Exosomes liquid biopsy in Lymphoma precision medicine: applications and challenges.

Introduction

Lymphoma is a heterogeneous group of malignant lymphoid neoplasms, classified into Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL), and is characterized by a high recurrence rate and complex clinical course [ 1 – 4 ]. Early and accurate diagnosis of malignant lymphoma is critical for reducing tumor dissemination and residual disease, thereby improving the cure rates and long-term survival. Currently, lymphoma diagnosis primarily depends on traditional imaging, tissue biopsy including MICM (Morphology, Immunology, Cytogenetics and Molecular Biology). However, traditional imaging and tissue biopsy lack the capability for dynamic assessment. In specific scenarios like the non-invasive diagnosis of primary central nervous system lymphoma (PCNSL) in patients with surgically inaccessible brain masses, it is highly difficult for clinicians to obtain tissue samples for biopsy [ 5 ]. The inability to obtain tissue samples again after therapy or surgery is another significant limitation of clinical tissue biopsy [ 6 ]. Furthermore, tissue biopsy is invasive and insensitive, particularly for the detection of measurable residual disease (MRD). The lack of specific and sensitive techniques for monitoring therapeutic efficacy and predicting prognosis further exacerbates this problem [ 7 ]. Interestingly, the fear and unwillingness of patients who resist surgery also bring hardship to this invasive detection method [ 8 ]. These limitations highlight the urgent need for novel, minimally invasive detection methods that enable serial sampling and dynamic monitoring of disease status. Therefore, achieving early and accurate diagnosis remains a significant clinical challenge.

In recent years, liquid biopsy has garnered widespread attention as a non-invasive, convenient, real-time and comprehensive tool for accurate diagnosis and prognosis in clinical treatment. Liquid biopsy detects a series of biomarkers, such as circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), and exosomes in body fluids to guide the diagnosis, prognosis and medication of cancer by tracking their molecular characterization [ 9 ]. Compared with CTCs and ctDNA, exosomes offer distinct advantages due to their cargo of comprehensive biomarkers containing highly specific bio-information, as well as their cellular origin as vesicles secreted by living cells, high abundance, and stability within circulation [ 10 ]. Consequently, exosomes can provide a broader picture of lymphoma alongside CTCs and ctDNAs.

Exosomes are small extracellular vesicles secreted by eukaryotic cells. They play a crucial role in intercellular communication within the tumor immune microenvironment (TME) [ 11 ]. Tumor-derived exosomes, which retain the molecular characteristics of their cells of origin, are widely detected in various biological fluids. Owing to their unique advantages including non-invasiveness, repeatable sampling, dynamic monitoring, and high specificity, they have emerged as promising biomarkers for liquid biopsy in tumors. Currently, the clinical applications of exosomes have been extensively studied in solid tumors such as lung cancer and colorectal cancer [ 12 ]. In hematologic malignancies like lymphoma, where tumor cells are in direct contact with the circulatory system, exosomes may possess more direct and unique clinical value as liquid biopsy markers. However, systematic reviews summarizing the clinical application value of exosomes in lymphoma remain relatively scarce. In lymphoma, exosomes serve as “real-time” messengers that facilitate non-invasive disease grading, monitoring of minimal residual disease (MRD), and even the discovery of new therapeutic targets, thereby offering a more dynamic and patient-friendly approach to clinical management [ 12 – 15 ] (Table 1 ).

Existing literature has exhaustively reviewed exosome applications spanning biomarkers, drug delivery systems, and targeted therapies. However, their role in liquid biopsy, while increasingly explored, has not yet been comprehensively summarized in the literature [ 30 ]. In this review, we highlight the importance of exosome biopsy in the clinical management of lymphoma, outline current detection methods, and provide an in-depth look at the role of exosomes within the immune microenvironment of lymphoma. Particular emphasis is placed on three key application areas of exosome biopsy in lymphoma: diagnosis, prognosis and therapeutic response evaluation, and drug-guided treatment, aiming to inform future research and clinical applications (Fig. 1 ).

Mechanistic roles of exosomes in TME

Composition and biogenesis of exosomes

Exosomes are nanoscale extracellular vesicles (EVs) secreted by cells, typically ranging from 30 to 150 nanometers in diameter. Enclosed by lipid bilayers, they carry a diverse array of biomolecules, including proteins, nucleic acids (e.g., mRNA, miRNA), lipids, etc. Exosome biogenesis begins with cellular endocytosis, resulting in the formation of early endosomes [ 31 ]. These subsequently mature and fuse with the plasma membrane, thereby releasing exosomes into the TME. This process is regulated by several key protein complexes, including endosomal sorting complex required for transport (ESCRT) machinery, ALG-2-interacting protein X (ALIX), tumor susceptibility gene 101 (TSG101), and targeting GTP hydrolases (Rab GTPase). The release pathway that relies on ESCRT participation throughout the entire process is termed the ESCRT mechanism. However, studies have revealed an ESCRT-independent mechanism that does not depend on this molecule. Through their surface proteins and molecular cargo, exosomes mediate intercellular communication by transferring proteins and RNAs, thereby reprogramming the function of recipient cells (Fig. 2 ).

The composition and biogenesis of exosomes are generally similar in each kind of tumor, including haematological malignancies, while their differences are primarily determined by the specific cargo they carry [ 33 , 34 ]. Compared with exosomes derived from solid tumors, lymphoma-derived exosomes contain distinctive lymphoma-associated biomarkers such as miR-125b-5p, miR-99a-5p, while certain molecules including miR-181a-5p, miR-181b-5p, miR-181d-5p are relatively reduced. Unfortunately, a comprehensive characterization of these differences has not yet been fully described in the literature, and existing studies lack an overview of protein-level differences. Moreover, few investigations have systematically compared exosomes among different lymphoma subtypes. These gaps underscore the need for further exploration in clinical research.

The role and mechanism of exosomes in the TME

Cancer promotion

The most prominent function of exosomes within the TME is cancer promotion [ 39 ]. This process is typically driven by two key factors: the surface proteins carried on exosomes and their internal cargo, with the latter receiving greater attention in current research [ 40 – 42 ].

Exosomes fuse with target cells, releasing their molecular cargo, thereby initiating a cascade of biochemical reactions that promote cancer progression [ 43 , 44 ]. These vesicles transport a wide spectrum of biomolecules, including miRNA, mRNA, lipids, proteins, etc. Notably, miRNAs and proteins are closely related to the role of exosomes in promoting tumors [ 45 , 46 ].

Mediated by exosomal miRNA

The main mechanisms of miRNA include the following [ 47 , 48 ] (Fig. 3 ): (A) Inhibition of specific gene expression in downstream target cells. For instance, miR-21 in diffuse large B-cell lymphoma (DLBCL)-derived exosomes promotes tumor growth and dissemination by suppressing phosphatase and tensin homolog (PTEN) expression [ 49 ]. (B) Induction of specific proteins expression in downstream target cells. For example, exosomes enriched with Epstein–Barr virus (EBV)-derived miRNA stimulate tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) production in monocytes [ 50 ], thereby driving tumor-associated macrophages (TAM)-like phenotype and significantly promoting lymphoma progression [ 51 ]. (C) Regulation of specific molecules in target cells. In multiple myeloma (MM)-derived exosomes, miR-135b inhibits hypoxia-inducible factor 1 (HIF-1) in recipient endothelial cells, enhancing angiogenesis under hypoxic conditions and promoting MM progression [ 52 ]. Decreased levels of miR-7e-5p in exosomes derived from follicular lymphoma (FL) relieve the inhibition of macrophage apoptosis factors, resulting in impaired anti-tumor immunity and ultimately facilitating tumor growth [ 53 ].

Mediated by exosomal proteins

The mechanisms of protein function are relatively straightforward and can be categorized into two primary components: (A) Activation of signaling pathways: this function is usually mediated by cytokines. For instance, lymphoma cell-derived exosomes carrying mutant myeloid differentiation factor 88 (MYD88) can activate endogenous pro-inflammatory signaling pathways, including activation of NF-κB and production of inflammatory cytokines/chemokines in macrophages and mast cells. These foster a microenvironment that supports tumor growth [ 54 ]. Activation of the TGF-β/SMAD pathway by exosomal transforming growth factor-β (TGF-β) type III receptors, which induces the production of cancer-associated fibroblasts (CAF), also falls within this category of mechanism [ 55 , 56 ]. (B) Tumor invasion and metastasis are promoted by hydrolytic enzymes. For example, matrix metalloproteinases (MMPs) contained in tumor-derived exosomes play a pivotal role in facilitating cancer invasion. This intricate interplay of pathways underscores the complexity of tumor progression and highlights potential therapeutic targets [ 51 ] (Fig. 4 ).

