Extracellular vesicles: a mailcoach from mast cell to other cell species

Extracellular vesicles: a mailcoach from mast cell to other cell species | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL This is a preprint and has not been peer reviewed. Data may be preliminary. 28 April 2025 V1 Latest version Share on Extracellular vesicles: a mailcoach from mast cell to other cell species Authors : Bingqi Zhang 0009-0004-9470-032X , Yueshan Sun , Tao Jiang , Runmin Long , and Yuanbiao Guo 0000-0002-7571-8864 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.174584647.70120541/v1 Published Immunity, Inflammation and Disease Version of record Peer review timeline 254 views 142 downloads Contents Abstract Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Mast cells play a crucial role not only in allergic responses but also in innate and adaptive immune responses. They release extracellular vesicles (EVs) containing proteins, lipids, and genetic material that facilitate intercellular communication. Mast cell-derived EVs (MC-EVs) have different characteristics depending on the state of the mast cells, and they are involved in various processes such as dendritic cell maturation, antigen presentation, T lymphocyte activation, macrophage polarization, and the modulation of epithelial cell permeability and epithelial-melancholy transition. EVs released by mast cells can interact with tumor cells and have dual roles in immune responses and disease pathogenesis. The RNA cargo in these EVs holds potential as diagnostic biomarkers and therapeutic targets in diseases including cancers, such as mastocytosis. Overall, these findings provide new insights into the role of EVs in mast cell function and inter-cellular communication. Extracellular vesicles: a mailcoach from mast cell to other cell species Bingqi Zhang 1 , Yueshan Sun 1 , Tao Jiang 2 , Runmin Long 1 , Yuanbiao Guo 1* Affiliation 1：The Third People’s Hospital of Chengdu/ School of Medicine, Southwest Jiaotong University ; Affiliation 2：North Sichuan Medical College * Correspondence： [email protected] ; Tel：61318719. ORCID:0000-0002-7571-8864 Current address: Wuhou Clinical Laboratory Center (No. 3 Yujie East Street), Chengdu, Sichuan Province, Postcode: 610041. Abstract: Mast cells play a crucial role not only in allergic responses but also in innate and adaptive immune responses. They release extracellular vesicles (EVs) containing proteins, lipids, and genetic material that facilitate intercellular communication. Mast cell-derived EVs (MC-EVs) have different characteristics depending on the state of the mast cells, and they are involved in various processes such as dendritic cell maturation, antigen presentation, T lymphocyte activation, macrophage polarization, and the modulation of epithelial cell permeability and epithelial-melancholy transition. EVs released by mast cells can interact with tumor cells and have dual roles in immune responses and disease pathogenesis. The RNA cargo in these EVs holds potential as diagnostic biomarkers and therapeutic targets in diseases including cancers, such as mastocytosis. Overall, these findings provide new insights into the role of EVs in mast cell function and inter-cellular communication. Keywords: Mast cells; Extracellular vesicles; Dendritic cells; T cells; Macrophage; Epithelial cell; Tumor cells Introduction Mast cells (MCs), unique tissue-resident immune cells of the myeloid lineage, exhibit heterogeneity in their granule composition and transcriptomes 1, 2 This diversity leads to the identification of distinct subsets across various tissues, primarily divided into two main categories: tryptase-rich MCs found in mucosa (MCT), and MCs in connective tissue (MCTC) rich in both tryptase and chymase 3 . Their heterogeneity, growth, differentiation and maturation are influenced by interactions with resident stromal cells and the local anatomical niches 3, 4 . As gatekeepers in inflammation and immune responses, MCs help maintain homeostasis and participate in pathogen infections, wound healing, vascularization, tissue remodeling, fibrosis, and autoimmune disease 2, 5 . It’s widely recognized that MCs act as facilitators of communication between the innate and adaptive immune systems