enable long-term live-cell imaging and ultrasensitive detection of viral particles, which in turn can potentially provide a practical means for detailed analysis of the underlying molecular mechanisms of virus entry. in this study, we report a general method of labeling adeno-associated virus serotype 2 (aav2) with qds for enhanced visualization of the intracellular behavior of viruses in living target cells. it was found that th ... | 2011 | 21473596 |
| intravenous gene therapy with pim-1 via a cardiotropic viral vector halts the progression of diabetic cardiomyopathy through promotion of prosurvival signaling. | rationale: studies in transgenic mice showed the key role of (pim-1) (proviral integration site for moloney murine leukemia virus-1) in the control of cardiomyocyte function and viability. objective: we investigated whether pim-1 represents a novel mechanistic target for the cure of diabetic cardiomyopathy, a steadily increasing cause of nonischemic heart failure. methods and results: in streptozotocin-induced type 1 diabetic mice, pim-1 protein levels declined during progression of cardiomyopat ... | 2011 | 21474815 |
| comparative analysis of the effects of ultrasound-targeted microbubble destruction on recombinant adeno-associated virus- and plasmid-mediated transgene expression in human retinal pigment epithelium cells. | ultrasound-targeted microbubble destruction (utmd) has been utilized to deliver a drug/gene into cells in both in vitro and in vivo studies. this study was performed to investigate the feasibility of utmd-enhanced recombinant adeno-associated virus- (raav) and plasmid-mediated transfection into the human retinal pigment epithelium (rpe) cell line arpe-19. additionally, the transfection efficiency of raav and plasmid was compared in order to choose the appropriate gene vector or strategy to be us ... | 2009 | 21475924 |
| optimizing promoters for recombinant adeno-associated virus-mediated gene expression in the peripheral and central nervous system using self-complementary vectors. | abstract with the increased use of small self-complementary adeno-associated viral (aav) vectors, the design of compact promoters becomes critical for packaging and expressing larger transgenes under ubiquitous or cell-specific control. in a comparative study of commonly used 800-bp cytomegalovirus (cmv) and chicken β-actin (cba) promoters, we report significant differences in the patterns of cell-specific gene expression in the central and peripheral nervous systems. the cmv promoter provides h ... | 2011 | 21476867 |
| liver gene transfer with vectors based on adeno-associated virus 8 in non-human primates: impact of pre-existing immunity. | vectors based on the primate derived adeno-associated virus serotype 8 (aav8) are being evaluated in pre-clinical and clinical models. natural infections with related aavs activate memory b cells that produce antibodies capable of modulating the efficacy and safety of the vector. we have evaluated the biology of aav8 gene transfer in macaque liver with a focus on assessing the impact of pre-existing humoral immunity. twenty one macaques with varying levels of aav neutralizing antibody (nab) were ... | 2011 | 21476868 |
| efficient mutagenesis of the rhodopsin gene in rod photoreceptor neurons in mice. | dominant mutations in the rhodopsin gene, which is expressed in rod photoreceptor cells, are a major cause of the hereditary-blinding disease, autosomal dominant retinitis pigmentosa. therapeutic strategies designed to edit such mutations will likely depend on the introduction of double-strand breaks and their subsequent repair by homologous recombination or non-homologous end joining. at present, the break repair capabilities of mature neurons, in general, and rod cells, in particular, are unde ... | 2011 | 21478169 |
| selective optical control of synaptic transmission in the subcortical visual pathway by activation of viral vector-expressed halorhodopsin. | the superficial layer of the superior colliculus (ssc) receives visual inputs via two different pathways: from the retina and the primary visual cortex. however, the functional significance of each input for the operation of the ssc circuit remains to be identified. as a first step toward understanding the functional role of each of these inputs, we developed an optogenetic method to specifically suppress the synaptic transmission in the retino-tectal pathway. we introduced enhanced halorhodopsi ... | 2011 | 21483674 |
