Journal of extracellular biology最新文献

Extra-large extracellular vesicles (XLEVs), with diameters > 600 nm, are increasingly recognised as mediators of specialized modes of intercellular communication; however, the molecular mechanisms governing their biogenesis and functional regulation remain poorly understood. Here, we show that PI3K–Rab18-GDP signalling promotes the secretion of XLEVs from human mesenchymal stem cells (hMSCs) and fibroblasts. These vesicles are highly enriched in sonic hedgehog (SHH) and display potent pro-angiogenic activity. We further demonstrate that Rab18 functions as a key regulator of this pathway specifically in its GDP-bound form, which can be enriched by the Rab inhibitor CID1067700 or by pharmacological activation of PI3K using SF1670. Rab18-GDP preferentially accumulates in the perinuclear region, where it promotes the formation of SHH-XLEV precursors from endosomal compartments. Mechanistically, PI3K–Rab18-GDP signalling recruits heat shock protein 90α (Hsp90α) and neutral sphingomyelinase 2 (nSMase2), facilitating polarized release of SHH-XLEVs from the perinuclear–plasma membrane interface, accompanied by an Hsp90α-enriched extracellular assembly. Together, these findings identify a PI3K–Rab18-GDP–dependent secretory pathway for SHH-XLEVs and provide a framework for understanding how XLEV biogenesis is coupled to SHH-associated angiogenic signalling in developmental and regenerative contexts.

Extracellular vesicles (EVs) are key mediators of maternal–foetal communication, regulating placental function and foetal development through the transfer of bioactive molecules. Although placental EVs play a crucial role in placental function during pregnancy, their contribution to foetal development, notably foetal brain, remains poorly understood. Human cytomegalovirus (HCMV) is the most common virus transmitted in utero and a leading cause of infectious brain malformations. Although certain central nervous system lesions caused by HCMV are explained, the neuropathogenesis of congenital infection remains poorly understood. In this study, we demonstrate that EVs from healthy placentas promote neurogenesis. However, EVs from HCMV-infected placentas lose this neurogenic potential, impairing differentiation and migration of neural stem cells, perturbations that may contribute to the neurodevelopmental defects observed in congenital HCMV infections. miRNA profiling revealed profound infection-induced changes, including the incorporation of viral miRNAs and dysregulation of host miRNAs involved in neurogenesis. These findings highlight the critical role of placental EVs in foetal brain development and their contribution to HCMV neuropathogenesis.

Human milk extracellular vesicles (HMEVs), secreted by mammary epithelial cells, are enriched in bioactive molecules that support intestinal epithelial integrity. Among these, oxylipins, that is, lipid mediators derived from polyunsaturated fatty acids, are gaining interest for their immunomodulatory and neuroprotective functions in breastfed infants. However, current workflows for oxylipin profiling in HMEVs often lack sensitivity or breadth, limiting mechanistic insights. This study presents an optimised workflow for comprehensive oxylipin profiling in HMEVs. HMEVs were isolated via size-exclusion chromatography and ultracentrifugation, followed by characterisation using attenuated total reflectance–Fourier transform infrared spectroscopy, Western blotting, Exoview immunocapture, tunable resistive pulse sensing and transmission electron microscopy. The influence of different pre-analytical protocols on HMEV recovery was assessed. Cryolysis with liquid nitrogen was employed for vesicle lysis before targeted oxylipin quantification using ultra-performance liquid chromatography–tandem mass spectrometry. The analysis of 10 human milk samples revealed 9,10-DiHOME, 12,13-DiHOME and 11,12-EET as the most abundant oxylipins, with concentrations ranging from 0.5 to 3.7, 0.8 to 4.5 and 0.1 to 0.3 nM, respectively. This refined pipeline enables in-depth oxylipin profiling in HMEVs and serves as a robust platform for future in vitro and in vivo investigations into EV-mediated lipid signalling.

Regenerative vascular medicine research has positioned mesenchymal stromal cells (MSCs) as a leading candidate to treat ischemic diseases. Recent studies have highlighted the emerging role of small extracellular vesicles (sEVs) produced by MSCs in their own potential. This study explores a strategy to improve the angiogenic potential of MSCs through the acquisition of endothelial features. Umbilical cord Wharton's jelly MSCs were cultured in fetal bovine serum-free endothelial growth medium under hypoxic conditions (SH-MSCs). sEVs were characterised by a multimodal approach: visualisation, count and particle size distribution, sEVs surface antigen, proangiogenic potential and ability to internalise into recipient cells. Compared with MSCs, SH-MSCs exhibited significant morphological and phenotypical change characterised by the up-regulation of CD31 and CD144 mRNA as well as a marked increase in sEVs secretion. MSC- and SH-MSC-derived sEVs had the capacity to internalise into endothelial cells, myoblasts and macrophages; exhibited a strong proangiogenic effect in vitro, particularly in promoting endothelial cell proliferation and pseudotube formation, likely due to an enriched cargo of angiogenic factors. These results highlight the dual benefit of hypoxia conditioning and endothelial differentiation of MSCs to optimise the angiogenic potential of their secreted sEVs, thus paving the way for innovative regenerative therapies in ischemic diseases.