Mediated by lipids and glycoconjugates

In addition to miRNAs and proteins mentioned above, lipids and glycoconjugates also contribute to cancer promotion. Their primary mechanism involves facilitating tumor invasion into surrounding tissues rather than activating a specific pathway [ 34 ]. A typical example is hyaluronic acid, which can promote cancer growth. Multiple cell types naturally possessing hyaluronan synthesis activity release large numbers of vesicles originating from the plasma membrane. These vesicles contain hyaluronan synthase-3 (HAS3). Such microvesicles possess the potential to deliver hyaluronan synthase to other cells [ 57 ]. The literature indicates that the mechanism by which HAS3 mediates cancer metastasis remains largely unexplored. The prevailing consensus holds that tumor-derived exosomes can transfer HAS3 to the TME target cells, while HAS3 promotes the secretion of exosomes by tumor cells.

Mediated by surface proteins

There is generally less literature focused on the mechanism of surface proteins compared to intracellular contents. Nevertheless, these mechanisms are crucial, as they involve specific binding to target molecules on recipient cell surfaces, triggering cascades of biochemical reactions that facilitate tumor growth or metastasis [ 58 ]. For instance, exosomal programmed cell death ligand-1 (PD-L1) binds specifically to programmed death-1 (PD-1) on the surface of T lymphocytes, suppressing T cell activation and enabling tumor cells to evade immune surveillance [ 58 ]. Similarly, cytotoxic T-lymphocyte–associated antigen-4 (CTLA-4), another surface protein that has received considerable attention, interacts with the B7 molecule to induce T cell anergy. This interaction effectively diminishes the anti-tumor immune response and promotes tumor progression and metastasis [ 59 ].

Mediated by the exosome as a whole

We will review the pro-carcinogenic pathways mediated by exosomes themselves in lymphoma TME. Delivery of specific gene-signaling molecules from lymphoma-derived exosomes into mesenchymal stromal cells (MSCs), which subsequently activate the corresponding signaling pathways [ 60 , 61 ]. EL-4 lymphoma-derived exosomes are endocytosed by bone marrow (BM)-MSCs, leading to the recruitment of a large number of macrophages via CC chemokine receptor 2 (CCR2) ligands to promote tumor hyperplasia—a mechanism that has been widely reviewed. In addition, adult T‐cell leukemia/lymphoma (ATL) has been reported to exploit an exosomal Tax-MSCs pathway, as described in several studies. Cells derived from tumor tissues secrete exosomes carrying key ALK signaling molecules. These exosomes can be absorbed by bone marrow stromal cells, leading to fibroblast activation and altering the cytokine profile of the tumor microenvironment (TME), which may contribute to tumor invasiveness [ 62 , 63 ].

Exosomes contribute to the remodeling of the TME by delivering biomolecules, including miRNAs, proteins, lipids and glycoconjugates, which drive tumor progression through multiple mechanisms such as immune evasion, angiogenesis, and macrophage phenotypic reprogramming. Targeting exosomes as a novel therapeutic strategy is particularly promising, as these vesicles can provide valuable insights for precision medicine. However, their specific mechanism of action and clinical potential remain unclear.

Drug resistance induction

Drug resistance is mainly mediated by exosome cargo, including miRNAs and proteins [ 64 ]. The underlying mechanisms can be categorized as follows: (A) Transfer of exosomal contents to target cells, conferring resistance. A study mentioned that tumor cell-derived exosomes can deliver specific miRNAs to recipient cells, inducing or enhancing drug resistance, but the specific types of miRNAs were not mentioned [ 65 ]. Notably, serum-derived exosomes from patients with B-cell lymphoma and cervical cancer can transfer P-glycoprotein (P-gp) into tumor cells. These exosomes not only facilitate the efflux of chemotherapeutic agents and their metabolites from tumor cells but also contribute to the acquisition of multidrug resistance [ 66 , 67 ]. (B) Exosome cargo can participate in a series of intracellular biochemical processes that drive the development of drug resistance. For instance, miR-584-5p carried by exosomes derived from DLBCL cells has been shown to target and modulate the cytochrome P450 (CYP)-related metabolic pathway in recipient cells [ 68 ]. By altering CYP-mediated metabolism, this miRNA promotes drug resistance in DLBCL cells. These findings underscore the complex role of exosomes in mediating drug resistance through multiple mechanisms and highlight the need for further exploration of their molecular contents and functions [ 69 ].

The process by which exosomes mediate drug resistance in lymphomas is similar to that observed in certain solid tumors. Previous studies have reported that the transfer of CD20 via membrane-bound exosomes derived from activated B-cell-like DLBCL (ABC-DLBCL) mediates the development of drug resistance.

Cancer inhibition

The tumor-suppressive role of exosomes remains one of the least explored topics in the literature, with current evidence suggesting that their contents can modulate gene expression in target cells. For instance, exosomal miR-665 has been reported to induce apoptosis in DLBCL cells and inhibit tumor growth and metastasis by downregulating its downstream target gene LIM and SH3 protein 1 (LASP1) [ 70 ]. In addition, exosomes may inhibit tumor progression by blocking oncogenic signaling pathways through the transfer of specific regulatory molecules. For example, exosomes derived from hepatocellular carcinoma can interact with receptors on the surface of T cells, upregulate IFN-γ levels, and downregulate IL-10 and TGF-β levels, which significantly improve the TME [ 71 , 72 ]. Nevertheless, studies in this field are still remarkably limited. Future investigations could focus on the tumor-suppressive effects of exosomes, which hold considerable promise for the development of exosome-based immunotherapies [ 73 ].

In summary, the mechanisms by which exosomes act within TME primarily involve the transfer of bioactive molecules—including surface proteins and internal cargo—to specific recipient cells. This transfer can occur through endocytosis, direct membrane fusion with target cells, or activation of biochemical signaling cascades via binding to cognate receptors. Fundamentally, exosomes function as mediators of intercellular communication by transporting molecules such as cytokines, nucleic acids, and proteins, thereby modulating the behavior of recipient cells. These processes are crucial for the establishment and maintenance of the TME [ 74 , 75 ]. Insights into these mechanisms have already begun to be applied to the exosome-based strategies for the treatment of solid tumors [ 76 ]. However, methods for detecting and characterizing the bioactive molecules that mediate these effects remain immature, highlighting the need for further technological innovation and translational research [ 77 ].

Feasibility of exosomes as biomarkers for liquid biopsy

As a result, exosomes play a pivotal role in various pathological processes, which are associated with their increased release. Owing to their unique advantages including stability, richness of tumor-associated composition, non-invasive and repeatable examination, they are considered ideal biomarkers for liquid biopsy [ 35 , 36 ]. The rationale is summarized as follows: (A) Exosomes carry abundant molecular contents, with both the type and amount sufficient to meet the requirements of liquid biopsy [ 37 ]. (B) The cargo of exosomes, especially RNA, is often closely related to the disease progression, and alterations in their levels or composition usually indicate disease onset or progression, and liquid biopsy analysis can therefore provide clinically relevant information [ 38 ]. (C) Exosomes are secreted into the blood circulation and can be obtained without invasive procedures at the lesion site, making them suitable biomarkers for liquid biopsy.

Exosome biopsy in lymphoma

Purification techniques

Due to exosomes’ unique biogenesis and specific cargo-sorting processes, the heterogeneity of exosomes in both size and molecular composition has driven the development of different purification techniques [ 78 – 80 ]. To date, multiple methods have been established, which differ significantly in terms of the yield and purity of exosomes separation [ 81 – 83 ]. The earliest and most common separation technique is ultracentrifugation-based separation [ 84 , 85 ]. However, this method is time-consuming, has limited efficiency, and may cause vesicle disruption, leading to reduced accuracy and reproducibility. Consequently, it has been increasingly complemented or replaced by emerging technologies. Emerging technologies offer improvements in specificity, recovery, and operational simplicity, although they often fall short of the high purity achieved by conventional ultracentrifugation.