| preclinical differences of intravascular aav9 delivery to neurons and glia: a comparative study of adult mice and nonhuman primates. | other labs have previously reported the ability of adeno-associated virus serotype 9 (aav9) to cross the blood-brain barrier (bbb). in this report, we carefully characterized variables that might affect aav9's efficiency for central nervous system (cns) transduction in adult mice, including dose, vehicle composition, mannitol coadministration, and use of single-stranded versus self-complementary aav. we report that aav9 is able to transduce approximately twice as many neurons as astrocytes acros ... | 2011 | 21487395 |
| the influence of epileptic neuropathology and prior peripheral immunity on cns transduction by raav2 and raav5. | adeno-associated virus (aav) provides a promising platform for clinical treatment of neurological disorders owing to its established efficacy and lack of apparent pathogenicity. to use viral vectors in treating neurological disease, however, transduction must occur under neuropathological conditions. previous studies in rodents have shown that aav5 more efficiently transduces cells in the hippocampus and piriform cortex than aav2. using the kainic acid (ka) model of temporal lobe epilepsy and aa ... | 2011 | 21490684 |
| investigation of the peak action wavelength of light-activated gene transduction. | light-activated gene transduction (lagt) is an approach to localize gene therapy via preactivation of cells with uv light, which facilitates transduction by recombinant adeno-associated virus vectors. previous studies demonstrated that uvc induces lagt secondary to pyrimidine dimer formation, whereas uva induces lagt secondary to reactive-oxygen species (ros) generation. however, the empirical uvb boundary of these uv effects is unknown. thus, we aimed to define the action spectra for uv-induced ... | 2011 | 21490685 |
| limb-girdle muscular dystrophy 2a. | limb-girdle muscular dystrophy type 2a (lgmd2a) is caused by mutations in the gene capn3 located in the chromosome region 15q15.1-q21.1. to date more than 300 mutations have been described. this gene encodes for a 94-kda nonlysosomal calcium-dependent cysteine protease and its function in skeletal muscle is not fully understood. it seems that calpain-3 has an unusual zymogenic activation that involves, among other substrates, cytoskeletal proteins. calpain-3 is thought to interact with titin and ... | 2011 | 21496626 |
| therapeutic in vivo gene transfer for genetic disease using aav: progress and challenges. | in vivo gene replacement for the treatment of inherited disease is one of the most compelling concepts in modern medicine. adeno-associated virus (aav) vectors have been extensively used for this purpose and have shown therapeutic efficacy in a range of animal models. successful translation to the clinic was initially slow, but long-term expression of donated genes at therapeutic levels has now been achieved in patients with inherited retinal disorders and haemophilia b. recent exciting results ... | 2011 | 21499295 |
| [dendritic cells infected by recombinant adeno-associated virus with cea gene to induce antigen-specific ctl response to cd44(+)cd24(-/low) cells from mcf-7 breast cancer cell line]. | to infect dendritic cells (dc) by recombinant human adeno-associated virus (rh-aav) vector with cea gene to generate antigen-specific ctl cells in vitro and to assay the cea specific cytotoxic t lymphocyte(ctl), response to cd44(+)cd24(-/low) breast cancer stem cells. | 2011 | 21503107 |
| recombinant aav-directed gene therapy for type i glycogen storage diseases. | introduction: glycogen storage disease (gsd) type ia and ib are disorders of impaired glucose homeostasis affecting the liver and kidney. gsd-ib also affects neutrophils. current dietary therapies cannot prevent long-term complications. in animal studies, recombinant adeno-associated virus (raav) vector-mediated gene therapy can correct or minimize multiple aspects of the disorders, offering hope for human gene therapy. areas covered: a summary of recent progress in raav-mediated gene therapy fo ... | 2011 | 21504389 |