Extracellular vesicles (EVs) have gained significant attention as emerging tools in diagnostics and therapeutics. Using the Gartner Hype Cycle framework, this commentary examines the current trajectory of EV research, from initial enthusiasm to growing concerns about reproducibility, standardization and clinical translation. We highlight key challenges, including EV heterogeneity, methodological inconsistencies and publication bias, which risk stalling progress. Ongoing efforts by the International Society for Extracellular Vesicles (ISEV), including Minimal information for studies of extracellular vesicles (MISEV) guidelines and the extracellular vesicle-transparent reporting and centralizing knowledge (EV-TRACK) database, have been crucial for advancing the field. We tackle actionable priorities to support rigorous, transparent and clinically meaningful EV research that would prompt the actual translation.

The placenta is a vital mediator of maternal–foetal communication, and extracellular vesicles (EVs) derived from placental tissue have gained attention as promising biomarkers of pregnancy health. Accurate molecular profiling of placental EVs is critical for advancing their diagnostic and mechanistic applications. However, how different EV isolation methods influence their composition remains poorly understood. This study directly compared EVs isolated from mouse placental tissue using two common approaches, enzymatic digestion and explant culture, evaluating their structural features, size distribution and proteomic content. Both methods successfully isolated small EVs (sEVs) with canonical markers (CD63, TSG101 and HSC70) and characteristic EV morphology. The digestion method produced a higher yield of larger EVs with a broader size range. Proteomic profiling showed substantial overlap but also revealed method-specific enrichment. Explant-derived EVs were enriched in RNA-binding proteins, translation factors and proteins related to post-transcriptional regulation and stress responses. In contrast, digestion-derived EVs were enriched for extracellular matrix (ECM) proteins and ER- and mitochondrial-associated proteins. These EVs also demonstrated stronger enrichment for placental-specific proteins. Density gradient purification confirmed that canonical EV markers localized to expected fractions. However, the ER protein GRP94 was also present, indicating possible vesicle association, although its intracellular versus extracellular origin remains unclear. Together, our findings show that the tissue dissociation strategy significantly shapes placental EV composition. Enzymatic digestion may improve the recovery of matrix-embedded EVs, but it increases the likelihood of capturing intracellular components. An explant culture approach yields a more selective EV population, potentially influenced by prolonged ex vivo conditions. These results underscore the importance of aligning EV isolation methods with specific experimental objectives and highlight key considerations for placental EV biomarker discovery and translational applications.

Extracellular vesicles (EVs) promote intercellular communication, playing a key role in the secondary immune-related pathologies driven by chronic inflammation in people living with HIV (PLWH). To identify molecular components within large EVs (lEVs) from PLWH's plasma that may influence immune function and contribute to the pathological process. PLWH were classified using clinical data, cellular immunophenotyping, and plasma mediator profiling. lEVs were characterized using transcriptomic, proteomic, and interactome analysis. Their functional impact on immune cells was also assessed. PLWH showed signs of chronic basal inflammation. Compared to the control group, lEVs from PLWH carried the miR-4433b-3p, 31 long non-coding RNAs and 45 proteins differentially expressed. Key proteins—FBXO7, C3, SUGT1 and DTX3L—were linked to the miR-4433b-3p regulatory network, suggesting their involvement in inflammation. Interactome and pathway enrichment analysis associated these molecules to critical pathways, including NF-kappa B signalling and PI3K-AKT signalling. Finally, lEVs from PLWH more effectively modulated the production of inflammatory mediators in bystander immune cells. This study underscores the role of lEVs in shaping immune response during chronic HIV infection. By identifying specific molecular components, it provides valuable insights into potential therapeutic targets and candidate biomarkers for disease progression monitoring.

Extracellular vesicles (EVs) are increasingly recognized as universal mediators of communication in nature across all domains of life and as versatile tools with roles spanning a wide range of industries. Although EVs have been extensively studied in biomedicine, mainly in diagnostic and nanotherapy of cancer and neurodegenerative diseases, their potential applications in other impactful society areas are only beginning to be explored. Microbial EVs contribute significantly to biofilm formation, virulence and the transmission of antibiotic resistance, highlighting their importance in pathogenicity and infection control. In the aesthetic and dermatological sectors, EVs are gaining traction as innovative agents for skin regeneration, anti-ageing and inflammation modulation, with applications extending to cosmetic dermatology and non-invasive treatments. Veterinary medicine is also exploring EVs for diagnostics and therapeutic delivery, while in agriculture, they show promise in improving crop resilience, acting as natural biopesticides and supporting plant–microbe interactions. Inter-species and interkingdom EV communication understanding, potentially help pest control and disease prevention. Moreover, EVs are being investigated as biosensors for environmental pollution and as agents in soil and water remediation. In the food industry, EVs are explored for their functional benefits in promoting gut and systemic health. However, to fully realize their potential, challenges in large-scale production, quality control and regulatory approval must be addressed. In this article, innovative solutions and potential of EVs across other health issues, environment, agriculture and biotechnology have been revised and discussed.

D. Mladenović, J. Brealey, B. Peacock, K. Koort, and N. Zarovni, “Quantitative Fluorescent Nanoparticle Tracking Analysis and Nano-Flow Cytometry Enable Advanced Characterization of Single Extracellular Vesicles,” Journal of Extracellular Biology 4 (2025): e70031, https://doi.org/10.1002/jex2.70031.

In the originally published article, the number of particles for plasma EVs in Table 2 should be 5e10 instead of 5e9. The corrected table is shown below.

We apologize for this error.