Currently, there is no universally accepted method for the enrichment of lymphoma-derived exosomes. Some studies suggest that the polyethylene glycol (PEG)-based precipitation is advantageous in experiments requiring a high exosome yield, while ultracentrifugation remains more suitable for functional experiments due to its ability to better preserve exosome integrity. In addition, commercial kits such as the exoEasy kit are often employed in exploratory or pilot studies because of their convenience. Clinical liquid biopsies usually require larger sample sizes, so the polyethylene glycol (PEG) precipitation is frequently prioritized in most cases due to its efficiency in processing higher input volumes. Meanwhile, immunoaffinity enrichment and magnetic separation are considered more expensive enrichment methods with poor accessibility in laboratories unable to afford the markers, and precipitation with commercial kits similarly has poor accessibility due to the high equipment costs [ 89 – 94 ]. Therefore, the clinical application of these methods is limited. In the future, their accessibility can be improved by developing new biomarkers or devices with lower costs, thereby leveraging the advantages of these approaches (Table 2) .

Detection techniques

Detection techniques for exosomal biomolecules are generally categorized into protein-based and nucleic acid-based approaches, given the distinct physicochemical properties of these molecules. In fact, several review articles have comprehensively summarized this area. Dan et al. [ 10 ] systematically outlined current liquid biopsy methods and related information, including detection methods, biomarker sources, and key references in tabular form. Conventional protein detection techniques, such as Western blotting and enzyme-linked immunosorbent assay (ELISA), remain widely used due to their well-established protocols. However, their relatively low sensitivity and labor-intensive procedures have prompted the development of novel methods with higher sensitivity and specificity [ 102 – 104 ]. Similarly, nucleic acid-based techniques have evolved beyond traditional approaches such as quantitative reverse transcription polymerase chain reaction (qRT-PCR), with emerging technologies offering improved accuracy, sensitivity, and scalability [ 105 , 106 ] (Tables 3 and 4 ).

The current landscape of exosome-based liquid biopsy offers a range of advantages for detecting proteins and nucleic acids through various methods [ 137 ]. Techniques such as thermophoretic detection are favored for their operational effectiveness and convenience, while other methods, like the DNA tetrahedron probe demonstrate superior specificity and sensitivity [ 138 ]. However, no single methodology currently integrates all these advantages. Additionally, existing literature has not provided conclusive evidence regarding the most suitable detection technique for lymphoma-derived exosomes [ 139 ]. This knowledge gap largely reflects the early stage of exosomal liquid biopsy in lymphoma. Future research could focus on establishing optimized and standardized methodologies, which could serve as a foundation for advancing both mechanistic studies and clinical translation in this field.

Applications and limitations

Application

Supplementary Diagnosis

DLBCL-derived exosomes have been shown to carry multiple miRNAs that hold potential as biomarkers. Studies have highlighted that serum exosomes hsa-miR-12,136, hsa-miR-150-5p, hsa-miR-25-3p, hsa-miR-342-3p, hsa-miR-4516, hsa-miR-576-5p, hsa-miR-116-5p can serve as supplementary diagnostic biomarkers for DLBCL, as their reduced expression is closely related to poor OS or PFS [ 140 , 141 ]. Notably, miR-451a has been frequently referenced in prognostic studies and demonstrated moderate diagnostic efficacy for DLBCL ( p < 0.01) [ 6 ]. Furthermore, the correlation between serum exosomal miRNAs and IPI scores underscores the potential clinical utility of exosome-based liquid biopsy. Besides DLBCL [ 6 ], Ryu et al. found that elevated levels of miR-4454, miR-21-5p and miR-320e in serum exosomes were significantly associated with worse overall survival (OS) in extranodal NK/T-cell lymphoma (ENKTL) [ 142 ]. In addition to serving as an adjunct to diagnosis of lymphoma, exosome-based liquid biopsy may also facilitate early detection of disease progression. For instance, the presence of miR-19b-enriched exosomes has been proposed as a predictor of Richter syndrome, marking the transformation of B-cell chronic lymphocytic leukemia (B-CLL) into aggressive lymphoma [ 143 ].

In addition to miRNA, other nucleic acids also hold significant diagnostic value. For example, the U6 small nuclear-1 (RNU6-1) in serum exosomes has been suggested as a biomarker for distinguishing primary central nervous system lymphoma from glioblastoma [ 6 ]. By contrast, other exosomal biomolecules such as lipids and polysaccharides have not been reported in the literature of lymphoma liquid biopsy. We speculate that this may be attributed to the following reasons: (A) Lipids and polysaccharides do not directly participate in tumor pathological processes like nucleic acids and proteins; (B) Methods for detecting these two substances have not been sufficiently developed; (C) Researchers have primarily focused on studies related to nucleic acid and protein detection.

Prognosis and efficacy evaluation

Similar to their application in diagnosis, prognostic assessments and evaluations of therapeutic response are commonly performed by analyzing nucleic acids carried by exosomes. Among them, serum exosomal miR-99a-5p and miR-125b-5p in DLBCL patients have frequently been mentioned as ideal biomarkers of prognosis and chemotherapy response, as their elevated levels have been associated with shorter PFS [ 144 , 145 ]. Another frequently investigated group includes exosomal miR-155, let-7 g and let-7i, which are considered to serve as potential prognostic indicators for evaluating treatment response in DLBCL patients receiving standard chemotherapy [ 146 , 147 ].

Nucleic acids that are regarded as valuable diagnostic biomarkers can also serve as effective indicators for prognosis and therapeutic efficacy evaluation, such as miR-451a, which was highlighted in the diagnostic context [ 148 , 149 ]. Additionally, several other exosomal nucleic acids demonstrate distinct clinical significance. For example, exosomal miR-200c and miR-141 have been mentioned as biomarkers for monitoring the chemotherapy efficacy in PCNSL, as their levels tend to increase in patients with PCNSL [ 144 ]. Exosomal BCL-6 and C-MYC mRNA have been associated with poor prognostic outcomes in patients with B-cell lymphoma at the time of diagnosis [ 150 ]. Elevated serum levels of miR-21-5p, miR-320e, and miR-4454 have similarly been correlated with poor prognosis in patients with ENKTL [ 142 ]. In a study, exosomal miR-24-3p, miR-127-3p, miR-21-5p, miR-155-5p and let-7a-5p have been mentioned to contribute to prognostic stratification of patients with Hodgkin lymphoma [ 6 , 151 ].

In addition to nucleic acids, exosomal proteins also have certain value in clinical assessments [ 152 ]. The proportion of PD-L1 + exosomes has been associated with the likelihood of achieving complete response (CR) in patients with DLBCL treated with first-line R-CHOP (Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone) [ 153 – 156 ]. Serum exosomal vascular endothelial growth factor (VEGF) has been reported as a potential prognostic biomarker, correlating with both OS and PFS in DLBCL patients [ 157 ]. Moreover, change in exosomal TGF-β1 levels may provide valuable insights into chemotherapy response [ 158 ] (Table 5 ).

Limitations

Despite the numerous advantages of exosome liquid biopsy, certain limitations remain. First, research on its clinical application in NHL remains limited. Most existing studies focus primarily on DLBCL, which restricts the ability to establish its diagnostic value across the broader spectrum of NHL, underscoring the need for further investigation [ 6 ]. Additionally, while studies have found that exosomes can mediate drug resistance, there is still a significant gap in research regarding the application of exosomal liquid biopsy for detecting drug resistance [ 147 ].

Practical limitations are mainly reflected in the shortcomings of various methods used in exosome liquid biopsy. The isolation and purification of exosomes remain a considerable technical challenge for us. For example, nanomembrane-based ultrafiltration has been shown to achieve high purity suitable for exosomal biomarker analysis in urine-derived EVs. However, ultrafiltration is not the primary method used to isolate EVs from plasma or serum, due to the large losses caused by non-specific membrane adsorption and the limited plasma volumes typically obtainable from cancer patients [ 159 ]. Similarly, co-precipitation of non-exosomal proteins, including high levels of albumin, may compromise exosome purity and specificity [ 159 ]. Consequently, certain exosomal liquid biopsy techniques remain difficult to standardize and implement in clinical practice [ 160 ].

