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Short-Chain Glycoceramides Promote Intracellular Mitoxantrone Delivery from Novel Nanoliposomes into Breast Cancer Cells

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Abstract

Purpose

To improve therapeutic activity of mitoxantrone (MTO)-based chemotherapy by reducing toxicity through encapsulation in nanoliposomes and enhancing intracellular drug delivery using short-chain sphingolipid (SCS) mediated tumor cell membrane permeabilization.

Methods

Standard (MTOL) and nanoliposomes enriched with the SCS, C8-Glucosylceramide or C8-Galactosylceramide (SCS-MTOL) were loaded by a transmembrane ammonium sulphate gradient and characterized by DLS and cryo-TEM. Intracellular MTO delivery was measured by flow cytometry and imaged by fluorescence microscopy. In vitro cytotoxicity was studied in breast carcinoma cell lines. Additionally, live cell confocal microscopy addressed the drug delivery mechanism by following the intracellular fate of the nanoliposomes, the SCS and MTO. Intratumoral MTO localization in relation to CD31-positive tumor vessels and CD11b positive cells was studied in an orthotopic MCF-7 breast cancer xenograft.

Results

Stable SCS-MTOL were developed increasing MTO delivery and cytotoxicity to tumor cells compared to standard MTOL. This effect was much less pronounced in normal cells. The drug delivery mechanism involved a transfer of SCS to the cell membrane, independently of drug transfer and not involving nanoliposome internalization. MTO was detected intratumorally upon MTOL and SCS-MTOL treatment, but not after free MTO, suggesting an important improvement in tumor drug delivery by nanoliposomal formulation. Nanoliposomal MTO delivery and cellular uptake was heterogeneous throughout the tumor and clearly correlated with CD31-positive tumor vessels. Yet, MTO uptake by CD11b positive cells in tumor stroma was minor.

Conclusions

Nanoliposomal encapsulation improves intratumoral MTO delivery over free drug. Liposome bilayer-incorporated SCS preferentially permeabilize tumor cell membranes enhancing intracellular MTO delivery.

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ACKNOWLEDGEMENTS AND DISCLOSURES

This work was financed by the Dutch Cancer Society. The authors thank Thomas Soullié for technical assistance with various aspects of histology and data processing and Sabine Barnert for performing cryo-TEM analyses.

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Authors and Affiliations

  1. Section Surgical Oncology, Department of Surgery, Laboratory Experimental Surgical Oncology, Room Ee151b, Erasmus MC, 3000CA, Rotterdam, PO Box 2040, The Netherlands

    Lília R. Cordeiro Pedrosa, Timo L. M. ten Hagen, Alexander M. M. Eggermont & Gerben A. Koning

  2. Department of Pharmaceutical Technology and Biopharmacy, Albert-Ludwigs University, D-79104, Freiburg, Germany

    Regine Süss

  3. Division of Biological Stress Response, he Netherlands Cancer Institute - Antoni van Leeuwenhoek Hospital, Amsterdam, 1066 CX, The Netherlands

    Albert van Hell & Marcel Verheij

  4. Institut de Cancerologie Gustave Roussy, Villejuif, Paris, 94800, France

    Alexander M. M. Eggermont

  5. Department of Radiotherapy, The Netherlands Cancer Institute - Antoni van Leeuwenhoek Hospital, Amsterdam, 1066 CX, The Netherlands

    Marcel Verheij

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  1. Lília R. Cordeiro Pedrosa
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Correspondence to Gerben A. Koning.

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Pedrosa, L.R.C., ten Hagen, T.L.M., Süss, R. et al. Short-Chain Glycoceramides Promote Intracellular Mitoxantrone Delivery from Novel Nanoliposomes into Breast Cancer Cells. Pharm Res 32, 1354–1367 (2015). https://doi.org/10.1007/s11095-014-1539-4

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  • DOI: https://doi.org/10.1007/s11095-014-1539-4

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