Search Results (3,538)

Background/Objectives: Pancreatic ductal adenocarcinoma (PDAC) is a devastating disease with limited therapeutic options, a high mortality rate, and poor overall survival, necessitating the development of new therapeutic and diagnostic strategies. This study investigated the potential of plasma-derived small extracellular vesicles (sEVs) as a source of molecular biomarkers associated with the treatment response. Methods: Plasma samples were obtained from patients with locally advanced and borderline resectable PDAC at baseline and following neoadjuvant chemotherapy, either FOLFIRINOX (5-FU [fluorouracil], leucovorin, oxaliplatin, and irinotecan) or GEM-ABRAX ( gemcitabine plus nab-paclitaxel), followed by stereotactic body radiation therapy (SBRT). sEVs were isolated from plasma at baseline, after neoadjuvant chemotherapy, and following SBRT, and were characterized by nanoparticle tracking analysis (NTA), transmission electron microscopy (TEM), nano-flow cytometry, and real-time PCR (RT-PCR). Results: The isolated sEVs exhibited an average size of <200 nm, expressed canonical exosome markers (CD63 and CD9), and exhibited pancreatic cancer (PanC)-associated markers, including cholecystokinin A receptor (CCK-AR) and carbohydrate antigen 19-9 (CA19-9). The sEV cargo included several PanC-associated microRNAs (miRNAs). Notably, the expression profiles of these miRNAs demonstrated interpatient variability, though a subset of miRNAs showed statistically significant changes following treatment. Conclusions: These findings support the feasibility of sEV isolation and molecular profiling from patient plasma and warrant further investigation as a potential source of biomarkers in pancreatic cancer. Full article

Ovarian cancer (OC) remains one of the most lethal gynaecological malignancies, which is mainly due to late diagnosis, high frequency of metastasis, and the risk of developing resistance to systemic therapy. In recent years, exosomes—small extracellular vesicles (EVs) secreted by cancer cells and components of the tumour microenvironment (TME)—have been identified as potential mediators of OC progression. Exosomes participate in intercellular communication and enable the transfer of RNA, proteins, and lipids. These vesicles may modulate the immune response, promote angiogenesis, remodel the extracellular matrix, and drive epithelial–mesenchymal transitions. Exosomes also appear to play a role in the development of drug resistance via direct transfer of resistance factors or indirect modification of TME. In this review article, we summarise current knowledge on the biological role of exosomes in OC pathogenesis. We also discuss their possible diagnostic, prognostic, and therapeutic relevance. The properties and composition of exosomes make them promising noninvasive liquid biomarkers and convenient carriers for anticancer drugs. However, to fully exploit their potential, further large-scale preclinical and clinical studies are required, which should focus primarily on standardising research methods and assessing the safety and efficacy of exosome-based diagnostic and therapeutic methods. Full article

Extracellular vesicle (EV) transcriptomic data provides a high-dimensional representation of cellular states but remains challenging to interpret due to noise, redundancy, and limited sample sizes. Most existing approaches rely on supervised differential expression analyses, which can be biased and may fail to capture latent structure in small datasets. In this study, we propose an unsupervised machine learning framework based on non-negative matrix factorization (NMF) to identify latent expression programs from EV mRNA profiles. A structured preprocessing pipeline combining expression filtering, variance selection, ANOVA-based feature selection, and correlation pruning was used to reduce dimensionality and improve signal quality prior to matrix factorization. NMF was applied to decompose the data into interpretable gene modules and sample-specific activation patterns. Model selection was performed using reconstruction error and component stability across multiple initializations. Candidate features were prioritized using a composite ranking score integrating module loadings, group-level expression differences, and model stability. The approach identified a stable low-rank representation capturing dominant patterns in the data and a compact set of informative features. These results demonstrate that unsupervised matrix factorization can effectively extract structured, interpretable signals from small-scale transcriptomic datasets and provide a robust framework for feature prioritization and representation learning in high-dimensional biological data. Full article

