Journal of Extracellular Vesicles最新文献

Kim EH, Son JP, Oh GS, et al. 2025. “Clinical Scale MSC-Derived Extracellular Vesicles Enhance Poststroke Neuroplasticity in Rodents and Non-Human Primates”. Journal of Extracellular Vesicles 14, no. 6: e70110. https://doi.org/10.1002/jev2.70110

In the originally published article, the funding entity for Eun Hee Kim, Gyun Sik Oh and Suji Park was given incorrectly as S&E bio, Inc. The correct funding information is given below.

Funding: S&E bio Co., Ltd., provided support for this study in the form of salaries for Eun Hee Kim, Gyun Sik Oh, and Suji Park.

We apologize for this error.

Epidermal permeability barrier defects are associated with several skin diseases, including atopic dermatitis (AD). Using an AD mouse model, we previously demonstrated that topically administered small extracellular vesicles (sEVs) (prepared following the International Society of Extracellular Vesicles recommendations) from human adipose tissue-derived mesenchymal stem cells (ASC) ameliorate skin inflammation and normalize barrier function in parallel with increased ceramide (a key barrier lipid) production. To elucidate how ASC-sEVs alleviate these AD skin abnormalities, we characterized lipids and ceramide metabolic enzymes in ASC-sEVs versus donor ASCs. Our study revealed that free fatty acid, ceramide, and sphingomyelin are enriched in ASC-sEVs versus donor ASCs, while the synthetic enzymes of ceramide (and acidic sphingomyelinase), and sphingosine-1-phosphate (sphingosine kinase) are significantly higher in ASC-sEVs versus donor ASCs. Conversely, ceramide (ceramidase), and sphingosine-1-phosphate hydrolytic enzymes (sphingosine-1-phosphate lyase and sphingosine-1-phosphate phosphatase) are lower in ASC-sEVs, suggesting that ceramide and sphingosine-1-phosphate levels could elevate in cells that receive ASC-sEVs. ASC-sEV-mediated increases in sphingosine-1-phosphate suppress pro-inflammatory cytokine production in AD-model human keratinocytes. Additionally, keratinocyte differentiation, which is required for a competent epidermal permeability barrier, was restored in AD-model human keratinocytes treated with ASC-sEVs. Taken together, cells that endocytose ASC-sEVs can normalize epidermal permeability barrier function as well as alleviate inflammation by stimulating a sphingosine-1-phosphate signalling pathway.

Lung cancer is the most prevalent malignancy worldwide, with the majority of fatalities attributed to metastasis. Recent studies have demonstrated the pivotal role of extracellular vesicles (EVs) and glycoproteins in tumor progression. In this study, we compared the glycoproteome of EVs from 95C (low metastatic) and 95D (high metastatic) lung cancer cells to discover key targets in metastasis. Through coupling lectin affinity chromatography with quantitative proteomics, 1562 glycoproteins were identified. Compared to 95C EVs, 23 glycoproteins were significantly upregulated more than 20-fold in 95D EVs, including CDCP1, TNC, NCAM2, and ITGA4. CUB-domain containing protein 1 (CDCP1) was upregulated 143-fold in 95D EVs, which is significantly correlated with poor prognosis of lung cancer patients in the TCGA database. We subsequently performed site-specific glycoform profiling of CDCP1 using intact glycopeptide enrichment. Then we generated CDCP1 knockout (KO) 95D cell lines and revealed that the absence of CDCP1 reduced cell migration ability, which was also confirmed by EVs and cell co-culture experiments. We further performed Ti⁴⁺-IMAC-based phosphoproteomic analysis to investigate the changes in signaling pathways in CDCP1 KO cell lines. 147 differentially expressed phosphoproteins were revealed. Verification experiments confirmed that the levels of phosphorylated SRC and JUN proteins, markers of ErbB signaling pathway, were decreased 5.5-fold and 4.2-fold, respectively. Glycosylation site mutagenesis identified N339 and N386 as critical functional determinants of CDCP1. Collectively, our data demonstrate that glycoprotein CDCP1 was selectively packed into EVs and potentially contributed to cancer metastasis, which is a critical target for anti-metastasis research and cancer therapy.

