Journal of extracellular biology最新文献

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.

Therapeutic proteins and peptides have revolutionized modern biomedicine, but their oral delivery is limited by gastrointestinal degradation and barriers. Small extracellular vesicles (sEVs), which are resistant to biochemical degradation and capable of traversing mucus and cellular barriers, hold great promise as next-generation oral delivery vehicles. Oral semaglutide, the first approved oral GLP-1 receptor agonist (GLP-1RA), employs vesicle-mediated transcellular transport, highlighting the potential of sEVs as an effective delivery vehicle. In this study, we demonstrate the successful oral delivery of two GLP-1RAs, semaglutide and previously unexplored tirzepatide, using milk-derived sEVs. Both peptides were efficiently loaded onto sEVs in vitro, and their oral administration effectively reduced blood glucose levels in diabetic db/db mice. Compared with the current SNAC technology, which is limited exclusively to semaglutide, our sEV platform provides broader applicability and versatility for oral peptide drug delivery.

Neural stem cell (NSC)-based therapies offer a promising strategy to promote brain repair by delivering neurotrophic factors, supporting cell replacement, and stimulating endogenous neurogenesis following injury. While numerous studies have highlighted the protective and regenerative potential of NSCs and their extracellular vesicles (EVs), progress toward clinical translation remains hindered by limited molecular characterization of NSC lines and their EV cargo. To address this gap, we characterized two therapeutically relevant human fetal NSC lines, LMNSC01 and LMNSC02, both engineered to express the L-MYC gene, along with their corresponding EVs. LMNSC01 cells primarily differentiated into neurones with limited glial populations, whereas LMNSC02 cells gave rise to all three major neural lineages: neural, glial and oligodendrocyte progenitor cells (OPCs). scRNA-seq revealed distinct transcriptional profiles with minimal overlap between the two LMNSC lines. Using single extracellular vesicle nanoscopy, we observed that both lines released predominantly circular EVs, with LMNSC02-EVs exhibiting higher levels of tetraspanins (CD9, CD63, and CD81) and a larger average diameter than LMNSC01-EVs. Proteomic profiling revealed that LMNSC01-EVs are enriched in proteins involved in cell adhesion, migration, junction formation, and neuronal projection development, while LMNSC02-EVs are enriched in factors related to cytoplasmic translation initiation and biosynthesis. These LMNSC-EVs (collected from undifferentiated LMNSCs) demonstrated neuroprotective effects in a brain organoid model of methotrexate-induced toxicity when added to corresponding LMNSC01- or LMNSC02-derived brain organoids. LMNSC01- and LMNSC02-derived EVs restored neuronal and astrocytic populations but failed to rescue OPCs. These findings demonstrate the therapeutic potential of LMNSC-derived EVs to counter chemotherapy-induced neurotoxicity by preserving neurones and astrocytes, while highlighting the need for repeated or complementary interventions to restore oligodendrocyte populations.

HIV-1 proteins and RNA are incorporated into extracellular vesicles (EVs) via the EV biogenesis machinery. Due to their similar size and content, EVs and HIV-1 particles are hard to separate, and current purification methods often overlook EVs' effects on infectivity. This study co-characterized HIV-1 particles and three EV subtypes to assess their impact on infection. The HIV-infected Raji CD4 DCIR cells' supernatants were harvested 2 and 8 days after infection. The 2-day supernatant was treated with proteinase K to discard viral components outside the EVs. The supernatants were fractionated into three pellets by differential centrifugation: 3K, 17K and 100K. EVs and viral particles were co-characterized for their host and viral contents and the pellets obtained after 8 days post-infection were tested for infectivity. Proteinase K reduced HIV-1 RNA in EVs without affecting p24 concentration. The p24 protein was mostly found in the 17K pellet and HIV-1 RNA was the most abundant in the 100K pellet for both 2- and 8-day productions. Nevertheless, the 3K pellet had the highest infectivity when cells were infected with an equal quantity of virus. Each EV subtype were co-purified with functional virus and uniquely influenced HIV-1 infectivity, underscoring the importance of considering EVs in viral preparations.

Extracellular vesicles (EVs) from blood plasma are promising biomarkers, as they carry surface markers indicative of their cell of origin. Size-exclusion chromatography (SEC) is commonly employed for EV enrichment, but the choice of pore size and plasma volume can significantly impact the yield, purity, and composition of isolated EVs. In this study, we systematically compared Izon SEC columns with pore sizes of 35 and 70 nm, using either 500 µL plasma (qEVoriginal, “small” column) or 10 mL plasma (qEV10, “large” column). Due to limited material obtained from small columns, fractions had to be pooled for downstream analyses, precluding detailed characterization of individual fractions. In contrast, the larger columns provided sufficient material to analyse each fraction separately, across multiple platforms, including nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), single-EV flow cytometry, MACSPlex surface protein array, immunoblotting, and LC-MS/MS. These analyses consistently identified fractions 1–3 as “EV-rich,” characterized by enrichment of EV markers and reduced levels of abundant plasma proteins. Moreover, a comparison of pore sizes demonstrated that the 70 nm column yielded a higher EV recovery with improved purity compared to the 35 nm column, including a greater abundance of immune cell-derived markers. Together, these findings established that the large 70 nm SEC columns are optimal for isolating EV-rich fractions from plasma, maximizing both EV yield and purity, while minimizing non-EV contaminants.

The tumour microenvironment (TME) constitution is decisive for cancer outcome and is manifested in diffusion-weighted (DW) magnetic resonance imaging (MRI). We hypothesized that the TME metabolic state is reflected by mitochondrial DNA (mtDNA) secreted in extracellular vesicles (EVs) and examined whether plasma EV-mtDNA variants may divulge MRI-assessed TME attributes of rectal cancer aggressiveness. On the diagnostic MRI scans from 60 rectal cancer patients, the apparent diffusion coefficient (ADC) was calculated on DW images (n = 29), and tumour volume (n = 57) and extramural vascular invasion (EMVI; all patients) were determined on anatomical images. Plasma EVs (all patients) were isolated by size exclusion chromatography and verified for EV features. The EV-mtDNA was sequenced along with mtDNA in whole blood (WB; normal tissue) to calculate the EV/WB-mtDNA total variant number (TVN) and heteroplasmic variant number (HVN)—as a proxy for TME intracellular mtDNA variants expelled in EVs. Low EV/WB-mtDNA TVN and HVN, indicative of hampered clearance of mutated mtDNA via EVs, were associated with low ADC (high TME cell density; p = 0.018, p = 0.005) and a large tumour volume (p = 0.002, p = 0.003). Likewise, low EV/WB-mtDNA TVN and HVN were associated with positive EMVI (tumour infiltration in blood vessels; p = 0.002, p = 0.003) and histologic ypN stage 1–2 (lymph nodes with tumour cells surviving radiotherapy; p = 0.002, p = 0.005), both indicators of high tumour aggressiveness. High cellular density may hamper the clearance of pathogenic tumour mtDNA variants by EVs and thus promote rectal cancer aggressiveness.

Trial Registration: ClinicalTrials.gov: NCT01816607. Registered 22 March 2013, https://clinicaltrials.gov/ct2/show/NCT01816607