Adeno-Associated Viral Vectors Based on Serotype 3b Use Components of the Fibroblast Growth Factor Receptor Signaling Complex for Efficient Transduction

Messina, Emily L.; Nienaber, Jeffrey J.; Daneshmand, Mani A.; Villamizar, Nestor; Samulski, Jude; Milano, Carmelo A.; Bowles, Dawn E.

doi:10.1089/hum.2012.066

Cited by 9 publications

(9 citation statements)

References 55 publications

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“…2D, indicate that both mutants performed in a similar manner as the WT rAAV3 vectors, in which the low dose of heparin enhanced viral vector-mediated transduction efficiency, whereas the high dose dramatically reduced it. Our data are also consistent with a recent report suggesting that fibroblast growth factor receptor is involved in the low-dose heparin-mediated augmentation of WT rAAV3 vector transduction (Messina et al, 2012).…”

Section: Raav3 Vectors Selectively Target Human Liver Tumors In a Mursupporting

confidence: 93%

SelectiveIn VivoTargeting of Human Liver Tumors by Optimized AAV3 Vectors in a Murine Xenograft Model

et al. 2014

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Current challenges for recombinant adeno-associated virus (rAAV) vector-based cancer treatment include the low efficiency and the lack of specificity in vivo. rAAV serotype 3 (rAAV3) vectors have previously been shown to be ineffective in normal mouse tissues following systemic administration. In the present study, we report that rAAV3 vectors can efficiently target and transduce various human liver cancer cells in vivo. Elimination of specific surface-exposed serine and threonine residues on rAAV3 capsids results in further augmentation in the transduction efficiency of these vectors, without any change in the viral tropism and cellular receptor interactions. In addition, we have identified a potential chemotherapy drug, shikonin, as a multifunctional compound to inhibit liver tumor growth as well as to significantly enhance the efficacy of rAAV vector-based gene therapy in vivo. Furthermore, we also document that suppression of tumorigenesis in a human liver cancer xenograft model can be achieved through systemic administration of the optimized rAAV3 vectors carrying a therapeutic gene, and shikonin at a dose that does not lead to liver damage. Our research provides a novel means to achieve not only targeted delivery but also the potential for gene therapy of human liver cancer.

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Section: Raav3 Vectors Selectively Target Human Liver Tumors In a Mursupporting

confidence: 93%

SelectiveIn VivoTargeting of Human Liver Tumors by Optimized AAV3 Vectors in a Murine Xenograft Model

et al. 2014

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“…Heparin 7 is a tetrasaccharide of 1.3 kDa that is considerably shorter than natural low-molecular-mass heparin, with a mean mass of 5 kDa, translating into a 14-mer glycosaminoglycan. Previously pub- lished competition assays with short, size-fractionated heparins have shown only marginal inhibitory effects on AAV2 infection efficiencies (29). This is in line with the recent claim that a chain length of at least 13 would be needed to make full contact with the positively charged amino acids at the 3-fold symmetry axis on the capsid surface of AAV2 (38).…”

Section: Discussionsupporting

confidence: 69%

“…7D). The only partial neutralization of AAV3 by heparin may be taken as an indication of an alternative AAV3 entry mechanism, presumably by coreceptors, as described previously (29)(30)(31). As expected, AAV serotypes 1, 6, and rh.10 were resistant to increasing concentrations of natural heparin (Fig.…”

Section: Generation Of Fluorescently Labeled Vectors Of Various Aav Ssupporting

confidence: 74%

Differential Adeno-Associated Virus Serotype-Specific Interaction Patterns with Synthetic Heparins and Other Glycans

Mietzsch

Broecker

Reinhardt

et al. 2014

J Virol

113

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All currently identified primary receptors of adeno-associated virus (AAV) are glycans. Depending on the AAV serotype, these carbohydrates range from heparan sulfate proteoglycans (HSPG), through glycans with terminal ␣2-3 or ␣2-6 sialic acids, to terminal galactose moieties. Receptor identification has largely relied on binding to natural compounds, defined glycan-presenting cell lines, or enzyme-mediated glycan modifications. Here, we describe a comparative binding analysis of highly purified, fluorescent-dye-labeled AAV vectors of various serotypes on arrays displaying over 600 different glycans and on a specialized array with natural and synthetic heparins. Few glycans bind AAV specifically in a serotype-dependent manner. Differential glycan binding was detected for the described sialic acid-binding AAV serotypes 1, 6, 5, and 4. The natural heparin binding serotypes AAV2, -3, -6, and -13 displayed differential binding to selected synthetic heparins. AAV7, -8, -rh.10, and -12 did not bind to any of the glycans present on the arrays. For discrimination of AAV serotypes 1 to 6 and 13, minimal binding moieties are identified. This is the first study to differentiate the natural mixed heparin binding AAV serotypes 2, 3, 6, and 13 by differential binding to specific synthetic heparins. Also, sialic acid binding AAVs display differential glycan binding specificities. The findings are relevant for further dissection of AAV host cell interaction. Moreover, the definition of single AAV-discriminating glycan binders opens the possibility for glycan microarray-based discrimination of AAV serotypes in gene therapy.

