| aav vectors transduce hepatocytes in vivo as efficiently in cirrhotic as in healthy rat livers. | in liver cirrhosis, abnormal liver architecture impairs efficient transduction of hepatocytes with large viral vectors such as adenoviruses. here we evaluated the ability of adeno-associated virus (aav) vectors, small viral vectors, to transduce normal and cirrhotic rat livers. using aav serotype-1 (aav1) encoding luciferase (aav1luc) we analyzed luciferase expression with a ccd camera. aav1luc was injected through the hepatic artery (intra-arterial (ia)), the portal vein (intra-portal (ip)), di ... | 2011 | 21850051 |
| authentically phosphorylated α-synuclein at ser129 accelerates neurodegeneration in a rat model of familial parkinson's disease. | parkinson's disease (pd) is characterized by the loss of dopaminergic neurons in the substantia nigra (sn) and the appearance of fibrillar aggregates of insoluble α-synuclein (α-syn) called lewy bodies (lbs). approximately 90% of α-syn deposited in lbs is phosphorylated at serine 129 (ser129). in contrast, only 4% of total α-syn is phosphorylated in normal brain, suggesting that accumulation of ser129-phosphorylated α-syn is involved in the pathogenesis of pd. however, the role of ser129 phospho ... | 2011 | 22090514 |
| dna-binding activity of aav rep is required for itr-dependent complex formation with hsv icp8. | hsv helper functions for aav replication comprise hsv icp8 and helicase-primase ul5/52/8. here we show that n-terminal amino acids of aav rep78 that contact the rep-binding site within the aav-itr are required for ternary complex formation with icp8 on aav ssdna in vitro and for colocalization in nuclear replication domains in vivo. our data suggest that hsv-dependent aav replication is initiated by rep contacting the aav-itr and by cooperative binding of icp8 on aav ssdna. | 2011 | 22205745 |
| Differential virus restriction patterns of rhesus macaque and human APOBEC3A: implications for lentivirus evolution. | The human apolipoprotein B mRNA editing enzyme catalytic peptide-like 3 (APOBEC3; A3) family of proteins (A3A-H) are known to restrict various retroviruses and retroelements, but the full complement of rhesus macaque A3 proteins remains unclear. We report the isolation and characterization of the hA3A homologue from rhesus macaques (rhA3A) and show that the rhesus macaque and human A3 genes are orthologous. RhA3A is expressed at high levels in activated CD4+ T cells, is widely expressed in macaq ... | 2011 | 21868050 |
| AAV-Mediated Gene Transfer of Human X-Linked Inhibitor of Apoptosis Protects against Oxidative Cell Death in Human RPE Cells. | Purpose. To determine whether human X-linked inhibitor of apoptosis (XIAP) enhances the survival of cultured human retinal pigment epithelial cells exposed to H(2)O(2). Methods. ARPE-19 cells were exposed to H(2)O(2) to induce oxidative cell death. Intracellular reactive oxygen species (ROS) were measured using 2',7'-dichlorofluorescein diacetate. MTT assay was performed to quantify mitochondrial stress. Cell apoptosis was determined by TUNEL assay. Human XIAP was delivered with bicistronic exp ... | 2011 | 22039248 |
| influence of viral vector-mediated delivery of superoxide dismutase and catalase to the hippocampus on spatial learning and memory during aging. | abstract aims: studies employing transgenic mice indicate that overexpression of superoxide dismutase 1 (sod1) improves memory during aging. it is unclear whether the improvement is due to a lifetime of overexpression, decreasing the accumulation of oxidized molecules, or if increasing antioxidant enzymes in older animals could reduce oxidative damage and improve cognitive function. we used adeno-associated virus to deliver antioxidant enzymes (sod1, sod2, catalase [cat], and sod1+cat) to the ... | 2011 | 21942371 |
| transduction, tropism, and biodistribution of aav vectors in the lacrimal gland. | purpose. the lacrimal gland (lg) delivers defensive and metabolic factors to the ocular surface. these functions may be disrupted in several diseases, and for most of them there is no cure. the aim of this study is to investigate conditions and limitations for using adeno-associated virus (aav) vectors as gene transfer agents to lg. methods. eight-week-old balb/c mice were used to investigate route, gene expression, and time course of aav gene vector transfer to lg. aav vectors encoding firefly ... | 2011 | 22110082 |
| [dna damage caused by suicide gene therapy system under tet-on regulation in breast cancer cells]. | to determine the effect and molecular mechanism of dna damage caused by suicide gene therapy system hsv-tk/gcv under tet-on regulation in human breast cancer cell line mcf-7 infected by recombinant adeno-associated virus (raav). | 2011 | 21946198 |
| advances in gene delivery systems. | the transfer of genes into cells, both in vitro and in vivo, is critical for studying gene function and conducting gene therapy. methods that utilize viral and nonviral vectors, as well as physical approaches, have been explored. viral vector-mediated gene transfer employs replication-deficient viruses such as retro-virus, adenovirus, adeno-associated virus and herpes simplex virus. a major advantage of viral vectors is their high gene delivery efficiency. the nonviral vectors developed so far i ... | 2011 | 22200988 |
