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Engineering Exosomes for Therapeutic Applications: Decoding Biogenesis, Content Modification, and Cargo Loading Strategies

Bahadorani, Mehrnoosh, Nasiri, Mahboobeh, Dellinger, Kristen, Aravamudhan, Shyam, Zadegan, Reza·2024·International Journal of Nanomedicine
DOI 10.2147/ijn.s464249PMC PMC11268655
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Figure 1 The biogenesis, contents, and cell uptake methods of the exosome. Exosomes, identified by markers CD63, CD9, and CD81, form when multivesicular bodies (MVBs) fuse with the plasma membrane, encapsulating nucleic acids, proteins, lipids, metabolites, and small molecules. MVBs can either be digested by lysosomes or release exosomes. Once released by donor cells, exosomes are absorbed by recipient cells through various pathways: pinocytosis, clathrin- and caveolin-mediated endocytosis, lipid raft endocytosis, macropinocytosis, or membrane fusion. Within recipient cells, exosome contents are sorted into various compartments or induce specific cellular responses. The figure was drawn using pictures from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).
Figure 1 The biogenesis, contents, and cell uptake methods of the exosome. Exosomes, identified by markers CD63, CD9, and CD81, form when multivesicular bodies (MVBs) fuse with the plasma membrane, encapsulating nucleic acids, proteins, lipids, metabolites, and small molecules. MVBs can either be digested by lysosomes or release exosomes. Once released by donor cells, exosomes are absorbed by recipient cells through various pathways: pinocytosis, clathrin- and caveolin-mediated endocytosis, lipid raft endocytosis, macropinocytosis, or membrane fusion. Within recipient cells, exosome contents are sorted into various compartments or induce specific cellular responses. The figure was drawn using pictures from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).CC BY-NC · PubMed Central
Figure 2 Schematic diagram of exosome engineering for cargo loading and surface modification. Techniques include gene editing, passive incubation, surfactant treatment, electroporation, heat shock, sonication, freeze-thaw, extrusion, and dialysis. Liposome hybridization enables surface and cargo loading. Cell extrusion produces biomimetic exosomes. Surface engineering employs non-covalent agents or click chemistry. One part of the figure was drawn using an image from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).
Figure 2 Schematic diagram of exosome engineering for cargo loading and surface modification. Techniques include gene editing, passive incubation, surfactant treatment, electroporation, heat shock, sonication, freeze-thaw, extrusion, and dialysis. Liposome hybridization enables surface and cargo loading. Cell extrusion produces biomimetic exosomes. Surface engineering employs non-covalent agents or click chemistry. One part of the figure was drawn using an image from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).CC BY-NC · PubMed Central
Figure 3 Engineering of the exosome using its protein platform. (A) The exosomal CD9 protein was modified with Enhanced Green Fluorescent Protein (EGFP) and Cry Interacting Protein (CIBN). This enables attachment to the photoreceptor protein CRY2 under blue light, and release of the photoreceptor and cargo protein in its absence. Reproduced with permission from Yim N, Ryu SW, Choi K, et al. Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein-protein interaction module. Nature Commun. 2016:9.13 (B) Using the bacteriophage library, the peptide CP05 with the best affinity for CD63 was selected. CP05 was conjugated with the muscle-targeting peptide (M12) and phosphorodiamidate morpholino oligomer (PMO). CP05-M12 and CP05-PMO were attached to exosomes, enabling targeting of dystrophy mice models (mdx). “i.v”. stands for intravenous. From Gao X, Ran N, Dong X, et al. Anchor peptide captures, targets, and loads exosomes of diverse origins for diagnostics and therapy. Sci Transl Med. 2018;10(444). Reprintedwith permission from AAAS.103
Figure 3 Engineering of the exosome using its protein platform. (A) The exosomal CD9 protein was modified with Enhanced Green Fluorescent Protein (EGFP) and Cry Interacting Protein (CIBN). This enables attachment to the photoreceptor protein CRY2 under blue light, and release of the photoreceptor and cargo protein in its absence. Reproduced with permission from Yim N, Ryu SW, Choi K, et al. Exosome engineering for efficient intracellular delivery of soluble proteins using optically reversible protein-protein interaction module. Nature Commun. 2016:9.13 (B) Using the bacteriophage library, the peptide CP05 with the best affinity for CD63 was selected. CP05 was conjugated with the muscle-targeting peptide (M12) and phosphorodiamidate morpholino oligomer (PMO). CP05-M12 and CP05-PMO were attached to exosomes, enabling targeting of dystrophy mice models (mdx). “i.v”. stands for intravenous. From Gao X, Ran N, Dong X, et al. Anchor peptide captures, targets, and loads exosomes of diverse origins for diagnostics and therapy. Sci Transl Med. 2018;10(444). Reprintedwith permission from AAAS.103CC BY-NC · PubMed Central
Figure 4 Natural resources for engineering exosomes. These resources include autologous or donor exosomes, embryonic stem cells, induced pluripotent stem cells, placenta, or milk. Parts of the figure were drawn using images from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).
Figure 4 Natural resources for engineering exosomes. These resources include autologous or donor exosomes, embryonic stem cells, induced pluripotent stem cells, placenta, or milk. Parts of the figure were drawn using images from Servier Medical Art. Servier Medical Art by Servier is licensed under a Creative Commons Attribution (https://creativecommons.org/licenses/by/4.0/).CC BY-NC · PubMed Central

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Metadata

Authors
Bahadorani, M., Nasiri, M., Dellinger, K., Aravamudhan, S., & Zadegan, R.
Venue
International Journal of Nanomedicine
Publisher
Informa UK Limited
Published
2024-07
Volume
Volume 19
Pages
7137-7164
ISSN
1178-2013
DOI
10.2147/ijn.s464249
PMC
PMC11268655
Cited by
121
References
18
Language
en
Licence
creativecommons.org/licenses/by-nc/3.0/ (unspecified)

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