Extracellular vesicles (EVs) facilitate intercellular communication in glioblastoma (GBM) by transferring microRNAs (miRNAs). GBM is the most aggressive primary brain tumor in adults, and despite multimodal therapy, the median survival remains approximately 15 months. Current diagnostic approaches, including contrast-enhanced MRI, are insufficient to reliably distinguish true tumor progression from pseudoprogression. Moreover, therapeutic efficacy is limited by intratumoral heterogeneity, acquired resistance, and the restrictive nature of the blood-brain barrier (BBB). In this context, EV-associated miRNAs (EV-miRNAs) contribute to GBM progression by regulating proliferation, angiogenesis, invasion, therapeutic resistance, and immune evasion. Notably, several EV-miRNAs are dysregulated in both GBM and neurodegenerative diseases (NDDs), suggesting shared molecular pathways across central nervous system (CNS) disorders. Circulating tumor-derived EV-miRNAs represent promising liquid biopsy biomarkers for diagnosis, prognosis, and longitudinal treatment monitoring. Beyond their biomarker potential, EVs can be engineered as nanocarriers capable of crossing the BBB to deliver therapeutic cargo, including inhibitors of oncogenic miRNAs (e.g., miR-21) or tumor-suppressive miRNAs (e.g., miR-124). This review summarizes the molecular functions, biomarker applications, and therapeutic strategies of EV-miRNAs in GBM. We further discuss current challenges related to methodological standardization, scalable production, and clinical translation. Collectively, advancing the understanding and clinical implementation of EV-miRNAs may provide new opportunities for precision diagnostics and therapeutic innovation in GBM.

Metabolic dysfunction–associated steatotic liver disease (MASLD) is the most common chronic liver disorder and can progress to steatohepatitis and fibrosis; although approved pharmacotherapies for metabolic dysfunction–associated steatohepatitis (MASH) with fibrosis remain limited. Autologous chemically induced liver progenitor (CLiP) cells, generated from mature hepatocytes without genetic modification, have shown therapeutic promise in rodents, but their efficacy has not been tested in large animals. Six female Clawn miniature pigs (15–42 kg) were fed a high-fat, high-cholesterol diet to induce MASLD with biopsy-proven fibrosis (Brunt stage ≥1). Animals were assigned to CLiP transplantation ( n = 3) or saline control ( n = 3). Autologous CLiPs (5 × 10 7 ) were generated from laparoscopically resected liver wedges, expanded ex vivo , and infused intraportally. Safety was assessed by monitoring, liver function tests, and lipid profiles. Efficacy was evaluated 1 month later by blinded histology and immunohistochemistry. CLiP transplantation was feasible and well tolerated. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) showed minimal changes in both groups, but total cholesterol and triglycerides decreased in treated pigs and increased in controls. Histologically, two of three CLiP-treated livers regressed from Brunt stage 1 to 0, with resolution of steatosis and reduced stellate cell activation, whereas controls showed no regression. These findings support CLiP therapy as a regenerative option for MASLD.

Abdominal aortic aneurysm (AAA) is a life-threatening disease. Although AAA is generally asymptomatic, the mortality rate remains very high once rupture occurs, even with successful treatment. The pathophysiology of AAA involves inflammatory cell infiltration, smooth muscle cell apoptosis, and extracellular matrix degradation. However, there are various unclear aspects of pathophysiology due to cellular heterogeneity and multifactorial disease. Moreover, there are no blood biomarkers or available pharmacological drugs for AAA. Extracellular vesicles (EVs) are lipid bilayer particles released from every type of cell for intercellular communication. EVs include proteins, DNA, RNA (mRNA, microRNA), and lipids. EV cargos are delivered to recipient cells and modulate their biological effects. Although fewer studies have investigated EVs in AAA than in other cardiovascular diseases with similar molecular mechanisms, recent research indicates that EVs play a significant role in AAA development. Further research on EVs and AAA will contribute to the elucidation of AAA pathophysiology and the development of novel pharmacological drugs. In this review, we summarize the EV-associated pathophysiology, EV-based biomarkers, and EV-based treatment strategies in AAA. We also discuss the prospects for EVs research in AAA.

