Journal of extracellular biology最新文献

Systemic inflammatory state found in obesity increases the risk of developing numerous diseases. While endurance training seems effective to reduce this inflammation, the underlying mechanisms are not fully understood. Among those, extracellular vesicles (EVs) have been proposed to be actors in the anti-inflammatory intercellular crosstalk induced by exercise training. This study aimed to investigate how endurance training modulates the EV proteome in the context of an inflammatory state in adults with obesity. Thirteen lean sedentary adults and 10 sedentary adults with obesity participated in a 12-week endurance training programme. Skeletal muscle, abdominal subcutaneous adipose tissue and venous blood samples were taken prior to and after the training period. The systemic and adipose tissue inflammatory states were assessed, and plasma EVs were isolated by size exclusion chromatography. EV content was analysed by mass spectrometry. EVs isolated from the medium of myotubes stimulated by electrical pulse stimulation in vitro were quantified, and their content was analysed by western blot. After the endurance training, C-reactive protein (CRP) levels decreased in participants with obesity. In abdominal subcutaneous adipose tissue, the phosphorylation state of nuclear factor-kappa B (NF-κB) was not affected by training, but interleukin (IL)-6 and IL-1β protein levels were reduced after the 12 weeks in both groups. Conversely, interferon gamma (IFNγ) level reduction was exclusively found in the obesity group. Despite no changes in EV abundance, EV proteome was modified by training. Among the modified proteins in participants with obesity, the antioxidant enzyme peroxiredoxin (PRDX) 1 abundance was increased after training. Additionally, the PRDX1 content of EVs isolated from stimulated myotubes was increased compared to control conditions. In conclusion, our results suggest that the anti-inflammatory effects of exercise training are not directly mediated by EV anti-inflammatory proteome changes. However, exercise training increases circulating EV antioxidant content, possibly through contractile activity of skeletal muscle during repeated exercise.

Extracellular vesicles (EVs) are lipid nano-to-micro-sized vesicles increasingly studied for their role in intercellular communication and their potential as minimally invasive molecular indicators in various diseases. However, challenges remain in characterizing specific surface molecules on EVs due to cargo heterogeneity and the lack of convenient quantification methods. In this study, we show the isolation, characterization, detection, and quantification of Trop2-carrying EVs (EV-Trop2) in serum of prostate cancer patients. This work combines the unique advantages of our EV isolation method with ELISA to enable surface-protein-specific EV analysis directly from serum. This is, to our knowledge, the first demonstration to isolate and quantify EV-Trop2 from prostate cancer patient serum to study its expression patterns in relation to prostate cancer status. Analysis of serum samples from three patient groups: high-risk prostate cancer (n = 22), low-risk prostate cancer (n = 23), and cancer-free groups (n = 21), revealed significantly different levels of EV-Trop2 expression between the high-risk and low-risk patient groups (p = 0.0015) and between high-risk patient and cancer-free groups (p < 0.0001). Multivariate modeling further showed that EV-Trop2 contributed to improved classifier metrics across the three sample groups. These findings highlight a strategy for probing EV-associated surface targets and suggest broader applicability of this approach across multiple cancers.

Cerebral oxygenation differences in the neonatal period of human preterm infants, along with sex-specific differences in combating oxidative stress, can lead to disruption of normal oligodendrocyte maturation and function, which in turn can differentially affect neuronal development and activity in the male and female brains. Secretory proteins and extracellular vesicles (EVs) are increasingly recognized as important mediators of intercellular communication and stress response in the brain. Our analysis of the secretome from cell culture supernatants obtained after treating male and female derived primary mouse OPCs with hyperoxia (80% O₂) for a 24 h period showed prominent sex-specific protein signatures with only 6% intersection between sexes upon hyperoxia. A higher proportion of mitochondrial proteins was observed to be secreted by male cells upon hyperoxic stress. Among specific factors that could be identified exclusively in the hyperoxia-treated groups, FGF-2 was present in significantly higher amounts in the female supernatant. Functional assays on neuronal cells (male) revealed that treatment with supernatant from female hyperoxic OPCs resulted in increased neuronal viability, potentially due to elevated levels of FGF-2. This suggests that female-specific extracellular proteins may play a key role in sex specific stress response and are potential candidates for further investigation.

