Immunometabolism has emerged as a central regulator of immune responses, linking cellular metabolism to inflammatory signaling and tissue homeostasis. Among tricarboxylic acid (TCA) cycle-derived metabolites, itaconate has gained recognition as an important metabolic feedback regulator promoting inflammatory resolution. Mesenchymal stromal/stem cells (MSCs) are multipotent cells widely recognized for their immunomodulatory and regenerative properties, primarily mediated through paracrine signaling and metabolic adaptation. Increasing evidence indicates that MSC immunoregulatory function is closely associated with metabolic reprogramming involving glycolysis, mitochondrial activity, lipid metabolism, and amino acid pathways. Within this context, itaconate has emerged as a potential metabolic interface linking innate immune activation to MSC function. This narrative review summarizes current evidence supporting both direct and indirect interactions between itaconate signaling and MSC biology. Itaconate and its derivatives influence MSC viability, apoptosis resistance, differentiation potential, and redox balance, while indirectly modulating macrophage polarization and inflammatory microenvironment remodeling through extracellular vesicles and paracrine communication. Despite these advances, critical questions remain regarding endogenous itaconate production by MSCs and its effects on MSC secretome composition and immunoregulatory activity. A deeper understanding of the itaconate-MSC axis may enable metabolic preconditioning strategies aimed at enhancing MSC-based therapies for inflammatory and immune-mediated diseases.

Bacterial outer membrane vesicles (OMVs) are emerging as natural nanocarriers for cancer therapy due to their ability to deliver bioactive molecules and to penetrate the tumor microenvironment. This study investigated the combined effects of Escherichia coli-derived OMVs and Myristica fragrans extract on pancreatic ductal adenocarcinoma (PDAC) cells. OMVs (50 µg/mL) and M. fragrans extract (IC₅₀: 25 µg/mL) significantly reduced PANC-1 cell viability and increased LDH release after 24 h of treatment (p < 0.01). The combination treatment induced oxidative stress, evidenced by increased total oxidant status and decreased antioxidant capacity. Molecular analysis showed upregulation of PTEN expression, reduced phosphorylated AKT levels, and an increased IL-1β/IL-10 ratio. These changes elevated the Bax/Bcl-2 ratio and activated caspase-3, indicating induction of apoptosis (p < 0.01). Our findings suggest that OMVs can effectively deliver M. fragrans phytochemicals to cancer cells, modulating oxidative balance and triggering apoptosis through the PTEN/AKT signaling pathway. This study provides preclinical evidence that combinations of microbial vesicles and plant extracts may enhance therapeutic efficacy against PDAC.

We hypothesize that intestinal microbiome dysbiosis may contribute to Parkinson's disease (PD) pathogenesis. Our prior proof-of-concept clinical trial demonstrated that a precision prebiotic intervention improved microbiota dysbiosis and alleviated gastrointestinal and motor symptoms in PD patients. Building on this, we analyzed plasma extracellular vesicles (EVs) from participants to explore EVs as a dynamic PD biomarker and to assess the systemic effects of a microbiota-directed intervention. Using mass spectrometry-based proteomics of EVs from PD and healthy control (HC) participants, we identified distinct human and bacterial proteins in plasma-derived EV. Crucially, this offers a holistic systemic readout of the microbiota-gut-brain axis by quantifying both host and microbial components. We found that EV proteomic profiles differed between PD and HC samples as well as between unmedicated/mild and medicated/moderate PD participants. Furthermore, the microbiota-directed prebiotic intervention induced an acutely modifiable PD signature, shifting host and microbial EV proteomic profiles toward the HC profile. Using a combined 16-feature host-microbe signature, we built a multiple linear regression model that accurately distinguishes PD status from HC (R2 = 0.88) and successfully stratified disease severity (R2 = 0.72). Based on these findings, we suggest that: (1) a precision prebiotic mixture can modulate PD-associated proteomic signatures and (2) plasma EV proteomics may be a platform to capture these biological responses and to explore potential diagnostic and staging biomarkers in the context of microbiome-targeted interventions.