Endometriosis is a complex disorder associated with dysregulated immune, hormonal, and microenvironmental signaling. Extracellular vesicles (EVs) are important mediators of intercellular communication and may contribute to disease pathogenesis, biomarker discovery, and therapeutic targeting. Here, we systematically reviewed the literature on EVs in endometriosis, focusing on EV classification, isolation and characterization methods, and the functional relevance of EV-associated cargo. A total of 50 original studies were included and evaluated in the context of current International Society for Extracellular Vesicles (ISEV) recommendations. Our analysis revealed marked heterogeneity in EV nomenclature, biological sources, and methodological approaches. Although most studies used standard EV markers, the assessment of sample purity and inclusion of negative controls was inconsistent. Further studies using standardized workflows and well-characterized cohorts are needed to clarify their biological and clinical significance. Full article

Human follicular development depends on coordinated communication between granulosa cells (GCs) and oocytes through endocrine cues, direct contacts, and extracellular vesicles (EVs). Exosomes are key EV mediators of intrafollicular signaling, but their cargo and functions in gonadotropin-stimulated GCs remain poorly defined. The human granulosa-like tumor cell line KGN was used to investigate exosome secretion and protein composition following human chorionic gonadotropin (hCG) stimulation. Exosomes were isolated by ultracentrifugation, characterized via nanoparticle tracking analysis (NTA), Scanning Electron Microscopy (SEM) and Western blotting, and analyzed using high-resolution mass spectrometry. Comparative proteomics integrating exosomal profiles with the whole secretome were performed, followed by bioinformatic analyses of protein networks, gene ontology, and pathway enrichment. hCG reshaped exosomal cargo, identifying 59 proteins enriched in exosomes, including Integrin α3 (ITGα3), Galectin-3-binding protein (LGALS3BP), tetraspanins (CD63, CD151), and proteasome subunits. Functional enrichment indicated roles in extracellular matrix remodeling, integrin signaling, proteostasis, and steroidogenesis. Comparison with the secretome revealed distinct protein distributions, supporting selective exosomal packaging. Western blot confirmed increased ITGα3 and LGALS3BP levels in exosomes upon hCG treatment. In conclusion, hCG modulates exosome cargo composition in granulosa cells, uncovering a novel mechanism of extracellular regulation. Full article

In the human reproductive system, extracellular vesicles (EVs) have been recognized as playing a vital role in mediating cell–cell communication. They are considered critical for embryo development, implantation, gamete interaction, and fertilization. The various cargoes carried by EVs, depending on the physiological and pathological state of the cell, include proteins, lipids, nucleic acids, and mitochondrial components. EVs are recognized as critical carriers of redox-related signals and mitochondrial components, linking oxidative stress (OS) to reproductive failure and influencing gamete quality and embryo competence. Although considerable progress has been made, research remains poorly integrated, despite individual omics technologies providing valuable molecular insights. The use of multi-omics technologies, including transcriptomics, proteomics, metabolomics, and microbiome analysis, has been proposed as a global approach to understanding the complexities associated with EVs and discovering new biomarkers associated with infertility. ML and AI have been proposed to identify predictive signatures linked to ART effectiveness and reproductive outcomes, with a strong capacity to handle high-dimensional data. The review aims to provide an overview of current knowledge on EV-mediated redox–mitochondrial signaling in human reproduction, while highlighting the importance of emerging multi-omics and AI technologies for EV-mediated biomarker development. The review discusses the promise of EVs in the development of minimally invasive diagnostic approaches and therapeutic interventions, as well as the challenges in the standardization, integration, and clinical translation of EV-mediated research. In addition, the review proposes integrating computational approaches to better understand molecular pathways involved in the development of next-generation precision medicine in human reproduction. Full article