by interacting with various immune cells, such as T cells, B cells, macrophages, and dendritic cells, to regulate immune responses and maintain physiological equilibrium 6 . The role MCs in tumors is complex, involved activities such as the release of factors that promote new blood vessel growth within the tumor environment 7 . However, in the case of nasopharyngeal cancer, a specific subset of MCs with high expression of tumor necrosis factor (TNF) but low levels of vascular endothelial growth factor A (VEGFA) are linked with better outcomes 2 . These interactions, whether over short or long distances within the body, may be facilitated by extracellular vesicles (EVs) 8 9, 10 . EVs are double-membrane vesicular structures released by cells into the extracellular matrix. There are primarily three types: exosomes (with a diameter smaller than 150nm), microvesicles/shedding particles, and apoptotic bodies (both with diameters larger than 100nm) 11 . The formation of EVs is mediated by the endosomal sorting complex required for transport (ESCRT) machinery. It mainly consists of ESCRT-0, I, II, III, and vacuolar protein sorting-associated protein 4 (VPS4) 12 . EVs carry both mRNA and microRNA, capable of being transported to and functional in a different cellular environment. In addition，EVs secreted by cells carry a variety of proteins and other components to participate in intercellular communication, which can be taken up by other cells and change the function of recipient cells 13 . EVs mediate the communication between MCs and immune cells, tumor cells and other cells. They have great potential in the physiological response to diseases, but the specific mode of action is still unclear. This review delves into the biological characteristics of MC-EVs and their interactions with various cell types, including immune cells, tumor cells, epithelial cells, and others. These findings offer a reference for further research on MC-EVs to decipher disease mechanisms, develop vaccines, and explore biological therapies. Reorganization of the features and biological functions of MC-EVs MCs monitor and respond to changes in metabolism and immune status in their surrounding microenvironment. When stimulated, particularly during allergic responses, they undergo degranulation and release various inflammatory mediators, including tryptase, carboxypeptidase, histamine, and interleukin 14 . In recent years, studies have demonstrated that MCs can produce EVs not only in the activated/stimulated state (Active-MC-EVs), but also in the resting state (Rest-MC-EVs). These MC-EVs differ in terms of quantity, membrane proteins, constitutive lipids, luminal RNAs and proteins, and their respective functions (Table 1 Differences of MC-EVs in resting and activated state of MCs). Table 1 Differences of MC-EVs in resting and activated state of MCs Project Rest-MC-EVs Active-MC-EVs Mast cell type Activators References Amount less more BMMC DNP- IgE 15 PMC and SpMC 16 Human synovium MC human myeloma IgE 17 MC-specific proteins * less more Human synovium MC human myeloma IgE 17 MC-associated protein# less more BMMC DNP- IgE 15 EV-surface markers CD9 CD63 PMC and SpMC DNP- IgE 16 Lipid cholesterol PA(30:1), PC(36:1) and SM(42:3) PE(32:1), PI(36:2), BMP(40:6) PMC and SpMC DNP- IgE 16 Biological functions of enriching proteins Glycolytic process, positive regulation of cell adhesion and cell migration Redox reaction,lipid metabolic processes, signal peptide processing, and vesicle-mediated transport BMMC DNP- IgE 15 lncRNAs H19 Zfas1, Snhg20, Jpx, Snhg4, Neat1, Malat1 BMMC DNP- IgE 15 Average length of microRNA >60nt <40nt BMMC DNP- IgE 15 microRNAs miR-142a-5p,miR-350-5p,miR-29a-5p和miR-700-3p miR- 126a-3p, miR-21a-3p,miR-210-3p和miR-150-5p BMMC DNP- IgE 15 / miR23b-3p, miR103a-3p Human synovium MC human myeloma IgE 17 miR-7022-5p, miR-532-5p, etc. miR-409-3p, miR7234-5p, etc. Murine P815 LPS 18 *：FcεRI and KIT #：tryptase, carboxypeptidase A, and IL-4 Abbreviations used in this table: BMMC, Bone Marrow Mesenchymal Stem Cells; PMC, peritoneal MC; SpMC, spleen-derived MC; DNP-IgE, DNP-specific IgE; PA, phosphatidic acid; PC, phosphatidylcholine; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PS, phosphatidylserine; SM, sphingomyelin; BMP, bis(monoacylglycero) phosphate; LPS, Lipopolysaccharide