Specific tools for targeting and expression in Müller glial cells

Pellissier, Lucie P.; Hoek, Robert M.; Vos, Rogier M.; Aartsen, Wendy M.; Klimczak, R. R.; Hoyng, Stefan A.; Flannery, John G.; Wijnholds, Jan

doi:10.1038/mtm.2014.9

Cited by 50 publications

(66 citation statements)

References 42 publications

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“…S5a in the supplemental material). Both rAAV1 and rAAV6 capsids displayed a punctate expression pattern that clustered around the retinal vessels and that has been documented in other reports (23,24). Because of the homology between rAAV1 and rAAV6, only rAAV1 and rAAV1-E531K were further evaluated for possible differences in cell tropism.…”

Section: Hs Binding Increases the Ivit Transduction Of Other Raav Sermentioning

confidence: 73%

“…In rodent models, transduction with this serotype has been seen in occasional Müller glia, amacrine, and horizontal cells. In addition, rAAV6 expression in the RGC and inner nuclear layer (INL) has been seen in rodent models (23)(24)(25). Understanding viral trafficking and barriers to efficient IVit transduction provides opportunities to rationally design capsids to overcome current limitations.…”

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confidence: 99%

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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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Section: Hs Binding Increases the Ivit Transduction Of Other Raav Sermentioning

confidence: 73%

mentioning

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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“…The AAV6-derived capsid variant ShH10 and the AAV 2 RLBP1 GFP WPRE bGHpolyA vector with a Müller cell specific retinaldehyde binding protein 1 promoter 25 (kindly provided by Dr. Jan Wijnholds, Leiden University, The Netherlands) were used to overexpress the human sEH in murine Müller glial cells. Briefly, the myc- and His-tagged sEH used for the adenovirus was subcloned from the pcDNA3.1/myc-His vector (Thermo Fisher Scientific, Karslruhe, Germany) via EcoRI and AflII after T4 polymerase treatment in the AgeI digested and T4 polymerase blunted AAV 2 RLBP1 GFP WPRE bGHpolyA vector.…”

Section: Methodsmentioning

confidence: 99%

Inhibition of soluble epoxide hydrolase prevents diabetic retinopathy

Dziumbla

Lin

et al. 2017

Nature

129

121

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Diabetic retinopathy is an important cause of blindness in the adult population1,2 and is characterized by a progressive loss of vascular cells and slow dissolution of inter-vascular junctions resulting in vascular leak and retinal edema3. Later stages of the disease are characterized by inflammatory cell infiltration, tissue destruction and neovascularization4,5. Here we identify the soluble epoxide hydrolase (sEH) as a key enzyme that initiates the pericyte “drop off” and loss of endothelial barrier function by generating a diol from docosahexaenoic acid (DHA) i.e. 19,20-dihydroxydocosapentaenoic acid (19,20-DHDP). The expression of the sEH and the accumulation of 19,20-DHDP were elevated in diabetic murine and human retinas as well as in human vitreous. Mechanistically, the diol targeted the cell membrane to alter the localisation of cholesterol-binding proteins, and interfered with the association of presenilin 1 (PS1) with N-cadherin and VE-cadherin to compromise pericyte-endothelial cell as well as inter-endothelial cell junctions. Treating diabetic mice with a specific sEH inhibitor prevented the pericyte loss and vascular permeability that are characteristic of non-proliferative diabetic retinopathy. Overexpression of the sEH in the retinal Müller glial cells of non-diabetic mice, on the other hand, resulted in vessel abnormalities similar to those seen in diabetic animals with retinopathy. Thus, increased expression of the sEH is a determinant event in the pathogenesis of diabetic retinopathy and sEH inhibition can prevent the progression of the disease.

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“…Our results show that a single intraganglionic injection of AAVshH10 or AAVshH19 carrying a CMV‐EGFP cassette induced highly efficient, selective transduction of sensory neurons, but no significant enhancement in transduction of SGCs. This indicates that although the divergent capsid residues in AAVshH10 and AAVshH19 may act in concert to improve glia transduction in the visual system or brain glioma (Byrne et al, ; Klimczak et al, ; Koerber et al, ; Pellissier et al, ; Vacca et al, ), this shift in tropism does not occur when applied in DRG. Nonetheless, the capsid variants of shH10 and shH19 may be considered for use as viable alternatives to AAV6 and AAV2 for efficient neuronal transduction in DRG.…”

Section: Discussionmentioning

confidence: 99%

Glial fibrillary acidic protein promoter determines transgene expression in satellite glial cells following intraganglionic adeno‐associated virus delivery in adult rats

Xiang

Fan

et al. 2017

J of Neuroscience Research

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Recombinant adeno-associated viral (AAV)-mediated therapeutic gene transfer to dorsal root ganglia (DRG) is an effective and safe tool for treating chronic pain. However, AAV with various constitutively active promoters leads to transgene expression predominantly to neurons while glial cells are refractory to AAV transduction in the peripheral nervous system. The present study evaluated whether in vivo satellite glial cells (SGC) transduction in the DRGs can be enhanced by the SGC-specific GFAP promoter and by using shH10 and shH19 that are engineered capsid variants with Müller glia-prone transduction. Titer-matched AAV6 (as control), AAVshH10, and AAVshH19, all encoding the EGFP driven by the constitutively active CMV promoter, as well as AAV6-EGFP and AAVshH10-EGFP driven by a GFAP promoter (AAV6-GFAP-EGFP and AAVshH10-GFAP-EGFP) were injected into DRGs of adult male rats. Neurotropism of gene expression was determined and compared by immunohistochemistry. Results showed that injection of AAV6- and AAVshH10-GFAP-EGFP induces robust EGFP expression selectively in SGCs, whereas injection of either AAVshH10-CMV-EGFP or AAVshH19-CMV-EGFP into DRGs resulted in a similar in vivo transduction profile to AAV6-CMV-EGFP, all showing efficient transduction of sensory neurons without significant transduction of glial cell populations. Co-injection of AAV6-CMV-mCherry and AAV6-GFAP-EGFP induces transgene expression in neurons and SGCs separately. This report, together with our prior studies, demonstrates that the GFAP promoter rather than capsid tropism determines selective gene expression in SGCs following intraganglionic AAV delivery in adult rats. A dual AAV system, one with GFAP promoter and the other with CMV promoter, can efficiently express transgenes selectively in neurons vs. SGCs.

show abstract

Specific tools for targeting and expression in Müller glial cells

Cited by 50 publications

References 42 publications

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

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

Inhibition of soluble epoxide hydrolase prevents diabetic retinopathy

Glial fibrillary acidic protein promoter determines transgene expression in satellite glial cells following intraganglionic adeno‐associated virus delivery in adult rats

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