Journal of Extracellular Vesicles最新文献

Dendrogenin A (DDA) is a cholesterol-derived antitumour metabolite that promotes the secretion of immunogenic antitumour exosomes (DDA-sEV) enriched in bis(monoacylglycero)phosphate (BMP). BMP is a phospholipid specific to late endosomes and lysosomes, where it plays a crucial role in lipid degradation, regulates the fate of endosomal cholesterol, and contributes to intraluminal vesicle formation. Dysregulation of BMP biosynthesis is associated with multiple diseases. Here, we show that the DDA/LXRβ complex activates the transcription and activity of phospholipase D (PLD) and CLN5, two enzymes involved in BMP biosynthesis. Inhibition of PLD in DDA-treated tumour cells reduces BMP levels in DDA-sEV, impairs their release, and their antitumour immune activity. Blocking BMP on DDA-sEV with a specific antibody abolishes their antitumour reponse, prevents the recruitment of activated dendritic cells (DC) and T cells into tumours, and decreases mouse survival. This blockade also impairs DDA-sEV uptake by immature DC (iDC) and hinders DC maturation and Th1 T cell activation. Notably, neutralising the BMP-presenting receptor on iDC inhibits DDA-sEV uptake and DC maturation. Treatment of iDC with free BMP induces their functional maturation, confirming BMP as a key immune activator. Furthermore, BMP-containing DDA-sEV enhance the efficacy of anti-PD-1 therapy in melanoma. Targeting LXRβ with DDA represents an innovative strategy to stimulate anticancer immunity by increasing BMP levels in tumours and sEV.

Precision cancer medicine with small tyrosine kinase inhibitors (TKIs) directed toward oncogenic drivers, are important treatment regimens for solid tumours. The epidermal growth factor receptor (EGFR)-TKI osimertinib is a preferred therapy for patients with non-small cell lung cancer (NSCLC) driven by activating mutations in EGFR, unfortunately responses are heterogeneous. This calls for non-invasive methods to predict or monitor treatment response, for example, via biomarker analyses in blood. To reveal such putative biomarkers, we analysed the proteome of extracellular vesicles (EVs) from osimertinib resistant or responsive NSCLC cells in vitro and from EVs isolated from serum samples of NSCLC patients treated with osimertinib in second line within the phase II clinical trial TREM. The protein cargo of the EVs was analysed by mass spectrometry (MS) and proximity extension assay (PEA). Western blotting, ELISA and single vesicle analysis was performed to validate and further confirm the expression of certain proteins. MS profiling of the NSCLC cells and their released EVs revealed a protein signature associated with osimertinib refractoriness. Among them were CSPG4, HSPG2, MCAM, L1CAM, TAGLN, THBS1 and TNC. GO-pathway analysis related several of these proteins to the focal adhesion and proteoglycan in cancer pathways. Some of these proteins, including CSPG4, which when suppressed by transient siRNA transfection in NSCLC cells resulted in reduced cell viability, were expressed also in EVs from serum of the NSCLC patients. Moreover, PEA profiling of the serum-isolated EVs revealed signatures associated with immune cells, best response and/or progression-free survival, including PD-L1, CD73/NT5E, FR-alpha/FOLR1, LAMP3, FASLG1 and ANXA1. In summary, we demonstrate that protein profiling of EVs in relation to osimertinib refractoriness has the potential to identify possible biomarkers that can indicate osimertinib treatment resistance, for example, CSPG4, HSPG2, TAGLN, TNC, THBS1, ANXA1 and CD73/NT5E. Studies in expanded cohorts should be conducted to further validate these putative osimertinib biomarkers.

The intestine plays a crucial role in regulating metabolism and immunity, with functional decline occurring during injury and ageing. Stimulating the neogenesis of intestinal stem cells (ISCs) by activating the WNT/β-catenin signalling pathway represents a promising approach for intestinal tissue regeneration and injury repair. However, effective oral delivery of functional WNT signalling agonists to the gut remains challenging. Herein, we report a potent WNT/β-catenin signalling-inducing small extracellular vesicles (sEV) that can be administered orally and present remarkable therapeutic efficacy. We demonstrate that active R-spondin1 (RSPO1) protein can be loaded onto the surface of sEV via heparan sulfate proteoglycans. Notably, sEV-delivered RSPO1 (evRSPO1) effectively induces WNT/β-catenin signalling-inducing activity, enhances ISCs proliferation, and supports intestinal organoid growth in vitro. Importantly, oral administration of evRSPO1 activates the WNT/β-catenin signalling pathway in the cryptic stem cell niche, thereby accelerating tissue repair and regeneration in a radiation-induced intestinal injury model. Furthermore, evRSPO1 treatment induces ISCs proliferation and reverses the intestinal senescence phenotype in aged mice. Collectively, this study establishes evRSPO1 as a potential first-in-class, orally deliverable therapeutic that overcomes biological barriers to activate ISCs, enabling efficient intestinal tissue repair and rejuvenation.