Most research consistently indicates that activated MCs release a significantly higher quantity of EVs compared to their resting counterparts 15 16 17 . However, discrepancies exist regarding the particle size distribution of Rest-MC-EVs versus Active-MC-EVs. For instance, Groot Kormelink et al. demonstrated that IgE/anti-IgE-stimulated connective tissue MCs (CTMCs) and mucosal MCs (MCTs) produced Rest-MC-EVs with broader size heterogeneity compared to Active-MC-EVs. In contrast, studies on bone marrow-derived MCs (BMMCs) reported no significant differences in EV size between resting and activated states 15 17 . These conflicting observations may stem from the inherent heterogeneity of MC subsets across tissues, highlighting the need for standardized models to dissect MC-EV biogenesis. The lipid composition of EVs reliably mirrors the distinct functional and phenotypic characteristics of their parent MCs. Notably, MCs express the stem cell factor (SCF) receptor KIT and the high-affinity IgE receptor (FcεRI), which trigger activation and degranulation upon SCF binding or antigenic cross-linking 19-21 . Regardless of variations in the experiments and MC subsets (CTMCs or MMCs), the lipid composition of EVs derived from the same MCs significantly changes post-stimulation (Table 1). Given the essential role of several phospholipids in vesicle formation and stabilization 16 , it is plausible that these lipids may influence Active-MC-EVs secretion 22, 23 . Functionally, MCs express the stem cell factor (SCF) receptor KIT and the high-affinity receptor for IgE (FcεRI). When these receptors bind with SCF or undergo antigenic cross-linking, they trigger MC activation and degranulation 24 . Consequently, MC-EVs hold higher levels of FcεRI and KIT, particularly in Active-MC-EVs compared to Rest-MC-EVs 17 . This is also true for the EV marker protein CD63 16 25 , suggesting its association not just with the origin of endocytic vesicles, but also with MC degranulation. Correspondingly, Active-MC-EVs contain abundant inflammatory factors and the proteins involved with redox reaction, lipid metabolic processes, signal peptide processing, and vesicle-mediated transport. On the other hand, Rest-MC-EVs are significantly enriched in proteins involved in glycolytic process, positive regulation of cell adhesion and cell migration 15 . The RNA cargo of MC-derived EVs exhibits subtype-specific enrichment patterns that correlate with functional states. Rest-MC-EVs are markedly enriched in GC-rich long non-coding RNAs (lncRNAs) and longer miRNA isoforms, likely reflecting their role in maintaining cellular quiescence through epigenetic and post-transcriptional regulation 15 26 27 . In contrast, Active-MC-EVs preferentially package mature miRNAs associated with immune activation, such as miR-142-3p, which amplifies FcεRI-mediated degranulation by targeting negative regulators of the PI3K/Akt pathway 28 . Mechanistic studies further reveal stimulus-dependent miRNA rewiring. For instance, LPS stimulation induces dynamic remodeling of the EV miRNA landscape, with microarray profiling identifying 7 upregulated miRNAs (e.g., miR-409-3p, linked to NF-κB activation) and 14 downregulated species (e.g., miR-499, a suppressor of apoptosis 29 ) in mouse MC-EVs 18 . Critically, the GC-rich lncRNAs in Rest-MC-EVs may stabilize miRNA precursors through RNA-RNA interactions, while Active-MC-EVs utilize shorter, mature miRNAs for rapid signal amplification in recipient cells (28, 31). This dichotomy positions EV-associated miRNAs not merely as biomarkers but as tunable regulators of MCs plasticity, offering dual therapeutic opportunities—silencing pathogenic miRNAs (e.g., antagomiR-142-3p) or restoring homeostatic species (e.g., miR-499 mimics). Heterogeneity also exists in the EVs derived even from the same MCs. For instance, upon activation with stem cell factor (SCF), a ligand of KIT, the human MCs line LAD2 produces two subgroups: KIT + large EVs (with higher KIT) and KIT + small EVs (with lower KIT). The levels of EV protein markers such as CD81, CD9, and ARF6 decrease in both EVs, suggesting that SCF stimulation may alter their