Research Article

The transmission and toxicity of polymer-bound doxorubicin-containing exosomes derived from human adenocarcinoma cells

*Author for correspondence:

E-mail Address: kristyna.gunar@seznam.cz

Department of Biological Models, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Lenka Kotrchová

https://orcid.org/0000-0003-4338-9362

Department of Biomedical Polymers, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Marcela Filipová

https://orcid.org/0000-0003-3503-7973

Department of Biological Models, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Tereza Krunclová

https://orcid.org/0000-0002-5232-8214

Department of Biological Models, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Aneta Dydowiczová

https://orcid.org/0000-0002-4662-5837

Department of Biological Models, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Robert Pola

https://orcid.org/0000-0003-0025-2016

Department of Biomedical Polymers, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Eva Randárová

https://orcid.org/0000-0002-1586-9871

Department of Biomedical Polymers, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Tomáš Etrych

https://orcid.org/0000-0001-5908-5182

Department of Biomedical Polymers, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Olga Janoušková

https://orcid.org/0000-0002-1365-6274

Department of Biological Models, Institute of Macromolecular Chemistry, Czech Academy of Sciences, 162 00, Prague 6, Czech Republic

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Published Online:18 Oct 2022https://doi.org/10.2217/nnm-2022-0081

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Abstract

Background: Exosomes are extracellular vesicles with the ability to encapsulate bioactive molecules, such as therapeutics. This study identified a new exosome mediated route of doxorubicin and poly(N-(2-hydroxypropyl)methacrylamide) (pHPMA)-bound doxorubicin trafficking in the tumor mass. Materials & methods: Exosome loading was achieved via incubation of the therapeutics with an adherent human breast adenocarcinoma cell line and its derived spheroids. Exosomes were characterized using HPLC, nanoparticle tracking analysis (NTA) and western blotting. Results: The therapeutics were successfully loaded into exosomes. Spheroids secreted significantly more exosomes than adherent cells and showed decreased viability after treatment with therapeutic-loaded exosomes, which confirmed successful transmission. Conclusion: To the best of our knowledge, this study provides the first evidence of pHPMA-drug conjugate secretion by extracellular vesicles.

Graphical abstract

Cancerostatics delivered to the tumor site via the bloodstream facilitated by the enhanced permeability and retention effect are taken up by peripheral tumor cells. Inside the multivesicular bodies cancerostatics (free, polymer-bound or cleaved from polymer backbone) are loaded into exosomes, secreted into the extracellular space and taken up by cells deeper inside the tumor mass. Cancerostatics maintain toxicity throughout the process.

Plain language summary

Background: In cancer treatment, low-molecular-weight drugs (e.g., doxorubicin [DOX]) with a broad spectrum of side effects are commonly used. Through their conjugation with hydrophilic polymers – N-(2-hydroxypropyl)methacrylamide (HPMA) copolymers – for example, most of the side effects can be reduced. These drug–polymer conjugates are delivered via bloodstream into the tumor. This study aimed to identify a new exosome-mediated route of DOX and polyHPMA(pHPMA)–DOX conjugates trafficking inside the tumor mass. Exosomes are small lipid membrane vesicles constitutively released from most of the cell types, including the tumor cells. Exosomes are able to encapsulate low-molecular-weight drugs. Methods: Exosomes were loaded with DOX and pHPMA-DOX in vitro via coincubation with cancer cells. Exosomes were isolated from the conditioned-cultivation medium after their release from cells and characterized (size, numbers, protein marker profiles). Results: The therapeutics were successfully loaded into exosomes and transmitted to the tumor cells. To the best of our knowledge, this is the first evidence of the pHPMA–drug conjugate secretion by exosomes.

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Papers of special note have been highlighted as: • of interest

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Vol. 17, No. 19

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History

Received 27 March 2022

Accepted 26 August 2022

Published online 18 October 2022

Published in print August 2022

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Keywords

Author contributions

Conceptualization, K Gunár and O Janoušková; methodology, K Gunár, O Janoušková, L Kotrchová; validation, K Gunár, A Dydowiczová, L Kotrchová; formal analysis, K Gunár, L Kotrchová, O Janoušková; investigation, K Gunár, L Kotrchová, A Dydowiczová, M Filipová, T Krunclová; resources, E Randárová, R Pola, T Etrych; data curation, K Gunár, O Janoušková; writing – original draft preparation, K Gunár; writing – review and editing, O Janoušková, T Etrych, E Randárová; visualization, K Gunár; supervision, O Janoušková, T Etrych; project administration, K Gunár, O Janoušková; funding acquisition, O Janoušková, T Etrych.

Financial & competing interests disclosure

This work was supported by the Czech Academy of Sciences (project JSPS-22-01) and by the Ministry of Education, Youth and Sports of the Czech Republic within the Inter-excellence program (project LTAUSA18083). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

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