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Abstract

A significant number of cancer-related deaths are recorded globally each year, despite attempts to cure this illness. Medical science is working to develop new medication therapies as well as to find ways to identify this illness as early as possible, even using noninvasive techniques. Early detection of cancer can greatly aid its treatment. Studies into cancer diagnosis and therapy have recently shifted their focus to exosome (EXO) biomarkers, which comprise numerous RNA and proteins. EXOs are minuscule goblet vesicles that have a width of 30 to 140 nm and are released by a variety of cells, including immune, stem, and tumor cells, as well as bodily fluids. According to a growing body of research, EXOs, and cancer appear to be related. EXOs from tumors play a role in the genetic information transfer between tumor and basal cells, which controls angiogenesis and fosters tumor development and spread. To identify malignant activities early on, microRNAs (miRNAs) from cancers can be extracted from circulatory system EXOs. Specific markers can be used to identify cancer-derived EXOs containing miRNAs, which may be more reliable and precise for early detection. Conventional solid biopsy has become increasingly limited as precision and personalized medicine has advanced, while liquid biopsy offers a viable platform for noninvasive diagnosis and prognosis. Therefore, the use of body fluids such as serum, plasma, urine, and salivary secretions can help find cancer biomarkers using technologies related to EXOs.

Keywords

cancer exosome microRNAs biomarker diagnosis

Introduction

Recent years have seen an increase in reports on the relevance of exosomes (EXOs) to cancer biology.¹ EXOs are membrane sacs and nanoscale vehicles of endosomal origin produced by almost all normal and pathological cells and are found in all body fluids. They contain a wide range of biological molecules such as proteins, lipids, and nucleic acids (DNA and miRNA) and reflect the physiological conditions and secretory functions of the cell.^2,3

The microRNA (miRNA) is a single-stranded molecule approximately 18 to 25 nucleotides in length⁴ that is responsible for regulating gene expression at the posttranscriptional level. It also plays an important role in cell proliferation.⁵ In cancer, a series of genetic deviations occur, and the control mechanism of cell proliferation is out of regulation.⁶ Furthermore, metastasis is a cause of death in cancer patients, and it is not unthinkable that exosomal miRNAs (ExomiRs) play a role in metastasis and are the link between cancer and the host.⁷ Evidence has shown that miRNAs enclosed in EXOs can cross the blood–brain barrier and are safe from immune attacks.⁸ Because of their double-layered membrane and small size, high stability, and, correspondingly, a higher half-life are among their characteristics.⁹

Currently, imaging techniques and morphological analysis of tissues (histology) or cells (cytology) can detect cancer only when there is a visible change in the tissue; for example, when thousands of cancer cells have multiplied and even metastasized. Moreover, cancer biomarkers currently in use suffer from issues such as false negatives or false positives because of a lack of specificity. Therefore, ExomiRs related to the production and progression of tumors can be proposed as potential diagnostic, prognostic, and predictive biomarkers of cancer.¹⁰ The main advantage of using EXOs as biomarkers is their presence in various body fluids, for example, blood, urine, saliva, breast milk, etc, which makes them easily and noninvasively collectible and utilizable in clinical trials¹¹ (Figure 1). Due to their biocompatibility, EXOs can be measured in such a way that they can be used to diagnose cancer in the future.

Figure 1.

Exosomes isolation from body fluids, EXOs can be extracted from different body fluids such as urine, saliva, plasma, serum, CSF, and BAL. These nanoparticles contain various compounds, including miRNA, which can be used as biomarkers in medical diagnostics.

Definition and Biogenesis of EXOs

An EXO is a membranous sac released into the extracellular matrix (ECM) after fusing with a cell membrane. Its surface is made up of protein receptors, polysaccharides, and other lipids and has a double structure containing many biologically active substances.¹² miRNAs, proteins, and coding-protein mRNAs are all present in EXOs and represent the physiological and functional conditions of secretory cells. According to recent research, blood, saliva, cerebrospinal fluid, tumor cells, and other body fluids contain EXOs.^3,13

The biogenesis of EXOs occurs in 3 distinct phases:

Formation of endocytic vesicles (early endosome) through the invagination of the plasma membrane.

Formation of multivesicular bodies (MVBs) with intraluminal vesicles (ILVs) generated by cytoplasmic components budding inside the endosomal membrane.