Early cancer detection using minimally invasive biomarkers remains a significant challenge, particularly in early-stage disease, where circulating tumor DNA is often below the limit of detection. Extracellular vesicles (EVs), which are actively secreted by viable cancer cells and carry tumor-associated proteins, represent a promising alternative target for liquid biopsy. In this study, we developed EV-finder®, a conceptual framework for the direct detection of EV-associated proteins in serum using proximity extension assay (PEA) technology. Unlike conventional EV-based analytical methods that require prior EV isolation or enrichment, the EV-finder approach enables direct profiling of EV-associated proteins from small serum volumes without an EV isolation step, thereby simplifying the analytical workflow while preserving EV-derived molecular information. Using serum samples from patients with five cancer types (n = 193) and independent healthy controls (n = 138), we established a two-step supervised machine learning framework for cancer detection and tissue-of-origin prediction. The screening model demonstrated promising discriminative performance, with an AUC of 0.985, sensitivity of 0.929, and specificity of 0.957. Notably, no false positives were observed in an external Japanese control cohort, whereas 4 of 29 Korean control samples were classified as cancer-positive. Analysis of EV-associated protein profiles identified both pan-cancer and cancer-type-specific signatures, supporting their value for multi-cancer detection. Collectively, these findings demonstrate the potential feasibility of direct detection of EV-associated proteins from serum using PEA technology and highlight its potential as a scalable and minimally invasive strategy for multi-cancer screening.

Metastatic colonization is governed by the earliest interactions between disseminated tumor cells (DTCs) and the tissue microenvironment. Extracellular vesicles (EVs) have emerged as critical mediators of this early communication, conditioning distant organs before and after DTC arrival. Despite extensive characterization of cancer EV cargoes and functions, a fundamental question remains unresolved: when, where, and to which cell types cancer EVs are delivered during the initial stages of metastatic colonization. This knowledge gap stems from limitations of conventional in vitro and in vivo assays, which lack physiological architecture or sufficient spatiotemporal resolution. Recent advances in physiological metastatic models, including organotypic ex vivo models, together with innovative EV labeling and tracking technologies, now enable direct visualization of EV transfer within native-like tissue contexts. Here, we propose a conceptual framework in which metastatic colonization is “decoded” as a spatiotemporally orchestrated EV-mediated intercellular signaling, where EV distribution, tropism, and recipient-cell responses collectively define DTC fate. We discuss how integrating these emerging platforms can decode EV-mediated communication during metastatic colonization that leads to therapeutic development.

ABSTRACT Extracellular vesicles (EVs) offer compelling opportunities for diagnostics and therapeutics, yet translation toward routine clinical use and commercial readiness remains limited. In diagnostics, EV‐based assays may reach practice as laboratory‐developed tests, while in therapeutics unproven “exosome” offerings have prompted safety communications, together underscoring the need for fit‐for‐purpose regulatory science. Here, we organize implementation challenges into six recurring bottlenecks: Identity; Purity; Potency; Measurement comparability; Manufacturing control; and Safety. We summarize how international standardization efforts led by the International Society for Extracellular Vesicles (ISEV), including MISEV2023 and task‐force outputs, strengthen reproducibility and comparability, while highlighting remaining gaps that arise when descriptive guidance must be converted into operational specifications and submission‐ready evidence packages. Using Japan as a case example, we illustrate how regulatory‐authority discussions, professional‐society statements, and emerging implementation‐oriented guidance can clarify review questions, distinguish therapeutic EV products from conditioned medium, and support trust‐preserving translation. Finally, we propose a non‐binding, deliverables‐based roadmap across near‐, mid‐, and long‐term horizons to enable clinical and commercial readiness. The roadmap should be adapted to product context, risk profile, and jurisdiction, and it emphasizes precompetitive collaboration across academia, industry, and government to build shared infrastructure for measurement, standardization, reproducibility, and quality evaluation, thereby translating scientific consensus into operational development and review frameworks.

Gastrointestinal (GI) cancers represent a significant global health burden, being among the leading causes of cancer-related deaths. The prognosis for patients remains unsatisfactory, largely because most cancers are detected at advanced stages. Traditional diagnostic methods, such as radiological and histopathological examinations and serum tumor markers like AFP, CEA, CA-125, and CA-199, possess limitations in sensitivity and specificity, particularly for early screening. A major drawback of tissue biopsy is its inability to fully capture the inherent heterogeneity within tumors, as mutations can differ between primary and metastatic sites. In this context, liquid biopsy has emerged as a promising, minimally invasive alternative for detecting cancer-associated materials present in various body fluids. The concept of liquid biopsy, initially centered on circulating tumor cells, has expanded to encompass other critical biomarkers such as circulating tumor DNA, extracellular vesicles, and circulating tumor RNA. Analyzing these biomarkers using advanced techniques like next-generation sequencing or proteomics can unveil a wealth of potential information. Liquid biopsy offers numerous advantages, being less invasive, more convenient, potentially more cost-effective, and providing a dynamic, real-time snapshot of the entire tumor burden that reflects both intertumoral and intratumoral heterogeneity. This review provides an overview of key liquid biopsy biomarkers and their associated detection technologies, discusses their burgeoning clinical applications across various GI cancer types, and highlights the current challenges and future directions in this rapidly evolving field.