The highly metastatic triple-negative breast cancer (TNBC) relies on the tumour microenvironment (TME) to maintain phenotypic heterogeneity and progression. Extracellular vesicles from hypoxic TNBC (EVh) have been previously shown to facilitate tumoural invasion; however, their function in the TME remains unclear. We used a novel method to investigate the TME in vitro called multicellular circulating co-culture, to characterise how EVh interferes with tumoural and endothelial cells, fibroblasts, monocytes and macrophages. EVh promoted monocyte differentiation to M2-like macrophages and inhibited macrophage-derived phagocytosis in endothelial and tumoural cells. The protection of endothelial, tumoural and stromal cellular integrity by EVh increased pro-tumoural and pro-angiogenic signalling, collagen matrix synthesis and showed a potential differentiation to cancer-associated fibroblasts. Our findings highlight the critical role of EVh in protecting tumour cells, indicating its cooperation towards a protective TME, which was demonstrated by the multicellular circulating co-culture and conventional co-culture protocols. These findings lead to an adequate system with potential for investigating other tumour-related processes, including circulating tumour cells and metastasis.

Pregnancy and live birth rates are commonly used metrics to define fertility in humans and animals. The impact of aberrant microRNA (miRNA) expression on fertility-related genes represents a significant knowledge gap in understanding post-transcriptional regulatory mechanisms associated with reproductive dysfunction. Identifying subfertility markers is therefore critical to the success of fertility intervention strategies, particularly in agriculture, where sustainable farming practices are linked to overall economic performance. Here, we explore the expression patterns and association of blood plasma small extracellular vesicles (sEV)-derived miRNA with Holstein-Friesian dairy cow (Bos taurus) subfertility. Small RNA-seq identified 14 differentially expressed plasma sEV-derived miRNAs (FDR < 0.05 and −logFC > 2) between divergent low fertile (LF) and high fertile (HF) primiparous dairy cows (n = 10/group) with known reproductive outcomes. qRT-PCR miRNA assay validation of these plasma sEV miRNA candidates isolated from a different sample population of young heifers (10 month old) (LF and HF; n = 8/group) confirmed that the abundance of miR-181b-2-3p was significantly higher in LF sEVs compared to HF sEVs [p value = 0.0093, relative expression ratio = 2.665 (2^−ΔΔCT; LF = 19.6095, HF = 7.35636)]. Our results suggest that circulating sEV miRNA may contribute, in part, to fertility traits in dairy cows. The association of miR-181b-2-3p with the subfertility phenotype suggests that this miRNA may serve as a putative early indicator of LF status.

Fungal infections cause approximately 1.6 million deaths annually. Diagnosing and treating fungal infections is difficult due to limited access to diagnostic tests and rising antifungal resistance. Extracellular vesicles (EVs) facilitate interactions between fungal cells and hosts, significantly influencing the pathogen-host relationship. Owing to the complexity of fungal EVs and the lack of clinical studies on their roles in human infections, we analysed EVs from serum and urine samples of patients with infections caused by Candida albicans, Cryptococcus neoformans, and Paracoccidioides brasiliensis to determine their roles. Using mass spectrometry, we identified sterols, sphingolipids, and fatty acids as key metabolites in the EVs. We quantified cholesterol and ergosterol, confirming the presence of both host and fungal EVs in clinical samples. Our research investigated whether these EVs could modulate the host immune response. We observed a proinflammatory response in murine and human macrophages, characterized by increased cytokines, such as tumour necrosis factor-α, interferon-γ, and interleukin-6, and elevated expression of the inducible nitric oxide synthase gene, a marker of M1 macrophage response. Thus, circulating EVs in patients with fungal infections likely play a role in disease pathophysiology. These findings enhance our understanding of EVs in fungal infections, suggesting potential therapeutic targets for systemic mycoses.

