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Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer

Exosomes are physiologically secreted nanoparticles recently established as natural delivery systems involved in cell-to-cell communication and content exchange. Due to their inherent targeting potential, exosomes are currently being harnessed for the development of anti-cancer therapeutics. Clinica...

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Autores principales: Pisano, Simone, Pierini, Irene, Gu, Jianhua, Gazze, Andrea, Francis, Lewis Webb, Gonzalez, Deyarina, Conlan, Robert Steven, Corradetti, Bruna
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7528637/
https://www.ncbi.nlm.nih.gov/pubmed/33042993
http://dx.doi.org/10.3389/fcell.2020.553576
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author Pisano, Simone
Pierini, Irene
Gu, Jianhua
Gazze, Andrea
Francis, Lewis Webb
Gonzalez, Deyarina
Conlan, Robert Steven
Corradetti, Bruna
author_facet Pisano, Simone
Pierini, Irene
Gu, Jianhua
Gazze, Andrea
Francis, Lewis Webb
Gonzalez, Deyarina
Conlan, Robert Steven
Corradetti, Bruna
author_sort Pisano, Simone
collection PubMed
description Exosomes are physiologically secreted nanoparticles recently established as natural delivery systems involved in cell-to-cell communication and content exchange. Due to their inherent targeting potential, exosomes are currently being harnessed for the development of anti-cancer therapeutics. Clinical trials evaluating their effectiveness are demonstrating safety and promising outcomes. However, challenging large-scale production, isolation, modification and purification of exosomes are current limitations for the use of naturally occurring exosomes in the clinic. Exosome mimetics hold the promise to improve the delivery of bioactive molecules with therapeutic efficacy, while achieving scalability and increasing bioavailability. In this study, we propose the development of Immune Derived Exosome Mimetics (IDEM) as a scalable approach to target and defeat ovarian cancer cells. IDEM were fabricated from monocytic cells by combining sequential filtration steps through filter membranes with different porosity and size exclusion chromatography columns. The physiochemical and molecular characteristics of IDEM were compared to those of natural exosomes (EXO). Nanoparticle Tracking Analysis confirmed a 2.48-fold increase in the IDEM production yields compared to EXO, with similar exosomal markers profiles (CD81, CD63) as demonstrated by flow cytometry and ELISA. To exploit the prospective of IDEM to deliver chemotherapeutics, doxorubicin (DOXO) was used as a model drug. IDEM showed higher encapsulation efficiency and drug release over time compared to EXO. The uptake of both formulations by SKOV-3 ovarian cancer cells was assessed by confocal microscopy and flow cytometry, showing an incremental drug uptake over time. The analysis of the cytotoxic and apoptotic effect of DOXO-loaded nanoparticles both in 2D and 3D culture systems proved IDEM as a more efficient system as compared to free DOXO, unraveling the advantage of IDEM in reducing side-effects while increasing cytotoxicity of targeted cells, by delivering smaller amount of the chemotherapeutic agent. The high yields of IDEM obtained compared to natural exosomes together with the time-effectiveness and reproducibility of their production method make this approach potentially exploitable for clinical applications. Most importantly, the appreciable cytotoxic effect observed on ovarian cancer in vitro systems sets the ground for the development of compelling nanotherapeutic candidates for the treatment of this malady and will be further evaluated.
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spelling pubmed-75286372020-10-09 Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer Pisano, Simone Pierini, Irene Gu, Jianhua Gazze, Andrea Francis, Lewis Webb Gonzalez, Deyarina Conlan, Robert Steven Corradetti, Bruna Front Cell Dev Biol Cell and Developmental Biology Exosomes are physiologically secreted nanoparticles recently established as natural delivery systems involved in cell-to-cell communication and content exchange. Due to their inherent targeting potential, exosomes are currently being harnessed for the development of anti-cancer therapeutics. Clinical trials evaluating their effectiveness are demonstrating safety and promising outcomes. However, challenging large-scale production, isolation, modification and purification of exosomes are current limitations for the use of naturally occurring exosomes in the clinic. Exosome mimetics hold the promise to improve the delivery of bioactive molecules with therapeutic efficacy, while achieving scalability and increasing bioavailability. In this study, we propose the development of Immune Derived Exosome Mimetics (IDEM) as a scalable approach to target and defeat ovarian cancer cells. IDEM were fabricated from monocytic cells by combining sequential filtration steps through filter membranes with different porosity and size exclusion chromatography columns. The physiochemical and molecular characteristics of IDEM were compared to those of natural exosomes (EXO). Nanoparticle Tracking Analysis confirmed a 2.48-fold increase in the IDEM production yields compared to EXO, with similar exosomal markers profiles (CD81, CD63) as demonstrated by flow cytometry and ELISA. To exploit the prospective of IDEM to deliver chemotherapeutics, doxorubicin (DOXO) was used as a model drug. IDEM showed higher encapsulation efficiency and drug release over time compared to EXO. The uptake of both formulations by SKOV-3 ovarian cancer cells was assessed by confocal microscopy and flow cytometry, showing an incremental drug uptake over time. The analysis of the cytotoxic and apoptotic effect of DOXO-loaded nanoparticles both in 2D and 3D culture systems proved IDEM as a more efficient system as compared to free DOXO, unraveling the advantage of IDEM in reducing side-effects while increasing cytotoxicity of targeted cells, by delivering smaller amount of the chemotherapeutic agent. The high yields of IDEM obtained compared to natural exosomes together with the time-effectiveness and reproducibility of their production method make this approach potentially exploitable for clinical applications. Most importantly, the appreciable cytotoxic effect observed on ovarian cancer in vitro systems sets the ground for the development of compelling nanotherapeutic candidates for the treatment of this malady and will be further evaluated. Frontiers Media S.A. 2020-09-17 /pmc/articles/PMC7528637/ /pubmed/33042993 http://dx.doi.org/10.3389/fcell.2020.553576 Text en Copyright © 2020 Pisano, Pierini, Gu, Gazze, Francis, Gonzalez, Conlan and Corradetti. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Pisano, Simone
Pierini, Irene
Gu, Jianhua
Gazze, Andrea
Francis, Lewis Webb
Gonzalez, Deyarina
Conlan, Robert Steven
Corradetti, Bruna
Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title_full Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title_fullStr Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title_full_unstemmed Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title_short Immune (Cell) Derived Exosome Mimetics (IDEM) as a Treatment for Ovarian Cancer
title_sort immune (cell) derived exosome mimetics (idem) as a treatment for ovarian cancer
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7528637/
https://www.ncbi.nlm.nih.gov/pubmed/33042993
http://dx.doi.org/10.3389/fcell.2020.553576
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