REVIEW article

Front. Immunol., 01 August 2024

Sec. T Cell Biology

Volume 15 - 2024 | https://doi.org/10.3389/fimmu.2024.1408415

Composition, functions, and applications of exosomal membrane proteins

  • 1. Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing, China

  • 2. Integrated Chinese and Western Medicine Center, Beijing Youan Hospital, Capital Medical University, Beijing, China

  • 3. Beijing Youan Hospital, Capital Medical University, Beijing, China

Abstract

Exosomes play a crucial role in various biological processes, such as human development, immune responses, and disease occurrence. The membrane proteins on exosomes are pivotal factors for their biological functionality. Currently, numerous membrane proteins have been identified on exosome membranes, participating in intercellular communication, mediating target cell recognition, and regulating immune processes. Furthermore, membrane proteins from exosomes derived from cancer cells can serve as relevant biomarkers for early cancer diagnosis. This article provides a comprehensive review of the composition of exosome membrane proteins and their diverse functions in the organism’s biological processes. Through in-depth exploration of exosome membrane proteins, it is expected to offer essential foundations for the future development of novel biomedical diagnostics and therapies.

1 Introduction

In recent years, as our understanding of exosomes has advanced, the biological roles of exosome membrane proteins in cells and organisms have garnered increasing attention. As a primary constituent of exosomes, exosome membrane proteins not only play a role in the formation and release of exosomes (1–6) but also exhibit diverse functions, including targeting or adhering to receptor cells, anti-apoptotic activities, membrane fusion, signal transduction, metabolism, and structural dynamics (7). Therefore, comprehending the composition and functions of exosome membrane proteins is crucial for understanding the biological characteristics and mechanisms of action of exosomes.

The generation of exosomes involves the inward budding of the plasma membrane and the formation of intraluminal vesicles (ILVs) within multivesicular bodies (MVBs) in the cell. ILVs are eventually secreted as exosomes by the fusion of MVBs with the plasma membrane and released via exocytosis (8–12). The initial inward budding of the plasma membrane forms a cup-shaped structure containing cell surface and soluble proteins related to the extracellular environment. Subsequently, budding of the inner membrane forms ILVs within endosomes, which contain specific proteins, lipids, nucleic acids, and other molecules (13–17). The biogenesis of exosomes is driven by multiple protein-regulated mechanisms, including ESCRT protein complexes, Rab GTPases, Tetraspanins, etc (18). Finally, mature MVBs fuse with the plasma membrane, releasing ILVs as exosomes through exocytosis into the extracellular environment (1, 2). These released exosomes can facilitate intercellular signaling, modulate immune responses, and promote cell-cell communication (18, 19).

In this review, we systematically summarize the composition of exosome membrane proteins and explore their potential applications in mediating target cell recognition, immune regulation, and disease control.

2 Composition and classification of exosome membrane proteins

Exosome membrane proteins are classified based on membrane localization into transmembrane proteins, lipid-anchored membrane proteins, peripheral-associated membrane proteins, and inner-associated membrane proteins. According to the current exosome content database, Exocarta (http://www.exocarta.org), 9769 exosome proteins have been identified in exosomes from various cell types and organisms. With the continuous development of modern technology, the detection methods for extracellular vesicle membrane proteins are also constantly being updated. Currently used methods include Western blot, ELISA, Atomic Force Microscopy (AFM), etc. (20). Table 1 summarizes the common methods for detecting extracellular vesicle membrane proteins. Recently, Xiaoni Fang et al. (27), using the integrated GF/PMO platform, identified a total of 334 exosome proteins, including 111 membrane proteins. The GF/PMO platform is an innovative approach that integrates two nanomaterials with different surface properties: hydrophilic macroporous graphene foam (GF) and amphiphilic periodic mesoporous organosilica (PMO). This platform is used for the efficient separation of exosomes from human serum and effective protein analysis, aiding in the identification of more exosome-based disease biomarkers. This method of efficient and specific separation and analysis of exosome proteins holds significant application prospects in biomedical research. Table 2 summarizes some important and noteworthy proteins distributed within the inner membrane, outer membrane, and transmembrane region of exosomes. The arrangement of exosome membrane proteins is illustrated in Figure 1.

Table 1

MethodDescriptionReferences
Flow cytometryDetect and characterize exosome surface proteins(21)
Enzyme-linked immunosorbent assay (ELISA)Used for the detection and quantification of exosomal proteins. Common capture antibodies include CD63 and CD81(22)
Western blotUsed to detect the presence of proteins on extracellular vesicles (CD9, CD63, CD81)(23, 24)
Atomic Force Microscopy (AFM)Using a very sharp cantilever to scan the sample surface, software analysis can be used to identify specific receptor sites on the surface of extracellular vesicles, including membrane proteins(15)
Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS)Antibodies labeled with extracellular vesicle surface markers can be arranged on silicon chips to detect extracellular vesicle surface proteins(25)
Surface plasmon resonance (SPR)Label-free and real-time quantitative analysis techniques have a high sensitivity of up to 1 nM for specific protein binding of 20 kDa(26)

Commonly used methods for identifying exosomal proteins.

Table 2

Protein ClassificationExosome SourceMembrane Protein NameReference
Transmembrane ProteinsHEK293 CellCD9、CD63、CD81(28, 29)
B Lymphocyte、DCMHC-II(30–33)
DCICAM-1(34)
MCF-7 CellSDCs(35)
Mouse E0771、Mouse Pan02Integrins(36)
B LymphocyteMHC-I(37)
Melanoma CellPD-L1(38)
DC line D1CD86(39)
SW480BCAM、CD109、CD44、CD46、CD47、CD70、GPC4、IGSF8、ITGA5、LTGAV、ITGB5、LDLR、MMP14、TFRC、TSPAN1、TSPAN14、VAMP7(40)
Lipid-Anchored Outer Membrane ProteinsHT1376、CACO2、DU145、PC3、MCF7CD39、CD73(41)
ErythrocyteCD55、CD59(42)
MDA-MB-231GPC-1(43)
Peripheral Membrane ProteinsPancreatic ductal adenocarcinoma with pancreatic duct fluidTenascin C(44)
Sw71Fibronectin(45)
Colon cancer patient
plasma		ECM1(46)
COS-7 CellMfgE8(47)
B-cell LymphomaWnt(48)
SW480CLU、DCXR、DNM1L、EIF3L、FKBP1A、GANAB、LGALS3BP、RACK1、SEC23B、USO1(40)
Lipid-Anchored Inner Membrane ProteinsHIV-1 BaL StrainRab27a(49)
S2 CellARC(50)
Inner Membrane ProteinsRN Cell、T cell、Human Mesothelioma Cell LineERM(51–53)
CHO-K1 CellSyntenin-1(54)
DC Line D1HSC73、HSP84、Tsg101(39, 55)
HeLa Kyoto CellAlix(2)

Exosome membrane proteins.

Figure 1

A specific class of membrane proteins serves as exosome-specific markers, such as the tetraspanins CD9, CD63, and CD81 (2, 4, 28, 30, 56–63). These proteins have been demonstrated to regulate the transport and function of associated proteins through membrane compartmentalization (64). Lipid-anchored outer membrane proteins, including CD39, CD73, GPC-1, CD55, and CD59, with enzymatic activity, notably CD39 and CD73, have been shown to promote angiogenesis through adenosine A2B receptor signaling (65). Peripheral membrane proteins such as Tenascin C, Fibronectin, ECM1, MfgE8, and Wnt play crucial roles in the functional processes of exosomes. For example, exosomes derived from embryonic stem cells (ESCs) carrying Fibronectin contribute to maintaining their stem cell characteristics (66). Lipid-anchored inner membrane protein Rab27a regulates exosome formation and release (67). Inner membrane proteins Tsg101 and Alix serve as exosome markers and are involved in the biogenesis of multivesicular bodies (MVB) (68). The arrangement of exosome membrane proteins is illustrated in Figure 1.

Exosome membrane proteins vary among different cell sources; for instance, exosomes from antigen-presenting cells (APCs) are rich in transmembrane proteins such as MHC-I, MHC-II, and ICAM-1 (68, 69). The diversity of these membrane proteins determines the versatility of exosome functions (70). Therefore, a focused discussion on the composition and clinical applications of exosome membrane proteins is crucial for guiding future research directions.

3 Roles and functions of exosome membrane proteins

3.1 Diagnostic role of exosome membrane proteins in diseases

Currently, a substantial body of literature indicates that the molecular components of exosomes, particularly exosome proteins, serve as promising novel markers for the clinical diagnosis of various diseases (71–84). Their application prospects are considerable due to unique advantages: high sensitivity (85), high specificity (43), and high stability (85), making them a preferred option for liquid biopsy. The presence of exosomes can be detected in various bodily fluids (86).

In the current stage, many potential targets for cancer treatment are tumor-specific biological markers. Since exosomes derived from cancerous sources carry similar markers on their membrane surfaces, researching exosome membrane protein biomarkers is crucial for the development of targeted cancer therapies (87, 88). The primary component of exosome proteins, membrane proteins (27), offers a reliable choice for developing new disease diagnostic biomarkers. It is gradually becoming a focal point in exosome research. Table 3 summarizes exosome membrane proteins from different disease sources.

Table 3

System ClassificationDisease ClassificationMembrane ProteinsReferences
Respiratory SystemLung CancerCD171、CD151、Tetraspanin 8、CD317、EGFR、PD-L1(82, 89, 90)
Nasopharyngeal CarcinomaGalectin 9、LMP1、HLA-II(91, 92)
Digestive SystemLiver CancerCD26、CD81、S1C3A1、CD10、GPC3、PIGR、14–3-3ζ(93–96)
Chronic Hepatitis CCD81(97)
Pancreatic CancerGPC1、CD151、EphA2、CKAP4、CD133(43, 81, 98–100)
Colorectal CancerCD147、CD9(101)
Gastric CancerTetraspanin 8、HER-2 (neu)、CCR6(102)
Nervous SystemParkinson’s DiseaseLRRK2、L1CAM(84, 103)
Malignant GliomaEGFRvIII、EGFR、PDPN(104, 105)
Genitourinary SystemRenal Cell CarcinomaCAIX(106)
Diabetic NephropathyEGFR(107)
Bladder CancerCD36、CD44、MUC1,Integrin β1, IntegrinBα6,CD10,5T4,Basigin,CD73(108, 109)
Prostate CancerPSA、PSMA(110)
Ovarian CancerL1CAM、CD24、TSG101、Alix、ADAM10、EMMPRIN、Claudin-4、HSP70、HER2、CD47(57, 111–113)
Endocrine SystemThyroid CancerITGB2(114)
Breast CancerCD9、Annexin‐1、GPC1、PMSA、EGFR、CD81、CEA(115–117)
Skeletal SystemOsteosarcomaCD63(118)
Immune SystemSystemic Lupus ErythematosusBPI(119)

Exosome membrane proteins from various disease sources.

Mariantonia Logozzi and colleagues designed an internal sandwich ELISA (Exotest), revealing a significant increase in CD63 and Caveolin-1 in plasma-derived exosomes from melanoma patients. They described a novel non-invasive detection method for assessing the expression of exosome-specific membrane proteins in melanoma patients’ plasma, providing a potential diagnostic tool (120). In 2013, Yusuke Yoshioka and colleagues conducted a comparative analysis of exosome protein markers in different human cancer types. They found elevated levels of CD63 in exosomes derived from malignant cancer cells compared to those from non-cancerous cells, further supporting CD63 as a protein marker for cancer (29, 121). Bingqian Lin et al. developed a specific dual-ligand recognition system based on the exosome membrane, combined with droplet digital PCR (ddPCR) (TRACER), for quantifying tumor-derived exosome PD-L1 (Exo-PD-L1). The tumor-derived Exo-PD-L1 levels detected by TRACER could distinguish cancer patients from healthy blood donors (122). Research indicates that the lipid-anchored outer membrane protein GPC-1 is significantly overexpressed in plasma-derived exosomes from pancreatic ductal adenocarcinoma (PDAC) patients compared to healthy controls, confirming the potential utility of GPC-1 for early PDAC diagnosis (123).

