Mechanisms of AAV transduction in glaucoma‐associated human trabecular meshwork cells

Borrás, Teresa; Wang, Xue; Choi, Vivian W.; Bartlett, Jeffrey S.; Li, Guorong; Samulski, R. Jude; Chisolm, Sarah

doi:10.1002/jgm.886

Cited by 49 publications

(55 citation statements)

References 45 publications

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“…Thus, it is likely that secreted MMP-3, expressed by corneal endothelia, will be directed toward the outflow tissue and activated with the aid of existing endogenous MMP-3 after which it will then be available for remodelling a range of ECM components. Attaining exclusive AAV expression in the cornea was obtained by using a single stranded AAV-2/9, as although single stranded AAV may enter other cells such as the TM, self-complementary viruses are required for sufficient DNA replication, and hence transduction of these cells (72). …”

Section: Discussionmentioning

confidence: 99%

Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma

O’Callaghan

Crosbie

Cassidy

et al. 2017

Human Molecular Genetics

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Intraocular pressure (IOP) is maintained as a result of the balance between production of aqueous humour (AH) by the ciliary processes and hydrodynamic resistance to its outflow through the conventional outflow pathway comprising the trabecular meshwork (TM) and Schlemm’s canal (SC). Elevated IOP, which can be caused by increased resistance to AH outflow, is a major risk factor for open-angle glaucoma. Matrix metalloproteinases (MMPs) contribute to conventional aqueous outflow homeostasis in their capacity to remodel extracellular matrices, which has a direct impact on aqueous outflow resistance and IOP. We observed decreased MMP-3 activity in human glaucomatous AH compared to age-matched normotensive control AH. Treatment with glaucomatous AH resulted in significantly increased transendothelial resistance of SC endothelial and TM cell monolayers and reduced monolayer permeability when compared to control AH, or supplemented treatment with exogenous MMP-3.Intracameral inoculation of AAV-2/9 containing a CMV-driven MMP-3 gene (AAV-MMP-3) into wild type mice resulted in efficient transduction of corneal endothelium and an increase in aqueous concentration and activity of MMP-3. Most importantly, AAV-mediated expression of MMP-3 increased outflow facility and decreased IOP, and controlled expression using an inducible promoter activated by topical administration of doxycycline achieved the same effect. Ultrastructural analysis of MMP-3 treated matrices by transmission electron microscopy revealed remodelling and degradation of core extracellular matrix components. These results indicate that periodic induction, via use of an eye drop, of AAV-mediated secretion of MMP-3 into AH could have therapeutic potential for those cases of glaucoma that are sub-optimally responsive to conventional pressure-reducing medications.

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Section: Discussionmentioning

confidence: 99%

Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma

O’Callaghan

Crosbie

Cassidy

et al. 2017

Human Molecular Genetics

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“…With intravitreal administration, adeno-associated viruses (AAVs) transduce ganglion cells, trabecular meshwork cells and various cells of the inner nuclear layer [34][35][36][37][38].…”

Section: Discussionmentioning

confidence: 99%

Baculovirus is an efficient vector for the transduction of the eye: comparison of baculovirus‐ and adenovirus‐mediated intravitreal vascular endothelial growth factor D gene transfer in the rabbit eye

Kinnunen

Kalesnykas

Laidinen

et al. 2009

The Journal of Gene Medicine

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“…They are relatively difficult to produce but high titers can be achieved (Grieger et al, 2006). There is an established track record for AAV vectors with inherited eye diseases (Bainbridge et al, 2008) but the transduction efficiency of conventional AAV in TM is very low due to the inability to convert single-stranded DNA into double-stranded DNA, which requires DNA polymerase complexes in host cells (Borrás et al, 2006). Transduction efficiency varies among serotypes of scAAV and host species (Bogner et al, 2015).…”

Section: Established Methods Of Ocular Gene Therapymentioning

confidence: 99%

Gene transfer to the outflow tract

Dang

Loewen

Parikh

et al. 2017

Experimental Eye Research

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Elevated intraocular pressure is the primary cause of open angle glaucoma. Outflow resistance exists within the trabecular meshwork but also at the level of Schlemm's canal and further downstream within the outflow system. Viral vectors allow to take advantage of naturally evolved, highly efficient mechanisms of gene transfer, a process that is termed transduction. They can be produced at biosafety level 2 in the lab using protocols that have evolved considerably over the last 15 to 20 years. Applied by an intracameral bolus, vectors follow conventional as well as uveoscleral outflow pathways. They may affect other structures in the anterior chamber depending on their transduction kinetics which can vary among species when using the same vector. Not all vectors can express long-term, a desirable feature to address the chronicity of glaucoma. Vectors that integrate into the genome of the target cell can achieve transgene function for the life of the transduced cell but are mutagenic by definition. The most prominent long-term expressing vector systems are based on lentiviruses that are derived from HIV, FIV, or EIAV. Safety considerations make non-primate lentiviral vector systems easier to work with as they are not derived from human pathogens. Non-integrating vectors are subject to degradation and attritional dilution during cell division. Lentiviral vectors have to integrate in order to express while adeno-associated viral vectors (AAV) often persist as intracellular concatemers but may also integrate. Adeno- and herpes viral vectors do not integrate and earlier generation systems might be relatively immunogenic. Nonviral methods of gene transfer are termed transfection with few restrictions of transgene size and type but often a much less efficient gene transfer that is also short-lived. Traditional gene transfer delivers exons while some vectors (lentiviral, herpes and adenoviral) allow transfer of entire genes that include introns. Recent insights have highlighted the role of non-coding RNA, most prominently, siRNA, miRNA and lncRNA. SiRNA is highly specific, miRNA is less specific, while lncRNA uses highly complex mechanisms that involve secondary structures and intergenic, intronic, overlapping, antisense, and bidirectional location. Several promising preclinical studies have targeted the RhoA or the prostaglandin pathway or modified the extracellular matrix. TGF-β and glaucoma myocilin mutants have been transduced to elevate the intraocular pressure in glaucoma models. Cell based therapies have started to show first promise. Past approaches have focused on the trabecular meshwork and the inner wall of Schlemm's canal while new strategies are concerned with modification of outflow tract elements that are downstream of the trabecular meshwork.

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Mechanisms of AAV transduction in glaucoma‐associated human trabecular meshwork cells

Cited by 49 publications

References 45 publications

Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma

Therapeutic potential of AAV-mediated MMP-3 secretion from corneal endothelium in treating glaucoma

Baculovirus is an efficient vector for the transduction of the eye: comparison of baculovirus‐ and adenovirus‐mediated intravitreal vascular endothelial growth factor D gene transfer in the rabbit eye

Gene transfer to the outflow tract

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