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“…S8 in the supplemental material). The lack of expression with HS-binding rAAV3 when injected intravitreally (data not shown) was expected, because the serotype is inefficient in the transduction of most cell types (44) and may encounter additional barriers to efficient transduction of cells. rAAV6 is a serotype of interest for retinal transduction and has been modified to increase the specific transduction of Müller glia using the ShH10 capsid (45).…”

Section: Discussionmentioning

confidence: 92%

Heparan Sulfate Binding Promotes Accumulation of Intravitreally Delivered Adeno-associated Viral Vectors at the Retina for Enhanced Transduction but Weakly Influences Tropism

Woodard

Liang

Bennett

et al. 2016

J Virol

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Many adeno-associated virus (AAV) serotypes efficiently transduce the retina when delivered to the subretinal space but show limited success when delivered to the vitreous due to the inner limiting membrane (ILM). Subretinal delivery of AAV serotype 2 (AAV2) and its heparan sulfate (HS)-binding-deficient capsid led to similar expression, indicating transduction of the outer retina occurred by HS-independent mechanisms. However, intravitreal delivery of HS-ablated recombinant AAV2 (rAAV2) led to a 300-fold decrease in transduction compared to AAV2. Fluorescence in situ hybridization of AAV transgenes was used to identify differences in retinal trafficking and revealed that HS binding was responsible for AAV2 accumulation at the ILM. This mechanism was tested on human ex vivo retinas and showed similar accumulation with HS-binding AAV2 capsid only. To evaluate if HS binding could be applied to other AAV serotypes to enhance their transduction, AAV1 and AAV8 were modified to bind HS with a single-amino-acid mutation and tested in mice. Both HS-binding mutants of AAV1 and AAV8 had higher intravitreal transduction than their non-HS-binding parent capsid due to increased retinal accumulation. To understand the influence that HS binding has on tropism, chimeric AAV2 capsids with dual-glycan usage were tested intravitreally in mice. Compared to HS binding alone, these chimeric capsids displayed enhanced transduction that was correlated with a change in tropism. Taken together, these data indicate that HS binding serves to sequester AAV capsids from the vitreous to the ILM but does not influence retinal tropism. The enhanced retinal transduction of HS-binding capsids provides a rational design strategy for engineering capsids for intravitreal delivery. A deno-associated virus (AAV) is a small (25-nm), nonpathogenic virus that has been extensively studied as a vector for gene transfer applications (1-3). The virus consists of two parts: the viral genome and the protein capsid. The viral genome can be largely replaced with a desired transgene to create recombinant AAV (rAAV) vectors used for gene delivery. The protein capsid is responsible for cell attachment and entry via a variety of glycans and cell surface receptors. There are 11 naturally occurring serotypes of AAV, denoted AAV1 to AAV11. Glycans and receptors have been elucidated for several AAV serotypes. Heparan sulfate proteoglycan (HSPG) has been shown to be used for both rAAV2 and rAAV3 cell entry (4). rAAV6 displays dual-glycan interaction with HSPG and sialic acid; however, HSPG binding alone is insufficient for cellular entry (5). Various linkages of sialic acid are important for the transduction of the rAAV1, rAAV4, and rAAV5 serotypes (6,7). N-linked galactose is used for the transduction of the rAAV9 serotype (8). Glycans expressed on the cell surface dictate the tissue and cellular tropism observed with the various AAV capsids. In addition to attachment to these glycans, AAV serotypes interact with cell receptors, including human growth factor rec...

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Adeno-Associated Viral Vectors Based on Serotype 3b Use Components of the Fibroblast Growth Factor Receptor Signaling Complex for Efficient Transduction

Cited by 9 publications

References 55 publications

SelectiveIn VivoTargeting of Human Liver Tumors by Optimized AAV3 Vectors in a Murine Xenograft Model

SelectiveIn VivoTargeting of Human Liver Tumors by Optimized AAV3 Vectors in a Murine Xenograft Model

Differential Adeno-Associated Virus Serotype-Specific Interaction Patterns with Synthetic Heparins and Other Glycans

Heparan Sulfate Binding Promotes Accumulation of Intravitreally Delivered Adeno-associated Viral Vectors at the Retina for Enhanced Transduction but Weakly Influences Tropism

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