Exosome-associated AAV2 vector mediates robust gene delivery into the murine retina upon intravitreal injection

Wassmer, Sarah; Carvalho, Lívia S.; György, Bence; Vandenberghe, Luk; Maguire, Casey A.

doi:10.1038/srep45329

Cited by 116 publications

(111 citation statements)

References 45 publications

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“…13 The natural release of a fraction of recombinant AAV vectors associated with exosomes (exosome-associated AAVs [exo-AAVs]) in the cell medium during vector production has been recently described. 14 The use of exo-AAV vectors has been demonstrated to enhance transduction in the setting of gene transfer to the retina, 15 the nervous system, 16 and the inner ear. 17 In the present study, we tested the ability of exo-AAV vectors to transduce the liver and drive efficient correction of hemophilia B.…”

Section: Introductionmentioning

confidence: 99%

Enhanced liver gene transfer and evasion of preexisting humoral immunity with exosome-enveloped AAV vectors

et al. 2017

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Section: Introductionmentioning

confidence: 99%

Enhanced liver gene transfer and evasion of preexisting humoral immunity with exosome-enveloped AAV vectors

et al. 2017

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“…One possible explanation is that secreted viral particles may be inherently different than those found intracellularly. Several recent papers have identified secreted rAAV particles inside vesicular bodies such as exosomes [40][41][42][43][44][45][46][47]. The secreted rAAVs utilized in our study may be associated with exosomes and this association could potentially alter the way in which they interact with the target cell.…”

Section: Discussionmentioning

confidence: 90%

Utilizing minimally purified secreted rAAV for rapid and cost-effective manipulation of gene expression in the CNS

Goodwin

Croft

Futch

et al. 2020

Mol Neurodegeneration

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Background: Recombinant adeno-associated virus (rAAV) is widely used in the neuroscience field to manipulate gene expression in the nervous system. However, a limitation to the use of rAAV vectors is the time and expense needed to produce them. To overcome this limitation, we evaluated whether unpurified rAAV vectors secreted into the media following scalable PEI transfection of HEK293T cells can be used in lieu of purified rAAV. Methods:We packaged rAAV2-EGFP vectors in 30 different wild-type and mutant capsids and subsequently collected the media containing secreted rAAV. Genomic titers of each rAAV vector were assessed and the ability of each unpurified virus to transduce primary mixed neuroglial cultures (PNGCs), organotypic brain slice cultures (BSCs) and the mouse brain was evaluated.Results: There was~40-fold wide variance in the average genomic titers of the rAAV2-EGFP vector packaged in the 30 different capsids, ranging from a low~4.7 × 10 10 vector genomes (vg)/mL for rAAV2/5-EGFP to a high of2 .0 × 10 12 vg/mL for a capsid mutant of rAAV2/8-EGFP. In PNGC studies, we observed a wide range of transduction efficiency among the 30 capsids evaluated, with the rAAV2/6-EGFP vector demonstrating the highest overall transduction efficiency. In BSC studies, we observed robust transduction by wild-type capsid vectors rAAV2/6, 2/8 and 2/9, and by capsid mutants of rAAV2/1, 2/6, and 2/8. In the in vivo somatic brain transgenesis (SBT) studies, we found that intra-cerebroventricular injection of media containing unpurified rAAV2-EGFP vectors packaged with select mutant capsids resulted in abundant EGFP positive neurons and astrocytes in the hippocampus and forebrain of non-transgenic mice. We demonstrate that unpurified rAAV can express transgenes at equivalent levels to lysate-purified rAAV both in vitro and in vivo. We also show that unpurified rAAV is sufficient to drive tau pathology in BSC and neuroinflammation in vivo, recapitulating previous studies using purified rAAV. Conclusions: Unpurified rAAV vectors secreted into the media can efficiently transduce brain cells in vitro and in vivo, providing a cost-effective way to manipulate gene expression. The use of unpurified virus will greatly reduce costs of exploratory studies and further increase the utility of rAAV vectors for standard laboratory use.

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“…We utilized exo-AAV in this study has we have previously shown that this vector system has attributes such as enhanced transduction to CNS 21 , liver 32 , inner ear 33 , and retina 34 compared to conventional AAV. Exo-AAV also has a benefit of antibody evasion which may have clinical relevance in patients with pre-existing humoral immunity to AAV 20, 32 .…”

Section: Discussionmentioning

confidence: 99%

Virus vector-mediated genetic modification of brain tumor stromal cells after intravenous delivery

et al. 2018

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The malignant primary brain tumor, glioblastoma (GBM) is generally incurable. New approaches are desperately needed. Adeno-associated virus (AAV) vector-mediated delivery of anti-tumor transgenes is a promising strategy, however direct injection leads to focal transgene spread in tumor and rapid tumor division dilutes out the extra-chromosomal AAV genome, limiting duration of transgene expression. Intravenous (IV) injection gives widespread distribution of AAV in normal brain, however poor transgene expression in tumor, and high expression in non-target cells which may lead to ineffective therapy and high toxicity, respectively. Delivery of transgenes encoding secreted, anti-tumor proteins to tumor stromal cells may provide a more stable and localized reservoir of therapy as they are more differentiated than fast-dividing tumor cells. Reactive astrocytes and tumor-associated macrophage/microglia (TAMs) are stromal cells that comprise a large portion of the tumor mass and are associated with tumorigenesis. In mouse models of GBM, we used IV delivery of exosome-associated AAV vectors driving green fluorescent protein expression by specific promoters (NF-κB-responsive promoter and a truncated glial fibrillary acidic protein promoter), to obtain targeted transduction of TAMs and reactive astrocytes, respectively, while avoiding transgene expression in the periphery. We used our approach to express the potent, yet toxic anti-tumor cytokine, interferon beta, in tumor stroma of a mouse model of GBM, and achieved a modest, yet significant enhancement in survival compared to controls. Noninvasive genetic modification of tumor microenvironment represents a promising approach for therapy against cancers. Additionally, the vectors described here may facilitate basic research in the study of tumor stromal cells in situ.

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Exosome-associated AAV2 vector mediates robust gene delivery into the murine retina upon intravitreal injection

Cited by 116 publications

References 45 publications

Enhanced liver gene transfer and evasion of preexisting humoral immunity with exosome-enveloped AAV vectors

Enhanced liver gene transfer and evasion of preexisting humoral immunity with exosome-enveloped AAV vectors

Utilizing minimally purified secreted rAAV for rapid and cost-effective manipulation of gene expression in the CNS

Virus vector-mediated genetic modification of brain tumor stromal cells after intravenous delivery

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