Treatment of Gram-negative infections remains challenging due to limited compound penetration and the presence of efflux mechanisms. Pseudomonas aeruginosa, in particular, exhibits strong intrinsic resistance, leaving few effective therapeutic options. Here, we provide mechanistic insight into the activity of NV716, a polyaminoisoprenyl antibiotic adjuvant previously described in Gram-negative bacteria, by linking outer membrane perturbation to quantitative changes in intracellular antibiotic accumulation. In P. aeruginosa and Escherichia coli, NV716 increased intracellular antibiotic accumulation and potentiated selected antibiotics, with the most pronounced effects observed for doxycycline. Studies using efflux-deficient and porin-mutant strains indicate that NV716 perturbs outer membrane organization, thereby facilitating enhanced intracellular antibiotic accumulation. Population-scale assays revealed increased outer membrane vesicle (OMV) release, and high-resolution imaging visualized membrane-associated alterations and OMV formation. Truncation of the lipopolysaccharide core sensitized P. aeruginosa to NV716, consistent with increased accessibility of lipid A. Together, these findings establish a quantitative framework that links controlled outer membrane perturbation to intracellular antibiotic accumulation and antibiotic-class-dependent potentiation.

Understanding how genetic polymer sequences became coupled with the reproduction of protocellular compartments is a fundamental challenge in the study of the emergence of living systems. Here, we demonstrate that the coexistence of peptides with defined amino acid sequences and fatty acid vesicles can establish a primitive form of this coupling. We prepared systematically sequence-controlled peptides and examined how their sequences influence the growth rate (fitness) of fatty acid vesicles. The growth of fatty acid vesicles was estimated by dynamic light scattering (DLS) and cryogenic transmission electron microscopic techniques when fatty acid molecules and peptides were fed into a fatty acid vesicle suspension. The relationship between amino acid sequences of peptides and vesicle growth rate was visualized as a fitness landscape, which reveals that specific amino acid sequences promote vesicle growth significantly. Furthermore, we observed epistasis, where the effect of amino acid residue replacement on the fitness depends on the remaining amino acid sequence. Finally, we show that vesicle growth is thermodynamically driven by peptide-induced modulation of the chemical potential of fatty acid molecules. These findings provide direct experimental evidence that primitive sequence information can become spontaneously coupled to vesicle growth.

A myriad of diverse developmental and pro-death signals converge on the mitochondrial pathway of apoptosis, which is governed by the BCL‑2 family of proteins. Comprised of both pro- and anti-apoptotic family members, the BCL‑2 family functions to regulate mitochondrial outer membrane permeabilization (MOMP), often considered the "point of no return" in which a cell commits to an apoptotic outcome. Specifically, the effector BCL‑2 family proteins, BAX and BAK, are responsible for inducing MOMP and therefore investigations into their structural, cellular, and pharmacological regulation are critical to understanding the cellular commitment to apoptosis. A gold standard methodology for studying activation of BAX or BAK is the permeabilization of large unilamellar vesicles (LUVs), which are biochemically-defined model liposomes that mimic the major lipid composition of the outer mitochondrial membrane (OMM). Here, we provide a detailed protocol for generating LUVs containing a fluorescent dye/quencher pair to monitor real-time BAX activation and membrane permeabilization using a standard plate reader. Additionally, we detail example assay strategies to model interactions within the BCL‑2 family and provide a robust mathematical model for fitting and parameterizing kinetic LUV permeabilization data.

Alzheimer's disease (AD) is characterized by progressive cognitive decline, with synaptic dysfunction as the strongest correlate of clinical symptoms. The apolipoprotein E ε4 (ApoE4) allele is the most potent genetic risk factor for late-onset AD. Beyond its roles in amyloid-β aggregation and tau hyperphosphorylation, ApoE4 disrupts synaptic integrity by perturbing lipid metabolism, neuroimmune regulation, mitochondrial dynamics, and activity-dependent plasticity. These ApoE4-driven mechanisms impair presynaptic vesicle trafficking, destabilize postsynaptic receptor and scaffolding networks (including PSD-95, SynGAP, and Shank3), and accelerate complement- and microglia-mediated synaptic pruning. Collectively, these processes converge to destabilize neuronal circuits and drive early cognitive decline. In this review, we synthesize current evidence on the molecular mechanisms by which ApoE4 compromises synaptic function, with particular emphasis on lipid microdomain instability, mitochondrial failure, and the collapse of postsynaptic density proteins. We also discuss therapeutic strategies to enhance synaptic resilience, including modulation of glutamatergic transmission, restoration of lipid homeostasis, augmentation of neurotrophic signaling, and regulation of microglial activity. Targeting synaptic preservation in APOE ε4 carriers holds promise as a disease-modifying approach to mitigate cognitive decline in AD.