Compared to biomarkers detected directly in conventional specimens (such as serum or urine), exosome biomarkers offer higher specificity and sensitivity due to their superior stability (124). Exosome biomarkers, especially those from easily obtainable biological fluids like saliva, show great potential for clinical applications. In conclusion, exosome biomarkers are still in the early stages of discovery and development, and their potential value in clinical diagnostics requires further exploration. Therefore, if certain membrane proteins are specifically expressed by a particular tumor (125), their expression on circulating exosomes can be utilized as an early diagnostic signal for cancer. The diagnostic potential of exosome membrane proteins in different diseases is depicted in Figure 2.

Figure 2

3.2 Remote regulatory role of exosome membrane proteins

Current data suggest that exosome membrane proteins can exert regulatory effects on recipient cells (17, 126–132). They identify target cells by binding to surface proteins on recipient cells (133), leading to changes in the recipient cells. Kun Zhao et al. (134) found that exosome tetraspanin protein Tspan8 and CD151 derived from tumor cells can activate the PI3K/Akt signalling pathway by binding to GPCR and RTK proteins on recipient cells, promoting tumor angiogenesis. Similarly, Shi Du et al. demonstrated that tumor cell-derived exosomes carrying tyrosine kinase 2 (TIE2) with an immunoglobulin and epidermal growth factor homology domain deliver TIE2 protein to macrophages. Macrophages carrying TIE2 (TEMs) interact with angiopoietin-2 (ANG2), ultimately promoting cervical cancer angiogenesis (135).

Furthermore, a study detected exosomes in the serum of osteosarcoma patients with lung metastasis and those without lung metastasis. The results revealed a significant expression of PD-L1 and N-cadherin in exosomes from serum of osteosarcoma patients with lung metastasis. This study suggests that exosomes derived from osteosarcoma and carrying PD-L1 and N-cadherin reach the lungs through the circulatory system. The osteosarcoma cells at the lung metastatic site further internalize these exosomes, ultimately promoting the migration and progression of metastatic tumors (136). The regulatory mechanism involves two steps. Firstly, osteosarcoma cells stimulate epithelial cells to transition from an adhesive epithelial state to an active mesenchymal state through the epithelial-mesenchymal transition (EMT) mechanism. This mechanism facilitates the spread of cancer cells at metastatic sites. Secondly, metastatic osteosarcoma cells internalize exosomes derived from primary osteosarcoma, which carry PD-L1 and N-cadherin, promoting lung metastasis. A comprehensive understanding of the complex regulatory mechanisms of exosome membrane proteins in diseases can deepen our understanding of disease development and provide stronger support for the development of innovative treatment methods.

3.3 The role of exosomal membrane proteins in epithelial-mesenchymal transition

EMT is a cellular process that drives the differentiation of epithelial cells into mesenchymal cells. Through specific programs, epithelial cells acquire mesenchymal characteristics, including reduced cell adhesion, loss of cell polarity, and increased cell migration (137–140). Notably, cancer cells that have undergone EMT not only gain distinct molecular characteristics but also develop resistance to chemotherapy and immunotherapy (141–143). Proteins in exosomes significantly influence chemotherapy resistance. Based on their mechanisms of inducing resistance, exosomal proteins are mainly classified into enzymes, transcription factors, membrane proteins, and secreted proteins (144). Laura J. Vellade et al. (145) demonstrated that exosomes carrying PDGFRβ interact with receptors on melanoma cells, leading to dose-dependent activation of the PI3K/AKT signaling pathway and bypassing BRAF inhibition in the MAPK pathway, ultimately resulting in reduced drug sensitivity in melanoma cells.

Reports indicate that tumor-derived exosomes (TEX) carry proteins that promote epithelial-mesenchymal transition, including EMT inducers such as TGF-β, HIF1α, β-catenin, Caveolin-1, and Vimentin. These proteins can enhance the invasion and migration capabilities of recipient cells and contribute to stromal remodeling and the formation of the pre-metastatic niche (146, 147). Research by Mohammad A. Rahman et al. (147) demonstrated that exosomes derived from lung cancer activate the migration process of human bronchial epithelial cells (HBECs) by enhancing their metastatic properties. TEX were isolated from the supernatants of non-metastatic and metastatic lung cancer cell lines via ultracentrifugation, and these exosomes carried epithelial (E-cadherin, ZO-1) and mesenchymal (N-cadherin, Vimentin) markers. Among these, E-cadherin and N-cadherin serve as membrane protein markers.

Furthermore, the exosomal membrane protein CD44 can promote cell migration and invasion by binding to hyaluronic acid and activating EMT-related signaling pathways (148). A recent study by Nakamura and colleagues showed that exosomes derived from ovarian cancer transfer CD44 to human peritoneal mesothelial cells (HPMC), thereby promoting cancer invasion (149). Research by Yao Li et al. (150) found that exosomes carrying the PSGR membrane protein enhanced the migration, invasion, and EMT of low-invasive prostate cancer cells (LNCaP and RWPE-1) and reshaped the mRNA profiles of these cells. Although the morphological, phenotypic, and functional changes associated with EMT have been well described, the molecular and genetic mechanisms by which exosomal membrane proteins drive this process require further investigation (151–154).

3.4 Therapeutic role of exosome membrane proteins

Existing studies indicate that exosome membrane proteins play a crucial role in mediating various disease treatments (125, 133, 155–169). CD47 is usually upregulated on the surface of tumor cells, binding to signal-regulatory protein alpha (SIRPα) on phagocytic cells and inhibiting their phagocytic function, creating a “don’t eat me” signal. Eunee Koh et al. (170) designed engineered exosomes with surfaces carrying SIRPα, disrupting the CD47-SIRPα interaction between cancer cells and macrophages, enhancing the efficiency of phagocytosis of tumor cells. Similarly, Eunji Cho et al. (171) found that exosomes containing SIRPα could more effectively counteract CD47 on cancer cells, enhancing phagocytosis of tumor cells by macrophages and inhibiting the metastatic growth of tumors, offering a new approach to cancer treatment (Figure 3A). Lydia Alvarez-Erviti et al. (172) achieved therapeutic effects for Alzheimer’s disease by modifying exosomes from dendritic cells to deliver therapeutic siRNA drugs, specifically knocking down the expression of beta-amyloid precursor protein 1 (BACE1). LAMP2B fused with a neuron-specific RVG3 peptide mediated the treatment of neurodegenerative diseases, as shown in Figure 3B.

Figure 3

Additionally, Yan Lin et al. (173) fused HSTP1 with exosome membrane protein LAMP2B and expressed it on the surface of exosomes through genetic engineering. Engineered exosomes (HSTP1-Exos) were more efficiently internalized by hepatic stellate cells (HSC-T6). HSTP1 is a reliable targeting peptide that specifically binds to activated hepatic stellate cells (aHSC). Exosomes modified with HSTP1 achieved precise treatment of aHSC in complex liver tissues, providing a new approach for the clinical treatment of liver fibrosis (Figure 3C). Currently, preclinical studies on the use of exosomal membrane proteins for disease treatment have achieved many successes (174–179), laying a solid foundation for the further development of clinical trials (178–187). Benjamin Besse et al. conducted a phase II clinical trial using dendritic cell-derived exosomes carrying MHC-I and MHC-II and loaded with IFN-γ (IFN-γ-Dex) to treat non-small cell lung cancer (NSCLC) patients, confirming the ability of Dex to enhance NK cell anti-tumor immunity in advanced NSCLC patients (188). Shengming Dai et al. conducted a phase I clinical trial using exosomes with surface-expressed MHC molecules and heat shock proteins (HSPs) derived from autologous ascites (Aex) combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) to treat colorectal cancer, showing that Aex combined with GM-CSF can induce specific anti-tumor cytotoxic T lymphocyte (CTL) responses (189). Table 4 lists the clinical trials involving exosomal membrane proteins (190, 191).

Table 4

Study TitleConditionsStudy TypeStart datePhaseNCT Number
LRRK2 and other novel exosome proteins in Parkinson's disease: biomarkers associated with Parkinson's disease susceptibility and/or progression in exosome-proteomes derivedParkinson's DiseaseObservational2013–01-01Not ApplicableNCT01860118
Study to measure the expression of the HER2-HER3 dimer in tumor and blood (exosomes) samples from patients with HER2 positive breast cancer receiving HER2 targeted therapiesHER2-positive Breast CancerObservational2019–12-20Not ApplicableNCT04288141
Pilot study with the aim to quantify a stress protein in the blood and in the urine for the monitoring and early diagnosis of malignant solid tumors: concentration of HSP70 exosomes in the blood and urineCancerInterventional2015–12-15Not ApplicableNCT02662621
Identification in blood sample of new diagnostic protein markers derived from circulating tumor exosomes for colorectal cancerColorectal CancerObservational2021–01-07Not ApplicableNCT04394572
Exosomes and Immunotherapy in Non-Hodgkin B-cell Lymphomas (ExoReBLy)Lymphoma, B-cell, Aggressive Non-Hodgkin (B-NHL)Interventional2019–07-02Not ApplicableNCT03985696
Analysis of Circulating Exosomes in Melanoma Patients (EXOMEL1)MelanomaObservational2019–03-01Not ApplicableNCT05744076
Safety and efficacy of EXO-CD24 in preventing clinical deterioration in patients with mild–moderate acute respiratory distress syndromeARDSInterventional2023–07-04Phase 2NCT05947747
Safety and Efficacy of Exosomes Overexpressing CD24 in Two Doses for Patients with Moderate or Severe COVID‐19Covid19Interventional2021–06-09Phase 2NCT04902183
Evaluation of the Safety of CD24‐Exosomes in Patients With COVID‐19 InfectionSARS-CoV-2Interventional2020–09-25Phase 1NCT04747574
A Phase II Randomized, Double‐blind, Placebo‐controlled Study to Evaluate the Safety and Efficacy of Exosomes Overexpressing CD24 to Prevent Clinical Deterioration in Patients with Moderate or Severe COVID‐19 InfectionCOVID-19 DiseaseInterventional2021–07-11Phase 2NCT04969172

Clinical trials using exosomal membrane proteins as primary outcome measures from 2013 to 2024.

Additionally, before the clinical application of exosomal membrane proteins, issues related to exosome isolation and comprehensive characterization must be addressed (192–194). The lack of standardized procedures for exosome isolation, proper quality control, and consistent characterization methods can hinder the clinical development of exosomes and limit their analysis in standard clinical laboratories (192, 194, 195). Table 5 lists some commonly used methods for exosome isolation and supplements these methods with their advantages and disadvantages.

Table 5

MethodApproachAdvantagesDisadvantagesReferences
Density-gradient ultracentrifugation (dUC)Combining centrifugal force and density gradient media, including iodixanol, to separate exosomes based on buoyant density. Centrifugation is typically performed at 100,000–120,000 g for 16 hoursCan handle large sample volumesPotential loss of exosomes may occur(20, 196, 197)
Immunoaffinity-based captureUsing particles with bound antibodies to specifically bind exosomesHigh specificityLack of standardization, requiring specific exosomal markers(198, 199)
Polymer based
precipitation		Employing polymer particles, such as polyethylene glycol (PEG), to isolate exosomes from the solutionImproves separation efficiency with commercially available instrumentsCo-precipitation of non-exosomal materials(200)
Size exclusion
chromatography (SEC)		Utilizing the elution time of substances in a column to separate exosomes based on sizeGood integrity of isolated exosomes, low cost using chromatography columnsNon-specific isolation leading to contamination by non-exosomal substances(201, 202)
Tangential-flow filtration (TFF) for exosome isolationCapturing exosomes by passing exosome-containing fluids through filters with membrane poresSupernatant can be concentrated and filtered simultaneously, and has been used for 3D cultureSecondary filtration needed to improve yield(200, 203, 204)
Ultra-centrifugationUsing an ultracentrifuge (100,000–110,000 g, 16–18 hours) to extract exosomes from the supernatantProcesses large sample volumes, simple operationTime-consuming, protein precipitation may disrupt exosome structure(205–207)
Hydrostatic Filtration Dialysis (HFD)Placing the supernatant in a dialysis membrane (1000 kPa) to be separated based on hydrostatic pressure differencesIsolates intact exosomes from highly diluted solutions without the need for ultracentrifugationTime-consuming, costly, with potential exosome loss(208)
Microfluidic-Based IsolationIncluding immunoaffinity capture of exosomes, nanoporous membrane filtration, or microcolumn nanocapture of exosomesHigh specificity, reproducibility, short separation time, low separation costComplex operation(209)
Antibody-coated magnetic beadsAttaching monoclonal antibodies to the surface of immunomagnetic beads to specifically bind exosomesCan select and extract specific subpopulations from samples based on specific marker expression, regardless of particle sizeDifficult separation of exosomes from magnetic beads, requiring appropriate analytical tools for exosome analysis(18, 210)

Common exosome isolation methods and their advantages and disadvantages.