Acid suppression with proton pump inhibitors (PPIs) or a potassium-competitive acid blocker (P-CAB) is the first-line treatment for gastroesophageal reflux disease (GERD); however, a significant proportion of patients exhibit symptomatically refractory GERD. Here, we investigated whether distinct serum miRNA profiles exist in individuals with persistent symptoms despite acid suppression, given that serum miRNAs are promising biomarkers for cancer detection and various pathophysiological conditions. We also examined the relationship between anxiety/depression and serum miRNAs associated with symptomatic refractoriness. GERD patients taking PPIs/P-CAB for at least 8 weeks were enrolled and were classified into symptomatically refractory and symptomatically responsive groups using a GerdQ questionnaire. Comprehensive serum miRNA profiling was performed for all participants. Logistic LASSO/Elastic Net regression analysis was used to identify miRNAs associated with symptomatic refractoriness to PPIs/P-CAB, and a discriminant model was developed. Associations of the identified miRNAs with the presence of reflux esophagitis (RE) and items on the Hospital Anxiety and Depression Scale (HADS) were assessed. A combination of four serum miRNAs (miR-4294, miR-4725-3p, miR-3679-5p, and miR-6893-5p) was able to distinguish the symptomatically refractory group from the responsive group (sensitivity, 0.83; specificity, 0.66; AUC, 0.79). This AUC was higher than that for the presence of RE. The serum levels of three of the four miRNAs were associated with items on the HADS-A subscales. Distinct serum miRNA profiles are associated with symptom persistence despite acid suppression, potentially reflecting neuropsychological factors. These findings may provide new insights into the complex pathophysiology of symptomatically refractory GERD.

Mice exhibit complete scarless wound healing, including restoration of skin texture, until embryonic day 13 (E13), whereas wounds created on or after embryonic day 17 (E17) result in scar formation. Regeneration is thought to depend on dermal fibroblasts at E13, whereas fascia fibroblasts at E17 contribute to scar formation. Therefore, we examined whether extracellular vesicles (EVs) could be isolated from E13 dermis and whether these EVs could be utilized as an anti-scarring therapy. The addition of EVs derived from E13 dermal fibroblasts (E13D EVs) exerted antifibrotic effects on transforming growth factor-β1 (TGF-β1)-stimulated E17 fascia fibroblasts. E13D EVs also suppressed the fibrotic phenotype of human keloid-derived fibroblasts. In contrast, EVs derived from E17 fascia fibroblasts (E17F EVs), adult mouse fascia fibroblasts (AdF EVs), and human keloid fibroblasts (keloid EVs) enhanced fibrotic responses. These findings suggest that E13 dermal fibroblasts possess intrinsic antifibrotic properties and that these factors may be functionally transferred via EVs. Further characterization of the microRNAs (miRNAs) and proteins contained in E13D EVs may lead to the development of new therapeutic strategies for human scar management and fibrotic skin diseases.

An aberrant six-base repeat in intron 1 of C9orf72 is the most frequent cause of solitary and familial amyotrophic lateral sclerosis and frontotemporal dementia. This mutation is a potential target for CRISPR/Cas9-based genome editing. However, the blood-brain barrier and limitations of current viral or nanoparticle-based delivery systems to neurons significantly restrict the clinical application of CRISPR-Cas9 in the brain. To address these challenges, we developed a drug delivery system using acerola-derived exosome-like nanoparticles (AELNs), which may overcome several limitations associated with human exosomes. AELNs stably form complexes with ribonucleoproteins (RNPs) comprised of Cas9 proteins and guide RNAs (gRNAs). We improved the delivery efficiency and selectivity of AELN/RNP complexes in GLP2-receptor-ex-pressing neurons by incorporating GLP2 peptides into the AELN/RNP complexes. Intranasal administration of peptide-tagged AELN/RNP complexes in vivo confirmed the successful genome editing of C9orf72, demonstrating the potential of this system for treating neurodegenerative diseases. This study presents a potentially innovative approach for in vivo genome editing using a noninvasive delivery system.