Infections caused by methicillin-resistant staphylococci (MRS), such as methicillin-resistant Staphylococcus aureus (MRSA), pose significant challenges to public health. The early detection of MRS infections and monitoring of antibiotic resistance profiles are critical for patient management and infection control strategies. Extracellular vesicles (EVs) have emerged as promising biomarkers in infectious disease. By combining single-particle nano-flow cytometry and immunoelectron microscopy (immuno-TEM), we discovered that PBP2a is present on the surface of EVs extracted from MRS. However, whether PBP2a can serve as an EVs-associated protein marker for diagnosing bacterial infections remains unexplored. Using MRSA as a model strain, mouse models of localized and systemic infections were established, alongside a clinical cohort study, to investigate the dynamics of PBP2a-positive (PBP2a⁺) EVs in plasma following bacterial infection, infection progression, and in response to antimicrobial therapy. In mouse infection models, PBP2a⁺ EVs were detected in plasma, with variable detection rates observed across different infection models. The study found a progressive correlation between increasing plasma PBP2a⁺ EVs levels and non-specific inflammation markers (CRP, IL-6) during infection progression. Antimicrobial therapies, however, inversely affected the ratio of PBP2a⁺ EVs. Furthermore, a clinical cohort study confirmed a direct association between the magnitude of PBP2a⁺ EVs in the plasma of patients with MRSA infection and the severity of infection. The investigation highlights the potential of PBP2a⁺ EVs as plasma biomarkers of MRSA antibiotic resistance, particularly during the early stages of resistant infections. Their persistence in plasma throughout the infectious episode makes them valuable indicators for monitoring disease progression and evaluating the efficacy of antibiotic treatments.

Systemic lupus erythematosus (SLE) has been linked to gut microbiome dysbiosis, notably an overabundance of Streptococcus anginosus; however, the impact of this microbial imbalance on disease pathogenesis remains unclear. Here, we investigated the contribution of S. anginosus-derived extracellular vesicles (SA-EVs) to SLE progression, with an emphasis on lupus nephritis (LN). Fifty-four SLE patients and 43 healthy controls (HC) were recruited. The faecal, blood and serum samples from participants were collected. SLE disease activity (SLEDA) was evaluated by the SLEDA Index (SLEDAI). Stool S. anginosus abundance was quantified by quantitative PCR, NK cell activation by flow cytometry and serum proinflammatory cytokines profile by ELISA. Lupus-prone MRL/lpr mice were orally administered SA-EVs to evaluate in vivo inflammatory responses, renal NK cell activation and renal histopathological changes. S. anginosus levels were significantly elevated in SLE patients relative to HC, positively correlated with SLEDAI scores and NK cell cytotoxicity. In vitro, SA-EVs stimulation of patient NK cells significantly heightened proinflammatory mediator production (granzyme B, TNF-α), increased cytotoxicity and downregulated inhibitory receptors (TIM-3, NKG2A, TIGIT) compared to control EVs from S. Salivarius (SS-EVs). Mechanistically, lipoteichoic acid (LTA) within SA-EVs engaged Toll-like receptor 2 (TLR2) on NK cells, activating MyD88/NF-κB signalling pathway. In MRL/lpr mice, SA-EVs treatment increased renal immune complex deposition, upregulated renal NK cell activation markers (NKp44, NKp46), and exacerbated LN pathology with greater immune cell infiltration and inflammatory cytokine levels. Furthermore, NK cell depletion with anti-NK1.1 antibodies significantly prolonged survival in SA-EVs administered mice. Thus, SA-EVs exacerbate SLE by hyperactivating NK cells via the TLR2-MyD88-NF-κB pathway, leading to amplified systemic inflammation and aggravated LN. These findings underscore the potential of targeting SA-EVs for therapeutic intervention in SLE.

Despite remarkable interest in the biomarker potential of urinary extracellular vesicles (uEVs) and the identification of numerous promising candidates, their clinical translation still presents multiple challenges. The opportunities for successful translation are obvious, yet the main roadblocks on the way have hardly been systematically considered and more coordinated approaches are needed to overcome them. In the present review article, we have identified the most relevant roadblocks of clinical translation of urinary EV-based biomarkers and discuss possible solutions to overcome them. These roadblocks are categorized as fundamental and technical but also related to development of novel biomarker assays and clinical acceptance. In addition, hurdles within the regulatory approval process are discussed. It is clear that various roadblocks to clinical translation of urinary EV biomarkers exist; however, they are addressable by promoting rigor and reproducibility as well as collaboration between basic and clinical scientists, clinicians, industry and regulatory bodies. Moreover, knowledge of obstacles for assay development and regulatory requirements should already be considered when developing a new biomarker to maximize the chance of successful translation. This review presents not only a status quo, but also a roadmap for the further development of the field.

Small extracellular vesicles (SEVs) are involved in diverse functions in normal and pathological situations, including intercellular communication, immunity, metastasis and neurodegeneration. Cell release of SEVs is assumed to occur passively right after multivesicular bodies of the endocytic pathway fuse with the plasma membrane. We show here that the completion of SEV release depends on membrane-bound ADAM10 and ADAM17 sheddases that promote the detachment of SEVs from the cell surface by catalysing the cleavage of adhesion proteins of the SEV membrane. The intensity of ADAM10/17-mediated release of SEVs depends on a balanced control of 3-phosphoinositide–dependent kinase 1 (PDK1) and ERK1/2 signalling pathways converging on 90-kDa ribosomal S6 kinase-2 (RSK2), which, in turn, fine-tunes ADAM17 bioavailability and ADAM10/17 enzymatic activities at the plasma membrane, according to a mechanism that relies, at least in part, on variation of the rhomboid-like pseudoprotease iRhom2 cell surface level. By identifying a new proteolytic step involved in the basal release of SEVs, our work may help understand how the deregulation of ADAM10/17-mediated discharge of SEVs contributes to several pathological states.