| adeno-associated virus serotype 9-mediated pulmonary transgene expression: effect of mouse strain, animal gender and lung inflammation. | gene therapy holds great potential for the treatment of various acquired and inherited pulmonary diseases. among various viral vectors, adeno-associated viral (aav) vectors have been most frequently used in different clinical trials of pulmonary gene therapy. in the present study, we examined the kinetics and duration of transgene expression, vector biodistribution and development of neutralizing antibodies (nab) in mice after pulmonary application of aav2/9 vector. the pulmonary route of applic ... | 2011 | 21512507 |
| increased expression of wild-type or a centronuclear myopathy mutant of dynamin 2 in skeletal muscle of adult mice leads to structural defects and muscle weakness. | dynamin 2 (dnm2) is a large gtpase implicated in many cellular functions, including cytoskeleton regulation and endocytosis. although ubiquitously expressed, dnm2 was found mutated in two genetic disorders affecting different tissues: autosomal dominant centronuclear myopathy (adcnm; skeletal muscle) and peripheral charcot-marie-tooth neuropathy (peripheral nerve). to gain insight into the function of dnm2 in skeletal muscle and the pathological mechanisms leading to adcnm, we introduced wild-ty ... | 2011 | 21514436 |
| pompe disease gene therapy. | pompe disease is an autosomal recessive metabolic myopathy caused by the deficiency of the lysosomal enzyme acid alpha-glucosidase and results in cellular lysosomal and cytoplasmic glycogen accumulation. a wide spectrum of disease exists from hypotonia and severe cardiac hypertrophy in the first few months of life due to severe mutations to a milder form with the onset of symptoms in adulthood. in either condition, the involvement of several systems leads to progressive weakness and disability. ... | 2011 | 21518733 |
| pnas plus: regeneration of axons in injured spinal cord by activation of bone morphogenetic protein/smad1 signaling pathway in adult neurons. | axon growth potential is highest in young neurons but diminishes with age, thus becoming a significant obstacle to axonal regeneration after injury in maturity. the mechanism for the decline is incompletely understood, and no effective clinical treatment is available to rekindle innate growth capability. here, we show that smad1-dependent bone morphogenetic protein (bmp) signaling is developmentally regulated and governs axonal growth in dorsal root ganglion (drg) neurons. down-regulation of the ... | 2011 | 21518886 |
| new adeno-associated virus strategies to support momentum in the clinic. | | 2011 | 21521072 |
| aav mediated gdnf secretion from retinal glia slows down retinal degeneration in a rat model of retinitis pigmentosa. | mutations in over 80 identified genes can induce apoptosis in photoreceptors, resulting in blindness with a prevalence of 1 in 3,000 individuals. this broad genetic heterogeneity of disease impacting a wide range of photoreceptor functions renders the design of gene-specific therapies for photoreceptor degeneration impractical and necessitates the development of mutation-independent treatments to slow photoreceptor cell death. one promising strategy for photoreceptor neuroprotection is neurotrop ... | 2011 | 21522134 |
| reprogramming virus nanoparticles to bind metal ions upon activation with heat. | we have reprogrammed the stimulus-responsive conformational change property of a virus nanoparticle (vnp) to enable the surface exposure of metal binding motifs upon activation with heat. the vnp is based on the widely investigated adeno-associated virus (aav). an intrinsic bioactive functionality of aav was genetically replaced with a hexahistidine (his) tag. the peptide domain with the inserted his tag is normally inaccessible. upon external stimulation with heat, the vnp undergoes a conformat ... | 2011 | 21528841 |
| radioprotective gene therapy. | introduction: radiation-induced myelosuppression or mucositis can limit the effectiveness of radiotherapy by requiring dose reduction or delaying treatment of tumour patients. the transfer of a radioprotective gene into normal tissue cells would provide the opportunity to reduce the risks associated with haematopoietic or intestinal toxicity after irradiation. areas covered: several potentially radioprotective genes like multidrug resistance 1 (mdr1), snail homolog 2 (snai2), and superoxide dism ... | 2011 | 21529309 |
| therapeutic effect of anti-amyloid β peptide single-chain antibody gene mediated by adeno-associated virus on animal alzheimer's disease. | to investigate the therapeutic effect of recombinant adeno-associated virus carrying anti-amyloid β peptide single-chain antibody gene on alzheimer's disease (ad) in animal models. | 2011 | 21529442 |