The development of novel treatments that restore brain function and improve patient outcomes for Alzheimer's disease (AD) is necessary, given the complications and lack of improvement in recently approved amyloid beta (Aβ)-targeting drugs. Cell-derived extracellular vesicles (EVs) have been found to improve cognitive function through reduced inflammation, oxidative stress, and apoptosis, restoring neuronal and blood-brain barrier function, and inhibiting Aβ and phosphorylated tau build-up in the brain. Given the recent emergence of EVs into clinical trials, it is essential to provide the field with an update on proposed mechanisms of action, gaps in knowledge for further study, and recommendations for producing EVs with high therapeutic efficacy to ensure success in subsequent clinical trials. This systematic review summarizes original research to date that reports effects of mammalian cell-derived EVs for the treatment of AD. Evidence of therapeutic benefits and reported mechanisms of action are discussed. Further, methods for engineering EVs to increase their therapeutic efficacy and produce high-quality EVs relevant to the AD field are outlined. The quality of evidence is discussed in terms of reporting guidelines from the Minimal Information for Studies of Extracellular Vesicles (MISEV). The review further discusses current preclinical AD models and provides direction to improve the quality of AD models for testing novel therapeutics.

Small extracellular vesicles (sEVs) derived from natural killer (NK) cells possess inherent anti-tumour activity and offer the advantages of cell-free therapy. In this study, we genetically engineered NK-sEVs to express interleukin 15 (IL15), an anti-tumour cytokine, and the monoclonal antibody cetuximab on their surface, creating a potent anti-tumour immunotherapy with enhanced tumour-targeting capabilities. These IL15- and cetuximab-tethered NK-sEVs (eEVs) were generated using lentivirus-based modification. eEVs selectively bound to EGFR⁺ cancer cells in vitro, confirming cetuximab-mediated targeting. Compared to control NK-sEVs, eEVs exhibited significantly enhanced cytotoxicity by directly inducing cancer cell death and promoting NK cell-mediated killing. In a lung cancer mouse model, eEVs selectively accumulated in tumours and exhibited significant anti-tumour efficacy. Notably, their administration, alone or in combination with anti-PD-1 antibody therapy, effectively suppressed tumour growth. Overall, our results indicate that genetically engineered NK-sEVs, equipped with IL15 and cetuximab, exhibit potent anti-tumour activity and tumour-targeting capabilities. These findings suggest that eEVs hold significant potential as a novel immunotherapeutic strategy for cancer treatment.

All approved RNA therapeutics require parenteral delivery. Here, we demonstrate an orally bioavailable formulation wherein synthetic noncoding (nc) RNA, packaged into lipid nanoparticles, is loaded into casein-chitosan (C2) micelles. We used the C2 formulation to deliver TY1, a 24-nucleotide synthetic ncRNA, which targets DNA damage and attenuates inflammation in macrophages. C2-formulated TY1 (TY1^C2) efficiently packages and protects TY1 against degradative enzymes. In healthy mice, oral TY1^C2 was well-tolerated and nontoxic. Oral TY1^C2 exhibited disease-modifying bioactivity in two models of tissue injury: (1) rat myocardial infarction, where a single oral dose of TY1^C2 was cardioprotective, on par with intravenously-delivered TY1; and (2) mouse acute lung injury, where a single dose of TY1^C2 attenuated pulmonary inflammation. Mechanistic dissection revealed that TY1^C2 is taken up by intestinal macrophages, namely those of the lamina propria and Peyer's patches. Afterwards, TY1 could be detected in circulating monocytes for up to 72 h post-ingestion. Unlike TY1, which acts on macrophages, an antisense oligonucleotide against Factor VII, which acts on hepatocytes, is not effective when administered in the C2 formulation. Thus, not all ncRNA drugs are bioactive when delivered by mouth. Oral delivery of macrophage-active RNA opens up a wide range of potential new therapeutic opportunities.

The gut microbiota, a collection of microorganisms residing within the human gastrointestinal tract, exerts profound effects on the health of the host. In recent years, studies have revealed that the gut microbiota influences not only the function of the digestive system but also has close associations with various systemic diseases, including cardiovascular diseases. Myocardial remodelling refers to the structural, functional and molecular changes in the myocardium that occur in response to alterations in load. This process encompasses changes such as myocardial hypertrophy, apoptosis, necrosis and myocardial fibrosis. Bacterial extracellular vesicles (BEVs) are small vesicles secreted by the gut microbiota that can carry bioactive substances such as proteins, lipids and nucleic acids, participating in intercellular communication. BEVs are capable of traversing the gut barrier and entering the bloodstream, thereby influencing the functional status of distant organs, including the heart. Under the condition of Myocardial remodelling, these BEVs may exert protective or detrimental effects on cardiomyocytes by modulating pathways such as inflammation, oxidative stress and apoptosis.