| an experimental rat model of sporadic alzheimer's disease and rescue of cognitive impairment with a neurotrophic peptide. | alzheimer's disease (ad) is multifactorial and, to date, no single cause of the sporadic form of this disease, which accounts for over 99% of the cases, has been established. in ad brain, protein phosphatase-2a (pp2a) activity is known to be compromised due to the cleavage and translocation of its potent endogenous inhibitor, i (2) (pp2a) , from the neuronal nucleus to the cytoplasm. here, we show that adeno- ... | 2012 | 22083255 |
| Comparison of AAV2 and AAV5 in gene transfer in the injured spinal cord of mice. | Recombinant adeno-associated virus (AAV) vectors are promising tools for gene therapy. In spinal cord injury where extensive damage occurs, vectors with high diffusion and transduction abilities are required. We compared the diffusion capacity and transduction efficiency of AAV2 and AAV5 vectors using a mouse spinal cord injury model. Our study demonstrates that AAV5 is more effective than AAV2 for delivering genes into the injured spinal cord tissue. AAV5 diffused 6.9 mm from the injection site ... | 2011 | 21879483 |
| Sustained induction of neuronal addition to the adult rat neostriatum by AAV4-delivered noggin and BDNF. | Intraventricular ependymal infection by adenoviruses expressing brain-derived neurotrophic factor (BDNF) and noggin is sufficient to induce the heterotopic recruitment of new medium spiny neurons to the adult neostriatum, from endogenous subependymal neural progenitor cells. This approach was found to slow disease progression and extend survival in an R6/2 mouse model of Huntington's disease (HD). However, the practical therapeutic value of this strategy is limited by the transient expression an ... | 2011 | 21918547 |
| Adeno-associated virus biology. | Adeno-associated virus (AAV) was first discovered as a contaminant of adenovirus stocks in the 1960s. The development of recombinant AAV vectors (rAAV) was facilitated by early studies that generated infectious molecular clones, determined the sequence of the genome, and defined the genetic elements of the virus. The refinement of methods and protocols for the production and application of rAAV vectors has come from years of studies that explored the basic biology of this virus and its interacti ... | 2011 | 22034024 |
| Phase 1 Gene Therapy for Duchenne Muscular Dystrophy Using a Translational Optimized AAV Vector. | Efficient and widespread gene transfer is required for successful treatment of Duchenne muscular dystrophy (DMD). Here, we performed the first clinical trial using a chimeric adeno-associated virus (AAV) capsid variant (designated AAV2.5) derived from a rational design strategy. AAV2.5 was generated from the AAV2 capsid with five mutations from AAV1. The novel chimeric vector combines the improved muscle transduction capacity of AAV1 with reduced antigenic crossreactivity against both parental s ... | 2011 | 22068425 |
| Formation of AAV single stranded DNA genome from a circular plasmid in Saccharomyces cerevisiae. | Adeno-associated virus (AAV)-based vectors are promising tools for targeted transfer in gene therapy studies. Many efforts have been accomplished to improve production and purification methods. We thought to develop a simple eukaryotic system allowing AAV replication which could provide an excellent opportunity for studying AAV biology and, more importantly, for AAV vector production. It has been shown that yeast Saccharomyces cerevisiae is able to replicate and form the capsid of many viruses. ... | 2011 | 21853137 |
| quantitative 3d tracing of gene-delivery viral vectors in human cells and animal tissues. | trafficking through a variety of cellular structures and organelles is essential for the interaction between gene-delivery vectors (i.e., adeno-associated virus (aav) and liposomes) and host cells/tissues. here, we present a method of computer-assisted quantitative 3d biodistribution microscopy that samples the whole population of fluorescently-labeled vectors and document their trafficking routes. using aav as a working model, we first experimentally defined numerical parameters for the singula ... | 2011 | 22108857 |
| novel random peptide libraries displayed on aav serotype 9 for selection of endothelial cell-directed gene transfer vectors. | we have demonstrated the potential of random peptide libraries displayed on adeno-associated virus (aav)2 to select for aav2 vectors with improved efficiency for cell type-directed gene transfer. aav9, however, may have advantages over aav2 because of a lower prevalence of neutralizing antibodies in humans and more efficient gene transfer in vivo. here we provide evidence that random peptide libraries can be displayed on aav9 and can be utilized to select for aav9 capsids redirected to the cell ... | 2011 | 21956692 |
| development and validation of novel aav2 random libraries displaying peptides of diverse lengths and at diverse capsid positions. | libraries based on the insertion of random peptide ligands into the capsid of adeno-associated virus type 2 (aav2) have been widely used to improve the efficiency and selectivity of the aav vector system. however, so far only libraries of 7-mer peptide ligands have been inserted at one well-characterized capsid position. here, we expanded the combinatorial aav2 display system to a panel of novel aav libraries, displaying peptides of 5, 7, 12, 19 or 26 amino acids in length at capsid position 588 ... | 2011 | 22171602 |
| suppression of endogenous pparγ increases vulnerability to methamphetamine-induced injury in mouse nigrostriatal dopaminergic pathway. | rationale: methamphetamine is a commonly abused drug and dopaminergic neurotoxin. repeated administration of high doses of methamphetamine induces programmed cell death, suppression of dopamine release, and reduction in locomotor activity. previous studies have shown that pretreatment with peroxisome proliferator-activated receptor gamma (pparγ) agonist reduced methamphetamine-induced neurodegeneration. objectives: ... | 2011 | 22160138 |