| adeno-associated virus loaded microbubbles as a tool for targeted gene delivery. | | 2010 | 21529628 |
| viral vector tropism for supporting cells in the developing murine cochlea. | gene-based therapeutics are being developed as novel treatments for genetic hearing loss. one roadblock to effective gene therapy is the identification of vectors which will safely deliver therapeutics to targeted cells. the cellular heterogeneity that exists within the cochlea makes viral tropism a vital consideration for effective inner ear gene therapy. there are compelling reasons to identify a viral vector with tropism for organ of corti supporting cells. supporting cells are the primary ex ... | 2011 | 21530627 |
| knockdown of npy expression in the dorsomedial hypothalamus promotes development of brown adipocytes and prevents diet-induced obesity. | hypothalamic neuropeptide y (npy) has been implicated in control of energy balance, but the physiological importance of npy in the dorsomedial hypothalamus (dmh) remains unclear. here we report that knockdown of npy expression in the dmh by adeno-associated virus-mediated rnai reduced fat depots in rats fed regular chow and ameliorated high-fat diet-induced hyperphagia and obesity. dmh npy knockdown resulted in development of brown adipocytes in inguinal white adipose tissue through the sympathe ... | 2011 | 21531339 |
| large-scale recombinant adeno-associated virus production. | since recombinant adeno-associated virus (raav) was first described as a potential mammalian cell transducing system, frequent reports purportedly solving the problems of scalable production have appeared. yet few of these processes have enabled the development of robust and economical raav production. two production platforms have emerged that have gained broad support for producing both research and clinical grade vectors. these processes differ fundamentally in several aspects. one approach i ... | 2011 | 21531790 |
| efficacious and safe tissue-selective controlled gene therapy approaches for the cornea. | untargeted and uncontrolled gene delivery is a major cause of gene therapy failure. this study aimed to define efficient and safe tissue-selective targeted gene therapy approaches for delivering genes into keratocytes of the cornea in vivo using a normal or diseased rabbit model. new zealand white rabbits, adeno-associated virus serotype 5 (aav5), and a minimally invasive hair-dryer based vector-delivery technique were used. fifty microliters of aav5 titer (6.5×10(12) vg/ml) expressing green flu ... | 2011 | 21533273 |
| vegf-c attenuates joint damage in chronic inflammatory arthritis by accelerating local lymphatic drainage. | objective.: to investigate if enhancement of joint lymphangiogenesis by injecting vegf-c adeno-associated virus (aav) into joints has therapeutic efficacy in chronic inflammatory arthritis in mice. methods.: tnf transgenic (tnf-tg) mice were used as a model of chronic inflammatory arthritis. human vegf-c was cloned into an aav expression vector to generate aav-vegf-c. aav-vegf-c or control aav-luc was injected into joints of tnf-tg mice. mri and lymphatic imaging were used during the 4-months fo ... | 2011 | 21538325 |
| direct injection into the dorsal root ganglion: technical, behavioral, and histological observations. | direct injection of agents into the dorsal root ganglia (drgs) offers the opportunity to manipulate sensory neuron function at a segmental level to explore pathophysiology of painful conditions. however, there is no described method that has been validated in detail for such injections in adult rats. we have found that 2μl of dye injected through a pulled glass pipette directly into the distal drg, exposed by a minimal foraminotomy, produces complete filling of the drg with limited extension int ... | 2011 | 21540055 |
| cardiac gene transfer of shrna directed against phospholamban effectively knocks down gene expression but causes cellular toxicity in canines. | derangements in calcium cycling have been described in failing hearts of several etiologies, and preclinical studies have suggested that therapies aimed at correcting this defect can lead to improvements in cardiac function and survival. one strategy to improve calcium cycling would be to inhibit phospholamban (plb), the negative regulator of serca2a that is upregulated in failing hearts. the goal of this study was to evaluate the safety and efficacy of using aav-mediated cardiac gene transfer o ... | 2011 | 21542669 |