Furthermore, as highlighted in the previous section, the range of markers available for exosome liquid biopsy in lymphomas is highly diverse, making it difficult to establish a uniform standard for clinical application.

Conclusion

Exosomes mediate intercellular communication by transporting cytokines and other molecules within lymphoma TME, thereby contributing to the shaping of the TME and becoming ideal biomarkers for liquid biopsy. They play significant roles in the diagnosis, prognosis, treatment monitoring, and therapeutic decision-making in lymphoma. However, their clinical implementation remains challenging, and the limited number of studies makes it difficult to fully confirm their clinical value. Further research is essential to advance their application in the precision management of lymphoma patients. In addition to further confirming their significance in NHL, future studies should also investigate the value of exosomal liquid biopsy in detecting drug resistance in lymphoma. At the methodological level, future work could centre on developing more efficient techniques for the purification and isolation of exosomes, as well as expanding the spectrum of biomolecules that can be used as biomarkers for liquid biopsy.

Acknowledgements

Not applicable.

Abbreviations

Extracellular vesicles

Tumor microenvironment

Morphology, Immunology, Cytogenetics and Molecular biology

Hodgkin lymphoma

Non-Hodgkin lymphoma

Primary central nervous system lymphoma

Measurable residual disease

Circulating tumor DNA

Circulating tumor cells

Endosomal sorting complex required for transport

ALG-2-interacting protein X

Tumor susceptibility gene 101

Targeting GTP hydrolases

Diffuse large B-cell lymphoma

Phosphatase and tensin homolog

Epstein–Barr virus

Tumor necrosis factor-α

Interleukin 10

Tumor-associated macrophages

Multiple myeloma

Hypoxia-inducible factor 1

Follicular lymphoma

Myeloid differentiation factor 88

Transforming growth factor-β

Cancer-associated fibroblasts

Matrix metalloproteinases

Hyaluronan synthase 3

Programmed cell death ligand 1

Programmed death 1

Cytotoxic T-lymphocyte–associated antigen-4

CRISPR associated protein 9

Mesenchymal stromal cells

Bone marrow

CC chemokine receptor 2

Adult T‐cell leukemia/lymphoma

P-glycoprotein

Cytochrome P450

Activated B-cell-type DLBCL

LIM and SH3 protein 1

Polyethylene glycol

Enzyme-linked immunosorbent assay

Quantitative reverse transcription polymerase chain reaction

Surface-enhanced Raman spectroscopy

Surface plasmon resonance

Overall survival

Extranodal NK/T-cell lymphoma

B-cell chronic lymphocytic leukemia

U6 small nuclear-1

Complete response

Retuximab, cyclophosphamide, doxorubicin, vincristine, and prednisone

Vascular endothelial growth factor

Progression-Free-Survival

Author contributions

Zailin Yang and Yao Liu provided direction and guidance throughout the preparation of this manuscript. Junxi Liu and Yu Peng wrote and edited the manuscript. Yu Peng, Zailin Yang, Yao Liu, Shuang Chen, Tingting Jiang, Can Lin, Xinyi Tang, Yakun Zhang, Huiyu Xiang and Yu Lu reviewed and made significant revisions to the manuscript. All authors read and approved the final manuscript.

Funding

This work was supported by Chongqing Technology Innovation and Application Development Special Key Project (CSTB2024TIAD-KPX0031), Chongqing Municipal Science and Health Joint Medical Research Major Project (2025DBXM002) and Chongqing Municipal Science and Health Joint Medical Research Project (2026QNXM043).

Data availability

No datasets were generated or analyzed during the current study.

Declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare no competing interests.

Footnotes

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Junxi Liu and Yu Peng contributed equally to this work.

References

1. Lewis WD, Lilly S, Jones KL. Lymphoma: Diagnosis and Treatment. Am Fam Physician. 2020;101(1):34–41.

2. Silkenstedt E, Salles G, Campo E, Dreyling M. B-cell non-Hodgkin lymphomas. Lancet. 2024;403(10438):1791–807.

3. Horwitz SM, Ansell S, Ai WZ, Barnes J, Barta SK, Brammer J, et al. T-Cell lymphomas, version 2.2022, NCCN clinical practice guidelines in oncology. J Natl Compr Canc Netw. 2022;20(3):285–308.

4. de Leval L, Jaffe ES. Lymphoma classification. Cancer J. 2020;26(3):176–85.

5. Rossi D, Spina V, Bruscaggin A, Gaidano G. Liquid biopsy in lymphoma. Haematologica. 2019;104(4):648–52.

6. Lv L, Liu Y. Clinical application of liquid biopsy in Non-Hodgkin Lymphoma. Front Oncol. 2021;11:658234.

7. Wang H, Fu BB, Gale RP, Liang Y. NK-/T-cell lymphomas. Leukemia. 2021;35(9):2460–8.

8. Huang Z, Fu Y, Yang H, Zhou Y, Shi M, Li Q, et al. Liquid biopsy in T-cell lymphoma: biomarker detection techniques and clinical application. Mol Cancer. 2024;23(1):36.

9. Kurma K, Eslami SZ, Alix-Panabières C, Cayrefourcq L. Liquid biopsy: paving a new avenue for cancer research. Cell Adh Migr. 2024;18(1):1–26.

10. Yu D, Li Y, Wang M, Gu J, Xu W, Cai H, et al. Exosomes as a new frontier of cancer liquid biopsy. Mol Cancer. 2022;21(1):56.

11. Jafari A, Babajani A, Abdollahpour-Alitappeh M, Ahmadi N, Rezaei-Tavirani M. Exosomes and cancer: from molecular mechanisms to clinical applications. Med Oncol. 2021;38(4):45.

12. Al-Madhagi H. The landscape of exosomes biogenesis to clinical applications. Int J Nanomedicine. 2024;19:3657–75.

13. Huang W, Paul D, Calin GA, Bayraktar R. miR-142: A master regulator in hematological malignancies and therapeutic opportunities. Cells. 2023. 10.3390/cells13010084

14. Li Y, Tang Y, Yang GY. Therapeutic application of exosomes in ischaemic stroke. Stroke Vasc Neurol. 2021;6(3):483–95.

15. Shen YQ, Sun L, Wang SM, Zheng XY, Xu R. Exosomal integrins in tumor progression, treatment and clinical prediction (Review). Int J Oncol. 2024. 10.3892/ijo.2024.5706.

16. Pote MS, Gacche RN. Exosomal signaling in cancer metastasis: molecular insights and therapeutic opportunities. Arch Biochem Biophys. 2025;764:110277.

17. Ryu KJ, Lee JY, Choi ME, Yoon SE, Cho J, Ko YH, Shim JH, Kim WS, Park C, Kim SJ. Serum-Derived Exosomal MicroRNA Profiles Can Predict Poor Survival Outcomes in Patients with Extranodal Natural Killer/T-Cell Lymphoma. Cancers (Basel). 2020;12(12):3548.

18. Mehryab F, Taghizadeh F, Goshtasbi N, Merati F, Rabbani S, Haeri A. Exosomes as cutting-edge therapeutics in various biomedical applications: an update on engineering, delivery, and preclinical studies. Biochimie. 2023;213:139–67.

19. Du Z, Huang Z, Chen X, Jiang G, Peng Y, Feng W, Huang N. Modified dendritic cell-derived exosomes activate both NK cells and T cells through the NKG2D/NKG2D-L pathway to kill CML cells with or without T315I mutation. Exp Hematol Oncol. 2022;11(1):36.

20. Kim SB. Function and therapeutic development of exosomes for cancer therapy. Arch Pharm Res. 2022;45(5):295–308.

21. Li D, Wang Y, Jin X, Hu D, Xia C, Xu H, et al. NK cell-derived exosomes carry miR-207 and alleviate depression-like symptoms in mice. J Neuroinflammation. 2020;17(1):126.

22. Liang Y, Duan L, Lu J, Xia J. Engineering exosomes for targeted drug delivery. Theranostics. 2021;11(7):3183–95.

23. Lopes D, Lopes J, Pereira-Silva M, Peixoto D, Rabiee N, Veiga F, et al. Bioengineered exosomal-membrane-camouflaged abiotic nanocarriers: neurodegenerative diseases, tissue engineering and regenerative medicine. Mil Med Res. 2023;10(1):19.