| mitotic catastrophe occurs in the absence of apoptosis in p53-null cells with a defective g1 checkpoint. | cell death occurring during mitosis, or mitotic catastrophe, often takes place in conjunction with apoptosis, but the conditions in which mitotic catastrophe may exhibit features of programmed cell death are still unclear. in the work presented here, we studied mitotic cell death by making use of a uv-inactivated parvovirus (adeno-associated virus; aav) that has been shown to induce a dna damage response and subsequent death of p53-defective cells in mitosis, without affecting the integrity of t ... | 2011 | 21853057 |
| Structural studies of adeno-associated virus serotype 8 capsid transitions associated with endosomal trafficking. | The single-stranded DNA (ssDNA) parvoviruses enter host cells through receptor-mediated endocytosis, and infection depends on processing in the early to late endosome as well as in the lysosome prior to nuclear entry for replication. However, the mechanisms of capsid endosomal processing, including the effects of low pH, are poorly understood. To gain insight into the structural transitions required for this essential step in infection, the crystal structures of empty and green fluorescent prote ... | 2011 | 21900159 |
| Targeted decorin gene therapy delivered with adeno-associated virus effectively retards corneal neovascularization in vivo. | Decorin, small leucine-rich proteoglycan, has been shown to modulate angiogenesis in nonocular tissues. This study tested a hypothesis that tissue-selective targeted decorin gene therapy delivered to the rabbit stroma with adeno-associated virus serotype 5 (AAV5) impedes corneal neovascularization (CNV) in vivo without significant side effects. An established rabbit CNV model was used. Targeted decorin gene therapy in the rabbit stroma was delivered with a single topical AAV5 titer (100 µl; 5×10 ... | 2011 | 22039486 |
| Injection of AAV2-BMP2 and AAV2-TIMP1 into the nucleus pulposus slows the course of intervertebral disc degeneration in an in vivo rabbit model. | BACKGROUND CONTEXT: Intervertebral disc degeneration (IDD) is a common cause of back pain. Patients who fail conservative management may face the morbidity of surgery. Alternative treatment modalities could have a significant impact on disease progression and patients' quality of life. PURPOSE: To determine if the injection of a virus vector carrying a therapeutic gene directly into the nucleus pulposus improves the course of IDD. STUDY DESIGN: Prospective randomized controlled animal study. MET ... | 2011 | 22023960 |
| Long-term Efficacy Following Readministration of an AAV Vector in Dogs With Glycogen Storage Disease Type Ia. | Glycogen storage disease type Ia (GSD-Ia) is the inherited deficiency of glucose-6-phosphatase (G6Pase), primarily found in liver and kidney, which causes life-threatening hypoglycemia. Dogs with GSD-Ia were treated with double-stranded adeno-associated virus (AAV) vectors encoding human G6Pase. Administration of an AAV9 pseudotyped (AAV2/9) vector to seven consecutive GSD-Ia neonates prevented hypoglycemia during fasting for up to eight hours; however, efficacy eventually waned between 2 and 30 ... | 2011 | 22185325 |
| The adeno-associated virus-mediated HSV-TK/GCV suicide system: a potential strategy for the treatment of bladder carcinoma. | Novel treatment strategies such as gene therapy are warranted in view of the failure of current treatment approaches to cure a high percentage of patients with advanced bladder cancers. The emergence of cancer gene therapy potentially offers a number of exciting treatments. The majority of approaches involve strategies to suppress the function of activated oncogenes to restore the expression of functional tumour suppressor genes or to initiate tumour self-destruction. One gene therapy approach a ... | 2011 | 22011935 |
| The innate antiviral factor APOBEC3G targets replication of measles, mumps, and respiratory syncytial virus. | The cytidine deaminase APOBEC3G (apolipoprotein B mRNA-editing enzyme-catalytic polypeptide 3G; A3G) exerts antiviral activity against retroviruses, hepatitis B virus, adeno-associated virus, and transposable elements. We assessed whether the negative-strand (-)RNA viruses measles (MV), mumps (MuV), and respiratory syncytial virus (RSV) might be affected by A3G, and found that their infectivity was reduced by 1-2 logs (90-99%) in A3G over-expressing Vero cells, and in T cell lines expressing A3G ... | 2011 | 22170635 |
| concerted nicking of donor and chromosomal acceptor dna promotes homology-directed gene targeting in human cells. | the exchange of genetic information between donor and acceptor dna molecules by homologous recombination (hr) depends on the cleavage of phosphodiester bonds. although double-stranded and single-stranded dna breaks (ssbs) have both been invoked as triggers of hr, until very recently the focus has been primarily on the former type of dna lesions mainly due to the paucity of ssb-based recombination models. here, to investigate the role of nicked dna molecules as hr-initiating substrates in human s ... | 2011 | 22189101 |
