Extracellular vesicles and circulating nucleic acids最新文献

The crosstalk between the skeletal muscles and the liver is receiving growing attention, as patients with chronic liver disease often develop a loss of skeletal muscle mass. In these patients, particularly those with metabolic dysfunction-associated steatotic liver disease, physical exercise improves insulin sensitivity and hepatic steatosis. However, excessive exercise may impair mitochondrial function, inflammation, and liver health. The study by Liu et al. demonstrates that overtraining promotes liver fibrosis through myocyte-derived small extracellular vesicles. Here, we comment on the novelty of these findings and areas to be developed in the future.

Aim: Extracellular vesicles (EVs) play a pivotal role in tumor progression, influencing the tumor microenvironment. Despite significant research, the targeted analysis of EVs directly derived from primary tumors remains limited, particularly in ovarian cancer. The majority of existing studies have focused on EVs derived from peritoneal fluid (ascites), which encompasses contributions from different cell types. This study aims to isolate and characterize EVs secreted specifically by ovarian cancer spheroids derived from primary patient ascites. Methods: A three-dimensional cell culture model was employed to cultivate tumor spheroids in a defined medium, with EVs purified via differential ultracentrifugation and size-exclusion chromatography. Purified EVs were characterized by nanoparticle tracking analysis, nanoflow cytometry, and electron microscopy prior to performing high-resolution mass spectrometry. Results: This approach allowed the identification of known cancer-associated proteins, including danger molecules, which are linked to poor prognosis. Moreover, enzyme-linked immunosorbent assay (ELISA) analysis demonstrated that the ascites abundance levels of novel candidates [RAB14 (Ras-related protein Rab-14), SCAMP3 (secretory carrier membrane protein 3), and FAM3C (FAM3 metabolism regulating signaling molecule C)] correlated with patients' progression-free survival, further validating their clinical relevance. Finally, we used the Gene Expression Profiling Interactive Analysis 2 (GEPIA2) database to compare our dataset with The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) data. Thereby, we revealed a signature of three TOP genes encoding proteins within our dataset (CORO1B, LAMP2, MSLN), which were differentially expressed in ovarian cancer patients compared to healthy individuals. Conclusion: This study provides the first proteomic profile of EVs derived directly from primary tumor spheroids, and paves the way for a better mechanistic understanding of EV-associated proteins and for the development of biomarkers or therapeutic strategies.

Extracellular vesicles (EVs) have significant potential as therapeutic agents and as sources of diagnostic, predictive, and prognostic nucleic acid biomarkers. However, variability in EV workflows and inadequate standardization of downstream analysis pose major obstacles to reproducibility. The MISEV (Minimal Information for Studies of Extracellular Vesicles) guidelines provide essential domain-specific recommendations for EV isolation, characterization, analysis, nomenclature, and reporting, but deliberately refrain from prescribing methods for the molecular quantification of EV cargo. Among the analytical platforms used in EV studies, quantitative reverse transcription PCR (RT-qPCR) is the most critical method for validating and quantifying EV-associated RNA. The recently revised MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) 2.0 guidelines offer a detailed foundation for ensuring analytical validity in RT-qPCR-based quantitative applications. The proposed model of harmonizing general and domain-specific guidelines provides a scalable blueprint for improving reproducibility across complex biomarker development workflows in molecular diagnostics.

In the rapidly developing field of skin care, non-surgical facial aesthetics are becoming increasingly favored by consumers. Plant-derived extracellular vesicles (PDEVs) have attracted much attention due to their low toxicity, cellular communication function, and ability to carry bioactive molecules, including proteins, lipids, nucleic acids, and small molecules with pharmacological activities. Recent in vitro studies have shown that PDEVs enhance the transdermal delivery of drugs and improve skin condition, suggesting promising applications in facial aesthetics. In this review, we provide a comprehensive overview of the application of PDEVs in anti-scarring, anti-aging, and anti-pigmentation therapies. We also discuss current limitations in their application and potential solutions to address these challenges. In conclusion, this review analyzes the roles and mechanisms of PDEVs in facial aesthetics and aims to support their future clinical application.

Aim: Liquid biopsies hold significant potential for the minimally invasive diagnosis of tumors and other diseases. While the clinical application of cell-free DNA (cfDNA) methodologies is emerging, the implementation of tumor-derived extracellular vesicles (EVs) as validated biomarkers is hindered by substantial preanalytical variations. In this work, we standardized the preanalytical procedures of blood collection for subsequent serial isolation of plasma cfDNA and EVs from a single blood collection tube. Methods: We compared the impact of blood preservation tubes and storage to enable proteomic profiling of resulting EVs in addition to cfDNA extraction and sequencing. Following a stringent method of large EV (lEV) and small EV (sEV) isolation, consisting of differential ultracentrifugation and size exclusion chromatography, we evaluated the protein concentration, particle number, quality and integrity of the isolated EVs. Subsequent proteomic analyses of EV isolates revealed the complexity of the respective tube-biased proteomes, allowing the interpretation of EV origins as well as contamination sources. Results: While ACD-A and Citrate tubes yield satisfactory results in the preservation of EV proteomes, only Streck RNA, Norgen, and PAX tubes can maintain high cfDNA purity for up to 7 days. When aiming for multiomics analyses, Streck RNA tubes showed the most stable performance across the tested parameters for both bioanalytes. Furthermore, we detected greater variability in protein composition in sEVs than in lEVs after 7 days of storage; thus, sEVs might be more susceptible to storage effects. Conclusion: Our clinically applicable workflow provides the basis for informed choice of liquid biopsy tubes along with a ready-to-use protocol to retrieve both genomic and EV proteomic biomarker information for multiomics biomarker-based liquid biopsy studies.

