Volume 7, Issue 2 (2026) – 26 articles
Cover Picture: The graphic on the cover summarizes the currently published review article "Quantification of EV-associated miRNA in liquid biopsies for biomarker signature development." The review describes the state of knowledge regarding miRNA, starting at the biogenesis of extracellular vesicles (EVs) to the packaging of their cargo, and illustrates the integrated workflow, from liquid biopsy collection and miRNA quantification to bioinformatic data analysis, culminating in clinically actionable EV-miRNA signatures. Over the last decade, EV-associated miRNAs have emerged as promising, minimally invasive biomarkers, as encapsulation within EVs protects miRNAs from degradation while preserving disease- and cell-type-specific expression patterns in various human body fluids. The review summarizes the wide range of EV sources used in liquid biopsy and molecular diagnostics, including blood, urine, saliva, milk, airway secretions, and cerebrospinal fluid, and highlights both the diagnostic utility and the key pre-analytical challenges associated with EV-miRNA analysis. Furthermore, current EV isolation techniques, such as differential ultracentrifugation, size-exclusion chromatography, affinity-based methods, microfluidic platforms, flow cytometry, and nanoparticle-based analyses, are compared in terms of yield, purity, and impact on downstream RNA profiling. In addition, various extraction and quantification methods for EV-associated RNA, including RNA sequencing and RT-qPCR or RT-dPCR, are critically evaluated. A key focus of the entire quantification process is on standardization approaches based on international guidelines, as mentioned MISEV, MIQE, EV-TRACK, and the EV Task Force guidelines for body fluids, all aimed at minimizing technical and biological variability. This underscores the importance of rigorous reporting standards, harmonized pre-analytical workflows, and multiparametric normalization strategies for ensuring reproducible detection and precise quantification. Finally, the discussion addresses emerging best practices and unresolved challenges regarding multivariate data modeling, analyses based on miRNA isoforms (isomiRs), and the in silico validation of target structures and regulatory signaling pathways. By combining technological advances with harmonized methodological frameworks, molecular diagnostics is well positioned to transition from exploratory research to clinically actionable, EV-miRNA-based diagnostic methods capable of significantly improving patient outcomes.
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Back Cover Picture: This cover artwork presents engineered bacterial extracellular vesicles (eBEVs) as next-generation precision postbiotics. The central luminous vesicle represents an engineered BEV with a semi-transparent lipid bilayer, golden targeting ligands, detoxified membrane regions, surface-displayed proteins, and glowing internal cargos, symbolizing targeted delivery, controlled release, programmable payload loading, and improved biosafety. The left-side progression illustrates the technological transition from traditional live probiotics to natural BEVs and finally to engineered BEVs, highlighting the shift from broad, heterogeneous microbial effects toward precise and controllable postbiotic nanomedicine. Surrounding the central vesicle, five application scenes - intestinal diseases, metabolic disorders, tumor therapy, central nervous system disorders, and skin regeneration - are linked by light trajectories, suggesting disease-oriented delivery and multi-system therapeutic potential. The bottom platforms represent genetic engineering, chemical modification, and membrane hybridization, the three key strategies enabling functional optimization of BEVs. Together, the deep-blue microenvironment, golden highlights, and radial delivery network depict eBEVs as programmable microbial messengers that integrate natural vesicle communication with the precision of modern nanomedicine.
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