Original Article
Development of exosome-encapsulated paclitaxel to overcome MDR in cancer cells

https://doi.org/10.1016/j.nano.2015.10.012Get rights and content

Abstract

Exosomes have recently come into focus as “natural nanoparticles” for use as drug delivery vehicles. Our objective was to assess the feasibility of an exosome-based drug delivery platform for a potent chemotherapeutic agent, paclitaxel (PTX), to treat MDR cancer. Herein, we developed different methods of loading exosomes released by macrophages with PTX (exoPTX), and characterized their size, stability, drug release, and in vitro antitumor efficacy. Reformation of the exosomal membrane upon sonication resulted in high loading efficiency and sustained drug release. Importantly, incorporation of PTX into exosomes increased cytotoxicity more than 50 times in drug resistant MDCKMDR1 (Pgp +) cells. Next, our studies demonstrated a nearly complete co-localization of airway-delivered exosomes with cancer cells in a model of murine Lewis lung carcinoma pulmonary metastases, and a potent anticancer effect in this mouse model. We conclude that exoPTX holds significant potential for the delivery of various chemotherapeutics to treat drug resistant cancers.

From the Clinical Editor

Exosomes are membrane-derived natural vesicles of ~40 - 200 nm size. They have been under extensive research as novel drug delivery vehicles. In this article, the authors developed exosome-based system to carry formulation of PTX and showed efficacy in the treatment of multi-drug resistant cancer cells. This novel system may be further developed to carry other chemotherapeutic agents in the future.

Graphical abstract

Exosomes released by autologous macrophages were loaded with paclitaxel (PTX) upon ultrasound treatment. The obtained formulation (exoPTX) showed a high loading capacity, sustained drug release, profound ability to accumulate in resistant cancer cells, and increased cytotoxicity compared to PTX in vitro and in vivo.

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Section snippets

Reagents

PTX and doxorubicin (DOX) was purchased from LC Laboratories (Woburn, MA). Lipophilic fluorescent dyes, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate (DIL), and 2-decanoyl-1-(O-(11-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl)amino)undecyl)-sn-glycero-3-phosphocholine (BODIPY-PC), were purchased from Invitrogen (Carlsbad, CA) and Molecular Probes (Eugene, OR), respectively. Rhodamine 123 (R123), 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI),

Manufacture and characterization of exosomal formulations of PTX (exoPTX)

Exosomes collected from the conditioned media of RAW 264.7 macrophages were characterized by size, charge, protein content, and morphology (Figure 1, A, B, and D). Exosomes showed elevated expression of exosome-associated proteins (Alix, TSG101, and Flotillin) as compared to cell lysate, which displayed greater levels of β-actin (Figure 1, B). Naive empty exosomes had a narrow size distribution, with an average particle diameter of 110.4 ± 4.2 nm and 70.8 ± 2.8 nm as revealed by NTA and DLS,

Discussion

Exosomal carriers can provide advantages of both cell-based drug delivery and nanotechnology for efficient drug transport capable of overcoming various biological barriers. However, several limitations need to be addressed before their use in the clinic. One of the difficulties is the efficient loading exosomes with a therapeutic agent without significant changes in the structure and content of exosomal membranes. In the present study, we utilized various methods for PTX incorporation into

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    This study was supported by the United States National Institutes of Health grants 1RO1 NS057748 and The Carolina Partnership, a strategic partnership between the UNC Eshelman School of Pharmacy and The University Cancer Research Fund through the Lineberger Comprehensive Cancer Center (to EVB), RR021937 (to AVK), and Ministry of Education and Science of Russian Federation grants 11.G34.31.0004, 02.740.11.5232, and RSF-14-13-00731 (both to AVK and NLK).

    No competing interests are present.

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