Metastasis is the main cause of cancer mortality for many types of cancer; however, difficulties remain in effectively preventing metastasis. It has been recently and widely reported that cancer-derived extracellular vesicles (EVs) contribute to cancer metastasis. Thus, therapeutic strategies targeting cancer-derived EVs hold great promise because of the possibility of EVs driving the cancer microenvironment toward metastasis. Here, we provide a novel strategy for therapeutic antibody treatment to target cancer-derived EVs and inhibit the metastasis of breast cancer in a mouse model, establishing a rationale for further clinical investigation. Treatment with human-specific anti-CD9 or anti-CD63 antibodies significantly decreased metastasis to the lungs, lymph nodes, and thoracic cavity, although no obvious effects on primary xenograft tumor growths were observed. In in vitro and in vivo experiments, the EVs incubated with the targeted antibodies were preferentially internalized by macrophages, suggesting that antibody-tagged cancer-derived EVs would be eliminated by macrophages. Our results suggested that therapeutic antibody administration effectively suppresses EV-triggered metastasis in cancer and that the removal of EVs could be a novel strategy for cancer therapy.

Bacterial extracellular vesicles (EVs) are emerging mediators of host-microbe communication; however, their role in human disease remains unclear. Here, we identify two bacterial small tRNAs (tsRNAs) enriched in gut commensal Klebsiella pneumoniae-derived EVs (KpEVs) that are markedly elevated in the serum of patients with hepatocellular carcinoma (HCC). These tsRNAs suppress the production of nitric oxide (NO) by macrophages, a critical antitumor molecule. Notably, KpEVs reach the liver more efficiently than bacterial cells, thus facilitating bacterial translocation from the gut by inducing distinctive immunosuppressive macrophages. Mechanistically, KpEVs drive an M2-like macrophage phenotype, enhance phagocytosis, and inhibit both NO production and caspase-1-dependent pyroptosis during infection. The results show that KpEVs shape a liver microenvironment promoting gut-liver bacterial translocation, which may also influence HCC progression. Our study uncovers a previously unrecognized strategy by which K. pneumoniae exploits EVs to modulate host immunity of distant organs, highlighting tsRNAs as potential biomarkers and therapeutic targets.

Compared with transplanted tumors, autochthonous tumors are difficult to cure using experimental radiation therapy in mice. Here we analyzed differences in immune-related gene expression profiles between mouse fibrosarcomas subcutaneously induced by 3-methylcholanthrene (3MC) and their corresponding transplanted tumors. The immune genes examined were Pd1, Pdl1, Pdl2, Cd3d, Cd8a, Cd8b, Ifnγ, Itga2, Gzmb, and Foxp3. Among 12 tumors, one was non-transplantable and showed a benign phenotype with an abundance of DX5+ natural killer cells and CD8+ T cells together with increased IFNγ expression and mRNA levels of all immune genes except for Itga2. The other 11 transplantable tumors showed increased expression of Pd1, Pdl1, Pdl2, Cd3d, Cd8b, and Ifnγ following transplantation into syngeneic mice. These effects of transplantation highlight the relevance of immune gene expression status to the curability of tumors.

Multiple system atrophy (MSA) is a fatal, rapidly progressive atypical parkinsonism that responds poorly to Parkinson’s disease (PD) medications, and diagnostic precision remains a critical unmet need. Earlier, accurate diagnosis would give patients realistic prognostic expectations and enable timely clinical-trial enrollment. We sought plasma biomarkers discriminating MSA from PD. Plasma small RNA sequencing and extracellular vesicle proteomics in discovery cohorts, filtered by the Biomedical Oriented Logistic Dantzig (BOLD) selector, nominated two microRNAs (hsa-miR-520a-5p, hsa-miR-22-3p) and two proteins (LCAT, kallistatin). Candidates were quantified in independent cohorts by XENO-Q qPCR and sandwich ELISA, and multivariate logistic regression models including all pairwise interactions were trained (N = 65) and validated in an independently recruited testing cohort (MSA N = 22; PD N = 16). The combined microRNA–protein model achieved test-set AUC 0.813 (95% CI 0.654–0.971), with sensitivity 0.818 and specificity 0.750, outperforming microRNA-only (AUC 0.599) and protein-only (AUC 0.747) models and indicating complementary rather than redundant information. The markers converge on pathways of glycerophospholipid metabolism, cholesterol homeostasis, myelin integrity, and inflammatory regulation, consistent with the oligodendrocyte pathology central to MSA. These exploratory findings show that a non-invasive plasma multi-analyte panel can differentiate MSA from PD with clinically meaningful accuracy, and support prospective validation in larger cohorts.