| Single-Chain Fv-Based Anti-HIV Proteins: Potential and Limitations. | The existence of very potent, broadly neutralizing antibodies against human immunodeficiency virus type 1 (HIV-1) offers the potential for prophylaxis against HIV-1 infection by passive immunization or gene therapy. Both routes permit the delivery of modified forms of IgGs. Smaller reagents are favored when considering ease of tissue penetration and the limited capacities of gene therapy vectors. Immunoadhesin (single-chain fragment variable [scFv]-Fc) forms of IgGs are one class of relatively s ... | 2012 | 22013046 |
| Novel adeno-associated viral vectors for retinal gene therapy. | Vectors derived from adeno-associated virus (AAV) are currently the most promising vehicles for therapeutic gene delivery to the retina. Recently, subretinal administration of AAV2 has been demonstrated to be safe and effective in patients with a rare form of inherited childhood blindness, suggesting that AAV-mediated retinal gene therapy may be successfully extended to other blinding conditions. This is further supported by the great versatility of AAV as a vector platform as there are a large ... | 2011 | 21993172 |
| IGF-1 delivery to CNS attenuates motor neuron cell death but does not improve motor function in type III SMA mice. | The efficacy of administering a recombinant adeno-associated virus (AAV) vector encoding human IGF-1 (AAV2/1-hIGF-1) into the deep cerebellar nucleus (DCN) of a type III SMA mouse model was evaluated. High levels of IGF-1 transcripts and protein were detected in the spinal cord at 2 months post-injection demonstrating that axonal connections between the cerebellum and spinal cord were able to act as conduits for the viral vector and protein to the spinal cord. Mice treated with AAV2/1-hIGF-1 and ... | 2012 | 21884794 |
| Viral-mediated overexpression of mutant huntingtin to model HD in various species. | Huntington's disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expansion of CAG repeats in the huntingtin (Htt) gene. Despite intensive efforts devoted to investigating the mechanisms of its pathogenesis, effective treatments for this devastating disease remain unavailable. The lack of suitable models recapitulating the entire spectrum of the degenerative process has severely hindered the identification and validation of therapeutic strategies. The discovery that the ... | 2011 | 21889981 |
| The recombinant Lectin-like domain of thrombomodulin inhibits angiogenesis through interaction with Lewis Y antigen. | Lewis Y antigen (LeY) is a cell surface tetrasaccharide that participates in angiogenesis. Recently, we demonstrated that LeY is a specific ligand of recombinant lectin-like domain of thrombomodulin (TM). However, the biological function of interaction between LeY and TM in endothelial cells has never been investigated. Therefore, the role of LeY in tube formation and the role of the recombinant lectin-like domain of TM -TM domain 1 (rTMD1) - in antiangiogenesis were investigated. The recombinan ... | 2011 | 22101897 |
| AAV6-ßARKct gene delivery mediated by molecular cardiac surgery with recirculating delivery (MCARD) in sheep results in robust gene expression and increased adrenergic reserve. | OBJECTIVE: Genetic modulation of heart function is a novel therapeutic strategy. We investigated the effect of molecular cardiac surgery with recirculating delivery (MCARD)-mediated carboxyl-terminus of the ß-adrenergic receptor kinase (ßARKct) gene transfer on cardiac mechanoenergetics and ß-adrenoreceptor (ßAR) signaling. METHODS: After baseline measurements, sheep underwent MCARD-mediated delivery of 10(14) genome copies of self-complimentary adeno-associated virus (scAAV6)-ßARKct. Four and 8 ... | 2011 | 22143102 |
| spinal delivery of aav vector restores enzyme activity and increases ventilation in pompe mice. | pompe disease is a form of muscular dystrophy due to lysosomal storage of glycogen caused by deficiency of acid α-glucosidase (gaa). respiratory failure in pompe disease has been attributed to respiratory muscle dysfunction. however, evaluation of spinal tissue from pompe patients and animal models indicates glycogen accumulation and lower motoneuron pathology. we hypothesized that restoring gaa enzyme activity in the region of the phrenic motor nucleus could lead to improved breathing in a muri ... | 2011 | 22008916 |
| Production of recombinant adeno-associated viral vectors and use in in vitro and in vivo administration. | Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit describes efficient methods to generate high-titer, research-grade, adenovirus-free recombinant single-stranded and self-complementary adeno-associated virus in various serotypes, along with methods to quantify the viral vectors. Two detailed metho ... | 2011 | 21971848 |
| Versatile and Efficient Genome Editing in Human Cells by Combining Zinc-Finger Nucleases With Adeno-Associated Viral Vectors. | Abstract Zinc-finger nucleases (ZFNs) have become a valuable tool for targeted genome engineering. Based on the enzyme's ability to create a site-specific DNA double-strand break, ZFNs promote genome editing by activating the cellular DNA damage response, including homology-directed repair (HDR) and nonhomologous end-joining. The goal of this study was (i) to demonstrate the versatility of combining the ZFN technology with a vector platform based on adeno-associated virus (AAV), and (ii) to as ... | 2011 | 21980922 |
| Preclinical safety evaluation of subretinal AAV2.sFlt-1 in non-human primates. | We report on the long-term safety of AAV2.sFlt-1 (a recombinant adeno-associated virus serotype 2 carrying the soluble form of the Flt-1 receptor) injection into the subretinal space of non-human primates. Levels of sFlt-1 protein were significantly higher (P<0.05) in the vitreous of four out of five AAV2.sFlt-1-injected eyes. There was no evidence of damage to the eyes of animals that received subretinal injections of AAV2.sFlt-1; ocular examination showed no anterior chamber flare, normal fund ... | 2011 | 22071974 |
