Intracellular tracking of protamine/antisense oligonucleotide nanoparticles and their inhibitory effect on HIV-1 transactivation

Dinauer, Norbert; Lochmann, Dirk; Demirhan, Ilhan; Bouazzaoui, Abdellatif; Zimmer, Andreas; Chandra, Angelika; Kreuter, Jörg; Briesen, Hagen von

doi:10.1016/j.jconrel.2004.02.020

Cited by 43 publications

(22 citation statements)

References 29 publications

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“…Recent discovery of the cell-penetrating, protein transduction domain (PTD) peptides [6][7][8][9][10][11][12][13][14][15] such as HIV-TAT [7,10,15] has provided insight into finally resolving the membrane barrier problem for intracellular drug delivery. Both cell culture and animal studies have shown that by covalently linking TAT to nearly any drug class including hydrophilic compounds and large protein molecules (MW >150 kDa), TAT was able to transduce the attached cargos into cells of all organ types including the brain [9,10].…”

Section: Introductionmentioning

confidence: 99%

PTD-modified ATTEMPTS system for enhanced asparaginase therapy: A proof-of-concept investigation

Kwon

Liang

et al. 2008

Journal of Controlled Release

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Macromolecular drugs such as proteins and gene products are presumably the most desirable therapeutic agents due to their unmatched substrate specificity and reaction efficiency. Yet, clinical use of these drugs has met with limited success, primarily due to the impermeable nature of the cell membrane that restricts cellular drug uptake to only small (<600 Da) and hydrophobic molecules. The recent discovery of the protein transduction domain (PTD) membrane-penetrating peptides, such as HIV-TAT, has finally offered the possibility of resolving this cell-membrane barrier for macromolecular drug delivery. Via covalent linkages, these PTD peptides have been shown to ferry the attached macromolecular species across membranes of all cell types, both in vitro and in vivo. Nevertheless, the lack of selectivity for PTD-mediated internalization restricts the application of this cell uptake method in clinical practice, due to concerns of inducing systemic toxicity caused by the carried drugs.Presented herein is a modified version of our previously established "ATTEMPTS" approach in delivery of macromolecular drugs, which integrates the cell-penetrating PTDs into a heparin/ protamine-regulated delivery system. In vitro findings using asparaginase (ASNase) as a model macromolecular anti-tumor agent were able to validate the feasibility of this delivery system. The chemically constructed TAT-ASNase conjugates not only were able to translocate into the MOLT-4 cells and elicit the cytotoxic effects, but also this PTD-mediated intracellular drug uptake could be regulated (with on/off control) by the addition of heparin and protamine. This modified ATTEMPTS system therefore presents a new avenue of treatment of various types of cancers and other diseases with macromolecular drugs. In vitro characterization and a preliminary proof-of-concept animal investigation that demonstrates the feasibility of this PTD-mediated ASNase therapeutic system is subsequently described.

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Section: Introductionmentioning

confidence: 99%

PTD-modified ATTEMPTS system for enhanced asparaginase therapy: A proof-of-concept investigation

Kwon

Liang

et al. 2008

Journal of Controlled Release

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“…These findings are in good agreement with those observed by other investigators, indicating that protamine and chitosan formed a condensed complex with pDNA and siRNA as effectively as any of the currently used cationic polymeric gene carriers. [18][19][20] …”

Section: Resultsmentioning

confidence: 99%

“…Complexation of siRNA with protamine and chitosan could reduce the number of accessible binding sites for nucleases and therefore provide protection against degradation. 21) Serum stability study was performed to detect whether or not nanoplexes could protect siRNA from serum nuclease attacks. 19) As shown in Fig.…”

Section: Stability Of Sirna In Serummentioning

confidence: 99%

The Development of Ternary Nanoplexes for Efficient Small Interfering RNA Delivery

Şalva

Turan

Akbuğa

2013

Biological & Pharmaceutical Bulletin

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Targeted posttranscriptional gene silencing by RNA interference (RNAi) has garnered considerable interest as an attractive new class of drugs for several diseases, such as cancer. Chitosan and protamine are commonly used as a vehicle to deliver and protect small interfering RNA (siRNA), but the strong interaction still remains to be modulated for efficient siRNA uptake and silencing. Therefore, in this study, ternary nanoplexes containing chitosan and protamine were designed to substantially enhance the siRNA efficiency. Binary and ternary nanoplexes were prepared at different the ratios of moles of the amine groups of cationic polymers to those of the phosphate ones of siRNA (N/P) ratios and characterized in terms of size, zeta potential, morphology and serum stability. The silencing efficiencies and cytotoxicities of prepared nanoplexes were evaluated by enzyme-linked immunosorbent assay (ELISA) (for human vascular endothelial growth factor; hVEGF) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays, respectively. The mean diameter of ternary nanoplexes ranged from 151 to 282 nm, depending on the weight ratio between polymers and siRNA. The gene silencing effect after transfection with ternary nanoplexes (chitosan/siRNA/protamine 83%) was significantly higher than that with binary nanoplexes (chitosan/siRNA 71% and protamine/siRNA 74%). Ternary nanoplexes showed the highest cellular uptake ability when compared with binary nanoplexes. Ternary nanoplexes did not induce substantial cytotoxicity. Serum stability and the lack of cytotoxicity of the nanoplexes provided advantages over other gene silencing studies. These results suggest ternary nanoplexes have the potential to be an effective siRNA carrier to study the gene silencing effect.

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“…5 For effi cient transfection of HIV-1 target cells, protamine was used to complex ODN and phosphorothioate (PTO) analogues to form nanoparticles with diameters of about 180 nm and surface charges in the range of −18 to +30 mV. 6 The uptake of these nanoparticles was signifi cantly improved as compared with naked oligonucleotides. Protamine/ODN nanoparticles showed release of the antisense compound leading to specifi c inhibition of tat-mediated HIV-1 transactivation.…”

Section: Protamine Nanoparticles For Gene Deliverymentioning

confidence: 99%

Protamine nanoparticles

Nimesh¹

2013

Gene Therapy

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Intracellular tracking of protamine/antisense oligonucleotide nanoparticles and their inhibitory effect on HIV-1 transactivation

Cited by 43 publications

References 29 publications

PTD-modified ATTEMPTS system for enhanced asparaginase therapy: A proof-of-concept investigation

PTD-modified ATTEMPTS system for enhanced asparaginase therapy: A proof-of-concept investigation

The Development of Ternary Nanoplexes for Efficient Small Interfering RNA Delivery

Protamine nanoparticles

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