Metastasis constitutes the principal factor leading to the unfavourable prognosis of osteosarcoma patients. Hypoxia, as the inherent microenvironment of osteosarcoma, can upregulate HIF-1α via multiple pathways, thereby facilitating osteosarcoma proliferation and metastasis. Our previous research indicated that the inwardly rectifying potassium channel subfamily J member 2 (KCNJ2) inhibits the degradation of HIF-1α in osteosarcoma. Concurrently, HIF-1α upregulates the expression of KCNJ2 through a positive feedback regulatory mechanism. This positive regulatory mechanism significantly promotes the proliferation and metastasis of osteosarcoma. Therefore, the development of a KCNJ2-targeted therapeutic strategy capable of disrupting this reciprocal regulatory loop represents a crucial intervention for impeding osteosarcoma progression. The CRISPR/Cas9 targeted gene editing technology has garnered extensive attention in the field of tumour treatment due to its high efficiency and low off-target rate. Nevertheless, the relative lag of the delivery systems has restricted its application. The extracellular vesicles (EVs) secreted by bone marrow mesenchymal stem cells (BMSCs) have a natural targeting specificity for osteosarcoma and possess superior biocompatibility, making them ideal carriers for in vivo delivery. However, it is essential to confirm whether the CRISPR/Cas9 system mediated by EVs can accurately function intracellularly. Hence, we developed a fluorescence-based Cas9 editing efficiency reporter system. When CRISPR/Cas9 system induces double-strand breaks at specific target sites and results in frameshift mutations, osteosarcoma cells will stably express GFP. This system enables the transformation of gene editing events into quantifiable fluorescence signals. Furthermore, we engineered radiolabelled EVs derived from BMSCs to deliver the CRISPR/Cas9 system targeting KCNJ2. Using this reporter system, we confirmed their efficient gene-editing capabilities in vitro. Additionally, leveraging their radiolabelling properties, we validated their targeted distribution in vivo. Subsequent investigations revealed that our constructed ¹²⁴I@EVs-Cas9 effectively suppresses the proliferation and metastasis of osteosarcoma by targeting the inhibition of KCNJ2 expression and promoting HIF-1α ubiquitin-dependent degradation (as depicted in Graphical Abstract).

Spinal cord injury (SCI) is a severe and complex condition that can lead to significant physical impairments and affect the life quality of patients. Neural stem cells (NSCs) transplantation holds as a promising therapeutic approach for SCI. However, the challenging post-SCI microenvironment limits NSCs effectiveness. Our current research has found that transplanted NSCs, though with lower survival and differentiation, still aided in injury repair. Hypoxia was identified as a stressor inducing the release of extracellular vesicles (EVs) from NSCs through HIF-1α/RAB17 enhancing SCI repair. By extracting and modifying these EVs derived from hypoxia treated NSCs with CAQK/Angiopep2 peptides, we were able to accurately deliver them to the injury site, enhancing recovery without relying on cell survival or differentiation. This study delved into the reparative role and underlying mechanisms of transplanted NSCs in SCI, focusing on their non-cellular contributions and developed an innovative, targeted strategy for the transplantation of EVs derived from NSCs, offering a cell-free, precision therapeutic intervention for the treatment of SCI.

Recent studies have confirmed that a biomolecular corona forms around extracellular vesicles (EVs) in biofluids. However, there is limited data on how this adsorbed corona affects the accessibility of EV surface molecules. Here, we investigated various potential corona-stripping conditions for their ability to affect the immune detection of EVs. First, we artificially formed an EV corona around nascent HEK293T-PalmGFP cell-derived large EVs (lEVs) by incubating them with Cy5-labelled human plasma proteins. The co-localisation rate of plasma proteins and lEVs decreased significantly upon high-salt washing with NaCl, LiCl and KCl solutions, suggesting a considerable removal of the corona components. Additional evidence for corona modification was a significantly increased fluorescent annexin V binding to plasma lEVs and annexin V affinity capture of both THP1- and blood plasma-derived lEVs upon high-salt washing. A similar effect of high ionic strength was observed when THP1 lEVs were separated from a serum-containing medium, which allowed for corona formation, but not when EVs were produced under serum-free conditions. Using a MACSPlex kit and high-salt washing for small EVs from plasma and THP1 conditioned medium, we also demonstrated significantly improved immunodetection of 15 and 9 out of 37 surface markers, respectively. In this Technical Note, we present evidence that modifying the protein corona around EVs can significantly affect the immune detection of specific EV markers.