C8-glycosphingolipids preferentially insert into tumor cell membranes and promote chemotherapeutic drug uptake.
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.
Laboratory Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC Cancer Center, Rotterdam 3000 CA, The Netherlands.
Lília R
LR
Cordeiro Pedrosa
Optical Imaging Center, Department of Pathology, Erasmus MC, Rotterdam 3000 CA, The Netherlands.
Wiggert A
WA
van Cappellen
Laboratory Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC Cancer Center, Rotterdam 3000 CA, The Netherlands.
Barbara
B
Steurer
Unidad de Biofísica (CSIC, UPV/EHU), P.O. Box 644, 48080 Bilbao, Spain; Departamento de Bioquímica, Universidad del País Vasco, P.O. Box 644, 48080 Bilbao, Spain.
Dalila
D
Ciceri
Laboratory Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC Cancer Center, Rotterdam 3000 CA, The Netherlands.
Timo L M
TL
ten Hagen
Laboratory Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC Cancer Center, Rotterdam 3000 CA, The Netherlands; Institut de Cancerologie Gustave Roussy, Villejuif, Paris 94800, France.
Alexander M M
AM
Eggermont
Division of Biological Stress Response, The Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital, Amsterdam 1066 CX, The Netherlands; Department of Radiotherapy, The Netherlands Cancer Institute-Antoni van Leeuwenhoek Hospital, Amsterdam 1066 CX, The Netherlands.
Marcel
M
Verheij
Unidad de Biofísica (CSIC, UPV/EHU), P.O. Box 644, 48080 Bilbao, Spain; Departamento de Bioquímica, Universidad del País Vasco, P.O. Box 644, 48080 Bilbao, Spain.
Felix María
FM
Goñi
Laboratory Experimental Surgical Oncology, Section Surgical Oncology, Department of Surgery, Erasmus MC Cancer Center, Rotterdam 3000 CA, The Netherlands. Electronic address: g.koning@erasmusmc.nl.
Gerben A
GA
Koning
Unidad de Biofísica (CSIC, UPV/EHU), P.O. Box 644, 48080 Bilbao, Spain; Departamento de Bioquímica, Universidad del País Vasco, P.O. Box 644, 48080 Bilbao, Spain; IKERBASQUE, Basque Foundation for Science, 48011 Bilbao, Spain. Electronic address: xabier.contreras@ehu.es.
F-Xabier
FX
Contreras
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002855:Chromatography, Thin Layer; D057930:Click Chemistry; D004317:Doxorubicin / Q000031:analogs & derivatives* / Q000378:metabolism; D005699:Galactosylceramides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D006367:HeLa Cells; D006801:Humans; D008051:Lipid Bilayers; D008081:Liposomes; D008563:Membrane Lipids / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D009369:Neoplasms / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000378:metabolism; D016062:Porosity; D013696:Temperature; D013997:Time Factors
D000903:Antibiotics, Antineoplastic; D005699:Galactosylceramides; D008051:Lipid Bilayers; D008081:Liposomes; D008563:Membrane Lipids; C046928:NBD-galactosylceramide; C506643:liposomal doxorubicin; D011092:Polyethylene Glycols; D004317:Doxorubicin; D009327:4-Chloro-7-nitrobenzofurazan
Doxorubicin; Liposome; Short-chain glycosphingolipid; Targeting tumor cell membrane; Tumor-cell membrane-permeability modulation