Utility of PEGylated recombinant adeno-associated viruses for gene transfer

Le, Hong T.; Yu, Qian; Wilson, James M.; Croyle, Maria A.

doi:10.1016/j.jconrel.2005.07.019

Cited by 69 publications

(57 citation statements)

References 50 publications

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“…To overcome AAV NAb inhibition of AAV vector transduction, several approaches have been exploited in the lab. One of them is to use a polymer to cover the AAV surface and block NAb recognition (11,32,33). The second approach is to use error-prone PCR to generate a library of AAV variants and select for NAb escape mutants in the presence of NAb in vitro (31,41,56).…”

Section: Discussionmentioning

confidence: 99%

“…These findings present a concern to the gene therapy community as to how we can avoid antagonistic NAb activity during systemic delivery of AAV vectors. To overcome this concern, several approaches have been exploited in our lab and by other groups, including (i) the utilization of polymers to cover the AAV capsid and block NAb recognition (11,32,33), (ii) the development of NAb escape mutants by error-prone PCR in vitro (31,41,56), (iii) the application of other types of AAV vectors (27,30,38,60,65), and (iv) the generation of chimeric types via AAV shuffling (2,36,37). In this study, we have systematically explored the possibility of using different types and AAV mutants as alternative vectors for intramuscular gene delivery in mice preimmunized against different AAV types.…”

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confidence: 99%

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Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

DiPrimio

Bowles

et al. 2012

J Virol

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Adeno-associated virus (AAV) vectors have the potential to promote long-term gene expression. Unfortunately, humoral immunity restricts patient treatment and in addition provides an obstacle to the potential option of vector readministration. In this study, we describe a comprehensive characterization of the neutralizing antibody (NAb) response to AAV type 1 (AAV1) through AAV5 both in vitro and in vivo. These results demonstrated that NAbs generated from one AAV type are unable to neutralize the transduction of other types. We extended this observation by demonstrating that a rationally engineered, muscle-tropic AAV2 mutant containing 5 amino acid substitutions from AAV1 displayed a NAb profile different from those of parental AAV2 and AAV1. Here we found that a single insertion of Thr from AAV1 into AAV2 capsid at residue 265 preserved high muscle transduction, while also changing the immune profile. To better understand the role of Thr insertion at position 265, we replaced all 20 amino acids and evaluated both muscle transduction and the NAb response. Of these variants, 8 mutants induced higher muscle transduction than AAV2. Additionally, three classes of capsid NAb immune profile were defined based on the ability to inhibit transduction from AAV2 or mutants. While no relationship was found between transduction, amino acid properties, and NAb titer or its cross-reactivity, these studies map a critical capsid motif involved in all steps of AAV infectivity. Our results suggest that AAV types can be utilized not only as templates to generate mutants with enhanced transduction efficiency but also as substrates for repeat administration.A deno-associated virus (AAV) vectors have been safely and successfully used in phase I clinical trials (39,40,54). In particular, therapeutic effects have been achieved in patients with Leber's congenital amaurosis and hemophilia B. Among 30 patients with Leber's congenital amaurosis, 28 demonstrated remarkably improved eyesight after AAV type 2 (AAV2) delivery of the rpe65 gene to the retina. Regarding the recent hemophilia B trial, all six patients had therapeutic factor IX (FIX) expression within a beneficial 1% to 8% of normal levels as long as 18 months after intravenous delivery of an AAV vector encoding an optimized FIX cassette (54).AAV is a nonenveloped, single-stranded DNA virus that requires a helper virus, such as adenovirus (Ad) or herpes simplex virus, for efficient replication. Despite the lack of inflammation or other AAV-associated complications following administration of AAV2 vectors in several organs, neutralizing antibody (NAb) titers against the AAV2 capsid were found to be significantly increased following vector administration, particularly in the lung, muscle, and liver (7, 8, 43-45, 49, 54, 63). NAbs in circulation are able to block AAV transduction after systemic administration. Recently, Manno et al. (44) performed a phase I study of AAV-mediated FIX transgene delivery in patients with hemophilia B, in which one patient with a higher preexisting NAb ti...