Blood-derived extracellular vesicle (EV) biomarkers have emerged as promising liquid-biopsy analytes for monitoring treatment response and prognosis in breast cancer. This scoping review mapped the clinical evidence on blood-derived EV in breast cancer and identified key barriers to clinical translation. Following the Joanna Briggs Institute framework and PRISMA-ScR guidelines, we searched PubMed, Embase, and Web of Science for eligible studies published through November 2025. After duplicate removal, title and abstract screening, and full-text assessment, 64 clinical studies were included. Research activity increased markedly from 2020 onward, accounting for 87.5% (56/64) of included studies. The literature was concentrated in East Asia, particularly China (51.6%, 33/64). RNA-based biomarkers predominated (60.9%, 39/64), especially microRNAs (39.1%, 25/64). Prognostic outcomes were evaluated in 89.1% (57/64) of studies, treatment response in 51.6% (33/64), and both endpoints in 40.6% (26/64). Triple-negative breast cancer was the most frequently studied subtype in isolation (15.6%, 10/64). Methodological heterogeneity was substantial, and kit-based precipitation was the most common EV isolation method (57.8%, 37/64). EV biomarkers show promise for non-invasive monitoring in breast cancer, but methodological standardization, compliance with Minimal Information for Studies of Extracellular Vesicles guidelines, and large prospective validation studies remain necessary before routine clinical implementation. Full article

In light of our previous publication, we hypothesized that chronic lymphocytic leukemia (CLL) cells also recruit monocytes to acquire survival advantage. To test this, we treated Buffy coat-driven monocytes with exosomes isolated from the peripheral blood of 45 treatment-naïve patients. The CLL-derived exosomes turned monocytes into IL-6-producing cells as an increase of 13-fold in the IL-6 levels was obtained in the growth medium of the exposed monocytes. Subsequently, we filtered out the monocytes and added CLL cells to this IL-6 enriched medium. As a result, the oncogene STAT3 became phosphorylated, and thus may have provided the cells with a survival advantage. A total of 67 phosphoproteins were upregulated in response to CLL-derived exosomal exposure in the recipient monocytes, with TFIIF being among the top scored proteins in this analysis. Transfection of monocytes with a TFIIF-containing vector increased the levels of IL-6 about 14-fold in the culture medium. Importantly, the CLL-derived exosomes induced the transformation of a portion of the recipient monocytes (45% compared to 30% of the unexposed cells) to become nurse-like fibrocyte cells. Taken together, CLL cells communicate with monocytes through the exosomes that they release. Once they are taken up by monocytes, they turn them into IL-6-producing cells, which provide a survival advantage to the neoplastic cells, creating a vicious circle that promotes disease progression. Full article

Intercellular mitochondrial trafficking has emerged as an important mechanism influencing tumor progression, metabolic adaptability, and cancer cell plasticity. Beyond their classical bioenergetic functions, mitochondria act as central regulators of redox homeostasis, signaling pathways, and epigenetic remodeling. Increasing evidence suggests that mitochondria can be transferred between tumor, stromal, and immune cells through tunneling nanotubes (TNTs), extracellular vesicles (EVs), gap junctions, and cell fusion within the tumor microenvironment. This dynamic excshange enables metabolically compromised cancer cells to restore oxidative phosphorylation, optimize energy production, and survive under hypoxia and therapeutic stress. Mitochondrial transfer has been increasingly associated with enhanced cellular plasticity and adaptive phenotypic transitions, including the acquisition of stem-like features that contribute to tumor heterogeneity, metastasis, and treatment resistance. In addition to bioenergetic restoration, transferred mitochondrial DNA and metabolites participate in retrograde signaling, linking metabolic state to epigenetic regulation and transcriptional reprogramming. This metabolic epigenetic interplay supports tumor cell adaptation to environmental stress and therapeutic pressure. Although significant progress has been made, the precise mechanisms governing mitochondrial integration and their long-term impact on cellular phenotypes remain incompletely understood. A deeper understanding of these processes may reveal novel therapeutic strategies to disrupt tumor adaptability and progression. Specifically, targeting intercellular mitochondrial trafficking and its associated metabolic and epigenetic effects could help limit tumor plasticity, overcome treatment resistance, reduce disease recurrence, and improve overall clinical outcomes in cancer patients. Full article

