AAV Biology, Infectivity and Therapeutic Use from Bench to Clinic

Vance, Melisa; Mitchell, Angela M.; Samulski, R. Jude

doi:10.5772/61988

Cited by 21 publications

(25 citation statements)

References 66 publications

(74 reference statements)

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“…Previous studies have shown that infection by different AAV serotypes is mediated by a variety of cell surface receptors (for review see 34 ). Specifically, cell surface N-linked sialic acid has been proposed as the primary receptor for AAV1 and AAV6 to infect and transduce cells.…”

Section: Aav Serotype Infection Is Mediated By Different Receptors Onmentioning

confidence: 99%

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Ambrosi

Sadananda

Klimas

et al. 2017

Preprint

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Aims: Adeno-associated viruses (AAVs) provide advantages in long-term, cardiac-specific gene expression. However, AAV serotype specificity data is lacking in cardiac models relevant to optogenetics. We aimed to identify the optimal AAV serotype (1, 6, or 9) in pursuit of scalable rodent and human models for cardiac optogenetics and elucidate the mechanism of virus uptake. Methods:In vitro syncytia of primary neonatal rat ventricular cardiomyocytes (NRVMs) and human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were infected with AAVs 1, 6, and 9 containing the transgene for eGFP or channelrhodopsin-2 (ChR2) fused to mCherry. In vivo adult rats were intravenously injected with AAV1 and 9 containing ChR2-mCherry.Results: Transgene expression profiles of rat and human cells in vitro revealed that AAV1 and 6 significantly outperformed AAV9. In contrast, systemic delivery of AAV9 in adult rat hearts yielded significantly higher levels of ChR2-mCherry expression and optogenetic responsiveness. We tracked the mechanism of virus uptake to purported receptor-mediators for AAV1/6 (cell surface sialic acid) and AAV9 (37/67kDa laminin receptor, LamR). In vitro desialylation of NRVMs and hiPSC-CMs with neuraminidase significantly decreased AAV1,6-mediated gene expression, but interestingly, desialylation of hiPSC-CMs increased AAV9-mediated expression. In fact, only very high viral doses of AAV9-ChR2-mCherry, combined with neuraminidase treatment yielded consistent optogenetic responsiveness in hiPSC-CMs. Differences between the in vitro and in vivo performance of AAV9 could be correlated to robust LamR expression in the adult and neonatal rat hearts, but no expression in vitro in cultured cells. The dynamic nature of LamR expression and its dependence on environmental factors was further corroborated in intact adult human ventricular tissue slices.

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Section: Aav Serotype Infection Is Mediated By Different Receptors Onmentioning

confidence: 99%

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Ambrosi

Sadananda

Klimas

et al. 2017

Preprint

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“…All AAV serotypes share at least 50% sequence homology. However, serotype AAV5 has the most divergent amino acid capsid sequence, and AAV4 also shows a considerable degree of divergence [50]. Surprisingly, this sequence diversity between serotypes is not scattered but primarily located in the looped out domains of the capsid protein [51].…”

Section: Aav Serotypes and Tissue Tropismmentioning

confidence: 99%

“…Close homology to all serotypes except AAV 4, 5, 11, and 12 [50] Low transduction efficiency [52] Liver, heart, muscle [52] AAV3 Low transduction efficiency Slow in targeting the tissues [52] Heart, liver [52] AAV4…”

Section: Aav1mentioning

confidence: 99%

“…Close homology to AAV11 (82%) and AAV12 (79%) serotypes [50] Low transduction efficiency Slow in targeting the tissues [52] Lung, heart, liver, central nervous system [52,134] AAV5…”