There is a critical need for development of non-invasive biomarkers of acute cellular rejection (ACR) in lung transplantation. We hypothesized that ACR induces changes in small extracellular vesicle (sEV) secretory output of T cells into the peripheral circulation, thereby providing a noninvasive method of diagnosis. First, we investigated this hypothesis in a mouse lung transplant model. In line with dense infiltration of lung allograft by T cells during ACR, peripheral blood total T cell and CD8+ T cell counts, as well as, T cell sEV RNA cargoes (Cd3, Cd8, Ifng, Tcrb, and microRNA (Mir)21a) were significantly upregulated in mice receiving allogeneic compared to syngeneic lungs at the postoperative day 7 time point. Second, translating these findings to clinical lung transplantation, we analyzed circulating T cell sEV RNA cargoes with time-matched surveillance transbronchial allograft biopsies in 20 patients. In eight patients with >A1 ACR, eight candidate RNA biomarkers (CD8, TCR, IFNG, HLA-DRA, CD38, MIR21, MIRLET7I, MIR101) were significantly upregulated compared to 12 patients without ACR. Collectively, these findings support further investigation of T cell sEVs as a novel biomarker for ACR diagnosis in lung transplantation.

Avian reovirus (ARV) poses a severe threat to the poultry industry owing to its immunosuppressive properties. Although current vaccine strategies have achieved certain efficacy, they still face challenges such as strain mismatch and immunosuppressive. Extracellular vesicles produced by probiotics, for instance those from Lactobacillus plantarum (Lp), are critically involved in modulating immune responses against bacterial and viral infections. However, the underlying molecular mechanisms remain unclear. This study demonstrated that Lactobacillus plantarum extracellular vesicles (LpEVs) could inhibit ARV replication in DF-1 (chicken embryo fibroblast) cells. We further demonstrated that treatment with LpEVs could promote the phosphorylation of STING (p-STING) and IRF3 (p-IRF3), leading to a significant up-regulation in the secretion of IFN-β and the transcription of interferon-stimulated genes (ISGs). Moreover, these changes primarily depended on STING, as its anti-ARV activity was significantly attenuated following treatment with STING siRNA. We also observed that LpEVs significantly alleviated damage induced by ARV in goslings, as evidenced by the lower rate of body weight loss, reduced viral shedding, and attenuated inflammatory responses in the tissue. Consistently, a significant activation of the STING-IRF3 pathway and an increase in IFN-β secretion were observed. In conclusion, LpEVs demonstrate anti-ARV activity by activating STING-mediated innate immunity, underscoring their considerable potential as novel antiviral postbiotics. This study lays a theoretical foundation for the development of new strategies for ARV prevention and control.

Co-infection with H9N2 subtype avian influenza virus (AIV) and avian pathogenic Escherichia coli (APEC) is common in poultry, yet no vaccine is currently available that simultaneously prevents both infections. Biomimetic bacterial vesicles (BBVs) derived from avian E. coli are a platform for delivering heterologous subunit antigens. In this study, an antigen display strategy was adopted, which involved the genetic fusion of the H9 hemagglutinin subunit HA1 to the outer membrane-anchored protein cytolysin A (ClyA), followed by high-pressure homogenization to prepare BBV-HA1. Intranasal immunization of specific-pathogen-free chickens with BBV-HA1 induced strong mucosal and systemic immunity. Compared with the PBS control, BBV-HA1 vaccination significantly increased the levels of serum IgY directed against outer-membrane proteins of APEC and the HA antigen of H9N2 AIV. It also enhanced the secretory IgA levels in nasal and tracheal lavages. Specifically, hemagglutination inhibition titers of 5.2 ± 0.84 log2 against H9N2 AIV were detected in the immunized sera of the BBV-HA1 group, along with improved in vitro bactericidal activity against APEC. Ex vivo re-stimulation of peripheral blood lymphocytes resulted in higher proliferation indices, and flow cytometry revealed an elevated CD4+ /CD8+ ratio in the BBV-HA1-immunized chickens. After the challenge, BBV-HA1 reduced the APEC O78 bacterial loads, alleviated air-sac and cardiac lesions, and accelerated body-weight recovery. Following the H9N2 challenge, oropharyngeal viral loading at day 7 was detected in 2/8 BBV-HA1-immunized chickens versus 6/8 PBS-inoculated chickens. These results demonstrate that ClyA-anchored HA1 displayed on BBVs induces coordinated mucosal and systemic responses and confers dual protection against H9N2 avian influenza virus and APEC O78.

