Journal of Extracellular Vesicles最新文献

Neuroinflammaging, a moderate, chronic, and sterile inflammation in the hippocampus, contributes to age-related cognitive decline. Neuroinflammaging comprises the activation of the nucleotide-binding domain, leucine-rich repeat family, and pyrin domain-containing 3 (NLRP3) inflammasomes, and the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway that triggers type 1 interferon (IFN-1) signalling. Studies have shown that extracellular vesicles from human induced pluripotent stem cell-derived neural stem cells (hiPSC-NSC-EVs) contain therapeutic miRNAs that can alleviate neuroinflammation. Therefore, this study examined the effects of late middle-aged (18-month-old) male and female C57BL6/J mice receiving two intranasal doses of hiPSC-NSC-EVs on neuroinflammaging in the hippocampus at 20.5 months of age. Compared with animals receiving vehicle treatment, the hippocampus of animals receiving hiPSC-NSC-EVs exhibited reductions in astrocyte hypertrophy, microglial clusters, and oxidative stress, along with elevated expression of antioxidant proteins and genes that maintain mitochondrial respiratory chain integrity. Moreover, hiPSC-NSC-EVs therapy decreased the levels of various proteins involved in the activation of the NLRP3 inflammasome, p38/mitogen-activated protein kinase, cGAS-STING-IFN-1, and Janus kinase and signal transducer and activator of transcription signalling pathways. Furthermore, in vitro assays using genetically engineered RAW cells and hiPSC-NSC-EVs, with or without targeted depletion of specific miRNAs, demonstrated that miRNA-30e-3p and miRNA-181a-5p, both present in hiPSC-NSC-EVs, can significantly inhibit the activation of the NLRP3 inflammasome and the STING pathway, respectively. Additionally, single-cell RNA sequencing conducted 7 days post-treatment revealed that hiPSC-NSC-EVs induce widespread transcriptomic changes in microglia, including increased expression of numerous genes that enhance oxidative phosphorylation and reduced expression of abundant genes that drive multiple proinflammatory signalling pathways. These changes mediated by hiPSC-NSC-EVs were also associated with improved cognitive and memory function. Thus, intranasal hiPSC-NSC-EVs therapy in late middle age can effectively diminish proinflammatory microglial transcriptome and signalling cascades that drive neuroinflammaging in the hippocampus, contributing to better brain function in old age.

Tissue-derived extracellular vesicles (Ti-EVs) play a crucial role in tumour progression, but their value as differential diagnostic markers for various renal cell carcinoma (RCC) subtypes remains uncertain. Through analysis of paired tumour and normal tissues, along with their corresponding Ti-EVs, we identified NEAT1 and MMP15 as markers for papillary RCC (pRCC); DSG2 and AC026888.1 for chromophobe RCC (chRCC); NDUFA4L2, SERPINA1, VEGFA, EGLN3, CPE and C6orf223 for low-grade clear cell RCC (low-grade ccRCC); and NDUFA4L2, APOC1, TGFBI and LINC00887 for high-grade ccRCC (high-grade ccRCC). In an external validation cohort, an area under the curve (AUC) of 0.922 for low-grade ccRCC detection and 0.874 for high-grade ccRCC detection was achieved, respectively, using urinary EVs. Furthermore, integrating single-cell sequencing data revealed that SERPINA1 and VEGFA in low-grade ccRCC, and APOC1 and TGFBI in high-grade ccRCC, were derived from tumour-associated macrophages, whereas NDUFA4L2 originated from cancer cells in both low- and high-grade ccRCC.

The capacity of host professional phagocytes to attenuate excessive inflammatory responses through pathogen cloaking during infection has been well-established. However, the involvement of non-professional phagocytes in this process remains unknown. Here, we identify a previously unrecognized mechanism by which lung epithelial cells (LECs) attenuate inflammatory responses during Staphylococcus aureus infection. S. aureus-challenged LECs rapidly shed extracellular vesicles (EVs) carrying surface receptors capable of binding invading bacteria and forming EV-bacteria complexes. The EV-bacteria complexes were internalized by LECs via RhoA-ROCK1-actin-driven endocytosis pathway, reducing free bacterial burden within the alveolar lumen. This EV-mediated pathogen cloaking conferred acute-phase protection, as demonstrated by mitigating early-stage pulmonary inflammation, and improving survival rates in infected mice. Paradoxically, this strategy permitted chronic bacterial persistence and sustaining low-grade inflammation. Our findings delineate a trade-off mechanism that non-professional phagocytes modulate acute bacterial infection and inflammatory responses via pathogen cloaking. This mechanistic perspective reframes non-professional phagocytes as active architects of infection outcomes based on EV-mediated host-pathogen interactions. Our work provides insights into the mechanism of bacterial cloaking during infection and suggests stage-specific therapeutic strategies.