protein composition. Furthermore, the KIT + large EVs is associated with actin filaments and the cytoskeleton, whereas the KIT + small EVs is associated with the ESCRT-I complex, which mediates the formation of EVs 30 . Nonetheless, it remains unclear whether this is a specific effect of SCF or if it is common to other MCs stimulations. MC-EVs steer the conversation between MCs and other immune cells MCs act as stimulators in innate immunity and as modulators and effectors in adaptive immunity, through direct cell-cell interactions and the secretion of immunomodulatory factors 31, 32 . They function in immune activation or tolerance by acting on various T cells 33,34, 35 . Accumulating evidence since 2020 has established EVs as key players in these communication processes (Table 2 Functional effects of exosomal components interacting with target cells). Table 2 Functional effects of exosomal components interacting with target cells Target Cells Exosomal Components Functional Effects Dendritic Cells (DCs) HSP60/HSC70, Antigen Complexes Promotes DC maturation and enhances antigen-presenting capacity. ILC2 Cells miR-103a-3p Enhances IL-5 secretion and promotes eosinophilic inflammation. Th2 Cells OX40L Induces differentiation and enhances Th2-type immune responses. B/T Cells MHC II, Adhesion Molecules Directly activates proliferation and cytokine secretion. 3.1. MC-EVs influence antigen presentation and maturation of dendritic cells MCs and dendritic cells (DCs) establish bidirectional communication not only through cytokine networks and direct membrane contacts 36 , but more importantly via EVs-mediated antigen transfer. Mechanistic studies using BMMCs reveal that endocytosed antigens (e.g., OVA) undergo proteasomal processing prior to being packaged into EVs. This EV-mediated antigen delivery is specifically mediated by molecular chaperones HSP60 and HSC70, which form stable complexes with antigens in MC-EVs through their substrate-binding domains. Notably, EVs from B cells or macrophages lack these HSPs (<5% abundance compared to MC-EVs by mass spectrometry), highlighting the unique immunogenic properties of MC-EVs. In vivo validation shows that MC-EV-educated DCs migrate to splenic T cell zones within 24 hr, inducing antigen-specific IgG1/IgG2a production and IFN-γ⁺ CD8⁺ T cell expansion. This mechanism allows T cells to respond effectively even when their MHC molecules don’t perfectly match, as proven by successful immune activation between mice with different genetic backgrounds (H-2b and H-2d strains). Emerging evidence highlights a bidirectional regulatory axis between MC-EVs and DC-derived EVs (DC-EVs). CD301b + DCs strategically positioned in perivascular niches act as sentinels for blood-borne allergens through their mannose receptors (MR). Upon allergen capture (e.g., TNP-OVA), these DCs dynamically release antigen-laden EVs via a VPS4-dependent ESCRT-III mechanism. These EVs serve as antigenic shuttles, delivering allergens to adjacent IgE-primed MCs with >80% degranulation efficiency (vs. 12% with free antigen). Notably, DC-EV release is antigen-dependent, contrasting with MCs’ constitutive EV secretion in steady-state conditions. Proteomic profiling reveals DC-EVs are enriched in MR cytoplasmic domain fragments that retain antigen-binding capacity, enabling sustained MC activation 37 . 3.2. MC-EVs induce lymphocyte activation T lymphocytes, including regulatory T cells (Tregs) and helper T cells (Th), are key protagonists in immune recognition. Th cells can differentiate into Th1 or Th2 cells, which mediate immune responses against intracellular and extracellular microorganisms, respectively 38 . MCs modulate T cell activity through multifaceted mechanisms: Antigen presentation via MHC-II complexes to effector T cells, promoting antigen-specific Treg expansion; Co-stimulatory interactions mediated by MC surface molecules such as ICAM-1 (CD54) and LFA-1 (CD11a), which stabilize T cell adhesion and activation; OX40L-OX40 signaling, a pathway critical for Th2 polarization. 