Highly Efficient Delivery of Adeno-Associated Viral Vectors to the Primate Retina

BoyeShannon, E; AlexanderJohn, J; Witherspoon, C. Douglas; BoyeSanford, L; PetersonJames, J; ClarkMark, E; SandeferKristen, J; Girkin, Christopher A.; Hauswirth, William W.; Gamlin, Paul D.

doi:10.1089/hum.2016.085

Cited by 67 publications

(54 citation statements)

References 60 publications

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“…We and others have shown transduction of macaque cones using AAV variants with ubiquitous promoters (16,(29)(30)(31)(32), but achieving cone transduction by vitreally administered AAV has only been possible at high doses, leading to inflammation (16,29). We reasoned that foveal cone targeting could be achieved if we use a strong cone-specific promoter at lower intravitreally injected AAV doses compatible with safety (29,33).…”

Section: Resultsmentioning

confidence: 99%

Noninvasive gene delivery to foveal cones for vision restoration

Khabou¹,

Garita-Hernández²,

Chaffiol³

et al. 2018

JCI Insight

101

105

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Section: Resultsmentioning

confidence: 99%

Noninvasive gene delivery to foveal cones for vision restoration

Khabou¹,

Garita-Hernández²,

Chaffiol³

et al. 2018

JCI Insight

101

105

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“…We were able to increase the amount of vector in the retina by adding an HS-binding motif to the non-HS-binding capsids. Currently, the methods for getting vector into the retina using high-titer virus are to either digest the ILM or perform a sub-ILM injection (19)(20)(21)43). It is easy to see how development of better capsids that are capable of transducing without these aids would be of interest to researchers and clinicians alike.…”

Section: Discussionmentioning

confidence: 99%

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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“…The greatest challenge when utilizing AAV vectors for in vivo labelling of RGCs is to overcome the inner limiting membrane, which acts as a physical barrier to achieving a high transduction rate. [54][55][56] Administering AAV vectors via intravitreal injection has been shown to be a useful method for longitudinal labelling and imaging ( Figure 5). 57 This technique provides the ability to quantify an estimate of cell density in a living animal and monitor changes in cell labelling.…”

Section: Adeno-associated Viral (Aav) Vector Labellingmentioning

confidence: 99%

Assessing retinal ganglion cell damage

Smith

Vianna

Chauhan

2017

Eye

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Retinal ganglion cell (RGC) loss is the hallmark of optic neuropathies, including glaucoma, where damage to RGC axons occurs at the level of the optic nerve head. In experimental glaucoma, damage is assessed at the axon level (in the retinal nerve fibre layer and optic nerve head) or at the soma level (in the retina). In clinical glaucoma where measurements are generally limited to non-invasive techniques, structural measurements of the retinal nerve fibre layer and optic nerve head, or functional measurements with perimetry provide surrogate estimates of RGC integrity. These surrogate measurements, while clinically useful, are several levels removed from estimating actual RGC loss. Advances in imaging, labelling techniques, and transgenic medicine are making enormous strides in experimental glaucoma, providing knowledge on the pathophysiology of glaucoma, its progression and testing new therapeutic avenues. Advances are also being made in functional imaging of RGCs. Future efforts will now be directed towards translating these advances to clinical care.

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Highly Efficient Delivery of Adeno-Associated Viral Vectors to the Primate Retina

Cited by 67 publications

References 60 publications

Noninvasive gene delivery to foveal cones for vision restoration

Noninvasive gene delivery to foveal cones for vision restoration

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

Assessing retinal ganglion cell damage

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