3.5 Immunomodulatory role of exosome membrane proteins

Previous literature has reported the role of exosomes in immune responses (211–221), primarily mediated by membrane proteins. For instance, the expression of PD-L1 on the surface of exosomes has been confirmed, and its abundance on exosomes is related to the progression of host tumors (38, 222–224). In 2022, Yunxing Lu et al. proposed an integrated microfluidic system for exosome isolation and detection (EXID system) to analyze the abundance of exosome PD-L1 protein markers. The study suggested that the abundance of PD-L1 reflects sensitivity to immune responses, and exosomes containing PD-L1 weaken anti-tumor immunity in the tumor microenvironment (225). Meizhang Li et al. indicated that exosomes derived from Wharton’s Jelly mesenchymal stem cells (WJMSCs) enhance T-cell inhibitory effects through the carried PD-L1, contributing to alleviating immune rejection in organ transplantation, as shown in Figure 4C (226). Furthermore, research results indicate that blocking exosome PD-L1 secretion significantly contributes to anti-tumor immune responses. Inhibiting exosome secretion combined with anti-PD-L1 therapy may enhance clinical anti-tumor effects (227).

Figure 4

Recently, Wei Zhang et al. (228) identified three classes of immunosuppressive membrane proteins expressed by syncytiotrophoblast-derived exosomes. These include NKG2D ligands (MICA/B, ULBP1–5/RAET1), oligomerization-induced apoptosis ligands (FASL, TRAIL), and immune checkpoint molecules interacting with PD-1 (PD-L1/B7-H1/CD274, PD-L2/B7-H2/CD273). The delivery of these immunosuppressive membrane protein signals by exosomes regulates the maternal immune system and promotes the development of maternal-fetal tolerance, as depicted in Figure 4A. Exosomes derived from dendritic cells express MHC-I, MHC-II, and immune co-stimulatory molecules CD80 and CD86 on their membrane surfaces, promoting T-cell activation and proliferation and regulating the body’s immune mechanisms (8), as shown in Figure 4B. Previous studies have indicated that MHC-II molecules transferred to recipient dendritic cells through exosomes activate CD4+ T cells. Similarly, MHC-I molecules transferred to dendritic cells through exosomes contribute to the activation of CD8+ T cells (229, 230). In addition, exosome membrane proteins derived from immune cells can influence the development of cancer cells (217), Figure 5. For immunosuppressive molecules expressed on the exosome membrane, blockade can be achieved by incorporating corresponding antibodies, while immune-activating molecules can be applied in clinical therapy.

Figure 5

3.6 Other functions of exosome membrane proteins

In addition to their role in diagnosing diseases, regulating the body’s immune system, and serving as biological carriers targeting receptor cells, exosome membrane proteins also possess other functionalities. Upon generation, exosomes interact with proteins circulating in the surrounding environment, leading to the formation of a “protein corona” (PC). This formation alters the properties of exosomes and influences their functionality within the body (231–233). The protein corona enhances the stability of exosomes, prolonging their circulation lifespan in the body. This protection shields exosomes from degradation and clearance, thereby increasing their survival time in vivo (234, 235).

Furthermore, the presence of the protein corona can impact the interaction between exosomes and target cells. Specific protein coronas may facilitate adhesion and uptake between exosomes and target cells, mediating the entry of biologically active substances released by exosomes into recipient cells (234, 236). In conclusion, research on exosome membrane proteins is ongoing, and the exploration of their functions is expected to deepen.

4 Summary and outlook

With the increasing understanding of exosome membrane proteins, more functionalities of these proteins are gradually coming to light. In addition to the roles mentioned in this article, such as diagnosis and immune regulation, exosome membrane proteins can be redesigned or modified, significantly enriching their functions. This diversity opens up vast potential applications for exosome membrane proteins in the future, making them a focal point of current research. Despite the extensive research on exosome membrane proteins, many proteins on the exosome membrane still have undetermined functions, requiring further in-depth investigation. Moreover, since exosome membrane proteins vary depending on the cell source, it is essential to study them in the context of their origin to obtain more accurate results (125, 133).

Furthermore, membrane proteins of exosomes have garnered significant interest in clinical trials for disease diagnosis and therapy. However, achieving a range of functions in clinical settings remains challenging for researchers (210, 237). To advance the clinical translation of exosomes, several key issues need to be addressed. These include: 1. The need for standardized methods to isolate, characterize, and quantify exosomes to ensure their stability and reproducibility; 2. Developing rigorous preclinical biosafety evaluation protocols to mitigate risks before human trials; 3. Conducting pilot clinical studies to demonstrate feasibility, biological distribution in humans, and preliminary efficacy before large-scale applications (13, 20, 24, 238).

Although researchers from different fields have explored exosome membrane proteins, gaining varying degrees of understanding of protein types and biological functions, the intricate environment within the body poses the need for further exploration and explanation of membrane protein-mediated mechanisms.

Statements

Author contributions

FX: Resources, Methodology, Formal analysis, Writing – original draft, Data curation. SL: Writing – original draft, Methodology. PL: Writing – review & editing, Formal analysis. CC: Investigation, Formal analysis, Writing – review & editing. WL: Writing – review & editing, Resources, Funding acquisition. CL: Visualization, Writing – review & editing.

Funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the Natural Science Foundation of Beijing (7212172), National Natural Science Foundation of China (82274447), High-Level Public Health Technical Talents Project of Beijing (2022–2-024), Beijing Municipal Public Welfare Development and Reform Pilot Project for Medical Research Institutes (JYY2021–10).

Acknowledgments

We sincerely appreciate the invaluable guidance and insightful suggestions provided by WL, CL, and CC during the writing process of this article. Their expertise and support greatly enhanced the quality of our work.

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Publisher’s note

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.

References

  • 1

    ColomboMMoitaCvan NielGKowalJVigneronJBenarochPet al. Analysis of ESCRT functions in exosome biogenesis, composition and secretion highlights the heterogeneity of extracellular vesicles. J Cell Sci. (2013) 126:5553–65. doi: 10.1242/jcs.128868

  • 2

    LariosJMercierVRouxAGruenbergJ. ALIX- and ESCRT-III-dependent sorting of tetraspanins to exosomes. J Cell Biol. (2020) 219. doi: 10.1083/jcb.201904113

  • 3

    OstrowskiMCarmoNBKrumeichSFangetIRaposoGSavinaAet al. Rab27a and Rab27b control different steps of the exosome secretion pathway. Nat Cell Biol. (2010) 12:19–30;sup pp 11-13. doi: 10.1038/ncb2000

  • 4

    WeiDZhanWGaoYHuangLGongRWangWet al. RAB31 marks and controls an ESCRT-independent exosome pathway. Cell Res. (2021) 31:157–77. doi: 10.1038/s41422-020-00409-1

  • 5

    LiXLiCZhangLWuMCaoKJiangFet al. The significance of exosomes in the development and treatment of hepatocellular carcinoma. Mol Cancer. (2020) 19:1. doi: 10.1186/s12943-019-1085-0

  • 6

    NooninCThongboonkerdV. Exosome-inflammasome crosstalk and their roles in inflammatory responses. Theranostics. (2021) 11:4436–51. doi: 10.7150/thno.54004

  • 7

    LiLHuangLHuangCXuJHuangYLuoHet al. The multiomics landscape of serum exosomes during the development of sepsis. J Adv Res. (2022) 39:203–23. doi: 10.1016/j.jare.2021.11.005

  • 8

    KalluriRLeBleuVS. The biology, function, and biomedical applications of exosomes. Science. (2020) 367. doi: 10.1126/science.aau6977

  • 9

    SunDZhuangXZhangSDengZBGrizzleWMillerDet al. Exosomes are endogenous nanoparticles that can deliver biological information between cells. Adv Drug Delivery Rev. (2013) 65:342–7. doi: 10.1016/j.addr.2012.07.002

  • 10

    VlassovAVMagdalenoSSetterquistRConradR. Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. Biochim Biophys Acta. (2012) 1820:940–8. doi: 10.1016/j.bbagen.2012.03.017

  • 11

    BangCThumT. Exosomes: new players in cell-cell communication. Int J Biochem Cell Biol. (2012) 44:2060–4. doi: 10.1016/j.biocel.2012.08.007

  • 12

    EL AndaloussiSMagerIBreakefieldXOWoodMJ. Extracellular vesicles: biology and emerging therapeutic opportunities. Nat Rev Drug Discov. (2013) 12:347–57. doi: 10.1038/nrd3978

  • 13

    MahlLOlligJSchweihoferVWiegandLTorklerPHaerteisSet al. Importance and implications of exosomes in nephrology and urology. Pflugers Arch. (2023) 475:153–66. doi: 10.1007/s00424-022-02771-y

  • 14

    PecheVSPietkaTAJacome-SosaMSamovskiDPalaciosHChatterjee-BasuGet al. Endothelial cell CD36 regulates membrane ceramide formation, exosome fatty acid transfer and circulating fatty acid levels. Nat Commun. (2023) 14:4029. doi: 10.1038/s41467-023-39752-3

  • 15

    Gebeyehu AKNMeckesDGJrSachdevaMS. Role of exosomes for delivery of chemotherapeutic drugs. Crit Rev Ther Drug Carrier Syst. (2021) 38:53–97. doi: 10.1615/CritRevTherDrugCarrierSyst

  • 16

    AlenquerMAmorimMJ. Exosome biogenesis, regulation, and function in viral infection. Viruses. (2015) 7:5066–83. doi: 10.3390/v7092862

  • 17

    JeppesenDKFenixAMFranklinJLHigginbothamJNZhangQZimmermanLJet al. Reassessment of exosome composition. Cell. (2019) 177:428–445.e418. doi: 10.1016/j.cell.2019.02.029

  • 18

    RaghavATripathiPMishraBKJeongGBBandaySGautamKAet al. Mesenchymal stromal cell-derived tailored exosomes treat bacteria-associated diabetes foot ulcers: A customized approach from bench to bed. Front Microbiol. (2021) 12:712588. doi: 10.3389/fmicb.2021.712588

  • 19

    GurungSPerocheauDTouramanidouLBaruteauJ. The exosome journey: from biogenesis to uptake and intracellular signalling. Cell Commun Signal. (2021) 19:47. doi: 10.1186/s12964-021-00730-1

  • 20

    LopezKLaiSWTLopez GonzalezEJR.G.DShuckSC. Extracellular vesicles: A dive into their role in the tumor microenvironment and cancer progression. Front Cell Dev Biol. (2023) 11:1154576. doi: 10.3389/fcell.2023.1154576

  • 21

    ChiangCYChenC. Toward characterizing extracellular vesicles at a single-particle level. J BioMed Sci. (2019) 26:9. doi: 10.1186/s12929-019-0502-4

  • 22

    HartjesTAMytnykSJensterGWvan SteijnVvan RoyenME. Extracellular vesicle quantification and characterization: common methods and emerging approaches. Bioengineering (Basel). (2019) 6:7-32. doi: 10.3390/bioengineering6010007