| aav8 vector expressing il24 efficiently suppresses tumor growth mediated by specific mechanisms in mll/af4-positive all model mice. | mixed-lineage leukemia (mll)/af4-positive acute lymphoblastic leukemia (all) is a common type of leukemia in infants, which is associated with a high relapse rate and poor prognosis. il24 selectively induces apoptosis in cancer cells and exerts immunomodulatory and antiangiogenic effects. we examined the effects of adeno-associated virus type 8 (aav8) vector-mediated muscle-directed systemic gene therapy in mll/af4-positive all using il24. in a series of in vitro studies, we examined the effects ... | 2011 | 22025528 |
| adeno-associated virus for cystic fibrosis gene therapy. | gene therapy is an alternative treatment for genetic lung disease, especially monogenic disorders such as cystic fibrosis. cystic fibrosis is a severe autosomal recessive disease affecting one in 2500 live births in the white population, caused by mutation of the cystic fibrosis transmembrane conductance regulator (cftr). the disease is classically characterized by pancreatic enzyme insufficiency, an increased concentration of chloride in sweat, and varying severity of chronic obstructive lung d ... | 2011 | 21952739 |
| Imaging gene delivery in a mouse model of congenital neuronal ceroid lipofuscinosis. | Adeno-associated virus (AAV)-mediated gene replacement for lysosomal disorders have been spurred by the ability of some serotypes to efficiently transduce neurons in the brain and by the ability of lysosomal enzymes to cross-correct among cells. Here, we explored enzyme replacement therapy in a knock-out mouse model of congenital neuronal ceroid lipofuscinosis (NCL), the most severe of the NCLs in humans. The missing protease in this disorder, cathepsin D (CathD) has high levels in the central n ... | 2011 | 21900963 |
| bifunctional anti-huntingtin proteasome-directed intrabodies mediate efficient degradation of mutant huntingtin exon 1 protein fragments. | huntington's disease (hd) is a fatal autosomal dominant neurodegenerative disorder caused by a trinucleotide (cag)(n) repeat expansion in the coding sequence of the huntingtin gene, and an expanded polyglutamine (>37q) tract in the protein. this results in misfolding and accumulation of huntingtin protein (htt), formation of neuronal intranuclear and cytoplasmic inclusions, and neuronal dysfunction/degeneration. single-chain fv antibodies (scfvs), expressed as intrabodies that bind htt and preve ... | 2011 | 22216210 |
| differential effects of brain-derived neurotrophic factor and neurotrophin-3 on hindlimb function in paraplegic rats. | we compared the effect of viral administration of brain-derived neurotrophic factor (bdnf) or neurotrophin 3 (nt-3) on locomotor recovery in adult rats with complete thoracic (t10) spinal cord transection injuries, in order to determine the effect of chronic neurotrophin expression on spinal plasticity. at the time of injury, bdnf, nt-3 or green fluorescent protein (gfp) (control) was delivered to the lesion via adeno-associated virus (aav) constructs. aav-bdnf was significantly more effective t ... | 2012 | 22211901 |
| zinc-finger nuclease-mediated gene correction using single aav vector transduction and enhancement by food and drug administration-approved drugs. | an emerging strategy for the treatment of monogenic diseases uses genetic engineering to precisely correct the mutation(s) at the genome level. recent advancements in this technology have demonstrated therapeutic levels of gene correction using a zinc-finger nuclease (zfn)-induced dna double-strand break in conjunction with an exogenous dna donor substrate. this strategy requires efficient nucleic acid delivery and among viral vectors, recombinant adeno-associated virus (raav) has demonstrated c ... | 2012 | 22257934 |
| high-throughput functional micrornas profiling by recombinant aav-based microrna sensor arrays. | micrornas (mirnas) are small and non-coding rnas which play critical roles in physiological and pathological processes. a number of methods have been established to detect and quantify mirna expression. however, method for high-throughput mirna function detection is still lacking. | 2012 | 22242174 |
| the aav vector toolkit: poised at the clinical crossroads. | the discovery of naturally occurring adeno-associated virus (aav) isolates in different animal species and the generation of engineered aav strains using molecular genetics tools have yielded a versatile aav vector toolkit. promising results in preclinical animal models of human disease spurred the much awaited transition toward clinical application, and early successes in phase i/ii clinical trials for a broad spectrum of genetic diseases have recently been reported. as the gene therapy communi ... | 2012 | 22273577 |
| effects of antioxidant gene therapy on retinal neurons and oxidative stress in a model of retinal ischemia/reperfusion. | retinal ischemia/reperfusion (i/r) results in neuronal death and generation of reactive oxygen species. the aim of this study was to investigate the neuroprotective effect of manganese superoxide dismutase (sod2) on retinal ganglion cells (rgcs) in an i/r-induced retinal injury model. one eye of each wistar rat was pretreated with recombinant adeno-associated virus containing the sod2 gene (aav-sod2) or recombinant aav containing the gfp gene (aav-gfp) by intravitreal injection 21days before ini ... | 2011 | 22240151 |
