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Quantitative Pharmacology of Intercellular Drug Shuttle by Exosomes
Author(s) -
Wang Jin,
Cui Minjian,
Yeung Bertrand Z.,
Lu Ze,
Wientjes Michael Guill,
Woo Sukyung,
Au Jessie L.S.
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.829.5
Subject(s) - microvesicles , exosome , pharmacology , extracellular , chemistry , microbiology and biotechnology , cancer research , biology , biochemistry , microrna , gene
Background Exosomes are extracellular vesicles (30–100 nm) that contain bioactive molecules, secreted by most cell types and play a role in intercellular communications. Tumor cells exosomes may affect tumor progression, metastasis, and therapeutic efficacy and resistance. The current study used in vitro experiments and in silico studies to investigate the roles of exosomes in intercellular drug transfer and the quantitative relationship between exosomes and pharmacodynamics of chemotherapy drugs in solid tumor cells. Methods Effects of paclitaxel (PTX) and doxorubicin (DOX) on exocytosis were studied in human breast (MCF7 and a lung metastatic subline of MDA‐MB‐231 cells, LM2) and ovarian (A2780 and OVCAR4) cancer cells. Inhibition of exocytosis was studied using omeprazole (proton pump inhibitor) and GW4869 (ceramide synthesis inhibitor). Time‐lapse laser confocal microscopy was used together with RFP‐transfected cells to monitor the release of exosomes into extracellular fluid. Extracellular exosomes were collected and quantified based on their acetylcholinesterase activity. Drug concentrations were measured using LC‐MS/MS. Pharmacodynamics of exosomes were measured in recipient cells, using the sulforhodamine B assay for anti‐proliferation and the wound‐healing assay for anti‐migration. Results All four cells in exponential growth phase maintained similar baseline exosome level (~220 exosomes/cell). PTX (2–1000 nM) or DOX (10–1000 nM) treatment for 24 or 48 h stimulated exosome production/secretion, in dose‐ and time‐dependent manner, by up to 10‐fold. After 24 h treatment, the amount of PTX in exosomes was 5–10% of that in donor cells, and these PTX‐containing exosomes inhibited cell viability and migration in vitro. Pretreatment for 24 h with omeprazole (10 μM) or GW4869 (10 μg/mL) reduced the production/secretion of, and the drug amount in, exosomes by >20%, and enhanced the exosome cytotoxicity in donor cells. Our quantitative pharmacology model, comprising 6 intracellular and intercellular compartments and 11 rate constants, captured and characterized the effect of exocytosis on intercellular transfer of drug‐containing exosomes and PTX pharmacodynamics in donor and recipient cells with good fitness (CV <19%). Conclusion Our results indicate (a) chemotherapeutic agents stimulate exosome production/secretion, (b) exocytosis is a mechanism of intercellular drug transfer responsible for conferring biological activities on neighboring recipient cells, and (c) successful application of quantitative pharmacology to depict the spatial and temporal effects of exosomes on chemotherapy pharmacodynamics. Support or Funding Information Supported in part by RO1CA163015 (GW, JA) from National Cancer Institute and RO1EB015253 (JA, GW) from National Institute of Biomedical Imaging and Bioengineering, DHHS and RSG‐16‐006‐01‐CCE (SW) from American Cancer Society.