39 . Studies have shown that co-culture of MC lines, P815 and MC/9, with spleen cells or injection of purified MC-EVs induced blast formation, proliferation, and production of IL-2 and IFN-γ. These responses were predominantly Th1-related, as IL-4 was not detected. The molecular mechanism of activation may involve adhesion-induced MC degranulation through LFA-1 and ICAM-1. The accumulation of MHC class II molecules in vesicles synthesized by MCs may enhance antigen presentation 40 , although this is not yet fully understood. Emerging studies reveal a nuanced role of MC-EVs in Th2-type immunity. BMMC-derived EVs (BMMC-EVs) were shown to synergize with IL-4 to promote naive CD4 + T cell differentiation toward Th2 phenotypes, a process mediated by OX40L-OX40 ligand-receptor engagement, However, the in vivo relevance and quantitative contribution of EV-associated OX40L to Th2 polarization remain debated, potentially reflecting tissue-specific microenvironmental cue 41 . Intriguingly, EV-mediated Th2 modulation extends beyond adaptive immunity. Type 2 innate lymphoid cells (ILC2s)—key producers of IL-5 and IL-13 under IL-25/IL-33 stimulation —exhibit selective IL-5 upregulation when exposed to miR-103a-3p-enriched EVs from human synovial MCs. This miRNA-specific effect stems from epigenetic reprogramming of GATA3—miR-103a-3p suppresses PRMT5 expression, thereby reducing arginine methylation of GATA3 and potentiating its transcriptional activation of IL5 over IL13 42 . Effects of MC-EVs on epithelial cells 4.1 MC-EVs drive mesenchymal transformation in epithelial cells The epithelial-mesenchymal transition (EMT) is a cellular process where stationary epithelial cells morph into mobile mesenchymal phenotype, characterized by a shift from protein expressing from E-cadherin to N-cadherin, and subsequent changes in cell polarity, migration, and invasion capabilities 43 . In airway epithelial cells (e.g., A549), MC-EVs drive EMT through TGF-β1/SMAD2 signaling—surface-bound TGF-β1 activates sustained SMAD2 phosphorylation, upregulating EMT transcription factors (TWIST1, MMP9) while suppressing E-cadherin and inducing N-cadherin . Concurrently, MC-EVs activate PI3K-AKT and HIF-1α pathways, synergizing to enhance metalloproteinase secretion (MMP-2/9 activity) and cytoskeletal remodeling 44 . However, in placental trophoblasts (HTR-8/SVneo), MC-EVs from preeclamptic patients exert anti-EMT effects via miR-181a-5p—this miRNA directly targets SNAI2, restoring E-cadherin while inhibiting N-cadherin /MMP-9, ultimately impairing cell migration 45 . These opposing outcomes stem from MC heterogeneity (pro-tumor vs. placental subtypes), signaling bias (TGF-β1 dominance vs. miRNA-mediated suppression), and receptor cell-specific programming (e.g., trophoblast miR-34c-5p counteracts EMT attenuation), underscoring the microenvironment’s pivotal role in dictating EV functionality. 4.2. Effects of MC-EVs on epithelial cell barrier MCs can be activated under physiological stresses in some pathogenic disorders, leading to increased permeability of the intestinal epithelial barrier. The degree of MC infiltration is associated with changes in epithelial barrier permeability 46,47 MC-EVs demonstrate divergent roles in modulating intestinal barrier integrity, contingent upon their cellular origin and pathophysiological context. Treatment with MC-EVs from the HMC-1 cell line significantly compromises epithelial barrier function in intestinal cell models (NCM460, HT-29, and CaCO2), a process mediated by miR-223 enrichment within these vesicles. This miRNA suppresses critical tight junction components, including zonula occludens-1 (ZO-1), occludin (OCLN), and claudin-8 (CLDN8), thereby destabilizing intercellular junctions 48 . Conversely, preliminary evidence suggests that MC-EVs isolated from duodenal mucosa—or those derived from eosinophils—may paradoxically enhance epithelial integrity and mitigate functional dyspepsia in rodent models. However, these protective effects lack mechanistic validation, with no delineation of cargo-specific contributions (e.g., anti-inflammatory miRNAs or reparative proteins) or receptor-cell signaling pathways 49 . MC-EVs mediate cellular interactions in tumors MCs infiltration is observed in various tumor types. 