Gene delivery by lentivirus vectors

Cockrell, Adam S.; Kafri, Tal

doi:10.1007/s12033-007-0010-8

Cited by 297 publications

(207 citation statements)

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“…However, both retroviral and lentiviral vectors undergo random integration into host chromosomal DNA, and there is persistent risk for insertional mutagenesis (Cockrell and Kafri, 2007;Nair, 2008). The development of leukemia in four treated children in an X-linked severe combined immunodeficiency clinical trial using retroviral vectors has re-emphasized the risks related to gene therapy (HaceinBey-Abina et al, 2003;Kohn et al, 2003;Nair, 2008).…”

Section: Discussionmentioning

confidence: 99%

“…However, it is of note that retroviruses and lentivirus are either the etiologic agents of, or are intimately associated with, malignant disorders (Weiss et al, 1984). Recombinant retro-and lentiviral vectors can also undergo random integration into host chromosome, thereby elevating the risk of insertional mutagenesis (Cockrell and Kafri, 2007;Nair, 2008). Thus, development of alternative vector systems, such as adeno-associated virus (AAV), needs to be pursued, given the proven safety of AAV vectors in several Phase I/II clinical trials (Bainbridge et al, 2008;Cideciyan et al, 2008;Hauswirth et al, 2008;Maguire et al, 2008).…”

Section: Introductionmentioning

confidence: 99%

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High-Efficiency Transduction of Fibroblasts and Mesenchymal Stem Cells by Tyrosine-Mutant AAV2 Vectors for Their Potential Use in Cellular Therapy

Jayandharan

et al. 2010

Human Gene Therapy

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Adeno-associated virus 2 (AAV2) vectors transduce fibroblasts and mesenchymal stem cells (MSCs) inefficiently, which limits their potential widespread applicability in combinatorial gene and cell therapy. We have reported that AAV2 vectors fail to traffic efficiently to the nucleus in murine fibroblasts. We have also reported that site-directed mutagenesis of surface-exposed tyrosine residues on viral capsids leads to improved intracellular trafficking of the mutant vectors, and the transduction efficiency of the single tyrosine-mutant vectors is *10-fold higher in human cells. In the current studies, we evaluated the transduction efficiency of single as well as multiple tyrosine-mutant AAV2 vectors in murine fibroblasts. Our results indicate that the Y444F mutant vectors transduce these cells most efficiently among the seven single-mutant vectors, with >30-fold increase in transgene expression compared with the wild-type vectors. When the Y444F mutation is combined with additional mutations (Y500F and Y730F), the transduction efficiency of the triple-mutant vectors is increased by *130-fold and the viral intracellular trafficking is also significant improved. Similarly, the triple-mutant vectors are capable of transducing up to 80-90% of bone marrow-derived primary murine as well as human MSCs. Thus, high-efficiency transduction of fibroblasts with reprogramming genes to generate induced pluripotent stem cells, and the MSCs for delivering therapeutic genes, should now be feasible with the tyrosine-mutant AAV vectors. Overview SummaryFibroblasts and mesenchymal stem cells (MSCs) are promising targets for gene and cell therapy. Recombinant adeno-associated virus 2 (AAV2) vectors are currently in use in several Phase I/II clinical trials for gene therapy. However, the existing data show that AAV2 vector-mediated transduction of fibroblasts and MSCs is inefficient. We observed that AAV2 vectors containing mutations in three surface-exposed tyrosine residues significantly increase transduction efficiency in fibroblasts, as well as in both human and murine primary MSCs. The increased transduction efficiency of these triple-mutant AAV2 vectors correlates well with improved viral intracellular trafficking in fibroblasts. These data provide strong evidence that high-efficiency gene delivery and transgene expression by AAV vectors are indeed feasible, which should facilitate the generation of induced pluripotent stem cells, as well as the use of MSCs in combinatorial gene and cell therapy.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

High-Efficiency Transduction of Fibroblasts and Mesenchymal Stem Cells by Tyrosine-Mutant AAV2 Vectors for Their Potential Use in Cellular Therapy

Jayandharan

et al. 2010

Human Gene Therapy

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“…Medium was changed to complete DMEM and expression was monitored at least 72 h post-infection. After infection with lentivirus, cells have stable integration of the transgene(s), allowing long-term monitoring of reporter gene expression [26]. Infection efficiency ranged from 40% to 80%.…”

Section: Lentiviral Vector Production and Infectionmentioning

confidence: 99%

Reciprocal modulation of adult beta cell maturity by activin A and follistatin

2010

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Aims/hypothesis The functional maturity of pancreatic beta cells is impaired in diabetes mellitus. We sought to define factors that can influence adult beta cell maturation status and function. Methods MIN6 cells labelled with a Pdx1 monomeric red fluorescent protein-Ins1 enhanced green fluorescent protein dual reporter lentivirus were used to screen candidate growth and/or differentiation factors using image-based approaches with confirmation by real-time RT-PCR and assays of beta cell function using primary mouse islets.Results Activin A strikingly decreased the number of mature beta cells and increased the number of immature beta cells. While activins are critical for pancreatic morphogenesis, their role in adult beta cells remains controversial. In primary islets and MIN6 cells, activin A significantly decreased the expression of insulin and several genes associated with beta cell maturity (e.g. Pdx1, Mafa, Glut2 [also known as Slc2a2]). Genes found in immature beta cells (e.g. Mafb) tended to be upregulated by activin A. Insulin secretion was also reduced by activin A. In addition, activin A-treated MIN6 cells proliferated faster than non-treated cells. The effects of endogenous activin A on beta cells were completely reversed by exogenous follistatin. Conclusions/interpretation These results suggest that autocrine and/or paracrine activin A signalling exerts a suppressive effect on adult beta cell maturation and function. Thus, the maturation state of adult beta cells can be modulated by external factors in culture. Interventions inhibiting activin or its signalling pathways may improve beta cell function. Understanding of maturation and plasticity of adult pancreatic tissue has significant implications for islet regeneration and for in vitro generation of functional beta cells.

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“…1,2 The glycoprotein G of vesicular stomatitis virus (VSV) has widely been used for pseudotyping. 3 Recently, we achieved pseudotyping of lentiviral vectors with the envelope proteins of a vaccine strain of measles virus (MV). 4 Precise truncations of the cytoplasmic tails of the MV attachment protein hemagglutinin (H) and the fusion protein (F) were crucial for efficient pseudotyping.…”

Section: Introductionmentioning

confidence: 99%