Postoperative surgical draining fluid (SDF) in oral squamous cell carcinoma (OSCC) was reported to contain circulating free DNA and extracellular vesicles (EVs). The potential role of SDF DNA as a cancer biomarker in OSCC patients is investigated. To determine whether EVs contribute to the DNA content of SDF, small EVs (sEVs) were isolated by size exclusion chromatography and compared with those in paired plasma. sEV obtained from a cohort of HPV(+) and HPV(−) patients were evaluated for the endocytic origin, size, vesicular morphology, and the DNA content. The sEV from HPV(+) and HPV(−) SDF were similar. The sEVs from SDF were larger and contained more DNA than the sEV from plasma. Treatments of sEV with the DNase/RNase cocktail established the presence of DNA on the surface and in the sEV lumen. Furthermore, 20 to 30% of the total SDF DNA was associated with sEV. Fragmentomic analyses identified the largest DNA (>10,000 bp) on the vesicle surface. The DNA in the vesicle lumen consisted of smaller (~5000 bp) DNA fragments. Electron microscopy of the enzyme-treated vesicles indicated that surface DNA/RNA may be involved in maintaining vesicular integrity and modulating the vesicle entry into cells. The sEV-carrying surface and luminal DNA emerge as significant contributors to the total SDF DNA and, following validation, might serve as useful biomarkers of disease presence/progression in OSCC. Full article

Boar fertility is negatively affected by subfertility and elevated temperatures, which alter seminal plasma (SP) composition and reduce semen quality. Extracellular vesicles (EVs) in SP transfer microRNAs (miRNAs) to sperm and may influence sperm function. This study aimed to identify SP-EV microRNAs associated with differences in boar semen quality during the summer season. Semen collected from Duroc boars was evaluated and classified as Passed (≥70%) or Failed (<70%) based on sperm quality parameters. SP-EVs were isolated and characterized, and small RNA sequencing was performed to profile miRNA content. SP-EVs ranged from 90 to 200 nm, with concentrations of 4.33 × 10¹⁰ particles/mL in the Passed group and 1.85 × 10¹¹ particles/mL in the Failed group. Western blotting confirmed the presence of EV surface markers CD9, CD63, and CD81. A total of 446 unique miRNAs were identified, with 28 downregulated and two upregulated miRNAs in Passed compared with Failed SP-EVs. Additionally, functional enrichment analysis revealed that target genes of upregulated miRNAs were involved in sperm-related biological processes and PI3K-Akt, regulation of actin cytoskeleton, and ErbB signaling pathways. These findings demonstrate that SP-EV miRNAs reflect physiological responses to changes in environmental conditions and may contribute to the regulation of boar semen quality. Full article

Extracellular vesicles (EVs) hold significant potential in cardiovascular diagnosis and treatment. However, their clinical applications are limited by challenges such as isolation efficiency, subpopulation heterogeneity, analytical standardization, and manufacturing scalability. Artificial intelligence (AI) and machine learning (ML) offer a computational framework to address these constraints through data-driven platform engineering. This review examines AI-assisted strategies in three interconnected EV platform pillars in cardiovascular medicine. These include: (i) isolation and processing platforms where ML algorithms optimize microfluidic separation and improve signal accuracy; (ii) analytical and diagnostic platforms where deep learning supports single vesicle phenotyping, multi-omics biomarker engineering, and biosensor interpretation; and (iii) therapeutic and manufacturing platforms where AI guides cargo loading, biodistribution estimation, and process control. We also assess key translational challenges, including MISEV2023 compliance, dataset bias, reproducibility, and regulatory alignment. This review positions artificial intelligence as the fundamental layer of the EV bioengineering process, providing a structured framework for advancing EV-based cardiovascular platforms from laboratory research to clinical application. Full article