  • 23

    TheryCWitwerKWAikawaEAlcarazMJAndersonJDAndriantsitohainaRet al. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. (2018) 7:1535750. doi: 10.1080/20013078.2018.1535750

  • 24

    ButreddyAKommineniNDudhipalaN. Exosomes as naturally occurring vehicles for delivery of biopharmaceuticals: insights from drug delivery to clinical perspectives. Nanomaterials (Basel). (2021) 11:1481-523. doi: 10.3390/nano11061481

  • 25

    DaaboulGGGagniPBenussiLBettottiPCianiMCretichMet al. Digital detection of exosomes by interferometric imaging. Sci Rep. (2016) 6:37246. doi: 10.1038/srep37246

  • 26

    MironRJZhangY. Understanding exosomes: Part 1-Characterization, quantification and isolation techniques. Periodontol 2000. (2024) 94:231–56. doi: 10.1111/prd.12520

  • 27

    FangXDuanYAdkinsGBPanSWangHLiuYet al. Highly efficient exosome isolation and protein analysis by an integrated nanomaterial-based platform. Anal Chem. (2018) 90:2787–95. doi: 10.1021/acs.analchem.7b04861

  • 28

    FordjourFKGuoCAiYDaaboulGGGouldSJ. A shared, stochastic pathway mediates exosome protein budding along plasma and endosome membranes. J Biol Chem. (2022) 298:102394. doi: 10.1016/j.jbc.2022.102394

  • 29

    SongZMaoJBarreroRAWangPZhangFWangT. Development of a CD63 aptamer for efficient cancer immunochemistry and immunoaffinity-based exosome isolation. Molecules. (2020) 25:5585-600. doi: 10.3390/molecules25235585

  • 30

    EscolaJMKleijmeerMJStoorvogelWGriffithJMYoshieOGeuzeHJ. Selective enrichment of tetraspan proteins on the internal vesicles of multivesicular endosomes and on exosomes secreted by human B-lymphocytes. J Biol Chem. (1998) 273:20121–7. doi: 10.1074/jbc.273.32.20121

  • 31

    PittJMAndreFAmigorenaSSoriaJCEggermontAKroemerGet al. Dendritic cell-derived exosomes for cancer therapy. J Clin Invest. (2016) 126:1224–32. doi: 10.1172/JCI81137

  • 32

    SkokosDBotrosHGDemeureCMorinJPeronetRBirkenmeierGet al. Mast cell-derived exosomes induce phenotypic and functional maturation of dendritic cells and elicit specific immune responses in vivo. J Immunol. (2003) 170:3037–45. doi: 10.4049/jimmunol.170.6.3037

  • 33

    ParkSAhnESKimY. Neuroblastoma SH-SY5Y cell-derived exosomes stimulate dendrite-like outgrowths and modify the differentiation of A375 melanoma cells. Cell Biol Int. (2015) 39:379–87. doi: 10.1002/cbin.10401

  • 34

    SeguraENiccoCLombardBVeronPRaposoGBatteuxFet al. ICAM-1 on exosomes from mature dendritic cells is critical for efficient naive T-cell priming. Blood. (2005) 106:216–23. doi: 10.1182/blood-2005-01-0220

  • 35

    RoucourtBMeeussenSBaoJZimmermannPDavidG. Heparanase activates the syndecan-syntenin-ALIX exosome pathway. Cell Res. (2015) 25:412–28. doi: 10.1038/cr.2015.29

  • 36

    HoshinoACosta-SilvaBShenTLRodriguesGHashimotoATesic MarkMet al. Tumour exosome integrins determine organotropic metastasis. Nature. (2015) 527:329–35. doi: 10.1038/nature15756

  • 37

    KleijmeerMJEscolaJMUytdeHaagFGJakobsonEGriffithJMOsterhausADet al. Antigen loading of MHC class I molecules in the endocytic tract. Traffic. (2001) 2:124–37. doi: 10.1034/j.1600-0854.2001.020207.x

  • 38

    ChenGHuangACZhangWZhangGWuMXuWet al. Exosomal PD-L1 contributes to immunosuppression and is associated with anti-PD-1 response. Nature. (2018) 560:382–6. doi: 10.1038/s41586-018-0392-8

  • 39

    TheryCBoussacMVeronPRicciardi-CastagnoliPRaposoGGarinJet al. Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles. J Immunol. (2001) 166:7309–18. doi: 10.4049/jimmunol.166.12.7309

  • 40

    XuRGreeningDWChenMRaiAJiHTakahashiNet al. Surfaceome of exosomes secreted from the colorectal cancer cell line SW480: peripheral and integral membrane proteins analyzed by proteolysis and TX114. Proteomics. (2019) 19:e1700453. doi: 10.1002/pmic.201700453

  • 41

    ClaytonAAl-TaeiSWebberJMasonMDTabiZ. Cancer exosomes express CD39 and CD73, which suppress T cells through adenosine production. J Immunol. (2011) 187:676–83. doi: 10.4049/jimmunol.1003884

  • 42

    RabesandratanaHToutantJ-PReggioHVidalM. Decay-accelerating factor (CD55) and membrane inhibitor of reactive lysis (CD59) are released within exosomes during in vitro maturation of reticulocytes. Blood. (1998) 91:2573–80. doi: 10.1182/blood.V91.7.2573

  • 43

    MeloSALueckeLBKahlertCFernandezAFGammonSTKayeJet al. Glypican-1 identifies cancer exosomes and detects early pancreatic cancer. Nature. (2015) 523:177–82. doi: 10.1038/nature14581

  • 44

    ZhengJHernandezJMDoussotABojmarLZambirinisCPCosta-SilvaBet al. Extracellular matrix proteins and carcinoembryonic antigen-related cell adhesion molecules characterize pancreatic duct fluid exosomes in patients with pancreatic cancer. HPB (Oxford). (2018) 20:597–604. doi: 10.1016/j.hpb.2017.12.010

  • 45

    AtaySGercel-TaylorCTaylorDD. Human trophoblast-derived exosomal fibronectin induces pro-inflammatory IL-1beta production by macrophages. Am J Reprod Immunol. (2011) 66:259–69. doi: 10.1111/aji.2011.66.issue-4

  • 46

    SantasusagnaSMorenoINavarroACastellanoJJMartinezFHernandezRet al. Proteomic analysis of liquid biopsy from tumor-draining vein indicates that high expression of exosomal ECM1 is associated with relapse in stage I-III colon cancer. Transl Oncol. (2018) 11:715–21. doi: 10.1016/j.tranon.2018.03.010

  • 47

    OshimaKAokiNKatoTKitajimaKMatsudaT. Secretion of a peripheral membrane protein, MFG-E8, as a complex with membrane vesicles. Eur J Biochem. (2002) 269:1209–18. doi: 10.1046/j.1432-1033.2002.02758.x

  • 48

    KochRDemantMAungTDieringNCicholasAChapuyBet al. Populational equilibrium through exosome-mediated Wnt signaling in tumor progression of diffuse large B-cell lymphoma. Blood. (2014) 123:2189–98. doi: 10.1182/blood-2013-08-523886

  • 49

    GerberPPCabriniMJancicCPaolettiLBanchioCvon BilderlingCet al. Rab27a controls HIV-1 assembly by regulating plasma membrane levels of phosphatidylinositol 4,5-bisphosphate. J Cell Biol. (2015) 209:435–52. doi: 10.1083/jcb.201409082

  • 50

    AshleyJCordyBLuciaDFradkinLGBudnikVThomsonT. Retrovirus-like gag protein arc1 binds RNA and traffics across synaptic boutons. Cell. (2018) 172:262–274.e211. doi: 10.1016/j.cell.2017.12.022

  • 51

    WubboltsRLeckieRSVeenhuizenPTSchwarzmannGMobiusWHoernschemeyerJet al. Proteomic and biochemical analyses of human B cell-derived exosomes. Potential implications for their function and multivesicular body formation. J Biol Chem. (2003) 278:10963–72. doi: 10.1074/jbc.M207550200

  • 52

    ShenBFangYWuNGouldSJ. Biogenesis of the posterior pole is mediated by the exosome/microvesicle protein-sorting pathway. J Biol Chem. (2011) 286:44162–76. doi: 10.1074/jbc.M111.274803

  • 53

    HegmansJPBardMPHemmesALuiderTMKleijmeerMJPrinsJBet al. Proteomic analysis of exosomes secreted by human mesothelioma cells. Am J Pathol. (2004) 164:1807–15. doi: 10.1016/S0002-9440(10)63739-X

  • 54

    LatyshevaNMuratovGRajeshSPadgettMHotchinNAOverduinMet al. Syntenin-1 is a new component of tetraspanin-enriched microdomains: mechanisms and consequences of the interaction of syntenin-1 with CD63. Mol Cell Biol. (2006) 26:7707–18. doi: 10.1128/MCB.00849-06

  • 55

    ChalminFLadoireSMignotGVincentJBruchardMRemy-MartinJPet al. Membrane-associated Hsp72 from tumor-derived exosomes mediates STAT3-dependent immunosuppressive function of mouse and human myeloid-derived suppressor cells. J Clin Invest. (2010) 120:457–71. doi: 10.1172/JCI40483

  • 56

    LaiRCArslanFLeeMMSzeNSChooAChenTSet al. Exosome secreted by MSC reduces myocardial ischemia/reperfusion injury. Stem Cell Res. (2010) 4:214–22. doi: 10.1016/j.scr.2009.12.003

  • 57

    RaimondoFMorosiLChinelloCMagniFPittoM. Advances in membranous vesicle and exosome proteomics improving biological understanding and biomarker discovery. Proteomics. (2011) 11:709–20. doi: 10.1002/pmic.201000422

  • 58

    HeCZhengSLuoYWangB. Exosome theranostics: biology and translational medicine. Theranostics. (2018) 8:237–55. doi: 10.7150/thno.21945

  • 59

    IwaiMHamataniTNakamuraAKawanoNKanaiSKangWet al. Membrane protein CD9 is repositioned and released to enhance uterine function. Lab Invest. (2019) 99:200–9. doi: 10.1038/s41374-018-0145-1

  • 60

    BardMPHegmansJPHemmesALuiderTMWillemsenRSeverijnenLAet al. Proteomic analysis of exosomes isolated from human Malignant pleural effusions. Am J Respir Cell Mol Biol. (2004) 31:114–21. doi: 10.1165/rcmb.2003-0238OC

  • 61

    SimpsonRJLimJWMoritzRLMathivananS. Exosomes: proteomic insights and diagnostic potential. Expert Rev Proteomics. (2009) 6:267–83. doi: 10.1586/epr.09.17

  • 62

    XiaoYZhongJZhongBHuangJJiangLJiangYet al. Exosomes as potential sources of biomarkers in colorectal cancer. Cancer Lett. (2020) 476:13–22. doi: 10.1016/j.canlet.2020.01.033

  • 63

    Ai YGCGarcia-ContrerasMSánchez BLSSafticsAShodubiORaghunandanSet al. Endocytosis blocks the vesicular secretion of exosome marker proteins. Sci Adv. (2024) 10:eadi9156. doi: 10.1126/sciadv.adi9156

  • 64

    CharrinSJouannetSBoucheixCRubinsteinE. Tetraspanins at a glance. J Cell Sci. (2014) 127:3641–8. doi: 10.1242/jcs.154906

  • 65

    LudwigNYerneniSSAzambujaJHGillespieDGMenshikovaEVJacksonEKet al. Tumor-derived exosomes promote angiogenesis via adenosine A(2B) receptor signaling. Angiogenesis. (2020) 23:599–610. doi: 10.1007/s10456-020-09728-8

  • 66

    HurYHFengSWilsonKFCerioneRAAntonyakMA. Embryonic stem cell-derived extracellular vesicles maintain ESC stemness by activating FAK. Dev Cell. (2021) 56:277–291.e276. doi: 10.1016/j.devcel.2020.11.017

