Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release. / Arouri, Ahmad; Trojnar, Jakub; Schmidt, Steffen; Hansen, Anders H.; Mollenhauer, Jan; Mouritsen, Ole G.

In: PLoS ONE, Vol. 10, No. 5, e0125508, 2015.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Arouri, A, Trojnar, J, Schmidt, S, Hansen, AH, Mollenhauer, J & Mouritsen, OG 2015, 'Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release', PLoS ONE, vol. 10, no. 5, e0125508. https://doi.org/10.1371/journal.pone.0125508

APA

Arouri, A., Trojnar, J., Schmidt, S., Hansen, A. H., Mollenhauer, J., & Mouritsen, O. G. (2015). Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release. PLoS ONE, 10(5), [e0125508]. https://doi.org/10.1371/journal.pone.0125508

Vancouver

Arouri A, Trojnar J, Schmidt S, Hansen AH, Mollenhauer J, Mouritsen OG. Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release. PLoS ONE. 2015;10(5). e0125508. https://doi.org/10.1371/journal.pone.0125508

Author

Arouri, Ahmad ; Trojnar, Jakub ; Schmidt, Steffen ; Hansen, Anders H. ; Mollenhauer, Jan ; Mouritsen, Ole G. / Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release. In: PLoS ONE. 2015 ; Vol. 10, No. 5.

Bibtex

@article{bf5cc25d3f8c44a98d36a3deee016495,
title = "Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release",
abstract = "The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies.",
author = "Ahmad Arouri and Jakub Trojnar and Steffen Schmidt and Hansen, {Anders H.} and Jan Mollenhauer and Mouritsen, {Ole G.}",
year = "2015",
doi = "10.1371/journal.pone.0125508",
language = "English",
volume = "10",
journal = "P L o S One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "5",

}

RIS

TY - JOUR

T1 - Development of a Cell-Based Bioassay for Phospholipase A2-Triggered Liposomal Drug Release

AU - Arouri, Ahmad

AU - Trojnar, Jakub

AU - Schmidt, Steffen

AU - Hansen, Anders H.

AU - Mollenhauer, Jan

AU - Mouritsen, Ole G.

PY - 2015

Y1 - 2015

N2 - The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies.

AB - The feasibility of exploiting secretory phospholipase A2 (sPLA2) enzymes, which are overexpressed in tumors, to activate drug release from liposomes precisely at the tumor site has been demonstrated before. Although the efficacy of the developed formulations was evaluated using in vitro and in vivo models, the pattern of sPLA2-assisted drug release is unknown due to the lack of a suitable bio-relevant model. We report here on the development of a novel bioluminescence living-cell-based luciferase assay for the monitoring of sPLA2-triggered release of luciferin from liposomes. To this end, we engineered breast cancer cells to produce both luciferase and sPLA2 enzymes, where the latter is secreted to the extracellular medium. We report on setting up a robust and reproducible bioassay for testing sPLA2-sensitive, luciferin remote-loaded liposomal formulations, using 1,2-distearoyl-sn-glycero-3-phosphatidylcholine/1,2-distearoyl-sn-glycero-3-phosphatidylglycerol (DSPC/DSPG) 7:3 and DSPC/DSPG/cholesterol 4:3:3 as initial test systems. Upon their addition to the cells, the liposomes were degraded almost instantaneously by sPLA2 releasing the encapsulated luciferin, which provided readout from the luciferase-expressing cells. Cholesterol enhanced the integrity of the formulation without affecting its susceptibility to sPLA2. PEGylation of the liposomes only moderately broadened the release profile of luciferin. The provided bioassay represents a useful tool for monitoring active drug release in situ in real time as well as for testing and optimizing of sPLA2-sensitive lipid formulations. In addition, the bioassay will pave the way for future in-depth in vitro and in vivo studies.

U2 - 10.1371/journal.pone.0125508

DO - 10.1371/journal.pone.0125508

M3 - Journal article

C2 - 25945937

AN - SCOPUS:84929180270

VL - 10

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 5

M1 - e0125508

ER -

ID: 230974457