| systemic aav-mediated gene therapy preserves retinal ganglion cells and visual function in dba/2j glaucomatous mice. | a slow progressive death of neurons is the hallmark of neurodegenerative diseases, such as glaucoma. a recently described therapeutic candidate, erythropoietin (epo), has shown promise in many models of these diseases; however it also causes polycythemia, a potentially lethal side effect. we have developed a novel mutant form of epo that is neuroprotective but no longer erythropoietic by altering a single amino acid (arginine to glutamate at position 76, r76e). we hypothesized that a single intr ... | 2011 | 21542676 |
| long-term in vivo resistin overexpression induces myocardial dysfunction and remodeling in rats. | we have previously reported that resistin induces hypertrophy and impairs contractility in isolated rat cardiomyocytes. to examine the long-term cardiovascular effects of resistin, we induced in vivo overexpression of resistin using adeno-associated virus serotype 9 injected by tail vein in rats and compared to control animals. ten weeks after viral injection, overexpression of resistin was associated with increased ratio of left ventricular (lv) weight/body weight, increased end-systolic lv vol ... | 2011 | 21549710 |
| significant inhibition of corneal scarring in vivo with tissue-selective targeted aav5 decorin gene therapy. | purpose: this study tested a hypothesis that tissue-selective targeted decorin gene therapy delivered to the stroma with adeno-associated virus serotype 5 (aav5) inhibits corneal fibrosis in vivo without significant side effects. methods: in vivo rabbit model of corneal fibrosis was used. targeted decorin gene therapy in the rabbit cornea was delivered by a single topical application of aav5 (100 μl; 6.5x10(12) vg/ml) onto the bare stroma for two minutes. the levels of corneal fibrosis were dete ... | 2011 | 21551414 |
| quantifying transduction efficiencies of unmodified and tyrosine capsid mutant aav vectors in vitro using two ocular cell lines. | with the increasing number of retinal gene-based therapies and therapeutic constructs, in vitro bioassays characterizing vector transduction efficiency and quality are becoming increasingly important. currently, in vitro assays quantifying vector transduction efficiency are performed predominantly for non-ocular tissues. a human retinal pigment epithelial cell line (arpe19) and a mouse cone photoreceptor cell line, 661w, have been well characterized and are used for many retinal metabolism and b ... | 2011 | 21552473 |
| a non membrane-targeted human soluble cd59 attenuates choroidal neovascularization in a model of age related macular degeneration. | age related macular degeneration (amd) is the most common cause of blindness amongst the elderly. approximately 10% of amd patients suffer from an advanced form of amd characterized by choroidal neovascularization (cnv). recent evidence implicates a significant role for complement in the pathogenesis of amd. activation of complement terminates in the incorporation of the membrane attack complex (mac) in biological membranes and subsequent cell lysis. elevated levels of mac have been documented o ... | 2011 | 21552568 |
| adeno-associated virus-mediated human acidic fibroblast growth factor expression promotes functional recovery of spinal cord-contused rats. | background: following spinal cord injury, delivery of neurotrophic factors to the injured spinal cord has been shown to promote axonal regeneration and functional recovery. in previous studies, we showed that acidic fibroblast growth factor (afgf) is a potent neurotrophic factor that promotes the regeneration of axotomized spinal cord or dorsal root ganglion neurons. methods: we constructed a recombinant adeno-associated virus (aav) vector to express human afgf and evaluated afgf expression and ... | 2011 | 21557400 |
| immune responses to aav in clinical trials. | findings in the first clinical trial in which an adeno-associated virus (aav) vector was introduced into the liver of human subjects highlighted an issue not previously identified in animal studies. upon aav gene transfer to liver, two subjects developed transient elevation of liver enzymes, likely as a consequence of immune rejection of transduced hepatocytes mediated by aav capsid-specific cd8+ t cells. studies in healthy donors showed that humans carry a population of antigen-specific memory ... | 2011 | 21557723 |