  • 67

    SongLTangSHanXJiangZDongLLiuCet al. KIBRA controls exosome secretion via inhibiting the proteasomal degradation of Rab27a. Nat Commun. (2019) 10:1639. doi: 10.1038/s41467-019-09720-x

  • 68

    PantSHiltonHBurczynskiME. The multifaceted exosome: biogenesis, role in normal and aberrant cellular function, and frontiers for pharmacological and biomarker opportunities. Biochem Pharmacol. (2012) 83:1484–94. doi: 10.1016/j.bcp.2011.12.037

  • 69

    GhossoubRCheryMAudebertSLeblancREgea-JimenezALLemboFet al. Tetraspanin-6 negatively regulates exosome production. Proc Natl Acad Sci U.S.A. (2020) 117:5913–22. doi: 10.1073/pnas.1922447117

  • 70

    NazarenkoIRanaSBaumannAMcAlearJHellwigATrendelenburgMet al. Cell surface tetraspanin Tspan8 contributes to molecular pathways of exosome-induced endothelial cell activation. Cancer Res. (2010) 70:1668–78. doi: 10.1158/0008-5472.CAN-09-2470

  • 71

    Wu FYZYangLFanJXuJJinYYuJet al. Smoking induced extracellular vesicles release and their distinct properties in non-small cell lung cancer. J Cancer. (2019) 10:3435–43. doi: 10.7150/jca.30425

  • 72

    Lyu QZXShiLQChenHYLuMMaXDRenL. Exosomes may be the carrier of acupuncture treatment for major depressive disorder. Front Behav Neurosci. (2023) 17:1107265. doi: 10.3389/fnbeh.2023.1107265

  • 73

    Hu SMJSuCChenYShuYQiZZhangBet al. Engineered exosome-like nanovesicles suppress tumor growth by reprogramming tumor microenvironment and promoting tumor ferroptosis. Acta Biomater. (2021) 135:567–81. doi: 10.1016/j.actbio.2021.09.003

  • 74

    Boukouris SMS. Exosomes in bodily fluids are a highly stable resource of disease biomarkers. Proteomics Clin Appl. (2015) 9:358–67. doi: 10.1002/prca.201400114

  • 75

    Penfornis PVKWhittJPochampallyR. Extracellular vesicles as carriers of microRNA, proteins and lipids in tumor microenvironment. Int J Cancer. (2016) 138:14–21. doi: 10.1002/ijc.29417

  • 76

    Cheng JWXYuanXLiuGChuQ. Emerging roles of exosome-derived biomarkers in cancer theranostics: messages from novel protein targets. Am J Cancer Res. (2022) 12:2226–48.

  • 77

    Araujo-AbadSSacedaMde Juan RomeroC. Biomedical application of small extracellular vesicles in cancer treatment. Adv Drug Delivery Rev. (2022) 182:114117. doi: 10.1016/j.addr.2022.114117

  • 78

    ChoiE-SFaruqueHAKimJ-HKimKJChoiJEKimBAet al. CD5L as an extracellular vesicle-derived biomarker for liquid biopsy of lung cancer. Diagnostics. (2021) 11:620-37. doi: 10.3390/diagnostics11040620

  • 79

    Khan NAAMEl-MenyarABiswasKHRizoliSAl-ThaniH. The evolving role of extracellular vesicles (exosomes) as biomarkers in traumatic brain injury: Clinical perspectives and therapeutic implications. Front Aging Neurosci. (2022) 14:933434. doi: 10.3389/fnagi.2022.933434

  • 80

    LiYZhangYQiuFQiuZ. Proteomic identification of exosomal LRG1: a potential urinary biomarker for detecting NSCLC. Electrophoresis. (2011) 32:1976–83. doi: 10.1002/elps.201000598

  • 81

    SakaueTKogaHIwamotoHNakamuraTIkezonoYAbeMet al. Glycosylation of ascites-derived exosomal CD133: a potential prognostic biomarker in patients with advanced pancreatic cancer. Med Mol Morphol. (2019) 52:198–208. doi: 10.1007/s00795-019-00218-5

  • 82

    Sandfeld-PaulsenBJakobsenKRBaekRFolkersenBHRasmussenTRMeldgaardPet al. Exosomal proteins as diagnostic biomarkers in lung cancer. J Thorac Oncol. (2016) 11:1701–10. doi: 10.1016/j.jtho.2016.05.034

  • 83

    Sandúa AAEGonzálezÁ. Exosomes in lung cancer: actors and heralds of tumor development. Cancers (Basel). (2021) 13:4330. doi: 10.3390/cancers13174330

  • 84

    XylakiMChopraAWeberSBartlMOuteiroTFMollenhauerB. Extracellular vesicles for the diagnosis of parkinson’s disease: systematic review and meta-analysis. Mov Disord. (2023) 38:1585–97. doi: 10.1002/mds.29497

  • 85

    ChenIHXueLHsuCCPaezJSPanLAndaluzHet al. Phosphoproteins in extracellular vesicles as candidate markers for breast cancer. Proc Natl Acad Sci USA. (2017) 114:3175–80. doi: 10.1073/pnas.1618088114

  • 86

    JiangLDongHCaoHJiXLuanSLiuJ. Exosomes in pathogenesis, diagnosis, and treatment of alzheimer’s disease. Med Sci Monit. (2019) 25:3329–35. doi: 10.12659/MSM.914027

  • 87

    LaiRCYeoRWTanKHLimSK. Exosomes for drug delivery - a novel application for the mesenchymal stem cell. Biotechnol Adv. (2013) 31:543–51. doi: 10.1016/j.biotechadv.2012.08.008

  • 88

    MinLZhuSChenLLiuXWeiRZhaoLet al. Evaluation of circulating small extracellular vesicles derived miRNAs as biomarkers of early colon cancer: a comparison with plasma total miRNAs. J Extracell Vesicles. (2019) 8:1643670. doi: 10.1080/20013078.2019.1643670

  • 89

    JakobsenKRPaulsenBSBaekRVarmingKSorensenBSJorgensenMM. Exosomal proteins as potential diagnostic markers in advanced non-small cell lung carcinoma. J Extracell Vesicles. (2015) 4:26659. doi: 10.3402/jev.v4.26659

  • 90

    ChangLWuHChenRSunXYangYHuangCet al. Microporous PdCuB nanotag-based electrochemical aptasensor with Au@CuCl(2) nanowires interface for ultrasensitive detection of PD-L1-positive exosomes in the serum of lung cancer patients. J Nanobiotechnology. (2023) 21:86. doi: 10.1186/s12951-023-01845-y

  • 91

    Keryer-BibensCPioche-DurieuCVillemantCSouquereSNishiNHirashimaMet al. Exosomes released by EBV-infected nasopharyngeal carcinoma cells convey the viral latent membrane protein 1 and the immunomodulatory protein galectin 9. BMC Cancer. (2006) 6:283. doi: 10.1186/1471-2407-6-283

  • 92

    HoualiKWangXShimizuYDjennaouiDNichollsJFioriniSet al. A new diagnostic marker for secreted Epstein-Barr virus encoded LMP1 and BARF1 oncoproteins in the serum and saliva of patients with nasopharyngeal carcinoma. Clin Cancer Res. (2007) 13:4993–5000. doi: 10.1158/1078-0432.CCR-06-2945

  • 93

    DengFMillerJ. A review on protein markers of exosome from different bio-resources and the antibodies used for characterization. J Histotechnol. (2019) 42:226–39. doi: 10.1080/01478885.2019.1646984

  • 94

    KoksalARThevenotPAydinYNunezKSandowTWidmerKet al. Impaired autophagy response in hepatocellular carcinomas enriches glypican-3 in exosomes, not in the microvesicles. J Hepatocell Carcinoma. (2022) 9:959–72. doi: 10.2147/JHC.S376210

  • 95

    ArbelaizAAzkargortaMSantos-LasoÁPerugorriaMJEriceOGonzalezEet al. Serum extracellular vesicles contain protein biomarkers for primary sclerosing cholangitis and cholangiocarcinoma. J Hepatol. (2017) 66:S207–8. doi: 10.1016/S0168-8278(17)30712-2

  • 96

    WangXShenHZhangyuanGHuangRZhangWHeQet al. 14-3-3zeta delivered by hepatocellular carcinoma-derived exosomes impaired anti-tumor function of tumor-infiltrating T lymphocytes. Cell Death Dis. (2018) 9:159. doi: 10.1038/s41419-017-0180-7

  • 97

    WelkerMWReichertDSusserSSarrazinCMartinezYHerrmannEet al. Soluble serum CD81 is elevated in patients with chronic hepatitis C and correlates with alanine aminotransferase serum activity. PloS One. (2012) 7:e30796. doi: 10.1371/journal.pone.0030796

  • 98

    CastilloJBernardVSan LucasFAAllensonKCapelloMKimDUet al. Surfaceome profiling enables isolation of cancer-specific exosomal cargo in liquid biopsies from pancreatic cancer patients. Ann Oncol. (2018) 29:223–9. doi: 10.1093/annonc/mdx542

  • 99

    FanJWeiQKoayEJLiuYNingBBernardPWet al. Chemoresistance transmission via exosome-mediated ephA2 transfer in pancreatic cancer. Theranostics. (2018) 8:5986–94. doi: 10.7150/thno.26650

  • 100

    KimuraHYamamotoHHaradaTFumotoKOsugiYSadaRet al. CKAP4, a DKK1 receptor, is a biomarker in exosomes derived from pancreatic cancer and a molecular target for therapy. Clin Cancer Res. (2019) 25:1936–47. doi: 10.1158/1078-0432.CCR-18-2124

  • 101

    YoshiokaYKosakaNKonishiYOhtaHOkamotoHSonodaHet al. Ultra-sensitive liquid biopsy of circulating extracellular vesicles using ExoScreen. Nat Commun. (2014) 5:3591. doi: 10.1038/ncomms4591

  • 102

    AnamiKOueNNoguchiTSakamotoNSentaniKHayashiTet al. TSPAN8, identified by Escherichia coli ampicillin secretion trap, is associated with cell growth and invasion in gastric cancer. Gastric Cancer. (2016) 19:370–80. doi: 10.1007/s10120-015-0478-z

  • 103

    Fraser KBMMAlcalayRNWestAB. Urinary LRRK2 phosphorylation predicts parkinsonian phenotypes in G2019S LRRK2 carriers. Neurology. (2016) 86:994–9. doi: 10.1212/WNL.0000000000002436

  • 104

    SkogJWurdingerTvan RijnSMeijerDHGaincheLSena-EstevesMet al. Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat Cell Biol. (2008) 10:1470–6. doi: 10.1038/ncb1800

  • 105

    ShaoHChungJBalajLCharestABignerDDCarterBSet al. Protein typing of circulating microvesicles allows real-time monitoring of glioblastoma therapy. Nat Med. (2012) 18:1835–40. doi: 10.1038/nm.2994

  • 106

    RaimondoFMorosiLCorbettaSChinelloCBrambillaPDella MinaPet al. Differential protein profiling of renal cell carcinoma urinary exosomes. Mol Biosyst. (2013) 9:1220–33. doi: 10.1039/c3mb25582d

  • 107

    LiTLiuTCLiuNLiMJZhangM. Urinary exosome proteins PAK6 and EGFR as noninvasive diagnostic biomarkers of diabetic nephropathy. BMC Nephrol. (2023) 24:291. doi: 10.1186/s12882-023-03343-7

  • 108

    WeltonJLKhannaSGilesPJBrennanPBrewisIAStaffurthJet al. Proteomics analysis of bladder cancer exosomes. Mol Cell Proteomics. (2010) 9:1324–38. doi: 10.1074/mcp.M000063-MCP201

  • 109

    WangYTShiTSrivastavaSKaganJLiuTRodlandKD. Proteomic analysis of exosomes for discovery of protein biomarkers for prostate and bladder cancer. Cancers (Basel). (2020) 12:2335-53. doi: 10.3390/cancers12092335