7, 50 . These cells are recruited to the tumor microenvironment (TME) via the interaction of stem cell factor (SCF) with its receptor KIT (CD117). However, their role in tumor progression remains controversial 51 . MCs promote tumor angiogenesis by releasing pro-angiogenic factors such as histamine, heparin, and tryptase 7 . Elevated levels of tryptase-positive MCs are frequently detected in solid tumors, including melanoma, colorectal cancer (CRC), and gastric cancer. 52 . EVs are abundant in the tumor microenvironment, attuning the behaviors of tumor cells. 53 In a study, it was demonstrated that EVs derived from BMMC have an impact on the proliferation, migration and invasion of mouse hepatoma cell Hepa1-6 53 . Additionally, when transferred protein KIT via MC-EVs, lung adenocarcinoma cells displayed an increase in cyclin D1 expression, leading to accelerated cell proliferation 54 . Likewise, the transfer of KIT through MC-EVs also promotes proliferation, migration, and invasion of colorectal cancer cells 55 . The underlying mechanisms involve KIT-induced activation of the PI3K/AKT pathway to regulate cyclin D1 expression in lung adenocarcinoma cells and activation of the MEK/ERK signaling pathway to induce epithelial-mesenchymal transition in colorectal cancer cells 55 . On the other hand, lung adenocarcinoma cells also produce EVs carrying SCF, which activate MCs to release tryptase. This in turn accelerates the proliferation and migration of endothelial cells, fostering angiogenesis 56 . Studies on the duty of MC-EVs in melanoma are incompatible. EVs originating from antigen-stimulated RBL2H3 MCs (Active-MC-EVs) can boost the expression of CCL2 (MCP1) in melanoma cells and lung macrophages, thereby increasing the carcinogenic and metastatic potential of melanoma cells 49 . This action is reliant on miR-154-5p, which is also plentiful in Active-MC-EVs. In contrast, Emerging evidence highlights the unconventional nuclear-targeting activity of mast cell (MC)-derived tryptase in melanoma. Melanoma cells secrete DNA-coated extracellular vesicles (EVs) that selectively capture tryptase released by neighboring MCs. Following endocytosis, tryptase translocates to the nucleus, where it induces dual disruptions: (1) epigenetic remodeling through histone cleavage, silencing oncogenes such as EGR1, and (2) structural damage to the nuclear envelope via degradation of lamina-associated proteins, ultimately triggering irreversible cell cycle arrest 57 . Therefore, it may be possible to propose a mechanism whereby the immune system, represented by MC, has the ability to hijack tumor cell-derived exosomes for anti-tumor purposes, since the number and variety of EVs increased significantly after MC degranulation, drug therapy that inhibits MCs degranulation may affect the release of MC -EVs and the inflammatory outcome 24 . EVs from neoplastic MCs affect the progression of mastocytosis Mastocytosis is a heterogeneous group of hematologic neoplasms characterized by the increased production and accumulation of clonal MCs. As the disease progresses, additional affected organs emerge, including the liver (hepatomegaly), spleen (splenomegaly), and bones (osteoporosis), among others 58 . Kim, DK observed that systemic mastocytosis patients, in comparison to healthy controls, exhibited a higher concentration of serum EVs, which contained MC proteins, such as KIT, FceRI, MRGPRX2, and tryptase, but not granule substances like histamine, heparin, or prohibitin. This suggests that malignant MCs in a resting state release substantial EVs in mastocytosis, which are positively correlated with hepatosplenomegaly. Furthermore, these EVs successfully deliver KIT into a human stellate cell, stimulating proliferation, cytokine production, and differentiation - processes linked to liver pathology. This effect can be tempered by KIT inhibition or neutralization, and is replicated by enforced expression of KIT or a constitutively active D816V-KIT, a gain-of-function mutation associated with mastocytosis. Additionally, these researchers discovered that EVs rising from mastocytosis also disrupt osteoblast maturation, negatively affecting trabecular bone volume and microarchitecture 59 . These effects are mediated by the delivery of miRNA-30a and miRNA-23a through the EVs released from neoplastic MCs, which target osteogenic transcription factors RUNX2 and SMAD1/5. In