Endosomolytic Activity of Cationic Liposomes Enhances the Delivery of Human Immunodeficiency Virus-1 Trans-activator Protein (TAT) to Mammalian Cells

Huang, Leaf; Farhood, Hassan; Serbina, Natalya V.; Teepe, Annette G.; Barsoum, James

doi:10.1006/bbrc.1995.2838

Cited by 21 publications

(11 citation statements)

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“…This property of DOPE can be invaluable in gene delivery as it enables better cell internalization and facilitates endosomal escape (Li et al, 2014). Hence DOPE has been used as a ''helper lipid'' along with cationic lipids in many gene delivery systems Huang et al, 1995). However, use of cationic lipids tends to increase cytotoxicity and trigger inflammatory response in biological systems and hence cationic lipid-free liposomes were explored in this study.…”

Section: Discussionmentioning

confidence: 99%

EpCAM-targeted liposomal si-RNA delivery for treatment of epithelial cancer

et al. 2014

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Background: RNA interference (RNAi) technology using short interfering RNA (si-RNA) has shown immense potential in the treatment of cancers through silencing of specific genes. Cationic non-viral vectors employed for gene delivery exhibit toxic effects in normal cells limiting their widespread use, therefore, site-specific delivery using benign carriers could address this issue. Objective: Design of a non-toxic carrier that enables site-specific delivery of si-RNA into the cancer cells is of prime importance to realize the promise of gene silencing. Methods: In the present study, non-cationic liposomes encapsulating si-RNA against epithelial cell adhesion molecule (EpCAM) were developed and characterized for encapsulation efficiency, colloidal stability, in vitro and in vivo gene silencing efficacy. Results: PEGylated liposomes containing phosphatidyl choline and phosphatidyl ethanolamine exhibited maximum si-RNA encapsulation efficiency of 47%, zeta potential of -21 mV, phase transition temperature of 51 C and good colloidal stability in phosphate-buffered saline (PBS) containing bovine serum albumin (BSA) and plasma protein (PP) at 37 C. Conjugation of epithelial cell adhesion molecule (EpCAM) antibody to the liposomes resulted in enhanced cell internalization and superior down-regulation of EpCAM gene in MCF-7 cell lines when compared with free si-RNA and the non-targeted liposomes. In vivo evaluation of immunoliposomes for their efficacy in regressing the tumor volume in Balb/c SCID mice showed about 35% reduction of tumor volume in comparison with the positive control when administered with an extremely low dose of 0.15 mg/kg twice a week for 4 weeks. Conclusion: Our results exhibit that the nanocarrier-mediated silencing of EpCAM gene is a promising strategy to treat epithelial cancers.

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Section: Discussionmentioning

confidence: 99%

EpCAM-targeted liposomal si-RNA delivery for treatment of epithelial cancer

et al. 2014

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“…The plasmid DNA-protein complexes were transfected into cells using cationic lipid-mediated transfection protocols. However, the delivery efficiency was very low, required chloroquine treatment (28) and the benefit of using the cationic lipid was unclear since the protein alone can spontaneously enter the cell without the aid of any delivery system (30,32). Cationic lipids have also been used to deliver protein into the intracellular processing pathway leading to antigen presentation, but the discrimination between inside and outside antigen presentation was difficult and the efficiency of functional intracellular protein delivery remains unclear (29).…”

Section: Discussionmentioning

confidence: 99%

Intracellular Delivery of Proteins with a New Lipid-mediated Delivery System

Zelphati¹,

Wang²,

Kitada³

et al. 2001

Journal of Biological Chemistry

209

180

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There are many very effective methods to introduce transcriptionally active DNA into viable cells but approaches to deliver functional proteins are limited. We have developed a lipid-mediated delivery system that can deliver functional proteins or other bioactive molecules into living cells. This delivery system is composed of a new trifluoroacetylated lipopolyamine (TFA-DODAPL) and dioleoyl phosphatidylethanolamine (DOPE). This cationic formulation successfully delivered antibodies, dextran sulfates, phycobiliproteins, albumin, and enzymes (␤-galactosidase and proteases) into the cytoplasm of numerous adherent and suspension cells. Two systems were used to demonstrate that the proteins were delivered in a functionally active form. First, intracellular ␤-galactosidase activity was clearly demonstrated within X-gal-stained cells after TFA-DODAPL:DOPE-mediated delivery of the enzyme. Second, the delivery system mediated delivery of several caspases (caspase 3, caspase 8, and granzyme B) into cultured cell lines and primary cells triggering apoptosis. Mechanistic studies showed that up to 100% of the protein mixed with the lipid formulation was captured into a lipid-protein complex, and up to 50% of the input protein associated with cells. This lipid-mediated transport system makes protein delivery into cultured cells as convenient, effective, and reliable as DNA transfection.

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“…13 Proteins and small molecules can also be delivered to cells as cationic complexes. [14][15][16][17] The mode of delivery of DNA to the nucleus by cationic lipids remains to be elucidated, although endocytosis is generally accepted as the major route of cellular uptake.…”

Section: Introductionmentioning

confidence: 99%

O‐ethylphosphatidylcholine: A metabolizable cationic phospholipid which is a serum‐compatible DNA transfection agent

MacDonald

Rakhmanova

Choi

et al. 1999

Journal of Pharmaceutical Sciences

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1,2-dioleoyl-sn-glycero-3-ethylphosphocholine was prepared in a one-step reaction from phosphatidylcholine by reaction with ethyl trifluoromethanesulfonate. This and related O-alkyl phosphatidylcholines constitute the first chemically stable triesters of biological lipid structures and the first cationic derivatives of phospholipids consisting entirely of biological metabolites linked with ester bonds. The complex of cationic phospholipid and plasmid DNA transfected cells with high efficiency. Maximum efficiency of transfection was obtained with complexes in which the positive charge was a few percent in excess over the negative charge. Modest stimulation of transfection of common cell lines was obtained by continuous culture in the presence of 10% serum. Incubation of the phospholipid complex for at least 2 h at 37 degrees C in nearly pure serum had no deleterious effects on transfection efficiency. The lipid has low toxicity; BHK cells tolerated amounts of 2 mg/2 x 10(6) cells at concentrations of 1 mg/mL. The lipid is biodegradable; it was hydrolyzed by phospholipase A(2) in vitro and was metabolized with a half-life of a few days in cells in culture. The synthetic route to cationic phospholipids is well suited to the preparation of derivatives that are tailor-made to have a wide variety of different properties.

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Endosomolytic Activity of Cationic Liposomes Enhances the Delivery of Human Immunodeficiency Virus-1 Trans-activator Protein (TAT) to Mammalian Cells

Cited by 21 publications

References 0 publications

EpCAM-targeted liposomal si-RNA delivery for treatment of epithelial cancer

EpCAM-targeted liposomal si-RNA delivery for treatment of epithelial cancer

Intracellular Delivery of Proteins with a New Lipid-mediated Delivery System

O‐ethylphosphatidylcholine: A metabolizable cationic phospholipid which is a serum‐compatible DNA transfection agent

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