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

DiPrimio

Bowles

et al. 2012

J Virol

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“…Importantly, PEGylation-that is, the process of attaching PEG-has been used previously to prevent neutralization of adenoviral vectors upon repeated vector administration (O'Riordan et al, 1999;Croyle et al, 2001Croyle et al, , 2002). It is not surprising then that several groups tested whether PEGylation or modification with other ligands of AAV could render AAV vectors resistant to NAbs (Le et al, 2005;Lee et al, 2005;Carlisle et al, 2008). The picture that emerged from these studies is that chemical surface modification is challenging and, overall, only resulted in moderate increase to neutralization by pre-existing antibodies (Le et al, 2005;Carlisle et al, 2008).…”

Section: Shielding Of Aav By Chemical Modificationmentioning

confidence: 99%

“…It is not surprising then that several groups tested whether PEGylation or modification with other ligands of AAV could render AAV vectors resistant to NAbs (Le et al, 2005;Lee et al, 2005;Carlisle et al, 2008). The picture that emerged from these studies is that chemical surface modification is challenging and, overall, only resulted in moderate increase to neutralization by pre-existing antibodies (Le et al, 2005;Carlisle et al, 2008). This can likely be attributed to the fact that, because of the small size and compactness of the AAV capsid, antigenic regions and regions essential for transduction are in close proximity.…”

Section: Shielding Of Aav By Chemical Modificationmentioning

confidence: 99%

Pre-existing Anti–Adeno-Associated Virus Antibodies as a Challenge in AAV Gene Therapy

Jeune

Joergensen

Hajjar

et al. 2013

Human Gene Therapy Methods

270

159

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Adeno-associated virus (AAV)-based vectors are promising tools for gene therapeutic applications, in part because AAVs are nonpathogenic viruses, and vectors derived from them can drive long-term transgene expression without integration of the vector DNA into the host genome. AAVs are not strongly immunogenic, but they can, nonetheless, give rise to both a cellular and humoral immune response. As a result, a significant fraction of potential patients for AAV-based gene therapy harbors pre-existing antibodies against AAV. Because even very low levels of antibodies can prevent successful transduction, antecedent anti-AAV antibodies pose a serious obstacle to the universal application of AAV gene therapy. In this review, we discuss the current knowledge of the role of anti-AAV antibodies in AAV-based gene therapy with a particular emphasis on approaches to overcome the hurdle that they pose.

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“…Coating of virus-like particles with polymers could have other benefits, such as protection of virus-like particles from neutralization by the immune system and improved infection efficiency. [28][29][30] It is important to know the mechanism of cell penetration and the mechanism of siRNA protection by the novel PEI-AAV2-VLPs. Investigation of these mechanisms in future studies will definitely provide more understanding of this new formulation.…”

Section: Discussionmentioning

confidence: 99%

A novel polyethyleneimine-coated adeno-associated virus-like particle formulation for efficient siRNA delivery in breast cancer therapy: preparation and in vitro analysis

Shao

Paul²,

Abbasi³

et al. 2012

IJN

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Background: Systemic delivery of small interfering RNA (siRNA) is limited by its poor stability and limited cell-penetrating properties. To overcome these limitations, we designed an efficient siRNA delivery system using polyethyleneimine-coated virus-like particles derived from adeno-associated virus type 2 (PEI-AAV2-VLPs). Methods: AAV2-VLPs were produced in insect cells by infection with a baculovirus vector containing three AAV2 capsid genes. Using this method, we generated well dispersed AAV2-VLPs with an average diameter of 20 nm, similar to that of the wild-type AAV2 capsid. The nanoparticles were subsequently purified by chromatography and three viral capsid proteins were confirmed by Western blot. The negatively charged AAV2-VLPs were surface-coated with PEI to develop cationic nanoparticles, and the formulation was used for efficient siRNA delivery under optimized transfection conditions. Results: PEI-AAV2-VLPs were able to condense siRNA and to protect it from degradation by nucleases, as confirmed by gel electrophoresis. siRNA delivery mediated by PEI-AAV2-VLPs resulted in a high transfection rate in MCF-7 breast cancer cells with no significant cytotoxicity. A cell death assay also confirmed the efficacy and functionality of this novel siRNA formulation towards MCF-7 cancer cells, in which more than 60% of cell death was induced within 72 hours of transfection. Conclusion: The present study explores the potential of virus-like particles as a new approach for gene delivery and confirms its potential for breast cancer therapy.

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Utility of PEGylated recombinant adeno-associated viruses for gene transfer

Cited by 69 publications

References 50 publications

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

Pre-existing Anti–Adeno-Associated Virus Antibodies as a Challenge in AAV Gene Therapy

A novel polyethyleneimine-coated adeno-associated virus-like particle formulation for efficient siRNA delivery in breast cancer therapy: preparation and in vitro analysis

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