Engineering adeno-associated virus vectors for gene therapy

Li, Chengwen; Samulski, R. Jude

doi:10.1038/s41576-019-0205-4

Cited by 837 publications

(746 citation statements)

References 189 publications

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“…The highly e cient transduction of LBOs with rAAV serotypes 2 and 6 is consistent with the observed strong positive staining for heparan sulphate (Figure 3g & h), a key entry receptor for these serotypes [1].…”

Section: Discussionsupporting

confidence: 80%

“…Figure 3c) compared with non-transduced organoids (Figure 3b) indicating the suitability of this vector to transduce the human lung parenchyma. To further understand the basis for rAAV transduction, which can depend on both primary glycan receptors and protein co-receptors [1], we also stained the LBOs for the universal AAV co-receptor (AAVR or KIAA0319L, Figure 3d), α-2,3-linked sialic acid (Figure 3e & f), and heparan sulphate (Figure 3g & h). Scattered cells staining positive for α-2,3-linked sialic acid were observed (Figure 3e & f), along with staining of subcellular structures characteristic of AAVR ( [10], Figure 3d) and the 'spotted' staining pattern commonly observed for heparan sulphate ( [11] Figure 3g & h).…”

Section: Resultsmentioning

confidence: 99%

“…Recombinant adeno-associated virus (rAAV) is a well-established vector for gene delivery, currently in use clinically for gene therapy, with multiple, naturally occurring serotypes and arti cial variants facilitating species-speci c cell and tissue tropisms [1]. Engineering of new AAV capsids has been the focus of extensive research, but capsids selected in animal models and cancer cell lines often translate poorly to large animal models and humans.…”

Section: Introductionmentioning

confidence: 99%

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Identification of AAV serotypes for lung gene therapy in human embryonic stem cell derived lung organoids.

Meyer‐Berg

Yang

Lucas

et al. 2020

Preprint

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Abstract Gene therapy is being investigated for a range of serious lung diseases, such as cystic fibrosis and emphysema. Recombinant adeno-associated virus (rAAV) is a well-established, safe, viral-vector for gene delivery with multiple natural and artificial serotypes available displaying alternate cell, tissue and species-specific tropisms. Efficient AAV serotypes for the transduction of the conducting airways have been identified for several species; however, efficient serotypes for human lung parenchyma have not yet been identified. Here, we screened the ability of multiple AAV serotypes to transduce lung bud organoids (LBOs) - a model of human lung parenchyma generated from human embryonic stem cells. Microinjection of LBOs allowed us to model transduction from the luminal surface, similar to dosing via vector inhalation. We identified the natural rAAV2 and rAAV6 serotypes, along with synthetic rAAV6 variants, as having tropism for the human lung parenchyma. Positive staining of LBOs for surfactant proteins B and C confirmed distal lung identity and suggested the suitability of these vectors for the transduction of alveolar type II cells. Our findings establish LBOs as a new model for pulmonary gene therapy and stress the relevance of LBOs as a viral infection model of the lung parenchyma as relevant in SARS-CoV-2 research.

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Section: Discussionsupporting

confidence: 80%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Identification of AAV serotypes for lung gene therapy in human embryonic stem cell derived lung organoids.

Meyer‐Berg

Yang

Lucas

et al. 2020

Preprint

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“…Recombinant vectors based on AAV (rAAV) are currently considered as an improved tool for safe gene transfer due to their unique properties, such as low immunogenicity and wide tissue tropism. 1 Thus, they have been frequently used in clinical trials. Three rAAV-based gene therapy drugs, Glybera, LUXTURNA, and ZOLGENSMA (formerly AVXS-101), have been approved to launch into the market for treating lipoprotein lipase deficiency, the vision loss caused by RPE65 gene mutations and spinal muscular atrophy, respectively.…”

Section: Introductionmentioning

confidence: 99%

Mechanism for enhanced transduction of hematopoietic cells by recombinant adeno‐associated virus serotype 6 vectors

Chen

Jia

et al. 2020

FASEB j.

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Hematopoietic gene delivery, such as hematopoietic stem/progenitor cells (HSPCs), is a promising treatment for both inherited and acquired diseases, such as hemophilia. Recently, a combined strategy to achieve more than 90% transduction efficiency was documented using recombinant adeno‐associated virus serotype 6 (rAAV6) vectors. However, the mechanisms of enhanced vector transduction efficiency in hematopoietic cells are largely unknown. In this manuscript, we first reported that proteasome inhibitors, which are well‐known to facilitate rAAV intracellular trafficking in various cell types, are not effective in hematopoietic cells. From the screening of small molecules derived from traditional Chinese medicine, we demonstrated that shikonin, a potential reactive oxygen species (ROS) generator, significantly increased the in vitro and ex vivo transgene expression mediated by rAAV6 vectors in hematopoietic cells, including human cord blood‐derived CD34 + HSPCs. Shikonin mainly targeted vector intracellular trafficking, instead of host cell entry or endonuclear single to double strand vector DNA transition, in a vector serotype‐dependent manner. Moreover, a ROS scavenger completely prevented the capability of shikonin to enhance rAAV6 vector‐mediated transgene expression. Taken together, these studies expand our understanding of rAAV6‐mediated transduction in hematopoietic cells and are informative for improving rAAV6‐based treatment of blood diseases.

