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Transferrin Receptor Targeted Lipopolyplexes for Delivery of Antisense Oligonucleotide G3139 in a Murine K562 Xenograft Model

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Abstract

Purpose

Transferrin (Tf) conjugated lipopolyplexes (LPs) carrying G3139, an antisense oligonucleotide for Bcl-2, were synthesized and evaluated in Tf receptor positive K562 erythroleukemia cells and then in a murine K562 xenograft model.

Materials and Methods

Particle size and Zeta potentials of transferrin conjugated lipopolyplexs containing G3139 (Tf-LP-G3139) were measured by Dynamic Light Scattering and ZetaPALS. In vitro and in vivo sample’s Bcl-2 downregulation was analyzed using Western blot and tumor tissue samples also exhibited by immunohistochemistry method. For athymic mice bearing with K562 xenograft tumors, tumor growth inhibition and survival rate were investigated. Nanoparticle distribution in 3-D cell cluster was observed by Laser scan confocal microscopy. IL-12 production in the plasma was measured by ELISA kit.

Results

In vitro, Tf-LP-G3139 was more effective in inducing down regulation of Bcl-2 in K562 cells than non-targeted LP-G3139, free G3139 and mismatched control ODN-G4126 in the same formulation. In vivo Tf-LP-G3139 was less effective than free G3139 in Bcl-2 down regulation. 3-D cell cluster model diffusion results indeed indicated limited penetration of the LPs into the cell cluster. Finally, the therapeutic efficacies of Tf-LP-G3139 and free G3139 were determined in the K562 xenograft model. Tf-LP-G3139 showed slower plasma clearance, higher AUC, and greater accumulation in the tumor compared to free G3139. In addition, Tf-LP-G3139 was found to be more effective in tumor growth inhibition and prolonging mouse survival than free G3139. This was associated with increased spleen weight and IL-12 production in the plasma.

Conclusion

The role of the immune system in the therapeutic response obtained with the Tf-LPs is necessary and in vitro 3-D cell cluster model can be a potential tool to evaluate the nanoparticle distribution.

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Abbreviations

AML:

Acute myelogenous leukemia

AUC:

Area under the plasma concentration versus time curve

Bcl-2:

B-cell leukemia/lymphoma 2 protein

DC-Chol:

3ß-[N-(N',N'-dimethylaminoethane)-carbamoyl] cholesterol hydrochloride

egg PC:

Egg phosphatidylcholine

EPR:

Enhanced permeability and retention

EtOH:

Ethanol

H&E:

Hematoxylin and eosin

LPs:

Lipopolyplexes

Mal-PEG-DSPE:

Maleimide-polyethyleneglycol (M.W. ~2000)–distearoylphosphatidylethanolamine

PEG-DSPE:

Methoxy-PEG2000-DSPE

TBST:

Tris-buffered saline/Tween-20

Tf:

Transferrin

TfR:

Transferrin receptor

Tf-LP-G3139:

Transferrin conjugated lipopolyplexs containing G3139

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Acknowledgment

This work was support in part by NSF grant EEC-0425626 and NIH grant R01 CA135243 and R21 CA131832. We wish to thank Genta Inc. for providing us with the ODNs G3139 and G4243.

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Authors and Affiliations

  1. NSF Nanoscale Science and Engineering Center (NSEC), The Ohio State University, Columbus, Ohio, 43210, USA

    Xulang Zhang, Robert J. Lee & L. James Lee

  2. Department of Chemical and Biomolecular Engineering, The Ohio State University, Columbus, Ohio, 43210, USA

    Chee Guan Koh, Bo Yu & L. James Lee

  3. Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, Ohio, 43210, USA

    Longzhu Piao & Robert J. Lee

  4. NCI Comprehensive Cancer Center (CCC), The Ohio State University, Columbus, Ohio, 43210, USA

    Shujun Liu, Guido Marcucci & Robert J. Lee

  5. 140 W 19th Ave., Columbus, Ohio, 43210, USA

    L. James Lee

  6. 500 W.12th Ave., Columbus, Ohio, 43210, USA

    Robert J. Lee

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  1. Xulang Zhang
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  2. Chee Guan Koh
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Correspondence to Robert J. Lee or L. James Lee.

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Zhang, X., Koh, C.G., Yu, B. et al. Transferrin Receptor Targeted Lipopolyplexes for Delivery of Antisense Oligonucleotide G3139 in a Murine K562 Xenograft Model. Pharm Res 26, 1516–1524 (2009). https://doi.org/10.1007/s11095-009-9864-8

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  • DOI: https://doi.org/10.1007/s11095-009-9864-8

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