Efficient gene transfer into human cord blood CD34+ cells and the CD34+CD38− subset using highly purified recombinant adeno-associated viral vector preparations that are free of helper virus and wild-type AAV

Nathwani, Amit C.; Hanawa, Hideki; Vandergriff, Jody; Kelly, P; Vanin, Elio F.; Nienhuis, Arthur W.

doi:10.1038/sj.gt.3301068

Cited by 71 publications

(47 citation statements)

References 59 publications

(45 reference statements)

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“…38 AAV8-G6Pase-a was packaged by cotransfection of 293 cells with UF11-G6Pase-a; 80-XX6, an adenoviral helper plasmid; 39 and a chimeric packaging plasmid pAAV8-2. 23 The AAV1-G6Pase-a or AAV8-G6Pase-a virions were obtained by freeze-thaw lysis of the transfected cells and purified by iodixanol gradient ultracentrifugation followed by HiTrap Q column chromatography (Pharmacia, Piscataway, NJ, USA).…”

Section: Methodsmentioning

confidence: 99%

Long-term correction of murine glycogen storage disease type Ia by recombinant adeno-associated virus-1-mediated gene transfer

Ghosh

et al. 2005

Gene Ther

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Glycogen storage disease type Ia (GSD-Ia) is caused by a deficiency in glucose-6-phosphatase-a (G6Pase-a), a ninetransmembrane domain, endoplasmic reticulum-associated protein expressed primarily in the liver and kidney. Previously, we showed that infusion of an adeno-associated virus (AAV) serotype 2 vector carrying murine G6Pase-a (AAV2-G6Pase-a) into neonatal GSD-Ia mice failed to sustain their life beyond weaning. We now show that neonatal infusion of GSD-Ia mice with an AAV serotype 1-G6Pase-a (AAV1-G6Pase-a) or AAV serotype 8-G6Pase-a (AAV8-G6Pase-a) results in hepatic expression of the G6Pase-a transgene and markedly improves the survival of the mice. However, only AAV1-G6Pase-a can achieve significant renal transgene expression. A more effective strategy, in which a neonatal AAV1-G6Pase-a infusion is followed by a second infusion at age 1 week, provides sustained expression of a complete, functional, G6Pase-a system in both the liver and kidney and corrects the metabolic abnormalities in GSD-Ia mice for the 57 week length of the study. This effective use of gene therapy to correct metabolic imbalances and disease progression in GSD-Ia mice holds promise for the future of gene therapy in humans. Gene Therapy (2005) 13, 321-329.

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Section: Methodsmentioning

confidence: 99%

Long-term correction of murine glycogen storage disease type Ia by recombinant adeno-associated virus-1-mediated gene transfer

Ghosh

et al. 2005

Gene Ther

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“…Highly purified stocks of a recombinant AAV vector containing the ␤-galactosidase (lacZ) reporter gene driven by the cytomegalovirus (CMV) immediate-early promoter (vCMVp-lacZ) were generated as described previously (14,15). Physical particle titers of recombinant vector stocks were determined by quantitative DNA slot blot analysis, as described previously (57).…”

Section: Methodsmentioning

confidence: 99%

Heat-shock Treatment-mediated Increase in Transduction by Recombinant Adeno-associated Virus 2 Vectors Is Independent of the Cellular Heat-shock Protein 90

Zhong

Qing

et al. 2004

Journal of Biological Chemistry

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Recombinant adeno-associated virus 2 (AAV) vectors transduction efficiency varies greatly in different cell types. We have described that a cellular protein, FKBP52, in its phosphorylated form interacts with the D-sequence in the viral inverted terminal repeat, inhibits viral second strand DNA synthesis, and limits transgene expression. Here we investigated the role of cellular heat-shock protein 90 (HSP90) in AAV transduction because FKBP52 forms a complex with HSP90, and because heat-shock treatment augments AAV transduction efficiency. Heat-shock treatment of HeLa cells resulted in tyrosine dephosphorylation of FKBP52, led to stabilization of the FKBP52-HSP90 complex, and resulted in ϳ6-fold increase in AAV transduction. However, when HeLa cells were pre-treated with tyrphostin 23, a specific inhibitor of cellular epidermal growth factor receptor tyrosine kinase, which phosphorylates FKBP52 at tyrosine residues, heat-shock treatment resulted in a further 18-fold increase in AAV transduction. HSP90 was shown to be a part of the FKBP52-AAV Dsequence complex, but HSP90 by itself did not bind to the D-sequence. Geldanamycin treatment, which disrupts the HSP90-FKBP52 complex, resulted in >22-fold increase in AAV transduction in heat-shock-treated cells compared with heat shock alone. Deliberate overexpression of the human HSP90 gene resulted in a significant decrease in AAV-mediated transduction in tyrphostin 23-treated cells, whereas down-modulation of HSP90 levels led to a decrease in HSP90-FKBP52-AAV D-sequence complex formation, resulting in a significant increase in AAV transduction following pre-treatment with tyrphostin 23. These studies suggest that the observed increase in AAV transduction efficiency following heat-shock treatment is unlikely to be mediated by HSP90 alone and that increased levels of HSP90, in the absence of heat shock, facilitate binding of FKBP52 to the AAV D-sequence, thereby leading to inhibition of AAV-mediated transgene expression. These studies have implications in the optimal use of recombinant AAV vectors in human gene therapy. Adeno-associated virus (AAV),1 type 2, a non-pathogenic human parvovirus, has gained attention as a vector for gene transfer and gene therapy (1-25). Although recombinant AAV vectors are currently in use in phase I/II clinical trials for gene therapy of cystic fibrosis and hemophilia B (1,5,8,11,22), and have been shown to transduce a wide variety of cells and tissues in vitro and in vivo (2-4, 6, 7, 9, 10, 12-21, 23-25), their transduction efficiency varies widely in different cell types. We and others have undertaken systematic studies to delineate various steps in the life cycle of AAV, which include viral binding and entry (26 -28), intracellular trafficking (29 -35), nuclear transport (29, 30, 34 -37), and viral second strand DNA synthesis (38 -47). The viral second strand DNA synthesis has been described by two independent laboratories to be the ratelimiting step, which leads to inefficient transduction by AAV vectors (38, 39). We have reported that ...

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“…Although some investigators have noted difficulty with long-term transduction of HSCs with rAAV vectors, we and others have demonstrated stable rAAV transduction and integration in human HSCs associated with long-term transgene expression, both in vitro and in the NOD/SCID xenograft model in vivo. [64][65][66][67][68] S. Chatterjee. personal communication.…”

Section: Ex Vivo Manipulation Of Target Cells For Gene Therapymentioning

confidence: 99%

Immune responses to adeno-associated virus and its recombinant vectors

Sun¹,

Anand-Jawa²,

et al. 2003

Gene Ther

190

116

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Efficient gene transfer into human cord blood CD34+ cells and the CD34+CD38− subset using highly purified recombinant adeno-associated viral vector preparations that are free of helper virus and wild-type AAV

Cited by 71 publications

References 59 publications

Long-term correction of murine glycogen storage disease type Ia by recombinant adeno-associated virus-1-mediated gene transfer

Long-term correction of murine glycogen storage disease type Ia by recombinant adeno-associated virus-1-mediated gene transfer

Heat-shock Treatment-mediated Increase in Transduction by Recombinant Adeno-associated Virus 2 Vectors Is Independent of the Cellular Heat-shock Protein 90

Immune responses to adeno-associated virus and its recombinant vectors

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