Specific iNOS-targeted antisense knockdown in endothelial cells

Hemmrich, Karsten; Suschek, Christoph V.; Lerzynski, Guido; Schnorr, Oliver; Kolb-Bachofen, Victoria

doi:10.1152/ajpcell.00045.2003

Cited by 6 publications

(2 citation statements)

References 25 publications

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“…The protocol for antisense treatment of monolayers was as described elsewhere (22). Cells (1.5 ϫ 10 5 /well) were placed into monolayer culture in 6-well culture plates.…”

Section: Methodsmentioning

confidence: 99%

iNOS initiates and sustains metabolic arrest in hypoxic lung adenocarcinoma cells: mechanism of cell survival in solid tumor core

Land

Rae

2005

American Journal of Physiology-Cell Physiology

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Nitric oxide (NO) modulates cellular metabolism by competitively inhibiting the reduction of O2 at respiratory complex IV. The aim of this study was to determine whether this effect could enhance cell survival in the hypoxic solid tumor core by inducing a state of metabolic arrest in cancer cells. Mitochondria from human alveolar type II-like adenocarcinoma (A549) cells showed a fourfold increase in NO-sensitive 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM) fluorescence and sixfold increase in Ca2+-insensitive NO synthase (NOS) activity during equilibration from Po2s of 100-->23 mmHg, which was abolished by N(omega)-nitro-L-arginine methyl ester-HCl (L-NAME) and the inducible NOS (iNOS) inhibitor, N6-(1-iminoethyl)-L-lysine dihydrochloride (L-NIL). Similarly, cytosolic and compartmented DAF-FM fluorescence increased in intact cells during a transition between ambient Po2 and 23 mmHg and was abolished by transfection with iNOS antisense oligonucleotides (AS-ODN). In parallel, mitochondrial membrane potential (deltapsi(m)), measured using 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolo-carbocyanine iodide (JC-1), decreased to a lower steady state in hypoxia without change in glycolytic rate, adenylate energy charge, or cell viability. However, L-NAME or iNOS AS-ODN treatment maintained deltapsi(m) at normoxic levels irrespective of hypoxia and caused a marked activation of glycolysis, destabilization energy charge, and cell death. Comparison with other cancer-derived (H441) or native tissue-derived (human bronchial epithelial; alveolar type II) lung epithelial cells revealed that the hypoxic suppression of deltapsi(m) was common to cells that expressed iNOS. The controlled dissipation of deltapsi(m), absence of an overt glycolytic activation, and conservation of viability suggest that A549 cells enter a state of metabolic suppression in hypoxia, which inherently depends on the activation of iNOS as Po2 falls.

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“…The protocol for antisense treatment of monolayers was as described elsewhere (22). Cells (1.5 ϫ 10 5 /well) were placed into monolayer culture in 6-well culture plates.…”

Section: Methodsmentioning

confidence: 99%

iNOS initiates and sustains metabolic arrest in hypoxic lung adenocarcinoma cells: mechanism of cell survival in solid tumor core

Land

Rae

2005

American Journal of Physiology-Cell Physiology

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“…The resulting high-output production of nitric oxide is often associated with cell damage or tissue destruction [37]. PLGA nanoparticles were loaded with both Nile Red (a lipophilic dye with inducible nitric oxide synthase (iNOS) as a result of pro-inflammatory cytokines.…”

Section: Antisense Oligonucleotide Delivery To the Posterior Segment mentioning

confidence: 99%

Polymeric Nanoparticles for Drug Delivery to the Posterior Segment of the Eye

Bejjani¹,

BenEzra²,

Gurny³

et al. 2005

Chimia

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Diseases of the posterior segment of the eye account for most cases of irreversible blindness worldwide. Drug delivery to this closed compartment remains a challenge because of the internal and external blood retinal barriers that selectively control drug penetration into the retina. Direct intraocular (intravitreal) delivery is currently used to achieve high drug concentration in the vitreous and the retina but is usually associated with several side effects. Alternatively topical, periocular, and systemic routes of administration can be used but are associated with low bioavailability and specific side effects. Therefore, intraocular sustained drug delivery systems are being designed to overcome these limitations. Polymeric nanoparticles loaded with therapeutic compounds are under investigation to provide new tools of administration to the eye. The first part of this paper will briefly review the barriers to ocular delivery of drugs and the advantages of using polymeric nanoparticle carriers as drug delivery systems. In the second part, the results in terms of preparation and characterization of polymeric nanoparticles loaded with nucleic acids, the study of the transretinal pathway of intravitreally injected nanoparticles and the assessment of their ability to efficiently deliver plasmids and oligonucleotides will be discussed.

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