| efficient growth suppression and apoptosis in human laryngeal carcinoma cell line hep-2 induced by an adeno-associated virus expressing human fas ligand. | background: apoptosis induced by fas/fasl system has been proposed as a gene therapy methold for various cancers. methods: we used adeno-associated virus-expressing enhanced green fluorescent protein (egfp)-human fasl (aav-egfp-hfasl) to deliver fasl into hep-2 cells, cytotoxicity was detected by mts assay , apoptosis was confirmed by flow cytometry. we also treated the xenograft of hep-2 tumor in nude mice with intratumoral injection of aav-egfp-hfasl. the size of the xenograft, the apoptosis i ... | 2012 | 22267220 |
| aav6.βarkct cardiac gene therapy ameliorates cardiac function and normalizes the catecholaminergic axis in a clinically relevant large animal heart failure model. | aimsg protein-coupled receptor kinase 2 (grk2), which is markedly upregulated in failing human myocardium, has been implicated as a contributing factor or consequence of heart failure (hf). importantly, cardiac-specific grk2 knockout mice have recently proved the pathological nature of grk2 in hf. targeted inhibition of grk2 is possible using a peptide inhibitor known as the βarkct, which has rescued several disparate small animal hf models. this study was designed to evaluate long-term βarkct e ... | 2012 | 22261894 |
| aav-based neonatal gene therapy for hemophilia a: long-term correction and avoidance of immune responses in mice. | hemophilia a gene therapy has been hampered by immune responses to vector-associated antigens and by neutralizing antibodies or inhibitors against the factor viii (fviii) protein; these 'inhibitors' more commonly affect hemophilia a patients than those with hemophilia b. a gene replacement strategy beginning in the neonatal period may avoid the development of these immune responses and lead to prolonged expression with correction of phenotype, thereby avoiding long-term consequences. a serotype ... | 2012 | 22241178 |
| delivery of aav2-cyp2j2 protects remnant kidney in the 5/6 nephrectomized rat via inhibition of apoptosis and fibrosis. | the cytochrome p450 epoxygenase, cyp2j2, converts arachidonic acid to four regioisomeric epoxyeicosatrienoic acids (eets) which are highly abundant in the kidney and considered reno-protective. accumulating evidence suggests that eets are important in regulating renal and cardiovascular function. further, eets have been confirmed to exert diverse biological activities including potent vasodilatory, fibrinolytic properties, anti-inflammatory, anti-apoptotic, and mitogenic effects. in the current ... | 2012 | 22260463 |
| taci-fc gene therapy improves autoimmune sialadenitis but not salivary gland function in non-obese diabetic mice. | oral diseases (2011) objective: patients with sjögren's syndrome (ss) show aberrant expression of the b cell-related mediators, b cell-activating factor (baff), and a proliferation-inducing ligand (april) in serum and salivary glands (sgs). we studied the biological effect of neutralizing these cytokines by local gene transfer of the common receptor transmembrane activator and caml interactor (taci) in an animal model of ss. material and methods: a recombinant serotype 2 adeno-associated virus ... | 2011 | 22212434 |
| rescue of dystrophic skeletal muscle by pgc-1α involves a fast to slow fiber type shift in the mdx mouse. | increased utrophin expression is known to reduce pathology in dystrophin-deficient skeletal muscles. transgenic over-expression of pgc-1α has been shown to increase levels of utrophin mrna and improve the histology of mdx muscles. other reports have shown that pgc-1α signaling can lead to increased oxidative capacity and a fast to slow fiber type shift. given that it has been shown that slow fibers produce and maintain more utrophin than fast skeletal muscle fibers, we hypothesized that over-exp ... | 2012 | 22253880 |
| effect of cigarette smoke exposure and structural modifications on the alpha-1 antitrypsin interaction with caspases. | a1at is a serpin with a major protective effect against cigarette smoke induced emphysema development, as patients with mutations of a1at display a markedly increased risk for developing emphysema. we reported that a1at protects lung endothelial cells from apoptosis and inhibits caspase-3 activity. it is not clear if cigarette smoking or a1at mutations alter the caspase-3 inhibitory activity of a1at and if this serpin alters the function of other caspases. we tested the hypothesis that the caspa ... | 2012 | 22245800 |
| acquisition of selective antitumoral effects of recombinant adeno-associated virus by genetically inserting tumor-targeting peptides into capsid proteins. | recombinant adeno-associated virus serotype 5 (raav5) is considered to be a promising gene transfer vehicle. however, preferential gene delivery to the tumor remains a requirement for cancer treatment. we generated raav5 mutants bearing tumor marker-binding peptides and analyzed their properties as viral vectors, as well as their transduction efficiencies and preferential antitumoral potencies. all of the mutants were successfully produced. transduction analyses showed that raav5 mutants harbori ... | 2011 | 22848276 |
| [establishment of a targeting protein for xenopus kinesin-like protein 2 c' terminal sbp-3 x flag tagged hct 116 colorectal cancer cell model]. | to develop a targeting protein for xenopus kinesin-like protein 2 (tpx2) c' terminal sbp-3 x flag-tagged hct 116 cell model. | 2011 | 22509544 |