In recent years, the prevalence of ocular diseases has increased considerably. However, timely diagnosis and treatment are hampered by the challenge of early detection since symptoms often appear in advanced stages. Emerging research highlights extracellular vesicles (EVs) as potential biomarkers for ocular diseases, with tear-derived EVs offering a minimally invasive source for early diagnosis. Tears play a crucial role in maintaining eye health and reflect the physiological state of the eye; thus, abnormalities in tear composition can provide valuable insight into inflammatory eye diseases. Studies have demonstrated the utility of tear-derived EVs in identifying biomarkers not only for inflammatory eye diseases but also for neurodegenerative disorders, as they carry molecular signatures (including proteins and various RNA species) reflective of their cells of origin. In this review, we discuss the potential of tear-derived EVs as biomarkers for early detection and monitoring of ocular and neurodegenerative diseases and highlight the importance of standardizing tear collection and EV isolation protocols to ensure reproducibility.

Neuroaging is a complex biological process in which the brain undergoes progressive functional decline marked by synaptic loss, neuroinflammation, and cognitive decline. At the molecular and cellular level, aging is driven by multiple interconnected hallmarks, including genomic instability, telomere attrition, epigenetic alterations, mitochondrial dysfunction, cellular senescence, stem cell exhaustion, and altered intercellular communication. Among these, cellular senescence, a state of irreversible cell cycle arrest, has emerged as a critical contributor to brain aging. Senescent cells accumulate with age, driven by the p53-p21 and p16-pRb pathways, and secrete pro-inflammatory factors via senescence-associated secretory phenotype (SASP), thereby exacerbating neurodegeneration, vascular dysfunction, and cognitive decline. Extracellular vesicles (EVs) are natural nanocarriers of proteins, lipids, and nucleic acids, and have emerged as key mediators of intercellular communication and therapeutics for aging and age-related conditions. EVs derived from various cell types, such as mesenchymal stem cells (MSCs), neural stem cells (NSCs), and induced pluripotent stem cells (iPSCs), can modulate senescence-related pathways, reduce inflammation, and promote tissue repair. Preclinical studies demonstrate that stem-cell-derived EVs can improve cognitive performance, enhance neurogenesis, reduce senescence phenotype, improve neuronal survival through neuroprotective miRNAs (miR-181a-2-3p), suppress neuroinflammation via inhibition of NLRP3 inflammasome, and support synaptic plasticity. Stem cell EVs possess natural biocompatibility, the ability to cross the blood-brain barrier (BBB), and targeted delivery mechanisms, making them promising candidates for anti-aging interventions. This review elaborates on the multifaceted role of stem cell EVs in mitigating brain aging, senescence, and age-associated chronic disease phenotype.

Kidney disease, encompassing both acute kidney injury (AKI) and chronic kidney disease (CKD), represents a major global health challenge. A pivotal aspect of the pathogenesis of these conditions is damage to renal tubular epithelial cells (TECs), which contributes to maladaptive repair mechanisms and fibrosis. Due to their essential role, TECs are regarded as a promising target for innovative therapeutic strategies. Mesenchymal stromal cell-derived extracellular vesicles (MSC-EVs) have attracted increasing attention for their therapeutic potential in kidney disease, with extensive literature documenting their beneficial effects on TEC damage through targeted mechanisms. In this review, we critically examine the existing literature on the targeting of TECs by MSC-EVs in both in vitro and in vivo settings. Furthermore, we highlight the limitations and potential of MSC-EV-based strategies for TEC targeting, aiming to provide insights for future clinical trials and therapeutic applications.

In the past decade, extracellular vesicles (EVs) have gained increasing attention in biomedical research. These membrane-bound particles are naturally secreted by cells under both physiological and pathological conditions, and they exhibit a wide range of sizes and molecular compositions. EVs transport bioactive molecules - such as proteins, nucleic acids, and lipids - making them ideal candidates for biomarker discovery. Consequently, their accurate characterization and quantification are critical for understanding their roles in intercellular communication and evaluating their potential in diagnostics, prognostics, disease monitoring, and therapeutic applications. Microfluidic technologies offer promising solutions for EV analysis, addressing key limitations of conventional methods by enabling precise and sensitive measurements with small sample volumes. While microfluidic devices have been predominantly used for EV separation and isolation, their application in EV quantification remains underexplored. Compared to traditional techniques like nanoparticle tracking analysis or flow cytometry, microfluidic systems can provide faster, more accessible alternatives for EV quantification. This review summarizes recent advances in microfluidic technologies for EV quantification, discussing their advantages, current limitations, and future prospects.