Section: Aav1mentioning

confidence: 99%

“…Low transduction efficiency [52] Liver [52] AAV6 Shares 99% homology with AAV1 serotype [50] Liver, heart, skeletal muscle [52] AAV7 Fast in targeting the tissues [52] Liver, skeletal muscle [52] AAV8 93% homology to AAV10 serotype [50] Heart, liver, brain, muscle (second most efficient serotype reaching the brain) [52] AAV9 Fast in targeting the tissues [52] Liver, heart, brain, lung, skeletal muscle (serotype with the broadest tissue tropism and most efficient in reaching the brain) [52] AAV10 93% homology to AAV8 serotype [50] Liver, heart, muscle, lung, kidney, uterus (with pseudotype AAV2/10) [135] AAV11 Close homology to AAV4 serotype [50] Muscle, kidney, spleen, lung, heart, stomach (with pseudotype AAV2/11) [135] AAV12 Close homology to AAV4 serotype [50] Muscle, salivary glands [136] Table 1. Characteristics and tissue tropism of AAV serotypes in the mouse.…”

Section: Aav1mentioning

confidence: 99%

See 2 more Smart Citations

Adeno-Associated Virus (AAV)-Mediated Gene Therapy for Disorders of Inherited and Non-Inherited Origin

Rajapaksha¹,

Angus²,

Herath³

2019

In Vivo and Ex Vivo Gene Therapy for Inherited and Non-Inherited Disorders

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Gene therapy is a novel promising approach for treating a spectrum of inherited and non-inherited disorders by delivering therapeutic genes to specific organs or tissues. Of the viral vectors that have been used to date to deliver the genes of interest, the adenoassociated viral (AAV) vector appears to be the most safe and effective vehicle and has the ability to maintain long-term gene and protein expression following a single injection of the vector. Gene therapy studies using AAV vector have shown significant progress not only in animal models but also in human gene therapy with no known pathogenicity. While success has been achieved in gene therapy using AAV vector to deliver the target genes for inherited disorders, however, clinical trials are yet to begin to see whether gene therapy has promise for treatment of non-inherited diseases. This chapter describes AAV biology, viral structure, and cell entry mechanisms, with special emphasis on AAV tissue tropism achieved by manipulating different serotypes and capsid engineering. This chapter also discusses successful application of the AAV vector for non-inherited disorders in animal models with particular reference to liver fibrosis, outlining advantages, disadvantages, and future challenges that this therapy may face.

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AAV Production Everywhere: A Simple, Fast, and Reliable Protocol for In‐house AAV Vector Production Based on Chloroform Extraction

et al. 2020

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Recombinant adeno‐associated virus (rAAV) is a mammalian virus that has been altered to be used as a gene delivery vehicle. Several changes to the viral genome have made them replication deficient so that this aspect of the viral infection cycle is under full control of the experimenter, while maintaining gene expression machinery. Over the last decades, rAAVs have become the gold standard for studying in vivo gene function and are especially favorable for gene transfer in the central nervous system. AAVs have been proven safe and provide stable gene expression over a long period of time. They are extensively used in preclinical experiments and show great potential for clinical applications. However, the use of AAVs in preclinical settings are often held back due to availability. Waiting lines are long at commercial production facilities, and in‐lab production is hindered due to lack of specific laboratory equipment needed. Here we present a novel production method that can be carried out in any molecular biology laboratory using standard laboratory equipment. We provide a simple, fast, and streamlined protocol for production that can result in titers comparable with the more time‐consuming iodixanol gradient ultracentrifugation method. The yield using this protocol is high enough for any type of study where AAV is the vector of choice. © 2020 The Authors.

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AAV Biology, Infectivity and Therapeutic Use from Bench to Clinic

Cited by 21 publications

References 66 publications

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Adeno-associated virus mediated gene delivery: Implications for scalablein vitroandin vivocardiac optogenetic models

Adeno-Associated Virus (AAV)-Mediated Gene Therapy for Disorders of Inherited and Non-Inherited Origin

AAV Production Everywhere: A Simple, Fast, and Reliable Protocol for In‐house AAV Vector Production Based on Chloroform Extraction

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