short, EVs from neoplastic MCs hold theranostic potential for mastocytosis. Effect of MCs - EVs on other cells MCs engage in brain disorders, such as increasing permeability of the blood-brain-barrier under acute stress 60 61 . A recent study 62 conducted a detailed analysis of cerebral malaria and found that intravenously injected MC-EVs (Rest-MC-EVs from murine MC P815) into a mouse model of experimental cerebral malaria aggravated brain vascular endothelial activation and disrupted the blood-brain barrier. In vitro experiments using brain microvascular endothelial cells showed that treatment with P815 cell-derived EVs resulted in decreased mRNA levels of Ang-1, ZO-1, and Claudin-5, and increased Ang-2, CCL2, CXCL1, and CXCL9. These findings suggest that MC-EVs can compromise the integrity of the blood-brain barrier and intensify brain histopathological damage. Further evidence from another study shows that EVs (Active-MC-EVs) from Lipopolysaccharide-stimulated P815 cell enhance cell migration, activation of microglia cells, and upregulation of CD86, IL-1β, IL-6 and TNF-α. This outcome is dependent on the EV-delivery of miR-409-3p, which activates the NF-κB pathway 18 . These findings suggest that MC-EVs can effectively penetrate the blood-brain barrier, reaching the central nervous system from the peripheral circulation, and thus represent potential therapeutic targets and drug delivery vehicles for brain disorders. In addition, MC-EVs augment the migration and activation of mesenchymal stem cells (MSC). Research has demonstrated that HMC-1-EVs can change the phenotype and function of human bone marrow-derived MSCs via TGFβ-1. Interestingly,TGFβ-1 associated with EVs has been found to be more potent than its free form in enhancing MSC cell migration in vitro via SMAD2 signalling 63 . Future research will likely uncover more intricate connections between various cell types and MC-EVs. Clinical potential of MC-EVs in allergic diseases The distinct role of MC-EVs in diseases, particularly their contested impact on allergic reactions, is a significant point of discussion within the broader acknowledgment of the importance of EVs in immune regulation and therapy. In a recent study on asthma, researchers discovered that upon activation of BMMCs by allergens leads to the release of EVs containing FcεRI. These EVs have the ability to bind and counteract free IgE in the bloodstream, consequently restraining MC activation by lowering its concentration 64 . This finding suggests a potential therapeutic approach for managing MCs hyperactivity in allergic conditions. Conversely, additional evidence reinforces the beneficial regulatory function of MC- EVs 40, 65, 66 . In one investigation 66 , after stimulation by an allergen, FcεRI complex was found in MC-EVs. When IgE or antigen attaches to FcεRI, these MC-EVs carry receptors, IgE, and antigens, working together with soluble antigens to activate additional MCs. This interaction amplifies the inflammatory response. The RNA transported via MC-EVs holds significant sway over various facets of human health and disease. MC-EVs are abundant in miR103a-3p and miR-142-3p, both of which actively encourage MC degranulation via the FcεRI receptor. 17, 28 . The tRNA-derived fragment tRF-Leu-AAG-001 detected in leukocyte-derived EVs from individuals with endometriosis originates specifically from MC situated within ectopic tissues. These findings underscore the potential therapeutic application of engineered MC-EVs. Future directions and Conclusions The following is a summary of the whole review, which contains the structure of the paper Figure 1. MC-EVs mediate intercellular communication in both physiological and pathological processes. MC-EVs facilitate communication between cells by amplifying the antigen presentation of dendritic cells, intensifying T cell activation, and guiding macrophage polarization towards M2. MC-EVs instigate numerous signaling events within epithelial cells, encouraging an epithelial-to-mesenchymal transition (EMT) for cell reprogramming while simultaneously disrupting cell barriers by inhibiting tight junction proteins and elevating epithelial permeability. Across cancer progression, MC-EVs interact within the tumor