  • 110

    NilssonJSkogJNordstrandABaranovVMincheva-NilssonLBreakefieldXOet al. Prostate cancer-derived urine exosomes: a novel approach to biomarkers for prostate cancer. Br J Cancer. (2009) 100:1603–7. doi: 10.1038/sj.bjc.6605058

  • 111

    LiangBPengPChenSLiLZhangMCaoDet al. Characterization and proteomic analysis of ovarian cancer-derived exosomes. J Proteomics. (2013) 80:171–82. doi: 10.1016/j.jprot.2012.12.029

  • 112

    ChanteloupGCordonnierMIsambertNBertautAMarcionGGarridoCet al. Membrane-bound exosomal HSP70 as a biomarker for detection and monitoring of Malignant solid tumours: a pilot study. Pilot Feasibility Stud. (2020) 6:35. doi: 10.1186/s40814-020-00577-2

  • 113

    ShimizuASawadaKKobayashiMYamamotoMYagiTKinoseYet al. Exosomal CD47 plays an essential role in immune evasion in ovarian cancer. Mol Cancer Res. (2021) 19:1583–95. doi: 10.1158/1541-7786.MCR-20-0956

  • 114

    Luo DZSXiaWHuangLGeWWangT. Proteomics study of serum exosomes from papillary thyroid cancer patients. Endocr Relat Cancer. (2018) 25:879–91. doi: 10.1530/ERC-17-0547

  • 115

    RishaYMinicZGhobadlooSMBerezovskiMV. The proteomic analysis of breast cell line exosomes reveals disease patterns and potential biomarkers. Sci Rep. (2020) 10:13572. doi: 10.1038/s41598-020-70393-4

  • 116

    WangXHuangJChenWLiGLiZLeiJ. The updated role of exosomal proteins in the diagnosis, prognosis, and treatment of cancer. Exp Mol Med. (2022) 54:1390–400. doi: 10.1038/s12276-022-00855-4

  • 117

    WangYGaoWSunMFengBShenHZhuJet al. A filter-electrochemical microfluidic chip for multiple surface protein analysis of exosomes to detect and classify breast cancer. Biosens Bioelectron. (2023) 239:115590. doi: 10.1016/j.bios.2023.115590

  • 118

    HanZPengXYangYYiJZhaoDBaoQet al. Integrated microfluidic-SERS for exosome biomarker profiling and osteosarcoma diagnosis. Biosens Bioelectron. (2022) 217:114709. doi: 10.1016/j.bios.2022.114709

  • 119

    ChuangHCChenMHChenYMYangHYCiouYRHsuehCHet al. BPI overexpression suppresses Treg differentiation and induces exosome-mediated inflammation in systemic lupus erythematosus. Theranostics. (2021) 11:9953–66. doi: 10.7150/thno.63743

  • 120

    LogozziMDe MilitoALuginiLBorghiMCalabroLSpadaMet al. High levels of exosomes expressing CD63 and caveolin-1 in plasma of melanoma patients. PloS One. (2009) 4:e5219. doi: 10.1371/journal.pone.0005219

  • 121

    YoshiokaYKonishiYKosakaNKatsudaTKatoTOchiya.T. Comparative marker analysis of extracellular vesicles in different human cancer types. J Extracell Vesicles. (2013) 2. doi: 10.3402/jev.v2i0.20424

  • 122

    LinBTianTLuYLiuDHuangMZhuLet al. Tracing tumor-derived exosomal PD-L1 by dual-aptamer activated proximity-induced droplet digital PCR. Angew Chem Int Ed Engl. (2021) 60:7582–6. doi: 10.1002/anie.202015628

  • 123

    YinTXuLGilBMeraliNSokolikovaMSGaboriauDCAet al. Graphene sensor arrays for rapid and accurate detection of pancreatic cancer exosomes in patients’ Blood plasma samples. ACS Nano. (2023) 17:14619–31. doi: 10.1021/acsnano.3c01812

  • 124

    LinJLiJHuangBLiuJChenXChenXMet al. Exosomes: novel biomarkers for clinical diagnosis. ScientificWorldJournal. (2015) 2015:657086. doi: 10.1155/2015/657086

  • 125

    SalunkheSDheerajBasakMChitkaraDMittalA. Surface functionalization of exosomes for target-specific delivery and in vivo imaging & tracking: Strategies and significance. J Control Release. (2020) 326:599–614. doi: 10.1016/j.jconrel.2020.07.042

  • 126

    LiangYDuanLLuJXiaJ. Engineering exosomes for targeted drug delivery. Theranostics. (2021) 11:3183–95. doi: 10.7150/thno.52570

  • 127

    TashimaT. Smart Strategies for Therapeutic Agent Delivery into Brain across the Blood-Brain Barrier Using Receptor-Mediated Transcytosis. Chem Pharm Bull (Tokyo). (2020) 68:316–25. doi: 10.1248/cpb.c19-00854

  • 128

    CreedenJFSevierJZhangJTLapitskyYBrunicardiFCJinGet al. Smart exosomes enhance PDAC targeted therapy. J Control Release. (2024) 368:413–29. doi: 10.1016/j.jconrel.2024.02.037

  • 129

    GhaffariKMoradi-HasanabadASobhani-NasabAJavaheriJGhasemiA. Application of cell-derived exosomes in the hematological Malignancies therapy. Front Pharmacol. (2023) 14:1263834. doi: 10.3389/fphar.2023.1263834

  • 130

    HermannDMPeruzzotti-JamettiLGiebelBPluchinoS. Extracellular vesicles set the stage for brain plasticity and recovery by multimodal signalling. Brain. (2024) 147:372–89. doi: 10.1093/brain/awad332

  • 131

    JangYParkJKimPParkEJSunHBaekYet al. Development of exosome membrane materials-fused microbubbles for enhanced stability and efficient drug delivery of ultrasound contrast agent. Acta Pharm Sin B. (2023) 13:4983–98. doi: 10.1016/j.apsb.2023.08.022

  • 132

    VandergriffAHuangKShenDHuSHensleyMTCaranasosTGet al. Targeting regenerative exosomes to myocardial infarction using cardiac homing peptide. Theranostics. (2018) 8:1869–78. doi: 10.7150/thno.20524

  • 133

    ChoiHChoiYYimHYMirzaaghasiAYooJKChoiC. Biodistribution of exosomes and engineering strategies for targeted delivery of therapeutic exosomes. Tissue Eng Regener Med. (2021) 18:499–511. doi: 10.1007/s13770-021-00361-0

  • 134

    ZhaoKWangZHackertTPitzerCZollerM. Tspan8 and Tspan8/CD151 knockout mice unravel the contribution of tumor and host exosomes to tumor progression. J Exp Clin Cancer Res. (2018) 37:312. doi: 10.1186/s13046-018-0961-6

  • 135

    DuSQianJTanSLiWLiuPZhaoJet al. Tumor cell-derived exosomes deliver TIE2 protein to macrophages to promote angiogenesis in cervical cancer. Cancer Lett. (2022) 529:168–79. doi: 10.1016/j.canlet.2022.01.005

  • 136

    WangJZhangHSunXWangXRenTHuangYet al. Exosomal PD-L1 and N-cadherin predict pulmonary metastasis progression for osteosarcoma patients. J Nanobiotechnology. (2020) 18:149-67. doi: 10.1186/s12951-020-00710-6

  • 137

    HongWXueMJiangJZhangYGaoX. Circular RNA circ-CPA4/let-7 miRNA/PD-L1 axis regulates cell growth, stemness, drug resistance and immune evasion in non-small cell lung cancer (NSCLC). J Exp Clin Cancer Res. (2020) 39:149. doi: 10.1186/s13046-020-01648-1

  • 138

    LuanWDingYXiHRuanHLuFMaSet al. Exosomal miR-106b-5p derived from melanoma cell promotes primary melanocytes epithelial-mesenchymal transition through targeting EphA4. J Exp Clin Cancer Res. (2021) 40:107. doi: 10.1186/s13046-021-01906-w

  • 139

    PaulIBolzanDYoussefAGagnonKAHookHKaremoreGet al. Parallelized multidimensional analytic framework applied to mammary epithelial cells uncovers regulatory principles in EMT. Nat Commun. (2023) 14:688. doi: 10.1038/s41467-023-36122-x

  • 140

    ZhouWJZhangJXieFWuJNYeJFWangJet al. CD45RO(-)CD8(+) T cell-derived exosomes restrict estrogen-driven endometrial cancer development via the ERbeta/miR-765/PLP2/Notch axis. Theranostics. (2021) 11:5330–45. doi: 10.7150/thno.58337

  • 141

    SinghASettlemanJ. EMT, cancer stem cells and drug resistance: an emerging axis of evil in the war on cancer. Oncogene. (2010) 29:4741–51. doi: 10.1038/onc.2010.215

  • 142

    MilmanNGininiLGilZ. Exosomes and their role in tumorigenesis and anticancer drug resistance. Drug Resist Update. (2019) 45:1–12. doi: 10.1016/j.drup.2019.07.003

  • 143

    BachDHHongJYParkHJLeeSK. The role of exosomes and miRNAs in drug-resistance of cancer cells. Int J Cancer. (2017) 141:220–30. doi: 10.1002/ijc.30669

  • 144

    LiSYiMDongBJiaoYLuoSWuK. The roles of exosomes in cancer drug resistance and its therapeutic application. Clin Transl Med. (2020) 10:e257. doi: 10.1002/ctm2.257

  • 145

    VellaLJBehrenAColemanBGreeningDWHillAFCebonJ. Intercellular resistance to BRAF inhibition can be mediated by extracellular vesicle-associated PDGFRbeta. Neoplasia. (2017) 19:932–40. doi: 10.1016/j.neo.2017.07.002

  • 146

    SynNWangLSethiGThieryJPGohBC. Exosome-mediated metastasis: from epithelial-mesenchymal transition to escape from immunosurveillance. Trends Pharmacol Sci. (2016) 37:606–17. doi: 10.1016/j.tips.2016.04.006

  • 147

    Rahman MABJLovatFGaoMOttersonGANana-SinkamP. Lung cancer exosomes as drivers of epithelial mesenchymal transition. Oncotarget. (2016) 7:54852–66. doi: 10.18632/oncotarget.v7i34

  • 148

    SkandalisSSKaralisTTChatzopoulosAKaramanosNK. Hyaluronan-CD44 axis orchestrates cancer stem cell functions. Cell Signal. (2019) 63:109377. doi: 10.1016/j.cellsig.2019.109377

  • 149

    NakamuraKSawadaKKinoseYYoshimuraATodaANakatsukaEet al. Exosomes promote ovarian cancer cell invasion through transfer of CD44 to peritoneal mesothelial cells. Mol Cancer Res. (2017) 15:78–92. doi: 10.1158/1541-7786.MCR-16-0191

  • 150

    LiYLiQLiDGuJQianDQinXet al. Exosome carrying PSGR promotes stemness and epithelial-mesenchymal transition of low aggressive prostate cancer cells. Life Sci. (2021) 264:118638. doi: 10.1016/j.lfs.2020.118638

  • 151

    WhitesideTL. The role of tumor-derived exosomes in epithelial mesenchymal transition (EMT). Transl Cancer Res. (2017) 6:S90–2. doi: 10.21037/tcr.2017.02.13

  • 152

    Jiao THYSunHYangL. Exosomal lnc-CDHR derived from human umbilical cord mesenchymal stem cells attenuates peritoneal epithelial-mesenchymal transition through AKT/FOXO pathway. Aging (Albany NY). (2023) 15:6921–32. doi: 10.18632/aging.v15i14

  • 153

    LinQChenXMengFOgawaKLiMSongRet al. ASPH-notch Axis guided Exosomal delivery of Prometastatic Secretome renders breast Cancer multi-organ metastasis. Mol Cancer. (2019) 18:156. doi: 10.1186/s12943-019-1077-0