Decreased percentage of Foxp3⁺ regulatory T cells (Tregs) in the lungs results in overwhelming inflammation and delayed recovery of acute lung injury (ALI) caused by acute respiratory distress syndrome (ARDS). Extracellular vesicles (EVs) in the pulmonary microenvironment significantly affect the immune system, but their underlying effects on Tregs are unclear. Here, we demonstrate increased endothelial cell-derived EVs (CD31⁺ EVs) in lipopolysaccharide (LPS)-induced ALI models by single-EV analysis. EVs from activated pulmonary endothelial cells (ECs) exhibit proinflammatory effects and suppress Treg induction. Exposure of Tregs to these EVs induces massive production of interleukin (IL)-21, which has been proven to reduce Foxp3 expression. Mechanistically, we find that Med1 enriched in these EVs can directly bind to the promoter region of the IL-21 gene thus activating IL-21 transcription in Tregs. Moreover, we confirm that suppressing Med1 accumulation in EVs from activated pulmonary ECs can reverse Treg differentiation, and alleviate lung inflammation. Finally, we observe a significant increase of EVs carrying Med1 in the BALF of patients with ARDS. Taken together, this study identifies that EV-mediated pulmonary endothelium-Treg communication is crucial for Treg suppression in ARDS and may provide potential therapeutic targets for the treatment of this fatal clinical syndrome.

Extracellular vesicles (EV) are emerging regulators of metabolic homeostasis through their bioactive cargo. This study first investigated the lipidomic profile and functional effects of plasma EV derived from adiponectin-knockout (KO) mice to identify EV-associated lipid signatures linked to metabolic dysfunction. Lipidomic profiling revealed that KO EV were enriched in sphingolipids and polyunsaturated phospholipids compared to wild-type (WT) EV. To evaluate functional consequences, recipient cell assays were conducted using macrophages, skeletal muscle cells, and pancreatic beta cells. KO EV showed an increased uptake in RAW 264.7 macrophages and induced elevated reactive oxygen species (ROS) and activation of NF-κB and IRF inflammatory pathways. In L6 skeletal muscle cells, WT EV increased ATP production, while KO EV failed to elicit this effect. Furthermore, KO EV impaired glucose-stimulated insulin secretion in INS-1 pancreatic beta cells. These findings suggested that altered lipid composition in EV from KO mice contributes to oxidative stress, inflammation, and impaired metabolic regulation in recipient cells. Next, translational relevance was established by documenting that plasma EV from patients with metabolic syndrome exhibited lipidomic remodeling features in parallel to the murine KO phenotype, in particular enriched PUFA-containing lipids. Together, these findings identify a conserved adiponectin-EV lipid composition axis regulating oxidative stress, inflammation, and impaired metabolic regulation. The new knowledge presented in this study has implications for biomarker discovery and therapeutic targeting in metabolic disease.

Daratumumab (DARA) is a human monoclonal antibody for the treatment of multiple myeloma (MM), an incurable hematologic malignancy characterised by the accumulation of malignant plasma cells, located in the bone marrow (BM). We previously reported that peripheral blood plasma (PB) extracellular vesicles (EVs), isolated from 57 MM patients treated with DARA contain elevated CD55, CD59 and CD147 relative to healthy PB EVs, and elevated PDL1 was associated with patient response to DARA. The aim of this study was to identify additional proteins altered in these patients in order to generate predictive MM EV protein signatures. Flow cytometry analysis revealed that CD31, CD36 and CD44 were significantly elevated in MM PB EVs relative to healthy PB EVs, while CD8 and LAT1 were significantly decreased. CD38, LAT1 and PDL1 were significantly higher in PB EVs of patients with a long-term response to DARA. Multivariate ROC curves revealed a diagnostic signature (MM panel) with a sensitivity 86.4% and specificity 91.6%, and a predictive signature (Response panel) with a sensitivity 80% and specificity 91.2%. In conclusion we identified two EV signatures that may have potential as a non-invasive liquid biopsy to complement or replace invasive BM sampling for monitoring patient response to DARA.

