Isolation and evaluation of novel adeno-associated virus sequences from porcine tissues

Bello, Alexander; Tran, Kaylie; Chand, Allan; Doria, Monica; Allocca, Mariacarmela; Hildinger, Markus; Beniac, Daniel R.; Kranendonk, Christiaan; Auricchio, Alberto; Kobinger, Gary P.

doi:10.1038/gt.2009.82

Cited by 47 publications

(51 citation statements)

References 37 publications

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“…At present, 13 primate AAVs designated AAV1-AAV13 have been identified from human or non-human primates (Atchison et al, 1965;Bantel-Schaal & zur Hausen, 1984;Chiorini et al, 1997Chiorini et al, , 1999Gao et al, 2002Gao et al, , 2003Gao et al, , 2004Melnick et al, 1965;Mori et al, 2004;Schmidt et al, 2008;Schnepp et al, 2005Schnepp et al, , 2009). In addition to primates, a number of phylogenetically distinct AAVs have also been identified from other animals such as cow (Schmidt et al, 2004), goat (Arbetman et al, 2005), pig (Bello et al, 2009), chicken (Bossis & Chiorini, 2003;Hess et al, 1995), waterfowl (Brown et al, 1995) and snake (Farkas et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Prevalence and genetic diversity of adeno-associated viruses in bats from China

Hon

et al. 2010

Journal of General Virology

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Section: Introductionmentioning

confidence: 99%

Prevalence and genetic diversity of adeno-associated viruses in bats from China

Hon

et al. 2010

Journal of General Virology

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“…There are more than 150 naturally occurring variants, and 13 nonhuman and human serotypes have been described (1)(2)(3)(4)(5)(6). Due to their nonpathogenicity, ability to package recombinant DNA, long-term transgene expression, and ability to transduce dividing and nondividing cells, the AAVs are being developed as gene delivery vectors (7,8).…”

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confidence: 99%

Characterization of the Adeno-Associated Virus 1 and 6 Sialic Acid Binding Site

Huang

Patel

et al. 2016

J Virol

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The adeno-associated viruses (AAVs), which are being developed as gene delivery vectors, display differential cell surface glycan binding and subsequent tissue tropisms. For AAV serotype 1 (AAV1), the first viral vector approved as a gene therapy treatment, and its closely related AAV6, sialic acid (SIA) serves as their primary cellular surface receptor. Toward characterizing the SIA binding site(s), the structure of the AAV1-SIA complex was determined by X-ray crystallography to 3.0 Å. Density consistent with SIA was observed in a pocket located at the base of capsid protrusions surrounding icosahedral 3-fold axes. Site-directed mutagenesis substitution of the amino acids forming this pocket with structurally equivalent residues from AAV2, a heparan sulfate binding serotype, followed by cell binding and transduction assays, further mapped the critical residues conferring SIA binding to AAV1 and AAV6. For both viruses five of the six binding pocket residues mutated (N447S, V473D, N500E, T502S, and W503A) abolished SIA binding, whereas S472R increased binding. All six mutations abolished or decreased transduction by at least 50% in AAV1. Surprisingly, the T502S substitution did not affect transduction efficiency of wildtype AAV6. Furthermore, three of the AAV1 SIA binding site mutants-S472R, V473D, and N500E-escaped recognition by the anti-AAV1 capsid antibody ADK1a. These observations demonstrate that common key capsid surface residues dictate both virus binding and entry processes, as well as antigenic reactivity. This study identifies an important functional capsid surface "hot spot" dictating receptor attachment, transduction efficiency, and antigenicity which could prove useful for vector engineering. IMPORTANCEThe adeno-associated virus (AAV) vector gene delivery system has shown promise in several clinical trials and an AAV1-based vector has been approved as the first gene therapy treatment. However, limitations still exist with respect to transduction efficiency and the detrimental effects of preexisting host antibodies. This study aimed to identify key capsid regions which can be engineered to overcome these limitations. A sialic glycan receptor recognition pocket was identified in AAV1 and its closely related AAV6, using X-ray crystallography. The site was confirmed by mutagenesis followed by cell binding and transduction assays. Significantly, residues controlling gene expression efficiency, as well as antibody escape variants, were also identified. This study thus provides, at the amino acid level, information for rational structural engineering of AAV vectors with improved therapeutic efficacy.

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“…The utilization of alternative AAV serotypes could facilitate an escape from preexisting neutralizing antibodies, which act against natural infection, or could be applicable prior to treatment with AAV-based vectors. The application of a nonprimate animal-derived novel AAV as a vector is expected to reduce the immune response and increase the transfection efficiency of rAAV vectors in vivo [15]. Bello et al [15] obtained new AAV capsid sequences from porcine tissues and successfully generated a recombinant vector (AAV2/po1) by transfection.…”

Section: Discussionmentioning

confidence: 99%

“…The application of a nonprimate animal-derived novel AAV as a vector is expected to reduce the immune response and increase the transfection efficiency of rAAV vectors in vivo [15]. Bello et al [15] obtained new AAV capsid sequences from porcine tissues and successfully generated a recombinant vector (AAV2/po1) by transfection. They indicated that the AAV2/po1 vector has a distinct antigenic profile, which is not cross-neutralized by antisera generated against all other commonly used AAVs (serotypes 1–8).…”

Section: Discussionmentioning

confidence: 99%

A Molecular Epidemiological Investigation of Carriage of the Adeno-Associated Virus in Murine Rodents and House Shrews in China

et al. 2018

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Objective: To investigate the prevalence of the adeno-associated virus (AAV) in murine rodents and house shrews in 4 provinces of China. Methods: A total of 469 murine rodents and 19 house shrews were captured between May 2015 and May 2017. Cap gene of AAV sequences was obtained to evaluate the genetic characteristics of rat AAV. Results: Rat AAVs were found in 54.7% (267/488) of throat swabs, 14.3% (70/488) of fecal samples, and 18.4% (41/223) of serum samples. Rat AAVs were detected in 3 species of murine rodents including Rattus norvegicus (34.8%), R. tanezumi (43.0%), and R. losea (2.3%), and house shrews (Suncus murinus) (26.1%) from the selected sampling sites. Fourteen near-full-length Cap gene sequences, ranging in length from 2,156 to 2,169 nt, were isolated from the fecal samples of R. norvegicus and R. tanezumi. These 14 sequences shared a high identity of 97.4% at the nucleotide level and 99.1% at the amino acid level. Phylogenetic analysis showed that the rat AAV formed a distinct clade, distinguishable from the AAV discovered in humans and in other animals. Conclusions: A high prevalence of rat AAV that was highly conserved within the Cap gene was found in 3 common murine rodents and house shrews in China.

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Isolation and evaluation of novel adeno-associated virus sequences from porcine tissues

Cited by 47 publications

References 37 publications

Prevalence and genetic diversity of adeno-associated viruses in bats from China

Prevalence and genetic diversity of adeno-associated viruses in bats from China

Characterization of the Adeno-Associated Virus 1 and 6 Sialic Acid Binding Site

A Molecular Epidemiological Investigation of Carriage of the Adeno-Associated Virus in Murine Rodents and House Shrews in China

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