24. Massey AE, Malik S, Sikander M, Doxtater KA, Tripathi MK, Khan S, et al. Clinical implications of exosomes: Targeted drug delivery for cancer treatment. Int J Mol Sci. 2021. 10.3390/ijms22105278.

25. Ghaffari K, Moradi-Hasanabad A, Sobhani-Nasab A, Javaheri J, Ghasemi A. Application of cell-derived exosomes in the hematological malignancies therapy. Front Pharmacol. 2023;14:1263834.

26. Kim J, Kim M, Han H, Kim S, Lahiji SF, Kim YH. Dual-delivery of exosome inhibitor and immune-activating gene via lipid nano-assemblies for tumor immune evasion inhibition. J Control Release. 2025;381:113569.

27. Kostyusheva A, Romano E, Yan N, Lopus M, Zamyatnin AA Jr., Parodi A. Breaking barriers in targeted Therapy: Advancing exosome Isolation, Engineering, and imaging. Adv Drug Deliv Rev. 2025;218:115522.

28. Rasihashemi SZ, Sahrai H, Rezazadeh-Gavgani E, Yazdani Y, Khalaji A, Lotfinejad P. Exosomes carrying immune checkpoints, a promising therapeutic approach in cancer treatment. Med Oncol. 2022;39(12):183.

29. Shi Y, Qiu B, Huang L, Lin J, Li Y, Ze Y, et al. Exosomes and ferroptosis: roles in tumour regulation and new cancer therapies. PeerJ. 2022;10:e13238.

30. Otsuka M, Kotani A. Recent advances in extracellular vesicles in gastrointestinal cancer and lymphoma. Cancer Sci. 2023;114(6):2230–7.

31. Krylova SV, Feng D. The machinery of exosomes: Biogenesis, release, and uptake. Int J Mol Sci. 2023. 10.3390/ijms24021337.

32. Shi Y, Qiu B, Huang L, Li Y, Ze Y, Yao Y. Exosomes in liquid biopsy: a nanotool for postradiotherapy cancer monitoring. Front Biosci (Landmark Ed). 2022;27(7):205.

33. Pandian SRK, Vijayakumar KK, Kunjiappan S, Babkiewicz E, Maszczyk P. Emerging role of exosomes in hematological malignancies. Clin Exp Med. 2023;23(4):1123–36.

34. Pegtel DM, Gould SJ. Exosomes. Annu Rev Biochem. 2019;88:487–514.

35. Wang S, Dong Y, Gong A, Kong H, Gao J, Hao X, et al. Exosomal circRNAs as novel cancer biomarkers: challenges and opportunities. Int J Biol Sci. 2021;17(2):562–73.

36. Wang X, Huang J, Chen W, Li G, Li Z, Lei J. The updated role of exosomal proteins in the diagnosis, prognosis, and treatment of cancer. Exp Mol Med. 2022;54(9):1390–400.

37. Lucien F, Gustafson D, Lenassi M, Li B, Teske JJ, Boilard E, et al. MIBlood-EV: Minimal information to enhance the quality and reproducibility of blood extracellular vesicle research. J Extracell Vesicles. 2023;12(12):e12385.

38. Kudo K, Miki Y, Carreras J, Nakayama S, Nakamoto Y, Ito M, et al. Secreted phospholipase A(2) modifies extracellular vesicles and accelerates B cell lymphoma. Cell Metab. 2022;34(4):615–e338.

39. Tuo B, Chen Z, Dang Q, Chen C, Zhang H, Hu S, et al. Roles of exosomal circRNAs in tumour immunity and cancer progression. Cell Death Dis. 2022;13(6):539.

40. Kok VC, Yu CC. Cancer-derived exosomes: their role in cancer biology and biomarker development. Int J Nanomedicine. 2020;15:8019–36.

41. Kumar A, Deep G. Hypoxia in tumor microenvironment regulates exosome biogenesis: Molecular mechanisms and translational opportunities. Cancer Lett. 2020;479:23–30.

42. Wang S, Li F, Ye T, Wang J, Lyu C, Qing S, et al. Macrophage-tumor chimeric exosomes accumulate in lymph node and tumor to activate the immune response and the tumor microenvironment. Sci Transl Med. 2021;13(615):eabb6981.

43. Abbaszade Dibavar M, Pourbagheri-Sigaroodi A, Asemani Y, Salari S, Bashash D. Extracellular vesicles (EVs): what we know of the mesmerizing roles of these tiny vesicles in hematological malignancies? Life Sci. 2021;271:119177.

44. Ferguson Bennit HR, Gonda A, Kabagwira J, Oppegard L, Chi D, Licero Campbell J, et al. Natural killer cell phenotype and functionality affected by exposure to extracellular survivin and lymphoma-derived exosomes. Int J Mol Sci. 2021. 10.3390/ijms22031255

45. Li J, Zhang G, Liu CG, Xiang X, Le MTN, Sethi G, et al. The potential role of exosomal circRNAs in the tumor microenvironment: insights into cancer diagnosis and therapy. Theranostics. 2022;12(1):87–104.

46. Li W, Wang X, Li C, Chen T, Yang Q. Exosomal non-coding RNAs: emerging roles in bilateral communication between cancer cells and macrophages. Mol Ther. 2022;30(3):1036–53.

47. Luongo M, Laurenziello P, Cesta G, Bochicchio AM, Omer LC, Falco G, et al. The molecular conversations of sarcomas: exosomal non-coding RNAs in tumor’s biology and their translational prospects. Mol Cancer. 2024;23(1):172.

48. Xu Z, Chen Y, Ma L, Chen Y, Liu J, Guo Y, et al. Role of exosomal non-coding RNAs from tumor cells and tumor-associated macrophages in the tumor microenvironment. Mol Ther. 2022;30(10):3133–54.

49. Jung BK, Song H, Shin H, Chai JY. Exosomal miRNA-21 from Toxoplasma gondii -infected microglial cells induces the growth of U87 glioma cells by inhibiting tumor suppressor genes. Sci Rep. 2022;12(1):16450.

50. Xu F, Luo S, Lu P, Cai C, Li W, Li C. Composition, functions, and applications of exosomal membrane proteins. Front Immunol. 2024;15:1408415.

51. Kawashima M, Higuchi H, Kotani A. Significance of trogocytosis and exosome-mediated transport in establishing and maintaining the tumor microenvironment in lymphoid malignancies. J Clin Exp Hematop. 2021;61(4):192–201.

52. Umezu T, Tadokoro H, Azuma K, Yoshizawa S, Ohyashiki K, Ohyashiki JH. Exosomal miR-135b shed from hypoxic multiple myeloma cells enhances angiogenesis by targeting factor-inhibiting HIF-1. Blood. 2014;124(25):3748–57.

53. Lou X, Fu J, Zhao X, Zhuansun X, Rong C, Sun M, et al. MiR-7e-5p downregulation promotes transformation of low-grade follicular lymphoma to aggressive lymphoma by modulating an immunosuppressive stroma through the upregulation of FasL in M1 macrophages. J Exp Clin Cancer Res. 2020;39(1):237.

54. Manček-Keber M, Lainšček D, Benčina M, Chen JG, Romih R, Hunter ZR, et al. Extracellular vesicle-mediated transfer of constitutively active MyD88(L265P) engages MyD88(wt) and activates signaling. Blood. 2018;131(15):1720–9.

55. Xin X, Cheng X, Zeng F, Xu Q, Hou L. The role of TGF-β/SMAD signaling in hepatocellular carcinoma: from mechanism to therapy and prognosis. Int J Biol Sci. 2024;20(4):1436–51.

56. Yi M, Wu Y, Niu M, Zhu S, Zhang J, Yan Y, et al. Anti-TGF-β/PD-L1 bispecific antibody promotes t cell infiltration and exhibits enhanced antitumor activity in triple-negative breast cancer. J Immunother Cancer. 2022. 10.1136/jitc-2022-005543

57. Rilla K, Pasonen-Seppänen S, Deen AJ, Koistinen VVT, Wojciechowski S, Oikari S, et al. Hyaluronan production enhances shedding of plasma membrane-derived microvesicles. Exp Cell Res. 2013;319(13):2006–18.

