Corneal endothelial cell nuclei are damaged after DNA transfer using a gene gun

Klebe, Sonja; Jw, Stirling; Williams, KJ

doi:10.1046/j.1442-9071.2000.00255.x

Cited by 13 publications

(7 citation statements)

References 3 publications

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“…Intrastromal injection of naked DNA Epithelium, stromal cells (in vivo) 5 [36] Ultrasound and microbubbles Stromal cells (in vivo) 1 4 [40] Electroporation Stromal cells (in vivo) 1 0 [37,38,41] Gene gun Epithelium (in vivo) 5 [42][43][44][45][46][47] Synthetic peptide (DNA/polylisinemolossin/fusogenic peptide) Endothelium (ex vivo) 7 [48] Dendrimers (polyamidoamine) Endothelium (ex vivo) 5-10 [49] Liposomes Epithelium, keratocytes and endothelium 7 [50][51][52][53] Antibody-targeted liposomes (transferrin and E-selectin) Endothelium (ex vivo) 10-15 [39,54] Dehydrated plasmids (p-bFGF-polyethylenimine)…”

Section: Methods Target Cells Duration Of Gene Expression (Days)mentioning

confidence: 99%

Gene therapy for corneal graft survival

Bárcia¹,

Kazlauskas²

2007

Expert Review of Ophthalmology

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Corneal transplantation is the most successful form of solid organ transplant but rejection still occurs in approximately 10% of cases, which often leads to blindness. The cornea is an ideal organ for gene therapy and, as such, has been the target of many gene transfer studies. Numerous nonviral and viral vectors have been tested in the cornea, together with different transgene approaches that modulate tolerance, inflammation and angiogenesis. Present and future efforts should enable the use of this technology in the clinic to improve corneal graft survival.

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Section: Methods Target Cells Duration Of Gene Expression (Days)mentioning

confidence: 99%

Gene therapy for corneal graft survival

Bárcia¹,

Kazlauskas²

2007

Expert Review of Ophthalmology

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“…The options for gene transfer to the cornea and sclera include the application or injection of naked DNA or plasmids, 42–44 the use of the gene gun, 45,46 electroporation 47 or iontophoresis, 48 dendrimer‐ 49,50 or liposome‐based 50 non‐viral vectors, antibody 51 or peptide‐targeting 52 vector systems, nanoparticle‐mediated delivery, 50,53 use of the femtosecond laser, 54 and viral vectors 17 (Table 2). The physical and non‐viral methods of gene transfer are generally somewhat inefficient, although there are a few notable exceptions, 26,55 and the majority produce only short‐lived transgene expression.…”

Section: The Mechanics Of Gene Transfer To the Cornea: Selection Of Amentioning

confidence: 99%

“…Because of the normal turnover of corneal epithelial cells and subsequent loss of the transgene, any such approach to gene therapy will necessarily be temporally limited. The use of a gene gun to transduce corneal endothelium was shown to cause potentially serious cell damage in one study 46 …”

Section: The Mechanics Of Gene Transfer To the Cornea: Selection Of Amentioning

confidence: 99%

Gene therapy for diseases of the cornea – a review

Williams

Coster

2010

Clinical Exper Ophthalmology

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The cornea is particularly suited to gene therapy. The cornea is readily accessible, normally transparent, and is somewhat sequestrated from the general circulation and the systemic immune system. The principle of genetic therapy for the cornea is to use an appropriate vector system to transfer a gene to the cornea itself, or to the ocular environs, or systemically, so that a transgenic protein will be expressed that will modulate congenital or acquired disease. The protein may be structural such as a collagen, or functionally active such as an enzyme, cytokine or growth factor that may modulate a pathological process. Alternatively, gene expression may be silenced by the use of modalities such as antisense oligonucleotides. Interestingly, despite a very considerable amount of work in animal models, clinical translation directed to gene therapy of the human cornea has been minimal. This is in contrast to gene therapy for monogenic inherited diseases of the retina, where promising early results of clinical trials for Leber's congenital amaurosis have already been published and a number of other trials are ongoing.

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“…Ballistic: the gene gun propels gold nanoparticles conjugated with genes at high velocity directly into cells. This may be efficient but is difficult to control and showed high damage to cells, especially CECs [ 14 ]. Electricity: electroporation by very short high voltage pulses triggers the formation of holes in cell membranes and electrophoresis of charged molecules.…”

Section: Introductionmentioning

confidence: 99%

Delivery of Molecules into Human Corneal Endothelial Cells by Carbon Nanoparticles Activated by Femtosecond Laser

Jumelle

Mauclair²,

Houzet³

et al. 2015

PLoS ONE

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Corneal endothelial cells (CECs) form a monolayer at the innermost face of the cornea and are the engine of corneal transparency. Nevertheless, they are a vulnerable population incapable of regeneration in humans, and their diseases are responsible for one third of corneal grafts performed worldwide. Donor corneas are stored in eye banks for security and quality controls, then delivered to surgeons. This period could allow specific interventions to modify the characteristics of CECs in order to increase their proliferative capacity, increase their resistance to apoptosis, or release immunosuppressive molecules. Delivery of molecules specifically into CECs during storage would therefore open up new therapeutic perspectives. For clinical applications, physical methods have a more favorable individual and general benefit/risk ratio than most biological vectors, but are often less efficient. The delivery of molecules into cells by carbon nanoparticles activated by femtosecond laser pulses is a promising recent technique developed on non-adherent cells. The nanoparticles are partly consummated by the reaction releasing CO and H2 gas bubbles responsible for the shockwave at the origin of cell transient permeation. Our aim was to develop an experimental setting to deliver a small molecule (calcein) into the monolayer of adherent CECs. We confirmed that increased laser fluence and time exposure increased uptake efficiency while keeping cell mortality below 5%. We optimized the area covered by the laser beam by using a motorized stage allowing homogeneous scanning of the cell culture surface using a spiral path. Calcein uptake reached median efficiency of 54.5% (range 50.3–57.3) of CECs with low mortality (0.5%, range (0.55–1.0)). After sorting by flow cytometry, CECs having uptaken calcein remained viable and presented normal morphological characteristics. Delivery of molecules into CECs by carbon nanoparticles activated by femtosecond laser could prove useful for future cell or tissue therapy.

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Corneal endothelial cell nuclei are damaged after DNA transfer using a gene gun

Cited by 13 publications

References 3 publications

Gene therapy for corneal graft survival

Gene therapy for corneal graft survival

Gene therapy for diseases of the cornea – a review

Delivery of Molecules into Human Corneal Endothelial Cells by Carbon Nanoparticles Activated by Femtosecond Laser

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