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“…This is highlighted by the recent FDA approval and clinical success of Zolgensma®, an intravenously dosed AAV vector delivering a functional copy of the SMN1 gene in children with Spinal Muscular Atrophy (SMA)(1). Further, the list of systemically dosed AAV-based gene therapies for rare disorders such as Hemophilia A & B, Duchenne Muscular Dystrophy (DMD), X-linked myotubularin myopathy (XLMTM) and Pompe disease amongst others continues to grow(2, 3). These promising clinical examples have concurrently highlighted important challenges that include manufacturing needs, patient recruitment, and the potential for toxicity at high AAV doses.…”

Section: Introductionmentioning

confidence: 99%

Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme

Elmore

Simon

et al. 2020

Preprint

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20Pre-existing humoral immunity to recombinant adeno-associated viral (AAV) vectors 21 restricts the treatable patient population and efficacy of human gene therapies. Approaches to 22 clear neutralizing antibodies (NAbs), such as plasmapheresis and immunosuppression are either 23 ineffective or cause undesirable side effects. Here, we describe a clinically relevant strategy to 24 rapidly and transiently degrade NAbs prior to AAV administration using an IgG degrading 25 enzyme (IdeZ). We demonstrate that recombinant IdeZ efficiently cleaves IgG in dog, monkey 26 and human antisera. Prophylactically administered IdeZ cleaves circulating, human IgG in mice 27 and prevents AAV neutralization in vivo. In macaques, a single intravenous dose of IdeZ rescues 28 AAV transduction by transiently reversing seropositivity. Importantly, IdeZ efficiently cleaves 29 NAbs and rescues AAV transduction in mice passively immunized with individual human donor 30 sera representing a diverse population. Our antibody clearance approach presents a new 31 paradigm for expanding the prospective patient cohort and improving efficacy of AAV gene 32 therapy. 34Human gene therapy using recombinant AAV vectors continues to advance steadily as a 37 treatment paradigm for rare, monogenic disorders. This is highlighted by the recent FDA approval 38 and clinical success of Zolgensma®, an intravenously dosed AAV vector delivering a functional 39 copy of the SMN1 gene in children with Spinal Muscular Atrophy (SMA)(1). Further, the list of 40 systemically dosed AAV-based gene therapies for rare disorders such as Hemophilia A & B, 41 Duchenne Muscular Dystrophy (DMD), X-linked myotubularin myopathy (XLMTM) and Pompe 42 disease amongst others continues to grow(2, 3). These promising clinical examples have 43 concurrently highlighted important challenges that include manufacturing needs, patient 44 recruitment, and the potential for toxicity at high AAV doses. One such challenge that limits the 45 recruitment of patients for gene therapy clinical trials and adversely affects the efficacy of AAV 46 gene therapy is the prevalence of pre-existing neutralizing antibodies (NAbs) to AAV capsids in 47 the human population. Such NAbs arise due to natural infection or cross-reactivity between 48 different AAV serotypes(4-7). NAbs can mitigate AAV infection through multiple mechanisms 49 by (a) binding to AAV capsids and blocking critical steps in transduction such as cell surface 50 attachment and uptake, endosomal escape, productive trafficking to the nucleus or uncoating and 51 (b) promoting AAV opsonization by phagocytic cells, thereby mediating their rapid clearance from 52 the circulation. Multiple preclinical studies in different animal models have demonstrated that pre-53 existing NAbs impede systemic gene transfer by AAV vectors(8-11). 54In humans, serological studies reveal a high prevalence of NAbs in the worldwide 55 population, with about 67% of people having antibodies against AAV1, 72% against AAV2, and 56 ~ 40% against AAV serotypes 5 thr...

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Engineering adeno-associated virus vectors for gene therapy

Cited by 837 publications

References 189 publications

Identification of AAV serotypes for lung gene therapy in human embryonic stem cell derived lung organoids.

Identification of AAV serotypes for lung gene therapy in human embryonic stem cell derived lung organoids.

Mechanism for enhanced transduction of hematopoietic cells by recombinant adeno‐associated virus serotype 6 vectors

Rescuing AAV gene transfer from antibody neutralization with an IgG-degrading enzyme

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