58. Zeng H, Wang J, Xu B, Deng H, Peng M, Deng R, et al. Exosomal PD‑L1 promotes the formation of an immunosuppressive microenvironment in gastric diffuse large b‑cell lymphoma. Oncol Rep. 2023. 10.3892/or.2023.8525

59. Agarwal S, Aznar MA, Rech AJ, Good CR, Kuramitsu S, Da T, et al. Deletion of the inhibitory co-receptor CTLA-4 enhances and invigorates chimeric antigen receptor T cells. Immunity. 2023;56(10):2388–e4079.

60. Hansen HP, Engels HM, Dams M, Paes Leme AF, Pauletti BA, Simhadri VL, et al. Protrusion-guided extracellular vesicles mediate CD30 trans-signalling in the microenvironment of Hodgkin’s lymphoma. J Pathol. 2014;232(4):405–14.

61. Hu YZ, Li Q, Wang PF, Li XP, Hu ZL. Multiple functions and regulatory network of miR-150 in B lymphocyte-related diseases. Front Oncol. 2023;13:1140813.

62. Chen W, Xie Y, Li F, Wen P, Wang L. EBV + B cell-derived exosomes promote EBV-associated T/NK-cell lymphoproliferative disease immune evasion by STAT3/IL-10/PD-L1 pathway. Immunol Res. 2024;72(6):1327–36.

63. Chioureas D, Beck J, Baltatzis G, Vardaki I, Fonseca P, Tsesmetzis N, et al. ALK+ anaplastic large cell lymphoma (ALCL)-derived exosomes carry ALK signaling proteins and interact with tumor microenvironment. Cancers (Basel). 2022. 10.3390/cancers14122939

64. Cariello M, Squilla A, Piacente M, Venutolo G, Fasano A. Drug resistance: The role of exosomal miRNA in the microenvironment of hematopoietic tumors. Molecules. 2022. 10.3390/molecules28010116

65. Huang HB, Shen JZ. [Research advances on abnormal secretion and function abnormality of exosomes derived from lymphoma cells caused by latent EB virus–review]. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2020;28(1):325–8.

66. Riou GF, Zhou D, Ahomadegbe J-C, Gabillot M, Duvillard P, Lhomme C. Expression of multidrug-resistance (MDR1) gene in normal epithelia and in invasive carcinomas of the uterine cervix. JNCI: Journal of the National Cancer Institute. 1990;82(18):1493–6. 10.1093/jnci/82.18.1493.

67. Shen H, Xu W, Luo W, et al. Upregulation of mdr1 gene is related to activation of the MAPK/ERK signal transduction pathway and YB-1 nuclear translocation in B-cell lymphoma. Exp Hematol. 2011;39(5):558–69.

68. Hou S, Bo J, Chen P, Guo M. Serum exo-miR584-5p mediates drug resistance in diffuse large B-cell lymphoma by upregulating the drug metabolism-CYP450 pathway, promoting high P450 expression, and remodling its intracellular localization. Blood. 2025;146:6823.

69. Shan X, Zhang L, Zhu DX, Zhou X, Zhang H, Liu QX, et al. Serum microRNA expression profiling revealing potential diagnostic biomarkers for lung adenocarcinoma. Chin Med J (Engl). 2020;133(21):2532–42.

70. Qu Z, Wu J, Wu J, Ji A, Qiang G, Jiang Y, et al. Exosomal miR-665 as a novel minimally invasive biomarker for hepatocellular carcinoma diagnosis and prognosis. Oncotarget. 2017;8(46):80666–78.

71. Li X, Li C, Zhang L, Wu M, Cao K, Jiang F, et al. The significance of exosomes in the development and treatment of hepatocellular carcinoma. Mol Cancer. 2020;19(1):1.

72. Santos P, Almeida F. Role of exosomal miRNAs and the tumor microenvironment in drug resistance. Cells. 2020. 10.3390/cells9061450

73. Kim J, Zhu Y, Chen S, Wang D, Zhang S, Xia J, et al. Anti-glioma effect of ginseng-derived exosomes-like nanoparticles by active blood-brain-barrier penetration and tumor microenvironment modulation. J Nanobiotechnol. 2023;21(1):253.

74. Li J, Tian T, Zhou X. The role of exosomal shuttle RNA (esRNA) in lymphoma. Crit Rev Oncol Hematol. 2019;137:27–34.

75. Fernandes M, Teixeira AL, Medeiros R. The opportunistic effect of exosomes on Non-Hodgkin Lymphoma microenvironment modulation. Crit Rev Oncol Hematol. 2019;144:102825.

76. Zhang M, Hu S, Liu L, Dang P, Liu Y, Sun Z, et al. Engineered exosomes from different sources for cancer-targeted therapy. Signal Transduct Target Ther. 2023;8(1):124.

77. Kalluri R, LeBleu VS. The biology, function, and biomedical applications of exosomes. Science. 2020. 10.1126/science.aau6977

78. Altıntaş Ö, Saylan Y. Exploring the Versatility of Exosomes: A Review on Isolation, Characterization, Detection Methods, and Diverse Applications. Anal Chem. 2023;95(44):16029–48.

79. Carvalho AS, Baeta H, Henriques AFA, Ejtehadifar M, Tranfield EM, Sousa AL, et al. Proteomic landscape of extracellular vesicles for Diffuse Large B-Cell Lymphoma subtyping. Int J Mol Sci. 2021. 10.3390/ijms222011004

80. Qian J, Zhang Q, Liu M, Wang Y, Lu M. A portable system for isothermal amplification and detection of exosomal microRNAs. Biosens Bioelectron. 2022;196:113707.

81. Cui H, Zheng T, Qian N, Fu X, Li A, Xing S, et al. Aptamer-functionalized magnetic Ti 3 C 2 based nanoplatform for simultaneous enrichment and detection of exosomes. Small. 2024;20(44):e2402434.

82. Dezhakam E, Khalilzadeh B, Mahdipour M, Isildak I, Yousefi H, Ahmadi M, et al. Electrochemical biosensors in exosome analysis; a short journey to the present and future trends in early-stage evaluation of cancers. Biosens Bioelectron. 2023;222:114980.

83. Yu X, Sha L, Dong L, Cao Y, Zhao J. Recent advances in bio-sensing methods for the detection of tumor exosomes. Crit Rev Anal Chem. 2022;52(2):356–74.

84. Kimiz-Gebologlu I, Oncel SS. Exosomes: large-scale production, isolation, drug loading efficiency, and biodistribution and uptake. J Control Release. 2022;347:533–43.

85. Yang D, Zhang W, Zhang H, Zhang F, Chen L, Ma L, et al. Progress, opportunity, and perspective on exosome isolation - efforts for efficient exosome-based theranostics. Theranostics. 2020;10(8):3684–707.

86. Johnstone RM, Adam M, Hammond JR, Orr L, Turbide C. Vesicle formation during reticulocyte maturation. Association of plasma membrane activities with released vesicles (exosomes). J Biol Chem. 1987;262(19):9412–20.

87. Alvarez ML, Khosroheidari M, Kanchi Ravi R, DiStefano JK. Comparison of protein, microRNA, and mRNA yields using different methods of urinary exosome isolation for the discovery of kidney disease biomarkers. Kidney Int. 2012;82(9):1024–32.

88. Böing AN, van der Pol E, Grootemaat AE, Coumans FA, Sturk A, Nieuwland R. Single-step isolation of extracellular vesicles by size-exclusion chromatography. J Extracell Vesicles. 2014;3.

89. Ding M, Wang C, Lu X, Zhang C, Zhou Z, Chen X, et al. Comparison of commercial exosome isolation kits for circulating exosomal microRNA profiling. Anal Bioanal Chem. 2018;410(16):3805–14.

90. Yang Q, Cheng L, Hu L, Lou D, Zhang T, Li J, et al. An integrative microfluidic device for isolation and ultrasensitive detection of lung cancer-specific exosomes from patient urine. Biosens Bioelectron. 2020;163:112290.

91. Zhang P, He M, Zeng Y. Ultrasensitive microfluidic analysis of circulating exosomes using a nanostructured graphene oxide/polydopamine coating. Lab Chip. 2016;16(16):3033–42.

92. He M, Crow J, Roth M, Zeng Y, Godwin AK. Integrated immunoisolation and protein analysis of circulating exosomes using microfluidic technology. Lab Chip. 2014;14(19):3773–80.

93. Zhao Z, Yang Y, Zeng Y, He M. A microfluidic ExoSearch chip for multiplexed exosome detection towards blood-based ovarian cancer diagnosis. Lab Chip. 2016;16(3):489–96.