Leishmaniasis caused by Leishmania infantum is a zoonotic disease endemic in many regions worldwide. The antigen-presenting dendritic cells (DCs) bridge the innate and adaptive immune response by activating T lymphocytes. Therefore, the present study examines whether T lymphocyte activation can be directed by autologous DCs primed by extracellular vesicles (EVs) derived from L. infantum. For this, lymphocytes were co-cultured with monocyte-derived DCs (moDCs) that were primed by EVs. moDC signaling and activation were examined by gene expression of toll-like receptors and cytokines. The antigen-presentation ability was analyzed through major histocompatibility complex molecules, and T cell subpopulations were explored by immunophenotyping. In co-cultures, EV-primed moDCs upregulated TLR2, TLR4, and TLR9, along with overexpression of MHC molecules. Co-cultures involving moDCs primed by EVs promoted the upregulation of both pro-inflammatory and regulatory cytokines associated with the expansion of non-conventional regulatory and central memory T cell subsets within the CD8⁺ T cell subpopulation. These findings suggest that activated moDCs can modulate cytotoxic lymphocytes, thereby promoting a balanced inflammatory microenvironment counterbalanced by a concurrent regulatory immune response. Thus, cell-based immune strategies using moDCs loaded with Leishmania-derived EVs represent a potential first step toward the development of innovative and personalized immune prophylactic and therapeutic approaches for leishmaniasis. Full article

Coronary artery bypass grafting (CABG) using the autologous saphenous vein (SV) remains widely performed for obstructive atherosclerosis; however, vein graft disease drives recurrent ischaemia through early thrombosis and progressive intimal hyperplasia, and accelerated atherosclerosis developing within the grafts. Extracellular vesicles (EVs) are membrane-bound particles that transfer proteins, lipids, and microRNAs between cells. They modulate endothelial dysfunction, vascular smooth muscle cell phenotypic switching, inflammation, and coagulation, which are core processes in vein graft remodelling. Arterialisation exposes the vein to abrupt rises in shear stress, cyclic stretch, and intraluminal pressure. These forces increase EV release and reshape EV cargo in experimental systems, suggesting a potential mechanism for amplifying early graft injury which warrants direct investigation in vein tissue. This review synthesises current evidence for cell-specific EV contributions from ECs, vascular smooth muscle cells, platelets, and macrophages, and appraises EV-associated microRNAs with biomarker potential relevant to graft failure pathways. We also review therapeutic strategies that may modulate EV signalling including antiplatelet therapy, statins, KCa3.1 inhibition, and pro-reparative mesenchymal stromal cell-derived EVs. No published clinical studies evaluate EV-based biomarkers specifically for saphenous vein graft patency, and none prospectively predict saphenous graft failure. CABG provides a well-defined time zero event that enables longitudinal sampling and risk stratification. Prospective studies linking EV phenotypes and miRNA signatures to imaging-defined graft outcomes are needed to support clinical translation. Full article

Safflower (Carthamus tinctorius L.) is an economically important crop, yet its production is severely threatened by fungal diseases including Botrytis cinerea. The molecular mechanism underlying disease resistance in safflower remains largely unclear. Extracellular vesicles (EVs), as vital carriers for cross-kingdom communication and transport, play crucial roles in plant antifungal defense. Lipid transfer proteins (LTPs), members of the pathogenesis-related protein 14 family, have also been shown to be key players in plant disease resistance. The promising resistance-related candidate gene CtLTP8 was previously identified via genome-wide association study (GWAS). In this study, a genome-wide analysis of the LTP gene family in safflower was performed. EVs were isolated from the apoplastic washing fluid of B. cinerea-infected safflower leaves, and proteomic analysis was performed. Numerous proteins associated with disease resistance, including CtLTP8, were detected by proteomic profiling. CtLTP8 was found to be present in EVs through molecular biological experiments. Moreover, stable overexpression of CtLTP8 in safflower significantly increased resistance to B. cinerea. In summary, this study characterized the disease resistance-related proteome of safflower EVs, and verified the presence of CtLTP8 in EVs and its antifungal function, providing valuable gene resources and theoretical support for safflower disease-resistance breeding and research on EV-mediated plant immune mechanisms. Full article