Microinflammation is a key driver of chronic kidney disease (CKD) progression, with interleukin-1β (IL-1β) playing a pivotal role. However, current anti-IL-1β antibody therapies face critical limitations, such as systemic side effects and substantial production costs, which hinder their therapeutic efficacy and clinical translation for CKD intervention. To address this, we developed Bacteroides fragilis-derived outer membrane vesicles (OMVs) encapsulating anti-IL-1β single-chain variable fragment (scFv) and conjugated with kidney-targeting peptides (KKEEE)₃K. Engineered OMV-(KKEEE)₃K-scFv's safety was evaluated in vitro (24, 48 h) and in vivo (2 months). Its anti-inflammatory efficacy was assessed in a high glucose-induced model in vitro, and in various kidney disease mouse models (streptozotocin-induced diabetic nephropathy, lupus nephritis, unilateral ureteral obstruction) in vivo. OMV-(KKEEE)₃K-scFv showed high stability, precisely delivered scFv to proximal renal tubules, had excellent safety, reduced the expression of inflammatory cytokines including tumour necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), decreased cell infiltration, and alleviated renal injury. In conclusion, engineered OMVs effectively deliver anti-IL-1β scFv, mitigating local inflammation and CKD-related renal damage.

Temporomandibular joint osteoarthritis (TMJ-OA) is a progressive degenerative disorder, for which therapeutic interventions remain limited. The disruption of metabolic homeostasis plays a critical role in the pathogenesis and advancement of TMJ-OA. However, it remains unclear whether extracellular vesicles (EVs) as cellular metabolites are correlated with the pathogenesis, treatment and diagnosis of TMJ-OA. In this study, we demonstrated that autologous circulating extracellular vesicles (C-EVs) possessed significant therapeutic potential for TMJ-OA through the targeted removal of senescent chondrocytes. In a randomized clinical trial (ChiCTR2200063153), C-EV administration was found to significantly enhance condylar bone regeneration and alleviate symptoms relative to hyaluronic acid controls, without eliciting any adverse effects. Comparative analysis revealed that joint cavity-derived EVs from TMJ-OA patients (OA-EVs) exhibited structural abnormalities, diminished expression of canonical EV markers, and pro-inflammatory characteristics. In contrast, C-EVs were significantly enriched with functional proteins C1q binding protein (C1QBP). And the level of C1QBP-positive EVs was positively correlated with therapeutic outcomes, thereby establishing C1QBP as a potential predictive biomarker for TMJ-OA. Furthermore, C-EVs reestablished joint homeostasis by regulating the immune microenvironment and tissue regeneration capacity. Mechanistically, C1QBP^high C-EVs upregulated the expression of membrane C1q on senescent chondrocytes, thereby initiating C1q–C1QBP binding, p14ARF translocation to mitochondria, and subsequent cytochrome C/caspase-3-dependent apoptosis. Our findings demonstrate that C-EVs serve a dual therapeutic role by facilitating the clearance of senescent cells via the C1QBP/C1q/p14ARF axis, while promoting tissue regeneration and regulating metabolites homeostasis, offering a novel biological strategy for TMJ-OA treatment.

Tumour development and progression are driven by intricate interactions among various cell types within the tumour microenvironment (TME). Targeting a single cell type often fails to eradicate cancer, highlighting the need for strategies to co-target multiple cell types. MicroRNA-21-5p, highly abundant in tumour cells and tumour-associated macrophages (TAMs), exerts strong cancer-promoting effects. Epidermal growth factor receptor (EGFR) is overexpressed in various cancer types, and programmed death-ligand 1 (PD-L1) is expressed predominantly by TAMs in multiple cancers. In this study, we developed a dual-targeted engineered milk-derived extracellular vesicles system (7D12/KN035-iEVs), decorated with 7D12 (an EGFR nanobody) and KN035 (a PD-L1 nanobody), to specifically deliver miR-21-5p inhibitors into EGFR⁺ and/or PD-L1⁺ tumour cells and TAMs, thereby inhibiting tumour progression while reprogramming immunosuppressive TME. Notably, this dual-targeting nanomedicine synergistically inhibits tumour growth when combined with immunotherapy and radiotherapy. In summary, this mEV-based nanomedicine represents a promising universal strategy for cancer treatment, offering a versatile platform for targeting multiple components of the TME.