microenvironment (TME), impacting cancer cell proliferation and migration. Additionally, MC-EVs target other receptor cells such as endothelial cells, mesenchymal stem cells (MSCs), and nerve cells. The field of EVs related with MCs is rapidly evolving, and there are several important areas that warrant further research. First, it is essential to explore the factors contributing to the heterogeneity of MC-EVs under varying conditions. The diversity of MC-EVs may emanate from both the inherent heterogeneity of MCs and the various pathological states in which they are implicated. The application of cutting-edge techniques such as single-cell transcriptomes, spatial cell-type classification, and comprehensive tissue imaging, which have been used to analyze the diversity of MCs, in conjunction with single EV analysis, can provide a profound understanding of the heterogeneity and functional diversity of MC-EVs in different physiological and pathological contexts. Second, it is imperative to understand the specific molecular components within MC-EVs to discern their effects on distinct cell types. Analytical techniques such as proteomic, genomic, and lipidomic analyses can be employed to identify these key elements. This insight could expedite the development of tailored therapies by manipulating the cargo of MC-EVs for targeted therapeutic applications. Third, decipher the mechanisms of MC-EV uptake by recipient cells to fully understand their mode of action. Fluctuations in membrane molecules on both MC-EVs and recipient cells can alter the endocytosis process and subsequent cellular responses. This, in turn, can potentially impact the selective uptake of MC-EVs by recipient cells and the resulting functional effects. Lastly, the development of therapeutic strategies based on MC-EVs itself shows great promise. MC-EVs have demonstrated potential as a tool for vaccine development and delivery, as they can modulate immune responses, reduce IgE levels, and enhance antigen presentation in the context of allergies. Modulating MC-EV release, cargo composition, and uptake by recipient cells through engineering and targeting approaches offer potential therapeutic interventions. However, developing these strategies requires a deep understanding of MC-EV biology and their interactions with recipient cells. In conclusion, MC-EVs exhibit diverse characteristics and are essential for intercellular communication, impacting immune responses, epithelial cell behavior, and other cellular functions. The role of MC-EVs in diseases is subject to debate, as they have the potential to both manage MCs hyperactivity and tumor malignancy, but also contribute to inflammatory responses. Further research in this area will enhance our understanding of MC-EVs, leading to the development of diagnostic tools and therapeutic interventions. Author Contributions: Conceptualization, Yueshan Sun and Yuanbiao Guo .; resources, Yueshan Sun and Yuanbiao Guo .; writing—original draft preparation, Bingqi Zhang .; writing—review and editing, Yuanbiao Guo .; . Funding: This research was supported by National Natural Science Foundation of China (81270465), the Foundation of Science and Technology Department of Sichuan province (24NSFSC1611, 2023JDRC0088, MZGC20240097). 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Keywords allergy mast cells/basophils Authors Affiliations Bingqi Zhang 0009-0004-9470-032X Chengdu Third People's Hospital View all articles by this author Yueshan Sun Chengdu Third People's Hospital View all articles by this author Tao Jiang Public Health Clinical Center of Chengdu View all articles by this author Runmin Long Chengdu Third People's Hospital View all articles by this author Yuanbiao Guo 0000-0002-7571-8864 [email protected] Chengdu Third People's Hospital View all articles by this author Metrics & Citations Metrics Article Usage 254 views 142 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Bingqi Zhang, Yueshan Sun, Tao Jiang, et al. Extracellular vesicles: a mailcoach from mast cell to other cell species. Authorea . 28 April 2025. DOI: https://doi.org/10.22541/au.174584647.70120541/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. 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