Iontophoretic Delivery of Oligonucleotide Derivatives into Mouse Tumor

Vlassov, V. V.; Nechaeva, M. V.; Karamyshev, V.N.; Yakubov, L. A.

doi:10.1089/ard.1994.4.291

Cited by 20 publications

(9 citation statements)

References 2 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…To control for sample loading, the membrane was reprobed with a monoclonal antibody against a-tubulin. uptake mechanisms and intracellular trafficking of PS-ODN in cell lines [29,30]. Therefore, we investigated the mechanisms which are involved in cellular uptake and trafficking of the PS-ODN-P in the presence and absence of LF in CD8 þ T cells.…”

Section: Involvement Of Different Mechanisms In the Uptake Of Ps-odnsmentioning

confidence: 99%

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

2003

View full text Add to dashboard Cite

Antisense oligodeoxyribonucleotides (ODNs) can specifically inhibit gene expression, but their application to fresh human CD8 þ T cells is limited by poor spontaneous uptake (<2%). We have examined and optimized the uptake of phosphorothioate-modified oligodeoxyribonucleotides (PS-ODNs) into these cells in an ex vivo expansion model. Optimal antisense treatments were found to be, for fresh CD8 þ T cells, 1 mM PS-ODNs complexed with lipofectin (LF), which resulted in 35% uptake and 10 mM PS-ODNs in the absence of LF, for cultured cells, which resulted in 95% uptake. The delivered antisenses were functional, as determined by the inhibition of protein expression. In this respect, partially phosphorothioate-modified ODNs (PS-ODNs-P) were twice as effective as completely modified (PS-ODNs-C), and the antisense specific for the cap site showed the highest protein suppression of those tested (68%). Uptake mechanisms were also investigated. To our knowledge, this is the first optimization of the delivery of antisense oligonucleotides into human CD8 þ T cells. This protocol could be used to study the function of a particular gene in cytotoxic T lymphocytes and also by those looking for a method to deliver short interfering RNA into cell lines to specifically suppress a gene of interest.

show abstract

Section: Involvement Of Different Mechanisms In the Uptake Of Ps-odnsmentioning

confidence: 99%

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

2003

View full text Add to dashboard Cite

show abstract

“…Penetration using physical enhancement techniques, such as iontophoresis (Brand and Iversen, 1996) and electroporation (Regnier et al, 1998), have been used to deliver PS-ODN successfully in vitro. Vlassov et al (1993Vlassov et al ( , 1994 demonstrated measurable tissue levels in mice of oligonucleotides after iontophoresis in vivo. However, functional evidence is lacking as to the effectiveness of these compounds when given transdermally.…”

Section: Introductionmentioning

confidence: 99%

Transdermal Delivery of Antisense Oligonucleotides Can Induce Changes in Gene ExpressionIn Vivo

Brand

Hannah

Norris

et al. 2001

Antisense and Nucleic Acid Drug Development

View full text Add to dashboard Cite

The potential for using antisense compounds as therapeutic agents has generated great enthusiasm. Strategies for delivery of these compounds are, therefore, of great interest. Transdermal iontophoresis has been used successfully as an enhancement technique for the transdermal delivery of these compounds in vitro. The effectiveness of using percutaneous penetration as a means to deliver therapeutic levels of these compounds in vivo, however, remains to be demonstrated. The purpose of this work was to demonstrate the ability of iontophoretically delivered compounds to alter enzyme levels in the intact rat. A C5 propyne-modified phosphorothioate oligonucleotide (PS-ODN) targeted to the cytochrome p450-3A2 (CYP3A2) mRNA translational start site and the reverse sequence, used as a control, were synthesized. A patch containing either an oligonucleotide or a buffer control was placed on the animal's back, and an iontophoretic current of 0.5 mA/cm2 was applied for 3.5 hours. Twenty-four hours later, CYP3A2 levels were measured noninvasively using the midazolam-induced sleeping rat model. Liver and small intestinal microsomes were made after completion of sleep studies and assayed for CYP3A2, CYP1A1/2, CYP2B1/2, and CYP2E1. Midozolam-treated animals with antisense to CYP3A2 slept significantly longer than did the controls (p < 0.05). CYP3A2 levels were significantly lower in liver microsomes from antisense-treated animals than in either buffer control (p < 0.001) or reverse sequence animals (p < 0.05). The reverse sequence was also significantly different from the buffer control (p < 0.01), indicating a nonspecific effect of the PS background. Nontarget cytochrome levels were not altered by treatment. There were no significant differences in small intestine CYP3A2 levels between treatment groups. These data demonstrate that transdermally delivered PS-ODN can reach concentrations sufficient to induce changes in specific target enzymes in vivo. Further studies are warranted to investigate potential uses for these molecules.

show abstract

“…In vivo studies have shown that antisense oligomers can enter into target organs (Vlassov et al, 1993;Vlassov et al, 1994), and induce changes in target enzymes (Brand et al, 2001;Arora et al, 2002). The transdermal delivery of neutral PMOs using chemical penetration enhancement techniques has yet to be investigated, though preliminary data has demonstrated that the flux of a PMO targeted to cytochrome P4503A2 in propylene glycol and linoleic acid was nearly 20 times greater than that reported with iontophoretic delivery of the same sequence with a phosphorothioate (PS) backbone (Brand and Iversen, 2000).…”

Section: Introductionmentioning

confidence: 99%

Transdermal delivery of phosphorodiamidate Morpholino oligomers across hairless mouse skin

Pannier

Arora

Iversen

et al. 2004

International Journal of Pharmaceutics

View full text Add to dashboard Cite

Iontophoretic Delivery of Oligonucleotide Derivatives into Mouse Tumor

Cited by 20 publications

References 2 publications

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

Transdermal Delivery of Antisense Oligonucleotides Can Induce Changes in Gene ExpressionIn Vivo

Transdermal delivery of phosphorodiamidate Morpholino oligomers across hairless mouse skin

Contact Info

Product

Resources

About

Iontophoretic Delivery of Oligonucleotide Derivatives into Mouse Tumor

Cited by 20 publications

References 2 publications

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8+ T‐Cell Ex Vivo Expansion Model

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8+ T‐Cell Ex Vivo Expansion Model

Transdermal Delivery of Antisense Oligonucleotides Can Induce Changes in Gene ExpressionIn Vivo

Transdermal delivery of phosphorodiamidate Morpholino oligomers across hairless mouse skin

Contact Info

Product

Resources

About

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model

Optimization of Functional Efficacy of Phosphorothioate‐Modified Oligonucleotides in a Human CD8⁺ T‐Cell Ex Vivo Expansion Model