| [enhancing effect of deoxynivalenol-mediated grp78 down-regulation on heavy chain secretion and bioactivity of two-chain fviii gene co-transfected cells]. | although two chain transfering separately could be used to overcome the volume limitation of adeno-associated virus vectors (aav) in coagulation factor viii (fviii) gene delivery, it leads to chain imbalance for inefficient heavy chain secretion. in this study we aimed to improve the efficacy of two chain strategy in fviii gene delivery through the degradation of glucose-regulated protein 78 (grp78) known as a protein chaperone in endoplasmic reticulum (er) by deoxynivalenol (don) to decrease gr ... | 2011 | 22375418 |
| [the development in the studies of aav-mediated gene therapy in retina]. | significant progress in understanding the molecular basis of retinal disorders has led to the development of gene therapies for treatment of these diseases. adeno-associated virus (aav) is a useful vector for the treatment of retinal diseases due to its low toxicity and immunogenicity, ability to transducer both dividing and non-dividing cells, and stable transgene expression. a variety of animal studies and clinical trials have proved the safety and effectivity of retinal aav-mediated gene ther ... | 2011 | 22336072 |
| gene therapy: light is finally in the tunnel. | after two decades of ups and downs, gene therapy has recently achieved a milestone in treating patients with leber's congenital amaurosis (lca). lca is a group of inherited blinding diseases with retinal degeneration and severe vision loss in early infancy. mutations in several genes, including rpe65, cause the disease. using adeno-associated virus as a vector, three independent teams of investigators have recently shown that rpe65 can be delivered to retinal pigment epithelial cells of lca pati ... | 2011 | 22231356 |
| glutamate transporter glt-1 upregulation attenuates visceral nociception and hyperalgesia via spinal mechanisms not related to anti-inflammatory or probiotic effects. | visceral pain is the most common reason for physician visits in us. glutamate is the major excitatory neurotransmitter and mediates visceral nociceptive neuro-transmission and hypersensitivity. removal of extracellular glutamate is predominantly mediated by glial glutamate transporter-1 (glt-1). the pharmacological approach to up-regulate glt-1 by 1 week administration of ceftriaxone (ctx) has been successful to mitigate visceral nociception. the present study shows that intrathecal delivery of ... | 2011 | 22220274 |
| adeno-associated virus-mediated gene transfer to renal tubule cells via a retrograde ureteral approach. | gene therapy involves delivery of exogenous dna to provide a therapeutic protein. ideally, a gene therapy vector should be non-toxic, non-immunogenic, easy to produce, and efficient in protecting and delivering dna into target cells. | 2011 | 22470395 |
| [recombinant adeno-associated virus type 8 mediated dual-luciferase gene expression in mouse]. | recombinant adeno-associated virus type 8 (raav8) mediating transgene expression in mice was investigated using co-expressed report gene of secreted gaussia princeps luciferase (gluc) and non-secreted firefly luciferase(fluc). | 2012 | 23627024 |
| long-term correction of type 1 and 2 diabetes by central leptin gene therapy independent of effects on appetite and energy expenditure. | adipocyte-derived leptin is a hormone associated with the regulation of energy homeostasis, including glucose metabolism. hyperleptinemia, induced by the consumption of energy-enriched diets, inhibits leptin transport across the blood-brain barrier, and thereby produces leptin insufficiency in the hypothalamus. as a result of sustained leptin insufficiency, the hypothalamic restraint on pancreatic insulin secretion is lost. additionally, both glucose metabolism and energy expenditure are also di ... | 2012 | 23565490 |
| [adeno-associated viral vectors: methods for production and purification for gene therapy applications]. | viral vectors based on adeno-associated virus (aav) are widely used in gene therapy protocols, because they have characteristics that make them valuable for the treatment of genetic and chronic degenerative diseases. aav2 serotype had been the best characterized to date. however, the aav vectors developed from other serotypes is of special interest, since they have organ-specific tropism which increases their potential for transgene delivery to target cells for performing their therapeutic effec ... | 2012 | 23544311 |
| deficiency in myd88 signaling results in decreased antibody responses to an adeno-associated virus vector in murine pompe disease. | we have previously shown that antibody and t cell responses limit the efficacy of an adeno-associated virus (aav) pseudotype 8 (2/8) vector containing the universally active cytomegalovirus enhancer/chicken β-actin regulatory cassette (aav2/8-cbhgaa) in treating murine pompe disease. however, the innate immune responses to aav2/8-cbhgaa are largely unknown. in this study, we investigated acute immune responses to aav2/8-cbhgaa and the role of myd88/trif signaling pathway in shaping adaptive immu ... | 2012 | 23514839 |
| [efficient and durable gene delivery of self complementary adeno-associated virus 6 vector and impact of pre-existing immunity]. | recombinant adeno-associated viral (raav) vectors are promising vectors for human gene therapy. however, aav-mediated gene transduction can be hampered because of the pre-existing neutralized natural antibodies (nabs) in primates. we evaluated transduction efficiency of raav6 expressing human alpha-1-anti-trypsin (haat) vectors in murine models, and found that these vectors showed stable and high levels of transgene expression. fluorescence imaging showed that aav6 expressing enhanced green fluo ... | 2012 | 23469547 |