94. Fang S, Tian H, Li X, Jin D, Li X, Kong J, et al. Clinical application of a microfluidic chip for immunocapture and quantification of circulating exosomes to assist breast cancer diagnosis and molecular classification. PLoS ONE. 2017;12(4):e0175050.

95. Wunsch BH, Smith JT, Gifford SM, Wang C, Brink M, Bruce RL, et al. Nanoscale lateral displacement arrays for the separation of exosomes and colloids down to 20 nm. Nat Nanotechnol. 2016;11(11):936–40.

96. Jiang S, Li Q, Wang C, Pang Y, Sun Z, Xiao R. In situ exosomal microRNA determination by target-triggered SERS and Fe(3)O(4)@TiO(2)-based exosome accumulation. ACS Sens. 2021;6(3):852–62.

97. Wan Y, Cheng G, Liu X, Hao SJ, Nisic M, Zhu CD, et al. Rapid magnetic isolation of extracellular vesicles via lipid-based nanoprobes. Nat Biomed Eng. 2017. 10.1038/s41551-017-0058.

98. Tayebi M, Yang D, Collins DJ, Ai Y. Deterministic Sorting of Submicrometer Particles and Extracellular Vesicles Using a Combined Electric and Acoustic Field. Nano Lett. 2021;21(16):6835–42.

99. Lee K, Shao H, Weissleder R, Lee H. Acoustic purification of extracellular microvesicles. ACS Nano. 2015;9(3):2321–7.

100. Liu C, Zhao J, Tian F, Cai L, Zhang W, Feng Q, et al. Low-cost thermophoretic profiling of extracellular-vesicle surface proteins for the early detection and classification of cancers. Nat Biomed Eng. 2019;3(3):183–93.

101. Tian F, Zhang S, Liu C, Han Z, Liu Y, Deng J, et al. Protein analysis of extracellular vesicles to monitor and predict therapeutic response in metastatic breast cancer. Nat Commun. 2021;12(1):2536.

102. Lin B, Tian T, Lu Y, Liu D, Huang M, Zhu L, et al. Tracing Tumor-Derived Exosomal PD-L1 by Dual-Aptamer Activated Proximity-Induced Droplet Digital PCR. Angew Chem Int Ed Engl. 2021;60(14):7582–6.

103. Liu WZ, Ma ZJ, Kang XW. Current status and outlook of advances in exosome isolation. Anal Bioanal Chem. 2022;414(24):7123–41.

104. Xie Z, Zhao S, Deng R, Tang X, Feng L, Xie S, et al. Logic-Measurer: A multienzyme-assisted ultrasensitive circuit for logical detection of exosomal microRNAs. ACS Nano. 2025;19(12):12222–36.

105. Ge W, Mu Z, Yang S, Zeng Y, Deng Y, Lin Y, et al. Biosensor-based methods for exosome detection with applications to disease diagnosis. Biosens Bioelectron. 2025;279:117362.

106. Guo Y, Tao J, Li Y, Feng Y, Ju H, Wang Z, et al. Quantitative localized analysis reveals distinct exosomal protein-specific glycosignatures: Implications in cancer cell subtyping, exosome biogenesis, and function. J Am Chem Soc. 2020;142(16):7404–12.

107. Shao H, Im H, Castro CM, Breakefield X, Weissleder R, Lee H. New technologies for analysis of extracellular vesicles. Chem Rev. 2018;118(4):1917–50.

108. Ueda K, Ishikawa N, Tatsuguchi A, Saichi N, Fujii R, Nakagawa H. Antibody-coupled monolithic silica microtips for highthroughput molecular profiling of circulating exosomes. Sci Rep. 2014;4:6232.

109. Xia Y, Liu M, Wang L, Yan A, He W, Chen M, et al. A visible and colorimetric aptasensor based on DNA-capped single-walled carbon nanotubes for detection of exosomes. Biosens Bioelectron. 2017;92:8–15.

110. Chen J, Xu Y, Lu Y, Xing W. Isolation and Visible Detection of Tumor-Derived Exosomes from Plasma. Anal Chem. 2018;90(24):14207–15.

111. Chen Z, Cheng SB, Cao P, Qiu QF, Chen Y, Xie M, et al. Detection of exosomes by ZnO nanowires coated three-dimensional scaffold chip device. Biosens Bioelectron. 2018;122:211–6.

112. Zhang P, Zhou X, Zeng Y. Multiplexed immunophenotyping of circulating exosomes on nano-engineered ExoProfile chip towards early diagnosis of cancer. Chem Sci. 2019;10(21):5495–504.

113. Tayebi M, Zhou Y, Tripathi P, Chandramohanadas R, Ai Y. Exosome Purification and Analysis Using a Facile Microfluidic Hydrodynamic Trapping Device. Anal Chem. 2020;92(15):10733–42.

114. Zhang Z, Tang C, Zhao L, Xu L, Zhou W, Dong Z, et al. Aptamer-based fluorescence polarization assay for separation-free exosome quantification. Nanoscale. 2019;11(20):10106–13.

115. !!! INVALID CITATION !!! 119.

116. Park J, Park JS, Huang CH, Jo A, Cook K, Wang R, et al. An integrated magneto-electrochemical device for the rapid profiling of tumour extracellular vesicles from blood plasma. Nat Biomed Eng. 2021;5(7):678–89.

117. Wang S, Zhang L, Wan S, Cansiz S, Cui C, Liu Y, et al. Aptasensor with expanded nucleotide using DNA nanotetrahedra for electrochemical detection of cancerous exosomes. ACS Nano. 2017;11(4):3943–9.

118. Dong S, Wang Y, Liu Z, Zhang W, Yi K, Zhang X, et al. Beehive-inspired macroporous SERS probe for cancer detection through capturing and analyzing exosomes in plasma. ACS Appl Mater Interfaces. 2020;12(4):5136–46.

119. Shin H, Oh S, Hong S, Kang M, Kang D, Ji YG, et al. Early-Stage Lung Cancer Diagnosis by Deep Learning-Based Spectroscopic Analysis of Circulating Exosomes. ACS Nano. 2020;14(5):5435–44.

120. Zhao X, Zhang W, Qiu X, Mei Q, Luo Y, Fu W. Rapid and sensitive exosome detection with CRISPR/Cas12a. Anal Bioanal Chem. 2020;412(3):601–9.

121. Xing S, Lu Z, Huang Q, Li H, Wang Y, Lai Y, et al. An ultrasensitive hybridization chain reaction-amplified CRISPR-Cas12a aptasensor for extracellular vesicle surface protein quantification. Theranostics. 2020;10(22):10262–73.

122. Guo K, Li Z, Win A, Coreas R, Adkins GB, Cui X, et al. Calibration-free analysis of surface proteins on single extracellular vesicles enabled by DNA nanostructure. Biosens Bioelectron. 2021;192:113502.

123. Wang H, Jiang D, Li W, Xiang X, Zhao J, Yu B, et al. Evaluation of serum extracellular vesicles as noninvasive diagnostic markers of glioma. Theranostics. 2019;9(18):5347–58.

124. Gandham S, Su X, Wood J, Nocera AL, Alli SC, Milane L, et al. Technologies and standardization in research on extracellular vesicles. Trends Biotechnol. 2020;38(10):1066–98.

125. Mateescu B, Kowal EJ, van Balkom BW, Bartel S, Bhattacharyya SN, Buzás EI, et al. Obstacles and opportunities in the functional analysis of extracellular vesicle RNA - an ISEV position paper. J Extracell Vesicles. 2017;6(1):1286095.

126. Jaluria P, Konstantopoulos K, Betenbaugh M, Shiloach J. A perspective on microarrays: current applications, pitfalls, and potential uses. Microb Cell Fact. 2007;6:4.

127. Wang R, Zhao X, Chen X, Qiu X, Qing G, Zhang H, et al. Rolling Circular Amplification (RCA)-Assisted CRISPR/Cas9 Cleavage (RACE) for Highly Specific Detection of Multiple Extracellular Vesicle MicroRNAs. Anal Chem. 2020;92(2):2176–85.

128. Thakur A, Qiu G, Ng SP, Guan J, Yue J, Lee Y, et al. Direct detection of two different tumor-derived extracellular vesicles by SAM-AuNIs LSPR biosensor. Biosens Bioelectron. 2017;94:400–7.

