C8-glycosphingolipids preferentially insert into tumor cell membranes and promote chemotherapeutic drug uptake.

Abstract

Insufficient drug delivery into tumor cells limits the therapeutic efficacy of chemotherapy. Co-delivery of liposome-encapsulated drug and synthetic short-chain glycosphingolipids (SC-GSLs) significantly improved drug bioavailability by enhancing intracellular drug uptake. Investigating the mechanisms underlying this SC-GSL-mediated drug uptake enhancement is the aim of this study. Fluorescence microscopy was used to visualize the cell membrane lipid transfer intracellular fate of fluorescently labeled C6-NBD-GalCer incorporated in liposomes in tumor and non-tumor cells. Additionally click chemistry was applied to image and quantify native SC-GSLs in tumor and non-tumor cell membranes. SC-GSL-mediated flip-flop was investigated in model membranes to confirm membrane-incorporation of SC-GSL and its effect on membrane remodeling. SC-GSL enriched liposomes containing doxorubicin (Dox) were incubated at 4°C and 37°C and intracellular drug uptake was studied in comparison to standard liposomes and free Dox. SC-GSL transfer to the cell membrane was independent of liposomal uptake and the majority of the transferred lipid remained in the plasma membrane. The transfer of SC-GSL was tumor cell-specific and induced membrane rearrangement as evidenced by a transbilayer flip-flop of pyrene-SM. However, pore formation was measured, as leakage of hydrophilic fluorescent probes was not observed. Moreover, drug uptake appeared to be mediated by SC-GSLs. SC-GSLs enhanced the interaction of doxorubicin (Dox) with the outer leaflet of the plasma membrane of tumor cells at 4°C. Our results demonstrate that SC-GSLs preferentially insert into tumor cell plasma membranes enhancing cell intrinsic capacity to translocate amphiphilic drugs such as Dox across the membrane via a biophysical process.

• Authors: Lília R Cordeiro Pedrosa, Wiggert A van Cappellen, Barbara Steurer, Dalila Ciceri, Timo L M ten Hagen, Alexander M M Eggermont, Marcel Verheij, Felix María Goñi, Gerben A Koning, F-Xabier Contreras
• Journal: Biochimica et biophysica acta (Biochim Biophys Acta) Vol. 1848 Issue 8 Pg. 1656-70 (Aug 2015) ISSN: 0006-3002 [Print] Netherlands
• PMID: 25917957 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier B.V. All rights reserved.

Chemical References

• Antibiotics, Antineoplastic
• Galactosylceramides
• Lipid Bilayers
• Liposomes
• Membrane Lipids
• NBD-galactosylceramide
• liposomal doxorubicin
• Polyethylene Glycols
• Doxorubicin
• 4-Chloro-7-nitrobenzofurazan

Topics

• 4-Chloro-7-nitrobenzofurazan (analogs & derivatives, chemistry, metabolism, pharmacology)
• Antibiotics, Antineoplastic (metabolism)
• Cell Membrane (drug effects, metabolism)
• Cell Membrane Permeability (drug effects)
• Chromatography, Thin Layer
• Click Chemistry
• Doxorubicin (analogs & derivatives, metabolism)
• Galactosylceramides (chemistry, metabolism, pharmacology)
• HeLa Cells
• Humans
• Lipid Bilayers
• Liposomes
• Membrane Lipids (chemistry, metabolism, pharmacology)
• Microscopy, Confocal
• Microscopy, Fluorescence
• Molecular Structure
• Neoplasms (metabolism)
• Polyethylene Glycols (metabolism)
• Porosity
• Temperature
• Time Factors