| rosuvastatin attenuates the elevation in blood pressure induced by overexpression of human c-reactive protein. | c-reactive protein (crp) has been shown to function as an inflammatory factor to induce endothelial dysfunction and hypertension in rats. the anti-inflammatory effects of statins suggest that they may attenuate crp-induced endothelial dysfunction and hypertension in sprague-dawley rats. male sprague-dawley rats were injected with an adeno-associated virus (aav) to induce overexpression of human crp (aav-hcrp) or green fluorescent protein (gfp) control (aav-gfp). at 2 months after injection, rats ... | 2011 | 21562509 |
| long-term vegf-a expression promotes aberrant angiogenesis and fibrosis in skeletal muscle. | vascular endothelial growth factor a (vegf-a) induces strong angiogenesis and it has been widely used in proangiogenic gene therapy studies. however, little is known about long-term effects of vegf-a expression in skeletal muscle. here the long- term effects of adeno-associated virus (aav) encoding human vegf-a(165) (aav-vegf-a) gene transfer in normal and ischemic rabbit hindlimb skeletal muscles were studied. aav-lacz was used as a control. in one-year follow-up, a remarkable increase in skele ... | 2011 | 21562595 |
| extending the activities of human nucleus pulposus cells with recombinant adeno-associated virus vector-mediated human telomerase reverse transcriptase(raav-htert) gene transfer. | even though human nucleus pulposus (hnp) cells have a limited replicative lifespan in vitro culture, the lifespan of ovine nucleus pulposus cells transferred by lipofectamine with human telomerase reverse transcriptase (htert) gene could be prolonged. but the lipofectamine is a transient expression system with an expressed duration of htert for less than 73 days. here we present a viral vector system to investigate whether recombinant adeno-associated virus vector-mediated htert gene ( raav-hter ... | 2011 | 21563861 |
| transduction of human adipose-derived mesenchymal stem cells by recombinant adeno-associated virus vectors. | human adipose-derived stem cells (ascs) are attractive targets for genetic manipulation and cellular therapies. however current methods of gene transfer are limited by lack of efficiency, toxicity, or safety concerns. recombinant adeno-associated virus (raav) has been extensively assessed as a gene therapy vector and has an excellent safety profile. this study reports the efficient transduction of well characterized, homogeneous cultures of human ascs by raav serotypes 2, 5, and 6. transduction ... | 2011 | 21563982 |
| percutaneous transendocardial delivery of aav6 results in highly efficient and global cardiac gene transfer in rhesus macaques. | heart disease is the leading cause of morbidity and mortality, and cardiac gene transfer has potential as a novel therapeutic approach. we previously demonstrated safe and efficient gene transfer to the canine heart using a percutaneous transendocardial injection procedure to deliver self-complementary (sc) aav6 vector. in the present study, we proceed with our vertical translation study to evaluate cardiac gene transfer in nonhuman primates (nhp). we screened approximately 30 adult male rhesus ... | 2011 | 21563985 |
| [mechanism of dads in the bystander effect of hsv-tk/gcv suicide gene therapy system in lens epithelial cells.] | objective to investigate the mechanism and effect of diallyl disulfide (dads) on the bystander effect of herpes simplex virus kinase/ganciclovir (hsv-tk/gcv) suicide gene therapy system which was mediated by recombinant adeno-associated virus (raav) in lens epithelial cells. methods immunohistochemistry method was used to determine the distribution of connexin 43 (cx43) protein in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by hsv-tk suicide gene. cx43 prot ... | 2011 | 21566285 |
| single intra-articular injection of adeno-associated virus results in stable and controllable in vivo transgene expression in normal rat knees. | objective: to test the hypothesis that in vivo transgene expression mediated by single intra-articular injection of adeno-associated virus serotype 2 (aav2) persists within intra-articular tissues 1 year post-injection and can be externally controlled using an aav2-based tetracycline-inducible gene regulation system containing the tetracycline response element (tre) promoter. methods: sprague dawley rats received intra-articular injections of aav2-cytomegalovirus (cmv)-enhanced green fluorescent ... | 2011 | 21571082 |