| apolipoprotein e-mediated cell cycle arrest linked to p27 and the cox2-dependent repression of mir221/222. | in addition to its effects on cholesterol levels, apoe3 has lipid-independent effects that contribute to cardiovascular protection; one of these effects is the ability to inhibit cell cycling in vsmcs. the goal of this study was to identify and characterize cell cycle-regulatory mechanisms responsible for the anti-mitogenic effect of apoe. | 2012 | 23294923 |
| serotype-independent method of recombinant adeno-associated virus (aav) vector production and purification. | a variety of gene transfer strategies have been developed to treat inherited, degenerative, and acquired diseases. among the different vector systems developed so far, recombinant adeno-associated viral (aav) vectors have shown notable benefits, including prolonged gene expression, transduction of both dividing and nondividing cells, and a lack of pathogenicity caused by wild-type infections. thanks to these features, the use of aav vectors as a gene transfer tool has increased dramatically duri ... | 2012 | 23291836 |
| focal expression of adeno-associated viral-mutant tau induces widespread impairment in an app mouse model. | adeno-associated virus serotype 6 (aav6) viral vectors encoding mutant and normal tau were used to produce focal tau pathology. two mutant forms of tau were used; the p301s tau mutation is associated with neurofibrillary tangle formation in humans, and the 3po mutation leads to rapid tau aggregation and is associated with pathogenic phosphorylation and cytotoxicity in vitro. we show that adeno-associated viral injection into entorhinal cortex of normal and tau knockout animals leads to local ove ... | 2012 | 23273572 |
| comparative analysis of dna nanoparticles and aavs for ocular gene delivery. | gene therapy is a critical tool for the treatment of monogenic retinal diseases. however, the limited vector capacity of the current benchmark delivery strategy, adeno-associated virus (aav), makes development of larger capacity alternatives, such as compacted dna nanoparticles (nps), critical. here we conduct a side-by-side comparison of self-complementary aav and ck30peg nps using matched itr plasmids. we report that although aavs are more efficient per vector genome (vg) than nps, nps can dri ... | 2012 | 23272225 |
| aav9.i-1c delivered via direct coronary infusion in a porcine model of heart failure improves contractility and mitigates adverse remodeling. | heart failure is characterized by impaired function and disturbed ca2+ homeostasis. transgenic increases in inhibitor-1 activity have been shown to improve ca2 cycling and preserve cardiac performance in the failing heart. the aim of this study was to evaluate the effect of activating the inhibitor (i-1c) of protein phosphatase 1 (i-1) through gene transfer on cardiac function in a porcine model of heart failure induced by myocardial infarction. | 2012 | 23271792 |
| biophysical and ultrastructural characterization of adeno-associated virus capsid uncoating and genome release. | we describe biophysical and ultrastructural differences in genome release from adeno-associated virus (aav) capsids packaging wild-type dna, recombinant single-stranded dna (ssdna), or dimeric, self-complementary dna (scdna) genomes. atomic force microscopy and electron microscopy (em) revealed that aav particles release packaged genomes and undergo marked changes in capsid morphology upon heating in physiological buffer (ph 7.2). when different aav capsids packaging ss/scdna varying in length f ... | 2012 | 23269804 |
| adenovirus-mediated overexpression of tcfe3 ameliorates hyperglycaemia in a mouse model of diabetes by upregulating glucokinase in the liver. | transcription factor e3 (tfe3) has been shown to increase insulin sensitivity by activating insulin-signalling pathways. however, the role of tfe3 in glucose homeostasis is not fully understood. here, we explored the possible therapeutic potential of tfe3 for the control of hyperglycaemia using a streptozotocin-induced mouse model of diabetes. | 2012 | 23269357 |
| development of novel recombinant aav vectors and strategies for the potential gene therapy of hemophilia. | recombinant vectors based on a non-pathogenic human parvovirus, the adeno-associated virus (aav), have gained attention as a potentially safe and useful alternative to the more commonly used retroviral and adenoviral vectors. aav vectors are currently in use in phase i/ii clinical trials for gene therapy of a number of diseases such as cystic fibrosis, α-1 antitrypsin deficiency, muscular dystrophy, batten's disease, and parkinson's disease, and have shown efficacy in patients with leber's conge ... | 2012 | 23264889 |
| optimal immunofluorescent staining for human factor ix and infiltrating t cells following gene therapy for hemophilia b. | immunofluorescent imaging is a valuable tool for investigating the outcome of gene therapy within the transduced tissue. with a multi-labeling technique, it is possible to both characterize local expression of the transgene and to evaluate the severity of the adaptive immune response through cytotoxic t cell infiltration. it is critical that the experimental parameters are optimal in order to prevent misinterpretation of important pathological events. to optimize this staining protocol, murine l ... | 2012 | 23264888 |