  • 154

    YaoYChenRWangGZhangYLiuF. Exosomes derived from mesenchymal stem cells reverse EMT via TGF-beta1/Smad pathway and promote repair of damaged endometrium. Stem Cell Res Ther. (2019) 10:225. doi: 10.1186/s13287-019-1332-8

  • 155

    Gao XRNDongXZuoBYangRZhouQMoultonHMet al. Anchor peptide captures, targets, and loads exosomes of diverse origins for diagnostics and therapy. Sci Transl Med. (2018) 10:1–14. doi: 10.1126/scitranslmed.aat0195

  • 156

    LiZZhouXWeiMGaoXZhaoLShiRet al. In vitro and in vivo RNA inhibition by CD9-huR functionalized exosomes encapsulated with miRNA or CRISPR/dCas9. Nano Lett. (2019) 19:19–28. doi: 10.1021/acs.nanolett.8b02689

  • 157

    WangXChenYZhaoZMengQYuYSunJet al. Engineered exosomes with ischemic myocardium-targeting peptide for targeted therapy in myocardial infarction. J Am Heart Assoc. (2018) 7:e008737. doi: 10.1161/JAHA.118.008737

  • 158

    WangCLiNLiYHouSZhangWMengZet al. Engineering a HEK-293T exosome-based delivery platform for efficient tumor-targeting chemotherapy/internal irradiation combination therapy. J Nanobiotechnology. (2022) 20:247. doi: 10.1186/s12951-022-01462-1

  • 159

    LiangYXuXXuLIqbalZOuyangKZhangHet al. Chondrocyte-specific genomic editing enabled by hybrid exosomes for osteoarthritis treatment. Theranostics. (2022) 12:4866–78. doi: 10.7150/thno.69368

  • 160

    ChenSYLinMCTsaiJSHePLLuoWTHerschmanHet al. EP(4) antagonist-elicited extracellular vesicles from mesenchymal stem cells rescue cognition/learning deficiencies by restoring brain cellular functions. Stem Cells Transl Med. (2019) 8:707–23. doi: 10.1002/sctm.18-0284

  • 161

    ElashiryMElsayedRElashiryMMRashidMHAraRArbabASet al. Proteomic characterization, biodistribution, and functional studies of immune-therapeutic exosomes: implications for inflammatory lung diseases. Front Immunol. (2021) 12:636222. doi: 10.3389/fimmu.2021.636222

  • 162

    FangYWuFShangGYinC. SCARA5 in bone marrow stromal cell-derived exosomes inhibits colorectal cancer progression by inactivating the PI3K/Akt pathway. Genomics. (2023) 115:110636. doi: 10.1016/j.ygeno.2023.110636

  • 163

    HuWHuangFNingLHaoJWanJHaoS. Enhanced immunogenicity of leukemia-derived exosomes via transfection with lentiviral vectors encoding costimulatory molecules. Cell Oncol (Dordr). (2020) 43:889–900. doi: 10.1007/s13402-020-00535-3

  • 164

    IyaswamyAThakurAGuanXJKrishnamoorthiSFungTYLuKet al. Fe65-engineered neuronal exosomes encapsulating corynoxine-B ameliorate cognition and pathology of Alzheimer’s disease. Signal Transduct Target Ther. (2023) 8:404. doi: 10.1038/s41392-023-01657-4

  • 165

    LiKXueWLuZWangSZhengJLuKet al. Tumor-derived exosomal ADAM17 promotes pre-metastatic niche formation by enhancing vascular permeability in colorectal cancer. J Exp Clin Cancer Res. (2024) 43:59. doi: 10.1186/s13046-024-02991-3

  • 166

    LiuCLiuDWangSGanLYangXMaC. Identification of the SNARE complex that mediates the fusion of multivesicular bodies with the plasma membrane in exosome secretion. J Extracell Vesicles. (2023) 12:e12356. doi: 10.1002/jev2.12356

  • 167

    LiuCXiaCXiaC. Biology and function of exosomes in tumor immunotherapy. BioMed Pharmacother. (2023) 169:115853. doi: 10.1016/j.biopha.2023.115853

  • 168

    RossiFNorenHJoveRBeljanskiVGrinnemoKH. Differences and similarities between cancer and somatic stem cells: therapeutic implications. Stem Cell Res Ther. (2020) 11:489. doi: 10.1186/s13287-020-02018-6

  • 169

    YinPJiangYFangXWangDLiYChenMet al. Cell-based therapies for degenerative musculoskeletal diseases. Adv Sci (Weinh). (2023) 10:e2207050. doi: 10.1002/advs.202207050

  • 170

    KohELeeEJNamGHHongYChoEYangYet al. Exosome-SIRPalpha, a CD47 blockade increases cancer cell phagocytosis. Biomaterials. (2017) 121:121–9. doi: 10.1016/j.biomaterials.2017.01.004

  • 171

    ChoENamGHHongYKimYKKimDHYangYet al. Comparison of exosomes and ferritin protein nanocages for the delivery of membrane protein therapeutics. J Control Release. (2018) 279:326–35. doi: 10.1016/j.jconrel.2018.04.037

  • 172

    Alvarez-ErvitiLSeowYYinHBettsCLakhalSWoodMJ. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat Biotechnol. (2011) 29:341–5. doi: 10.1038/nbt.1807

  • 173

    LinYYanMBaiZXieYRenLWeiJet al. Huc-MSC-derived exosomes modified with the targeting peptide of aHSCs for liver fibrosis therapy. J Nanobiotechnology. (2022) 20:432. doi: 10.1186/s12951-022-01636-x

  • 174

    LiuQWuJWangHJiaZLiG. Human infrapatellar fat pad mesenchymal stem cell-derived extracellular vesicles purified by anion exchange chromatography suppress osteoarthritis progression in a mouse model. Clin Orthop Relat Res. (2024). 482:1246-62. doi: 10.1097/CORR.0000000000003067

  • 175

    GaoDShangJSunRShiYJiangHMaMet al. Changes in the morphology, number, and protein levels of plasma exosomes in CADASIL patients. J Alzheimers Dis. (2021) 81:221–9. doi: 10.3233/JAD-210101

  • 176

    AlaM. The beneficial effects of mesenchymal stem cells and their exosomes on myocardial infarction and critical considerations for enhancing their efficacy. Ageing Res Rev. (2023) 89:101980. doi: 10.1016/j.arr.2023.101980

  • 177

    CookNHansenARSiuLLAbdul RazakAR. Early phase clinical trials to identify optimal dosing and safety. Mol Oncol. (2015) 9:997–1007. doi: 10.1016/j.molonc.2014.07.025

  • 178

    FuscoCDe RosaGSpatoccoIVitielloEProcacciniCFrigeCet al. Extracellular vesicles as human therapeutics: A scoping review of the literature. J Extracell Vesicles. (2024) 13:e12433. doi: 10.1002/jev2.12433

  • 179

    BarileLMarbanE. Injury minimization after myocardial infarction: focus on extracellular vesicles. Eur Heart J. (2024) 45:1602–9. doi: 10.1093/eurheartj/ehae089

  • 180

    NakamoriYParkEJShimaokaM. Immune deregulation in sepsis and septic shock: reversing immune paralysis by targeting PD-1/PD-L1 pathway. Front Immunol. (2020) 11:624279. doi: 10.3389/fimmu.2020.624279

  • 181

    AwalHBNandulaSRDominguesCCDoreFJKunduNBrichacekBet al. Linagliptin, when compared to placebo, improves CD34+ve endothelial progenitor cells in type 2 diabetes subjects with chronic kidney disease taking metformin and/or insulin: a randomized controlled trial. Cardiovasc Diabetol. (2020) 19:72. doi: 10.1186/s12933-020-01046-z

  • 182

    PropperDJGaoFSaundersMPSarkerDHartleyJASpanswickVJet al. PANTHER: AZD8931, inhibitor of EGFR, ERBB2 and ERBB3 signalling, combined with FOLFIRI: a Phase I/II study to determine the importance of schedule and activity in colorectal cancer. Br J Cancer. (2023) 128:245–54. doi: 10.1038/s41416-022-02015-x

  • 183

    AchourBAl-MajdoubZMGrybos-GajniakALeaKKilfordPZhangMet al. Liquid biopsy enables quantification of the abundance and interindividual variability of hepatic enzymes and transporters. Clin Pharmacol Ther. (2021) 109:222–32. doi: 10.1002/cpt.2102

  • 184

    AllegraAPetrarcaCDi GioacchinoMCasciaroMMusolinoCGangemiS. Exosome-mediated therapeutic strategies for management of solid and hematological Malignancies. Cells. (2022) 11:1128-60. doi: 10.3390/cells11071128

  • 185

    JohnsonJLawSQKShojaeeMHallASBhuiyanSLimMBLet al. First-in-human clinical trial of allogeneic, platelet-derived extracellular vesicles as a potential therapeutic for delayed wound healing. J Extracell Vesicles. (2023) 12:e12332. doi: 10.1002/jev2.12332

  • 186

    HuDYangRWangGLiHFanXLiangG. Emerging strategies to overcome current CAR-T therapy dilemmas - exosomes derived from CAR-T cells. Int J Nanomedicine. (2024) 19:2773–91. doi: 10.2147/IJN.S445101

  • 187

    ChengBLiCLiJGongLLiangPChenYet al. The activity and immune dynamics of PD-1 inhibition on high-risk pulmonary ground glass opacity lesions: insights from a single-arm, phase II trial. Signal Transduct Target Ther. (2024) 9:93. doi: 10.1038/s41392-024-01799-z

  • 188

    BesseBCharrierMLapierreVDansinELantzOPlanchardDet al. Dendritic cell-derived exosomes as maintenance immunotherapy after first line chemotherapy in NSCLC. Oncoimmunology. (2016) 5:e1071008. doi: 10.1080/2162402X.2015.1071008

  • 189

    DaiSWeiDWuZZhouXWeiXHuangHet al. Phase I clinical trial of autologous ascites-derived exosomes combined with GM-CSF for colorectal cancer. Mol Ther. (2008) 16:782–90. doi: 10.1038/mt.2008.1

  • 190

    PoinsotVPizzinatNOng-MeangV. Engineered and mimicked extracellular nanovesicles for therapeutic delivery. Nanomaterials (Basel). (2024) 14:639-71. doi: 10.3390/nano14070639

  • 191

    AljuhaniAAlbalawiOAlbalawiRAlsalamaRAlatawiSAltemaniRet al. Exosomes in COVID-19 infection: Focus on role in diagnosis, pathogenesis, immunity, and clinical trials. Cell Biol Int. (2023) 47:1049–67. doi: 10.1002/cbin.12014

  • 192

    KhanNAAsimMBiswasKHAlansariANSamanHSarwarMZet al. Exosome nanovesicles as potential biomarkers and immune checkpoint signaling modulators in lung cancer microenvironment: recent advances and emerging concepts. J Exp Clin Cancer Res. (2023) 42:221. doi: 10.1186/s13046-023-02753-7

  • 193

    VrablovaVKosutovaNBlsakovaABertokovaAKasakPBertokTet al. Glycosylation in extracellular vesicles: Isolation, characterization, composition, analysis and clinical applications. Biotechnol Adv. (2023) 67:108196. doi: 10.1016/j.biotechadv.2023.108196

  • 194

    TretonGSayerCSchurzMJaritschMMullerAMateaCTet al. Quantitative and functional characterisation of extracellular vesicles after passive loading with hydrophobic or cholesterol-tagged small molecules. J Control Release. (2023) 361:694–716. doi: 10.1016/j.jconrel.2023.08.010

  • 195

    VahabiMComandatoreACentraCBlandinoGMorelliLGiovannettiE. Thinking small to win big? A critical review on the potential application of extracellular vesicles for biomarker discovery and new therapeutic approaches in pancreatic cancer. Semin Cancer Biol. (2023) 97:50–67. doi: 10.1016/j.semcancer.2023.11.003

  • 196

    ZhangZWangCLiTLiuZLiL. Comparison of ultracentrifugation and density gradient separation methods for isolating Tca8113 human tongue cancer cell line-derived exosomes. Oncol Lett. (2014) 8:1701–6. doi: 10.3892/ol.2014.2373

