Milk-Derived Exosomes as Nanocarriers to Deliver Curcumin and Resveratrol in Breast Tissue and Enhance Their Anticancer Activity.

Antonio González-Sarrías,Alice Cattivelli,Adrián Cortés-Martín,Alberto Dávalos,Andrea Del Saz,Juan Carlos Espín,Fernando Vallejo,Lorena Del Pozo-Acebo,Carlos E Iglesias-Aguirre,María Carmen López De Las Hazas

doi:10.3390/ijms23052860

Abstract

Dietary (poly)phenols are extensively metabolized, limiting their anticancer activity. Exosomes (EXOs) are extracellular vesicles that could protect polyphenols from metabolism. Our objective was to compare the delivery to breast tissue and anticancer activity in breast cancer cell lines of free curcumin (CUR) and resveratrol (RSV) vs. their encapsulation in milk-derived EXOs (EXO-CUR and EXO-RSV). A kinetic breast tissue disposition was performed in rats. CUR and RSV were analyzed using UPLC-QTOF-MS and GC-MS, respectively. Antiproliferative activity was tested in MCF-7 and MDA-MB-231 breast cancer and MCF-10A non-tumorigenic cells. Cell cycle distribution, apoptosis, caspases activation, and endocytosis pathways were determined. CUR and RSV peaked in the mammary tissue (41 ± 15 and 300 ± 80 nM, respectively) 6 min after intravenous administration of EXO-CUR and EXO-RSV, but not with equivalent free polyphenol concentrations. Nanomolar EXO-CUR or EXO-RSV concentrations, but not free CUR or RSV, exerted a potent antiproliferative effect on cancer cells with no effect on normal cells. Significant (p < 0.05) cell cycle alteration and pro-apoptotic activity (via the mitochondrial pathway) were observed. EXO-CUR and EXO-RSV entered the cells primarily via clathrin-mediated endocytosis, avoiding ATP-binding cassette transporters (ABC). Milk EXOs protected CUR and RSV from metabolism and delivered both polyphenols to the mammary tissue at concentrations compatible with the fast and potent anticancer effects exerted in model cells. Milk EXOs enhanced the bioavailability and anticancer activity of CUR and RSV by acting as Trojan horses that escape from cancer cells’ ABC-mediated chemoresistance.

Highlights

Exosomes (EXOs) are a subset of extracellular vesicles secreted by cells that regulate intercellular communication in organisms [1,2]
EXOs were first isolated by ultracentrifugation (UC), loaded with polyphenols by either sonication, electroporation, or passive incubation, and further purified by size exclusion chromatography (SEC)
To the best of our knowledge, we have described the delivery of CUR and RSV to mammary tissue for the first time via administration of milk-derived EXO-CUR and EXORSV

Summary

Introduction

Exosomes (EXOs) are a subset of extracellular vesicles secreted by cells that regulate intercellular communication in organisms [1,2]. EXOs can transfer their cargo, consisting of different metabolites, lipids, functional proteins, and nucleic acids, and modulate the response of distant recipient cells [1–5]. EXOs can cross physical barriers, including the blood–brain barrier and placenta [10]. In this scenario, EXOs have emerged as molecule delivery vehicles, including for drugs, natural bioactives, and nucleic acids, with potential clinical applications [11–14]. It is possible to develop technology to modify EXOs’ surfaces to ensure tissue-specific biodistribution [17,18]. In this regard, milk is a scalable source of EXOs, highly recommended for therapeutic applications [19,20]. EXOs protect their cargo from gastrointestinal digestion, metabolism, and degradation, allowing its potential biodistribution in systemic tissues, including the brain [24–26]

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