Extracellular vesicle (EV) secretion is an important, though not fully understood, intercellular communication process. Lipid metabolism has been shown to regulate EV activity, though the impact of specific lipid classes is unclear. Through analysis of small EVs (sEVs), we observe aberrant increases in sEV release within genetic models of cholesterol biosynthesis disorders, where cellular cholesterol is diminished. Inhibition of cholesterol synthesis at multiple synthetic steps mimics genetic models in terms of cholesterol reduction and sEVs secreted. Further analyses of sEVs from cholesterol-depleted cells revealed structural deficits and altered surface marker expression, though these sEVs were also more easily internalized by recipient cells. Transmission electron microscopy of cells with impaired cholesterol biosynthesis demonstrated multivesicular and multilamellar structures potentially associated with autophagic defects. We further found autophagic vesicles being redirected towards late endosomes at the expense of autophagolysosomes. These findings were subsequently validated in cellular models of head and neck cancer, where cholesterol depletion induced sEV release and promoted late endosome-autophagosome fusion. Through CRISPR-mediated inhibition of autophagosome formation, we mechanistically determined that release of sEVs after cholesterol depletion is autophagy dependent. We conclude that cholesterol imbalance initiates autophagosome-dependent secretion of sEVs, which may have pathological relevance in diseases of cholesterol disequilibrium.

Extracellular vesicles (EVs) have emerged as powerful mediators of intercellular communication with significant potential across biomedical, veterinary, cosmetic, agricultural and environmental applications. However, the translation of EV-based discoveries into practical and commercially viable products remains constrained by scientific complexity, regulatory uncertainty and manufacturing challenges. To address these barriers, the International Society for Extracellular Vesicles (ISEV) established the Translation, Regulation and Advocacy Committee (ISEV-TRA). ISEV-TRA aims to catalyse the responsible advancement of EV technologies by fostering cross-sector collaboration, harmonising quality and regulatory frameworks and providing strategic advocacy to enhance market readiness. Through targeted initiatives—such as workshops and the development of translational resources and guidance—ISEV-TRA seeks to bridge the gap between research and real-world implementation. By promoting dialogue amongst academia, industry, investors and policymakers, ISEV-TRA positions itself as a central driver in shaping the global roadmap for EV translation and commercialisation.

Idiopathic pulmonary fibrosis (IPF) is a fatal fibrotic lung disease characterized by aberrant myofibroblast activation and excessive extracellular matrix deposition, with extracellular vesicles (EVs) playing a crucial role in this pathological process. We observed that EVs levels are significantly elevated in IPF and positively correlate with nestin expression, a known marker of lung myofibroblasts. These myofibroblast-derived EVs further amplify profibrotic responses, creating a self-perpetuating cycle. To elucidate the mechanisms driving increased EVs secretion, we conducted in vitro and in vivo experiments, demonstrating that nestin knockdown not only suppresses EVs release but also impairs their ability to promote TGF-β-induced myofibroblast differentiation. Mechanistically, nestin recruits TBC1D15 to inactivate Rab7, thereby inhibiting multivesicular body (MVB) degradation and enhancing EVs secretion. Importantly, pharmacological activation of Rab7 using ML-098 significantly attenuated pulmonary fibrosis in mouse models. Our findings establish the Nestin-Rab7 axis as a key regulator of EVs-mediated fibrotic signaling and highlight its therapeutic potential for IPF treatment.

Oncolytic adenoviruses (OVs) can directly eliminate cancer cells and subsequently activate immune responses, exhibiting potent antitumor therapeutics. However, it was observed that the immune cells can also be lysed during viral treatment, evidently dampening the OVs-mediated antitumor immune response. In this study, we develop a microneedle (MN)-based in situ tumor cell-derived extracellular nanovesicle (TDEV)-cloaked OVs platform to enhance cancer immunotherapy and reduce immune cell exhaustion. In this platform, tumor cells pre-infected with OVs are loaded into the upper reservoir of the MN device. Following the transdermal administration, the hollow MN would constantly facilitate the transport of in situ the generated TDEV-encapsulating OVs into the tumor site for sustained delivery of OVs, which could subsequently infect cancer cells selectively rather than immune cells. Enhanced antigens triggered by improved intratumoral OVs killing can be presented by non-exhausted dendritic cells, further evoking significant immunotherapeutic effects in both TC-1-hCD46 xenograft tumor-bearing mice and postoperative tumor recurrence mice models.

As a novel identified manner of cell death, NETosis is widely regarded as an effective approach to resist pathogen infection but mainly focused on vertebrates with systematic cell typing. Besides, the role of extracellular vesicles (EVs), which are essential tools for intercellular information exchange, in regulating NETosis during pathogen infection has yet to be addressed. Here, we found that viral mRNA wsv271 could be packaged by EVs secreted by haemocytes during WSSV infection in mud crab, and delivered to the neutrophil-like cells, followed by translation into viral protein, and then interacted with the TIR domain of Toll4 to recruit MyD88, so as to activate P38-MAPK signal pathway and further facilitate PAD4 phosphorylation and nuclear translocation to mediate histone-H3 citrullination, which eventually activated NETosis-like response in haemocytes to suppress the spread of viral infection. Therefore, our research not only identified neutrophil-like cells from the haemocytes of a crustacean based on single-cell transcriptomics but also revealed a novel NETosis induction mechanism mediated by EVs-derived viral nucleic acid delivery.

Tan, J., Y. Hu, L. Zheng, et al. 2025. “Microbead Encapsulation Strategy for Efficient Production of Extracellular Vesicles Derived From Human Mesenchymal Stem Cells.” Journal of Extracellular Vesicles 14, no. 4: e70053. https://doi.org/10.1002/jev2.70053.

The footnote of this article has been updated to read Jiayi Tan, Yunxia Hu, and Lijuan Zheng contributed equally to this work. This was left out of an earlier corrected version of this article. The online version of this article has been corrected.

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