Efficiency and Toxicity of Liposome-mediated Gene Transfer to Corneal Endothelial Cells

Pleyer, Uwe; Groth, Detlef; Hinz, Britta; Keil, Oliver; Bertelmann, Eckart; Rieck, Peter; Reszka, Regina

doi:10.1006/exer.2001.1005

Cited by 45 publications

(30 citation statements)

References 20 publications

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“…Indeed, such adaptation may be at the basis for seemingly discordant reports of capillary transcytosis of albumin in 2 different caveolin-1 knockout lines. 41,42 Transfection methods, on the other hand, are recognized as potentially toxic 24 as well as inefficient when applied to nonproliferating epithelial/endothelial cells. 25 And subjection of growing cells to transfection further posed the risk that confluence would not be achieved.…”

Section: Discussionmentioning

confidence: 99%

“…This qualification precluded introducing siRNA by transfection, because nonproliferating epithelial/endothelial monolayers resist nucleic acid uptake and the transfection process itself significantly damages endothelial integrity. 24,25 Hence, caveolin-1 knockdown was achieved by transduction of just subconfluent BMECs with recombinant adenovirus encoding a hairpin loop siRNA (Ad-siCav-1) that targeted the caveolin-1 transcript. This was feasible because mouse endothelial cells contain the requisite coxsackievirus and adenovirus receptor (CAR) that mediates viral attachment and infection.…”

Section: Introductionmentioning

confidence: 99%

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Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells

Song

Pachter

2006

Blood

152

127

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Recent evidence from this laboratory indicated that reduced expression of caveolin-1 accompanied the diminished expression of tight junction (TJ)-associated proteins occludin and zonula occludens-1 (ZO-1) following stimulation of brain microvascular endothelial cells (BMECs) with the chemokine CCL2 (formerly called MCP-1). Because attenuated caveolin-1 levels have also been correlated with heightened permeability of other endothelia, the objective of this study was to test the hypothesis that reduced caveolin-1 expression is causally linked to the action of CCL2 on BMEC junctional protein expression and barrier integrity. This was achieved using adenovirus to nondestructively deliver caveolin-1 siRNA (Ad-siCav-1) to BMEC monolayers, which model the blood-brain barrier (BBB). Treatment with siRNA reduced the caveolin-1 protein level as well as occludin and ZO-1. Additionally, occludin exhibited dissociation from the cytoskeletal framework. These changes were attended by comparable alterations in adherens junction (AJ)-associated proteins, VE-cadherin and ␤-catenin, increased BMEC paracellular permeability, and facilitated the ability of CCL2 to stimulate monocytic transendo- IntroductionElevated permeability of the normally highly restrictive bloodbrain barrier (BBB) accompanies a variety of central nervous system (CNS) afflictions, including inflammation, infection, ischemia, seizures, and trauma. [1][2][3][4][5][6] Nevertheless, mechanisms regulating BBB permeability in physiological and pathophysiological situations remain poorly defined. Among the many factors affecting BBB permeability, this laboratory recently reported that the beta-chemokine CCL2 (formerly known as monocyte chemoattractant protein-1 [MCP-1]), which is elevated in the CNS during a variety of neuroinflammatory conditions, 7-10 plays an influential role. 11 Specifically, it effects dissociation of tight junction (TJ)-associated proteins occludin and zonula occludens-1 (ZO-1) from the cytoskeletal framework of brain microvascular endothelial cells (BMECs) comprising the BBB as well as diminished expression of these proteins. Given the proposed role(s) of TJs in restricting solute and cellular passage across endothelial and epithelial barriers, 12-14 such action could conceivably lie, in part, at the basis of the altered BBB permeability and accumulation of leukocytes in the CNS observed in certain neuroinflammatory episodes.Aside from the loss of occludin and ZO-1 following BMEC exposure to CCL2, expression of caveolin-1 was also significantly down-regulated. 11 This additional chemokine-associated loss may further contribute to BBB alteration, because caveolin-1 is the major structural protein of caveolae, membrane microdomains critically involved in various aspects of vesicular trafficking and cell signaling. [15][16][17] Of particular significance in this regard, Nusrat et al 18 reported that both occludin and ZO-1 might be organized within TJs by association with caveolin-1 in detergent-insoluble glycolipid rafts, membrane specializations...

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells

Song

Pachter

2006

Blood

152

127

View full text Add to dashboard Cite

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“…Recent attempts have been made to formulate cationic liposomes from degradable amphiphilic chemicals that can be metabolized following uptake into cells (Choi et al 2001;Pleyer et al 2001). These contrast with the nonbiodegradable nature of lipids used in earlier formulations such as DOTMA.…”

Section: Toxicity Owing To Cationic Liposomes and Lipoplexes In Cell mentioning

confidence: 99%

Cytotoxicity issues pertinent to lipoplex-mediated gene therapy in-vivo

Dass

2002

Journal of Pharmacy and Pharmacology

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Cationic liposomes bind with nucleic acids such as plasmids and oligodeoxynucleotides to form complexes known as lipoplexes. Although these lipoplexes have several advantages over other forms of nucleic acid transfer methods in cell culture and in-vivo, toxicity remains a problem, especially in-vivo. Nevertheless, these carriers have been used in clinical trials against cystic brosis and cancer and their usage is attributed mainly to their versatility, especially when it comes to the range of routes available for administration of nucleic-acid-based drugs in-vivo.

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“…9,10 Non-viral vectors, although safe, are generally inefficient. 35 The few exceptions, for example the synthetic peptide-based Gene-modified corneal allografts KA Williams et al vector described by Fabre and co-workers, 36 produce only short-term gene expression, of the order of a few days up to a week, in ocular tissues. The majority of studies exploring gene transfer to corneal endothelium have used replication-deficient adenoviral vectors.…”

Section: Vectors For Gene Therapy Of the Corneal Endotheliummentioning

confidence: 99%

Prospects for genetic modulation of corneal graft survival

Williams

Brereton

Coster

2009

Eye

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Irreversible immunological rejection is the major cause of clinical corneal graft failure. Ex vivo gene therapy directed at the donor cornea has been shown to prolong orthotopic corneal allograft survival significantly in experimental animal models including the mouse, rat, rabbit, and sheep. Transgenes effective in prolonging corneal graft survival include immunomodulatory cytokines and cytokine receptors, an inhibitor of neovascularisation, a blocker of antigen-presenting cell-T cell co-stimulation, nerve growth factor, a dominant negative regulator of apoptosis, and the enzyme indoleamine 2,3-dioxygenase. Although many viral and non-viral vectors have been shown to transduce the corneal endothelium efficiently, allograft survival has so far been prolonged only following transduction of the donor cornea with adenoviral and lentiviral vectors. The degree of graft prolongation, although promising, is still insufficient for immediate translation to the clinic. Increasing the time that the therapeutic gene is expressed in the eye with an integrative, non-immunogenic viral vector is likely to be one way to achieve long-term graft survival. Simultaneous targeting of multiple pathways of graft rejection with more than one transgene is likely to be another. We suggest that the use of an adeno-associated viral or lentiviral vector combined with multiple transgenes may provide the key to future clinical trials.

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Efficiency and Toxicity of Liposome-mediated Gene Transfer to Corneal Endothelial Cells

Cited by 45 publications

References 20 publications

Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells

Caveolin-1 regulates expression of junction-associated proteins in brain microvascular endothelial cells

Cytotoxicity issues pertinent to lipoplex-mediated gene therapy in-vivo

Prospects for genetic modulation of corneal graft survival

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