129. Zhao J, Liu C, Li Y, Ma Y, Deng J, Li L, et al. Thermophoretic Detection of Exosomal microRNAs by Nanoflares. J Am Chem Soc. 2020;142(11):4996–5001.

130. Chen Y, Zhai LY, Zhang LM, Ma XS, Liu Z, Li MM, et al. Breast cancer plasma biopsy by in situ determination of exosomal microRNA-1246 with a molecular beacon. Analyst. 2021;146(7):2264–76.

131. Oliveira GP Jr., Zigon E, Rogers G, Davodian D, Lu S, Jovanovic-Talisman T, et al. Detection of Extracellular Vesicle RNA Using Molecular Beacons. iScience. 2020;23(1):100782.

132. He D, Wang H, Ho SL, Chan HN, Hai L, He X, et al. Total internal reflection-based single-vesicle in situ quantitative and stoichiometric analysis of tumor-derived exosomal microRNAs for diagnosis and treatment monitoring. Theranostics. 2019;9(15):4494–507.

133. Chen X, Jia M, Liu L, Qiu X, Zhang H, Yu X, et al. High-fidelity determination and tracing of small extracellular vesicle cargoes. Small. 2020;16(40):e2002800.

134. Wang C, Ding Q, Plant P, Basheer M, Yang C, Tawedrous E, et al. Droplet digital PCR improves urinary exosomal miRNA detection compared to real-time PCR. Clin Biochem. 2019;67:54–9.

135. Chen WW, Balaj L, Liau LM, Samuels ML, Kotsopoulos SK, Maguire CA, et al. BEAMing and Droplet Digital PCR Analysis of Mutant IDH1 mRNA in Glioma Patient Serum and Cerebrospinal Fluid Extracellular Vesicles. Mol Ther Nucleic Acids. 2013;2(7):e109.

136. Zhang P, Zhou X, He M, Shang Y, Tetlow AL, Godwin AK, et al. Ultrasensitive detection of circulating exosomes with a 3D-nanopatterned microfluidic chip. Nat Biomed Eng. 2019;3(6):438–51.

137. Mohammadi M, Zargartalebi H, Salahandish R, Aburashed R, Wey Yong K, Sanati-Nezhad A. Emerging technologies and commercial products in exosome-based cancer diagnosis and prognosis. Biosens Bioelectron. 2021;183:113176.

138. Hosseini K, Ranjbar M, Pirpour Tazehkand A, Asgharian P, Montazersaheb S, Tarhriz V, et al. Evaluation of exosomal non-coding RNAs in cancer using high-throughput sequencing. J Transl Med. 2022;20(1):30.

139. Villa M, Sharma GG, Manfroni C, Cortinovis D, Mologni L. New advances in liquid biopsy technologies for Anaplastic Lymphoma Kinase (ALK)-positive cancer. Cancers (Basel). 2021. 10.3390/cancers13205149

140. Xiao C, Calado DP, Galler G, Thai TH, Patterson HC, Wang J, et al. MiR-150 Controls B Cell Differentiation by Targeting the Transcription Factor c-Myb. Cell. 2016;165(4):1027.

141. Cui T, Bell EH, McElroy J, Becker AP, Gulati PM, Geurts M, et al. miR-4516 predicts poor prognosis and functions as a novel oncogene via targeting PTPN14 in human glioblastoma. Oncogene. 2019;38(16):2923–36.

142. Ryu KJ, Lee JY, Choi ME, Yoon SE, Cho J, Ko YH, et al. Serum-derived exosomal microRNA profiles can predict poor survival outcomes in patients with Extranodal Natural Killer/T-Cell Lymphoma. Cancers (Basel). 2020. 10.3390/cancers12123548

143. Reiners KS, Shatnyeva O, Vasyutina E, Bösl T, Hansen HP, Hallek M, et al. Extracellular vesicles released from chronic lymphocytic leukemia cells exhibit a disease relevant mRNA signature and transfer mRNA to bystander cells. Haematologica. 2017;102(3):e100–3.

144. Zhang H, Xia J, Wang X, Wang Y, Chen J, He L, et al. Recent Progress of Exosomes in Hematological Malignancies: Pathogenesis, Diagnosis, and Therapeutic Strategies. Int J Nanomed. 2024;19:11611–31.

145. Feng Y, Zhong M, Zeng S, Wang L, Liu P, Xiao X, et al. Exosome-derived miRNAs as predictive biomarkers for diffuse large B-cell lymphoma chemotherapy resistance. Epigenomics. 2019;11(1):35–51.

146. Liu J, Wang X. Focus on exosomes-From pathogenic mechanisms to the potential clinical application value in lymphoma. J Cell Biochem. 2019;120(12):19220–8.

147. Ofori K, Bhagat G, Rai AJ. Exosomes and extracellular vesicles as liquid biopsy biomarkers in diffuse large B-cell lymphoma: current state of the art and unmet clinical needs. Br J Clin Pharmacol. 2021;87(2):284–94.

148. Yazdanparast S, Huang Z, Keramat S, Izadirad M, Li YD, Bo L, et al. The Roles of Exosomal microRNAs in Diffuse Large B-Cell Lymphoma: Diagnosis, Prognosis, Clinical Application, and Biomolecular Mechanisms. Front Oncol. 2022;12:904637.

149. Cao D, Cao X, Jiang Y, Xu J, Zheng Y, Kang D, et al. Circulating exosomal microRNAs as diagnostic and prognostic biomarkers in patients with diffuse large B-cell lymphoma. Hematol Oncol. 2022;40(2):172–80.

150. Bang YH, Shim JH, Ryu KJ, Kim YJ, Choi ME, Yoon SE, et al. Clinical relevance of serum-derived exosomal messenger RNA sequencing in patients with non-Hodgkin lymphoma. J Cancer. 2022;13(5):1388–97.

151. Gargiulo E, Morande PE, Largeot A, Moussay E, Paggetti J. Diagnostic and therapeutic potential of extracellular vesicles in B-cell malignancies. Front Oncol. 2020;10:580874.

152. Hu C, Jiang W, Lv M, Fan S, Lu Y, Wu Q, et al. Potentiality of exosomal proteins as novel cancer biomarkers for liquid biopsy. Front Immunol. 2022;13:792046.

153. Li C, Li C, Zhi C, Liang W, Wang X, Chen X, et al. Clinical significance of PD-L1 expression in serum-derived exosomes in NSCLC patients. J Transl Med. 2019;17(1):355.

154. Lu MM, Yang Y. Exosomal PD-L1 in cancer and other fields: recent advances and perspectives. Front Immunol. 2024;15:1395332.

155. Lu Y, Ye L, Jian X, Yang D, Zhang H, Tong Z, et al. Integrated microfluidic system for isolating exosome and analyzing protein marker PD-L1. Biosens Bioelectron. 2022;204:113879.

156. Xu P, Liu J, Chen H, Shang L, Wang F, Zhu Y, et al. Clinical significance of plasma PD-L1(+) exosomes in the management of diffuse large B cell lymphoma. Ann Hematol. 2023;102(9):2435–44.

157. Negaard HF, Iversen N, Bowitz-Lothe IM, Sandset PM, Steinsvik B, Ostenstad B, et al. Increased bone marrow microvascular density in haematological malignancies is associated with differential regulation of angiogenic factors. Leukemia. 2009;23(1):162–9.

158. Litwińska Z, Łuczkowska K, Machaliński B. Extracellular vesicles in hematological malignancies. Leuk Lymphoma. 2019;60(1):29–36.

159. Khalife J, Sanchez JF, Pichiorri F. Extracellular vesicles in hematological malignancies: from biomarkers to therapeutic tools. Diagnostics (Basel). 2020. 10.3390/diagnostics10121065

160. Liu J, Ren L, Li S, Li W, Zheng X, Yang Y, et al. The biology, function, and applications of exosomes in cancer. Acta Pharm Sin B. 2021;11(9):2783–97.

161. Jiang S, Li H, Zhang L, et al. Generic Diagramming Platform (GDP): a comprehensive database of high-quality biomedical graphics. Nucleic Acids Res. 2025;53(D1):D1670–6.

Associated Data

Data Availability Statement

No datasets were generated or analyzed during the current study.