Cortical Actin Depolymerisation in 3D Cell Culture Enhances Extracellular Vesicle Secretion and Therapeutic Effects.

Yang, Z; Li, X; Lin, Q; Zhou, F; Liang, K; Li, JJ; Niu, Y; Meng, Q; Zhao, T; Li, H; Wang, D; Lin, J; Li, H; Wang, B; Liu, W; Du, Y; Lin, J; Xing, D

Cortical Actin Depolymerisation in 3D Cell Culture Enhances Extracellular Vesicle Secretion and Therapeutic Effects.

Yang, Z Li, X Lin, Q Zhou, F Liang, K Li, JJ Niu, Y Meng, Q Zhao, T Li, H Wang, D Lin, J Li, H Wang, B Liu, W Du, Y Lin, J Xing, D

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Publisher:: Wiley
Publication Type:: Journal Article
Citation:: J Extracell Vesicles, 2025, 14, (6), pp. e70109
Issue Date:: 2025-06

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Field	Value	Language
dc.contributor.author	Yang, Z
dc.contributor.author	Li, X
dc.contributor.author	Lin, Q
dc.contributor.author	Zhou, F
dc.contributor.author	Liang, K
dc.contributor.author	Li, JJ
dc.contributor.author	Niu, Y
dc.contributor.author	Meng, Q
dc.contributor.author	Zhao, T
dc.contributor.author	Li, H
dc.contributor.author	Wang, D
dc.contributor.author	Lin, J
dc.contributor.author	Li, H
dc.contributor.author	Wang, B
dc.contributor.author	Liu, W
dc.contributor.author	Du, Y
dc.contributor.author	Lin, J
dc.contributor.author	Xing, D
dc.date.accessioned	2025-07-04T05:18:23Z
dc.date.available	2025-05-19
dc.date.available	2025-07-04T05:18:23Z
dc.date.issued	2025-06
dc.identifier.citation	J Extracell Vesicles, 2025, 14, (6), pp. e70109
dc.identifier.issn	2001-3078
dc.identifier.issn	2001-3078
dc.identifier.uri	http://hdl.handle.net/10453/188082
dc.description.abstract	Three-dimensional (3D) culture systems have been shown to enhance cellular secretion of small extracellular vesicles (sEVs) compared to two-dimensional (2D) culture. However, the molecular mechanisms driving sEV secretion and influencing their potential for disease treatment have not been elucidated. In this study, we discovered the depolymerisation of cortical actin as a new mechanism that leads to increased sEV release, and that in 3D cultured mesenchymal stem cells (MSCs), this process was modulated by the downregulation of integrin-α1 (ITGA1) and subsequent inhibition of the RhoA/cofilin signalling pathway. Interestingly, the knockdown of Rab27A and Rab27B significantly reduced sEV secretion by MSCs to 0.5- and 0.1-fold, respectively. However, there was no difference in expression levels of Rab27A/B between MSCs cultured in 2D and 3D environments. In addition, sEVs derived from 3D cultured MSCs demonstrated enhanced therapeutic function both in vitro and in rat models of osteoarthritis (OA) and wound healing. Collectively, this study illustrates a new mechanism for enhanced secretion of sEVs, involving RhoA/cofilin pathway-dependent cortical actin depolymerisation, which is independent of Rab27A/B. These findings provide novel insights for optimising the yield of stem cell-derived sEVs, as well as their therapeutic efficacy for treating chronic diseases.
dc.format	Print
dc.language	eng
dc.publisher	Wiley
dc.relation.ispartof	J Extracell Vesicles
dc.relation.isbasedon	10.1002/jev2.70109
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	0601 Biochemistry and Cell Biology
dc.subject.classification	3101 Biochemistry and cell biology
dc.subject.mesh	Animals
dc.subject.mesh	Extracellular Vesicles
dc.subject.mesh	Mesenchymal Stem Cells
dc.subject.mesh	Actins
dc.subject.mesh	Rats
dc.subject.mesh	Humans
dc.subject.mesh	rhoA GTP-Binding Protein
dc.subject.mesh	Male
dc.subject.mesh	rab27 GTP-Binding Proteins
dc.subject.mesh	Cells, Cultured
dc.subject.mesh	Rats, Sprague-Dawley
dc.subject.mesh	Signal Transduction
dc.subject.mesh	rab GTP-Binding Proteins
dc.subject.mesh	Wound Healing
dc.subject.mesh	Cell Culture Techniques, Three Dimensional
dc.subject.mesh	Cells, Cultured
dc.subject.mesh	Mesenchymal Stem Cells
dc.subject.mesh	Animals
dc.subject.mesh	Humans
dc.subject.mesh	Rats
dc.subject.mesh	Rats, Sprague-Dawley
dc.subject.mesh	Actins
dc.subject.mesh	rab GTP-Binding Proteins
dc.subject.mesh	rhoA GTP-Binding Protein
dc.subject.mesh	Wound Healing
dc.subject.mesh	Signal Transduction
dc.subject.mesh	Male
dc.subject.mesh	Extracellular Vesicles
dc.subject.mesh	rab27 GTP-Binding Proteins
dc.subject.mesh	Cell Culture Techniques, Three Dimensional
dc.title	Cortical Actin Depolymerisation in 3D Cell Culture Enhances Extracellular Vesicle Secretion and Therapeutic Effects.
dc.type	Journal Article
utslib.citation.volume	14
utslib.location.activity	United States
utslib.for	0601 Biochemistry and Cell Biology
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
pubs.organisational-group	University of Technology Sydney/UTS Groups
pubs.organisational-group	University of Technology Sydney/UTS Groups/Centre for Health Technologies (CHT)
utslib.copyright.status	open_access	*
dc.rights.license	This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0). To view a copy of this license, visit https://creativecommons.org/licenses/by-nc/4.0/
dc.date.updated	2025-07-04T05:18:17Z
pubs.issue	6
pubs.publication-status	Published
pubs.volume	14
utslib.citation.issue	6

Abstract:

Three-dimensional (3D) culture systems have been shown to enhance cellular secretion of small extracellular vesicles (sEVs) compared to two-dimensional (2D) culture. However, the molecular mechanisms driving sEV secretion and influencing their potential for disease treatment have not been elucidated. In this study, we discovered the depolymerisation of cortical actin as a new mechanism that leads to increased sEV release, and that in 3D cultured mesenchymal stem cells (MSCs), this process was modulated by the downregulation of integrin-α1 (ITGA1) and subsequent inhibition of the RhoA/cofilin signalling pathway. Interestingly, the knockdown of Rab27A and Rab27B significantly reduced sEV secretion by MSCs to 0.5- and 0.1-fold, respectively. However, there was no difference in expression levels of Rab27A/B between MSCs cultured in 2D and 3D environments. In addition, sEVs derived from 3D cultured MSCs demonstrated enhanced therapeutic function both in vitro and in rat models of osteoarthritis (OA) and wound healing. Collectively, this study illustrates a new mechanism for enhanced secretion of sEVs, involving RhoA/cofilin pathway-dependent cortical actin depolymerisation, which is independent of Rab27A/B. These findings provide novel insights for optimising the yield of stem cell-derived sEVs, as well as their therapeutic efficacy for treating chronic diseases.

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http://hdl.handle.net/10453/188082