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Exogenous modification of EL-4 T cell extracellular vesicles with miR-155 induce macrophage into M1-type polarization

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Abstract

Extracellular vesicles (EVs) show promising potential to be used as therapeutics, disease biomarkers, and drug delivery vehicles. We aimed to modify EVs with miR-155 to modulate macrophage immune response that can be potentially used against infectious diseases. Primarily, we characterized T cells (EL-4) EVs by several standardized techniques and confirmed that the EVs could be used for experimental approaches. The bioactivities of the isolated EVs were confirmed by the uptake assessment, and the results showed that target cells can successfully uptake EVs. To standardize the loading protocol by electroporation for effective biological functionality, we chose fluorescently labelled miR-155 mimics because of its important roles in the immune regulations to upload them into EVs. The loading procedure showed that the dosage of 1 µg of miRNA mimics can be efficiently loaded to the EVs at 100 V, further confirmed by flow cytometry. The functional assay by incubating these modified EVs (mEVs) with in vitro cultured cells led to an increased abundance of miR-155 and decreased the expressions of its target genes such as TSHZ3, Jarid2, ZFP652, and WWC1. Further evaluation indicated that these mEVs induced M1-type macrophage polarization with increased TNF-α, IL-6, IL-1β, and iNOS expression. The bioavailability analysis revealed that mEVs could be detected in tissues of the livers. Overall, our study demonstrated that EVs can be engineered with miR-155 of interest to modulate the immune response that may have implications against infectious diseases.

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Acknowledgements

We thank Cheng lab members for constructive suggestion on the discussion.

Funding

This work was supported, in whole or in part, by the Key Program for International S&T Cooperation Projects of China (2021YFE0191600 to GC) and the National Natural Science Foundation of China (31950410564 to BG, 31672550 to GC).

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Authors and Affiliations

  1. Shanghai Tenth People’s Hospital, Tongji University School of Medicine, 500 Zhen-Nan Road, Shanghai, 200311, China

    Bikash R. Giri & Guofeng Cheng

  2. Department of Zoology, Utkal University, Bhubaneswar, 751004, India

    Bikash R. Giri

  3. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai, 200241, China

    Shun Li

  4. School of Life Sciences and Engineering, Foshan University, Foshan, Guangdong, 528225, China

    Shun Li

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  1. Bikash R. Giri
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  2. Shun Li
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Contributions

Bikash R. Giri: conceptualization, investigation, analysis, writing, funding acquisition; Li Shun: investigation, analysis; Cheng Guofeng: conceptualization, supervision, methodology, editing, funding acquisition.

Corresponding author

Correspondence to Guofeng Cheng.

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Animal experiments were performed in compliance with the institutional guidelines and by following the protocol approved by the Institutional Care and Use Committee of Tongji University School of Medicine, China (TJAA00621101).

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The authors declare no competing interests.

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Giri, B.R., Li, S. & Cheng, G. Exogenous modification of EL-4 T cell extracellular vesicles with miR-155 induce macrophage into M1-type polarization. Drug Deliv. and Transl. Res. 14, 934–944 (2024). https://doi.org/10.1007/s13346-023-01442-4

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