  • 197

    BrennanKMartinKFitzGeraldSPO’SullivanJWuYBlancoAet al. A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum. Sci Rep. (2020) 10:1039. doi: 10.1038/s41598-020-57497-7

  • 198

    BrettSILucienFGuoCWilliamsKCKimYDurfeePNet al. Immunoaffinity based methods are superior to kits for purification of prostate derived extracellular vesicles from plasma samples. Prostate. (2017) 77:1335–43. doi: 10.1002/pros.23393

  • 199

    Morales-KastresanaAMusichTAWelshJATelfordWDembergTWoodJCSet al. High-fidelity detection and sorting of nanoscale vesicles in viral disease and cancer. J Extracell Vesicles. (2019) 8:1597603. doi: 10.1080/20013078.2019.1597603

  • 200

    KuoWP. Extracellular vesicles methods and protocols. Methods Mol Biol. (2017) 1660:1–441. doi: 10.1007/978-1-4939-7253-1

  • 201

    PalmieriVLucchettiDGattoIMaioranaAMarcantoniMMaulucciGet al. Dynamic light scattering for the characterization and counting of extracellular vesicles: a powerful noninvasive tool. J Nanoparticle Res. (2014) 16:2583-90. doi: 10.1007/s11051-014-2583-z

  • 202

    Gamez-ValeroAMonguio-TortajadaMCarreras-PlanellaLFranquesaMBeyerKBorrasFE. Size-Exclusion Chromatography-based isolation minimally alters Extracellular Vesicles’ characteristics compared to precipitating agents. Sci Rep. (2016) 6:33641. doi: 10.1038/srep33641

  • 203

    BusattoSVilanilamGTicerTLinWLDicksonDWShapiroSet al. Tangential flow filtration for highly efficient concentration of extracellular vesicles from large volumes of fluid. Cells. (2018) 7:273-83. doi: 10.3390/cells7120273

  • 204

    PaternaARaoEAdamoGRaccostaSPicciottoSRomancinoDet al. Isolation of extracellular vesicles from microalgae: A renewable and scalable bioprocess. Front Bioeng Biotechnol. (2022) 10:836747. doi: 10.3389/fbioe.2022.836747

  • 205

    Momen-HeraviF. Isolation of extracellular vesicles by ultracentrifugation. Methods Mol Biol. (2017) 1660:25–32. doi: 10.1007/978-1-4939-7253-1_3

  • 206

    GardinerCDi VizioDSahooSTheryCWitwerKWWaubenMet al. Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey. J Extracell Vesicles. (2016) 5:32945. doi: 10.3402/jev.v5.32945

  • 207

    TakovKYellonDMDavidsonSM. Comparison of small extracellular vesicles isolated from plasma by ultracentrifugation or size-exclusion chromatography: yield, purity and functional potential. J Extracell Vesicles. (2019) 8:1560809. doi: 10.1080/20013078.2018.1560809

  • 208

    MusanteLTataruchDEHolthoferH. Use and isolation of urinary exosomes as biomarkers for diabetic nephropathy. Front Endocrinol (Lausanne). (2014) 5:149. doi: 10.3389/fendo.2014.00149

  • 209

    DaviesRTKimJJangSCChoiEJGhoYSParkJ. Microfluidic filtration system to isolate extracellular vesicles from blood. Lab Chip. (2012) 12:5202–10. doi: 10.1039/c2lc41006k

  • 210

    RashidiMBijariSKhazaeiAHShojaei-GhahrizjaniFRezakhaniL. The role of milk-derived exosomes in the treatment of diseases. Front Genet. (2022) 13:1009338. doi: 10.3389/fgene.2022.1009338

  • 211

    GehrmannUNaslundTIHiltbrunnerSLarssenPGabrielssonS. Harnessing the exosome-induced immune response for cancer immunotherapy. Semin Cancer Biol. (2014) 28:58–67. doi: 10.1016/j.semcancer.2014.05.003

  • 212

    RobbinsPDMorelliAE. Regulation of immune responses by extracellular vesicles. Nat Rev Immunol. (2014) 14:195–208. doi: 10.1038/nri3622

  • 213

    KurywchakPTavorminaJKalluriR. The emerging roles of exosomes in the modulation of immune responses in cancer. Genome Med. (2018) 10:23. doi: 10.1186/s13073-018-0535-4

  • 214

    Ayala-MarSDonoso-QuezadaJGonzalez-ValdezJ. Clinical implications of exosomal PD-L1 in cancer immunotherapy. J Immunol Res. (2021) 2021:8839978. doi: 10.1155/2021/8839978

  • 215

    FanYCheXQuJHouKWenTLiZet al. Exosomal PD-L1 retains immunosuppressive activity and is associated with gastric cancer prognosis. Ann Surg Oncol. (2019) 26:3745–55. doi: 10.1245/s10434-019-07431-7

  • 216

    HuangSHLiYZhangJRongJYeS. Epidermal growth factor receptor-containing exosomes induce tumor-specific regulatory T cells. Cancer Invest. (2013) 31:330–5. doi: 10.3109/07357907.2013.789905

  • 217

    LinWCLinYTChaoHCLinYYHwangWL. Haematopoietic cell-derived exosomes in cancer development and therapeutics: From basic science to clinical practice. Clin Transl Med. (2023) 13:e1448. doi: 10.1002/ctm2.1448

  • 218

    LiuJLiuJQinGLiJFuZLiJet al. MDSCs-derived GPR84 induces CD8(+) T-cell senescence via p53 activation to suppress the antitumor response. J Immunother Cancer. (2023) 11. doi: 10.1136/jitc-2023-007802

  • 219

    PoggioMHuTPaiCCChuBBelairCDChangAet al. Suppression of exosomal PD-L1 induces systemic anti-tumor immunity and memory. Cell. (2019) 177:414–427.e413. doi: 10.1016/j.cell.2019.02.016

  • 220

    QinSXuLYiMYuSWuKLuoS. Novel immune checkpoint targets: moving beyond PD-1 and CTLA-4. Mol Cancer. (2019) 18:155. doi: 10.1186/s12943-019-1091-2

  • 221

    XingCLiHLiRJYinLZhangHFHuangZNet al. The roles of exosomal immune checkpoint proteins in tumors. Mil Med Res. (2021) 8:56. doi: 10.1186/s40779-021-00350-3

  • 222

    TimanerMKotsofrukRRavivZMagideyKShechterDKanTet al. Microparticles from tumors exposed to radiation promote immune evasion in part by PD-L1. Oncogene. (2020) 39:187–203. doi: 10.1038/s41388-019-0971-7

  • 223

    Del ReMMarconciniRPasquiniGRofiEVivaldiCBloiseFet al. PD-L1 mRNA expression in plasma-derived exosomes is associated with response to anti-PD-1 antibodies in melanoma and NSCLC. Br J Cancer. (2018) 118:820–4. doi: 10.1038/bjc.2018.9

  • 224

    NingYShenKWuQSunXBaiYXieYet al. Tumor exosomes block dendritic cells maturation to decrease the T cell immune response. Immunol Lett. (2018) 199:36–43. doi: 10.1016/j.imlet.2018.05.002

  • 225

    LuYYeLJianXYangDZhangHTongZet al. Integrated microfluidic system for isolating exosome and analyzing protein marker PD-L1. Biosens Bioelectron. (2022) 204:113879. doi: 10.1016/j.bios.2021.113879

  • 226

    LiMSoderRAbhyankarSAbdelhakimHBraunMWTrinidadCVet al. WJMSC-derived small extracellular vesicle enhance T cell suppression through PD-L1. J Extracellular Vesicles. (2021) 10. doi: 10.1002/jev2.12067

  • 227

    YangYLiCWChanLCWeiYHsuJMXiaWet al. Exosomal PD-L1 harbors active defense function to suppress T cell killing of breast cancer cells and promote tumor growth. Cell Res. (2018) 28:862–4. doi: 10.1038/s41422-018-0060-4

  • 228

    ZhangWZhongWWangBYangJYangJYuZet al. ICAM-1-mediated adhesion is a prerequisite for exosome-induced T cell suppression. Dev Cell. (2022) 57:329–343.e327. doi: 10.1016/j.devcel.2022.01.002

  • 229

    TheryCDubanLSeguraEVeronPLantzOAmigorenaS. Indirect activation of naive CD4+ T cells by dendritic cell-derived exosomes. Nat Immunol. (2002) 3:1156–62. doi: 10.1038/ni854

  • 230

    AndreFChaputNSchartzNEFlamentCAubertNBernardJet al. Exosomes as potent cell-free peptide-based vaccine. I. Dendritic cell-derived exosomes transfer functional MHC class I/peptide complexes to dendritic cells. J Immunol. (2004) 172:2126–36. doi: 10.4049/jimmunol.172.4.2126

  • 231

    YangJZhangXLiuCWangZDengLFengCet al. Biologically modified nanoparticles as theranostic bionanomaterials. Prog Materials Sci. (2021) 118. doi: 10.1016/j.pmatsci.2020.100768

  • 232

    WuJ-YLiY-JWangJHuX-BHuangSLuoSet al. Multifunctional exosome-mimetics for targeted anti-glioblastoma therapy by manipulating protein corona. J Nanobiotechnology. (2021) 19:405-19. doi: 10.1186/s12951-021-01153-3

  • 233

    TothEATuriakLVisnovitzTCserepCMazloASodarBWet al. Formation of a protein corona on the surface of extracellular vesicles in blood plasma. J Extracell Vesicles. (2021) 10:e12140. doi: 10.1002/jev2.12140

  • 234

    HeidarzadehMZarebkohanARahbarghaziRSokulluE. Protein corona and exosomes: new challenges and prospects. Cell Commun Signal. (2023) 21:64. doi: 10.1186/s12964-023-01089-1

  • 235

    ChiesaMGargJKangYTChenG. Thermal conductivity and viscosity of water-in-oil nanoemulsions. Colloids Surfaces A: Physicochemical Eng Aspects. (2008) 326:67–72. doi: 10.1016/j.colsurfa.2008.05.028

  • 236

    ParkDJYunWSKimWCParkJELeeSHHaSet al. Improvement of stem cell-derived exosome release efficiency by surface-modified nanoparticles. J Nanobiotechnology. (2020) 18:178. doi: 10.1186/s12951-020-00739-7

  • 237

    GiovannelliLBariEJommiCTartaraFArmocidaDGarbossaDet al. Mesenchymal stem cell secretome and extracellular vesicles for neurodegenerative diseases: Risk-benefit profile and next steps for the market access. Bioact Mater. (2023) 29:16–35. doi: 10.1016/j.bioactmat.2023.06.013

  • 238

    ChenZXiongMTianJSongDDuanSZhangL. Encapsulation and assessment of therapeutic cargo in engineered exosomes: a systematic review. J Nanobiotechnology. (2024) 22:18. doi: 10.1186/s12951-023-02259-6

Summary

Keywords

exosomes, membrane proteins, functions, applications, immunoregulation

Citation

Xu F, Luo S, Lu P, Cai C, Li W and Li C (2024) Composition, functions, and applications of exosomal membrane proteins. Front. Immunol. 15:1408415. doi: 10.3389/fimmu.2024.1408415

Received

28 March 2024

Accepted

15 July 2024

Published

01 August 2024

Volume

15 - 2024

Edited by

Muhammad Babar Khawar, Chinese Academy of Sciences (CAS), China

Reviewed by

Paul Engeroff, Bern University Hospital, Switzerland

Anish Chakkumkal, Pharmaceutical Companies of Johnson and Johnson, Netherlands

Updates

Copyright

© 2024 Xu, Luo, Lu, Cai, Li and Li.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

*Correspondence: Chao Cai, ; Weihua Li, ; Chuanyun Li,

Disclaimer

All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.

Outline

Figures

Cite article

Copy to clipboard


Export citation file


Share article

Share on WeChat

Scan with WeChat to share this article

Article metrics