| [inhibition of infectious bursal disease virus replication in chicken embryos by mirnas delivered by recombinant avian adeno-associated viral vector]. | we studied the inhibition of infectious bursal disease virus (ibdv) replication in chicken embryos by recombinant avian adeno-associated virus (aaav)-delivered vp1- and vp2-specific micrornas (mirnas). | 2011 | 21574388 |
| unique characteristics of aav1, 2, and 5 viral entry, intracellular trafficking, and nuclear import define transduction efficiency in hela cells. | biological differences between recombinant adeno-associated virus (raav) serotypes define their efficiencies in expressing a transgene in a particular target cell. few studies have directly compared how differences in viral entry, intracellular trafficking, and nuclear import of raav serotypes influence the effectiveness of transduction in the same cell type. we evaluated these characteristics for three raav serotypes in hela cells, using biochemical techniques and fluorescence-based detection o ... | 2011 | 21574868 |
| the aav9 receptor and its modification to improve in vivo lung gene transfer in mice. | vectors based on adeno-associated virus (aav) serotype 9 are candidates for in vivo gene delivery to many organs, but the receptor(s) mediating these tropisms have yet to be defined. we evaluated aav9 uptake by glycans with terminal sialic acids (sas), a common mode of cellular entry for viruses. we found, however, that aav9 binding increased when terminal sa was enzymatically removed, suggesting that galactose, which is the most commonly observed penultimate monosaccharide to sa, may mediate aa ... | 2011 | 21576824 |
| capsid-specific t-cell responses to natural infections with adeno-associated viruses in humans differ from those of nonhuman primates. | hepatic adeno-associated virus serotype 2 (aav2)-mediated gene transfer failed to achieve sustained transgene product expression in human subjects. we formulated the hypothesis that rejection of aav-transduced hepatocytes is caused by aav capsid-specific cd8(+) t cells that become reactivated upon gene transfer. although this hypothesis was compatible with clinical data, which showed a rise in circulating aav capsid-specific t cells following injection of aav vectors, it did not explain that aav ... | 2011 | 21587208 |
| effect of soluble icam-1 on a sjögren's syndrome-like phenotype in nod mice is disease stage dependent. | intercellular adhesion molecule-1 (icam-1) is involved in migration and co-stimulation of t and b cells. membrane bound icam-1 is over expressed in the salivary glands (sg) of sjögren's syndrome (ss) patients and has therefore been proposed as a potential therapeutic target. to test the utility of icam-1 as a therapeutic target, we used local gene therapy in non obese diabetic (nod) mice to express soluble (s)icam-1 to compete with membrane bound icam-1 for binding with its receptor. therapy was ... | 2011 | 21589878 |
| overview of current scalable methods for purification of viral vectors. | as a result of the growing interest in the use of viruses for gene therapy and vaccines, many virus-based products are being developed. the manufacturing of viruses poses new challenges for process developers and regulating authorities that need to be addressed to ensure quality, efficacy, and safety of the final product. the design of suitable purification strategies will depend on a multitude of variables including the vector production system and the nature of the virus. in this chapter, we p ... | 2011 | 21590394 |
| adeno-associated viruses. | adeno-associated virus (aav) vectors have evolved over the past decade as a particularly useful gene -vector for in vivo applications. in contrast to oncoretro- and lentiviral vectors, this vector stays essentially episomal after gene transfer, making it safer because of the absence of insertional mutagenesis. aav's non-pathogenicity is a further advantage. for decades, this vector could only be produced at a small scale for research purposes and, eventually, used at very small doses for clinica ... | 2011 | 21590399 |