Plant-derived extracellular vesicles (PDEVs) are emerging as bioactive dietary particles with the capacity to modulate mammalian physiology. Here, we characterize the structure and functional properties of apple-derived extracellular vesicles (ADEVs) and evaluate their relevance to neuroimmune and gut-brain communication. ADEVs exhibit canonical PDEV features and elicit rapid IP₃-dependent Calcium (Ca²⁺) signaling in fibroblasts while preserving blood-brain barrier integrity. Neural assays reveal marked cell-type specificity: ADEVs are efficiently internalized by glial cells and activate glial Ca²⁺ signaling yet display minimal neuronal uptake and no detectable Ca²⁺ response in differentiated SH-SY5Y neurons. Consistent with this selectivity, ADEVs attenuate TNF-α-induced cytokine secretion in activated glia but remain inert in resting neural cells. Although capable of encapsulating L-DOPA efficiently, ADEVs fail to deliver functional neuroprotection against rotenone toxicity, indicating limited neuronal compatibility for dopaminergic cargo. In parallel, using advanced in vitro colon simulation platforms, ADEVs modulate the colonic microbiome in a dose-dependent manner, promoting carbohydrate fermentation and short-chain fatty acid production while reducing proteolytic metabolism under physiologically relevant conditions, with sustained, region-specific effects during prolonged exposure. In vivo, ADEV administration in dogs with chronic intestinal inflammation is associated with altered circulating serotonin levels, suggesting engagement of gut-brain neurochemical pathways with potential implications for mood regulation. Collectively, these findings identify ADEVs as biocompatible, glia-responsive plant vesicles with potential neuromodulatory activity, while delineating intrinsic constraints in their use as neuronal drug-delivery systems.

Alcohol consumption is a modifiable, dose‑dependent risk factor that accelerates cancer initiation, progression, and therapy resistance across breast, liver, colorectal, esophageal, and melanoma cancers. Alcohol metabolism generates acetaldehyde, a genotoxic metabolite that induce DNA damage and impairs repair; drives oxidative stress, hormonal imbalance and promoting a tumor‑supportive microenvironment. These effects are amplified by obesity and insulin resistance, which heighten aggressiveness and blunt treatment response. Alcohol reshapes the tumor immune microenvironment (TIME) and undermines therapies, including immunotherapy. It impairs dendritic cell antigen presentation and co‑stimulation, limits CD4⁺/CD8⁺ priming, and expands immunosuppressive MDSCs and M2‑like TAMs that inhibit cytotoxic T cells and express checkpoint ligands. Concurrent cytokine shifts (TNFα, IL‑6), persistent ROS-NF‑κB signaling, and stromal/vascular remodeling promote immune exclusion and weaken responses to checkpoint blockade, targeted agents, and chemotherapy. Alcohol‑related gut dysbiosis is linked to ICB responsiveness-dampens antitumor immunity. Alcohol increases exosome biogenesis; circulating vesicles enriched in hepatocyte and inflammatory miRNAs (e.g., miR‑122, miR‑192) reprogram immune signaling and metastasis, forming a vesicular axis linking exposure to TIME dysfunction and resistance. Risk is synergistic with tobacco and worsens with aging as detoxification and DNA repair decline. Sex‑specific pharmacokinetics and composition confer vulnerability in women, while socioeconomic barriers delay diagnosis and care. Personalized prevention strategies that incorporate alcohol reduction, smoking cessation, and nutritional support, with emerging biomarkers such as exosomal miRNAs and alcohol‑related metabolites for early detection and risk stratification, are needed to mitigate alcohol‑related carcinogenesis, given the lack of definitive human evidence on alcohol's independent effects on immunotherapy and chemotherapy efficacy.

Targeted intra-atrial delivery of GMP-grade human cardiac extracellular vesicles (EVs) was evaluated for prevention of postoperative atrial fibrillation (AF). EVs were manufactured under serum-free, xenogen-free GMP-compatible conditions and tested in rat and porcine sterile pericarditis models. In rats, intra-atrial EV delivery resulted in selective atrial retention without systemic toxicity. In pigs, EV administration was technically feasible, well tolerated, and completely prevented spontaneous AF while significantly reducing inducible AF. EV therapy also attenuated atrial inflammation and fibrosis without adverse end-organ effects. These findings support localized cardiac EV therapy as a first-in-human-ready, disease-modifying biologic strategy for postoperative AF prevention.