MVB fusion with the plasma membrane and extracellular release of ILVs as EXOs.^14–16

MVBs may potentially decay through fusion with lysosomes or autophagosomes.¹⁷ Many different proteins, including the endosomal sorting complexes required for transport (ESCRT) proteins, are involved in the development of MVBs and ILVs, which contain 4 protein complexes, ESCRT-0, -I, -II, and -III.^14,18,19 In addition to ESCRT proteins, the X protein that interacts with apoptosis-linked gene 2 (ALIX),²⁰ the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) proteins,²¹ the vesicle trafficking 1 (VTA1) protein,²² and guanosine triphosphatases (GTPases) are also thought to be important players in the biogenesis and secretion of EXOs.²³

Current Challenges in EXO Isolation

EXO application in medicine has technological difficulties, much like any other novel nanobiotechnology that is under development. One issue is the uniformity of EXO separation methods, which needs special consideration. EXO isolation has no industry-accepted gold standard, and new techniques are developed practically daily. EXO extraction using ultracentrifugation is by far the commonly most used approach, although it has several limitations, including the simultaneous isolation of nonexosomal contaminants, poor repeatability, low RNA yield, and probable EXO destruction. Additionally, the majority of separation techniques make it impossible to achieve EXO purity at all levels.^24–26 EXOs content, purity, and size vary depending on the technique employed to separate them from body fluids. Information suggests that the isolation technique may alter the miRNA profile in EXOs.²⁷

According to the findings, EXO separation techniques from plasma or body fluids are the most important factor for defining the molecular or genetic payload of EXOs. Therefore, it is important to be cautious when selecting the best procedure for isolating EXOs and to distinguish between the EXO's true composition and its contamination with nucleic acids or plasma proteins.²⁸ Additionally, a study of various separation methods revealed that there were differences in the amounts of numerous proteins, including tetraspanins, that are frequently employed as “markers” of EXOs. It comes from several tumor cells. EXO purity is greatly influenced by the separation technique and has an impact on how the results should be interpreted. The highest purity, however, typically comes at the expense of poor yield and may not be significant for studies in which biomarkers can be reliably detected, but it is significant for other applications, such as proteomic analysis.^29,30

Definition and Biogenesis of miRNAs

Much evidence has shown that miRNAs, a small part of ncRNAs, regulate posttranscriptional gene expression, intracellular transcription, and mRNA degradation to manage cancer cell proliferation, apoptosis, differentiation, metastasis, and stem cell features. It is estimated that miRNAs target more than 50% of all mRNAs, and each miRNA is expected to influence hundreds of target mRNAs.^31,32

Various studies have shown that miRNAs in body fluids maintain their stability under a variety of severe situations, including boiling, extremely low or high pH, multiple freeze-thaw cycles, and long-term room temperature storage.³³

A well-known process is responsible for the synthesis of these small single-strand RNAs; RNA polymerase II transcribes the miRNA genes in the nucleus to create a primary transcript, also known as a pri-miRNA. Exportin 5 is a protein that then transports the pre-miRNA to the cytoplasm, where the Dicer complex turns it into a mature miRNA duplex around 20 to 22 nucleotides in length. Once the strands have been separated, the passenger strand is destroyed, and the second strand is incorporated in the RNA-induced silencing complex (RISC), which targets the specific mRNA and inhibits gene expression through mRNA degradation or translational suppression based on the 3ʹ-untranslated region (3ʹUTR) and complementary sequence of the target.^34,35

Role of ExomiRs in Cancer

ExomiRs play an important role in controlling the development of cancer.³⁶ Cancer cells secrete at least 10-fold more EXOs than normal cells and tumor-derived EXOs (TDEs) can transport chemokines, ExomiRs, growth factors, and various small molecules, thereby enhancing intercellular communication.^37,38 EXOs are released and then ingested by both nearby and distant cells, where the miRNAs they contain regulate processes like sabotaging tumor immunity and the microenvironment, potentially promoting tumor proliferation, invasion, metastasis, angiogenesis, and drug resistance.³⁶ Furthermore, evidence suggests that the tumor microenvironment significantly participates in the metabolic rewiring of cancerous cells through extracellular vesicles, promoting complete nutrient exploitation and changing the microenvironment from a normal to a tumor-favorable state. This alteration allows for invasion, drug resistance, and tumor growth.³⁹

Additionally, ExomiRs produced by cancer help recruit and modify elements of the tumor environment.⁴⁰ These molecules unquestionably play a part in cancer as tumor suppressors and promoters, influencing angiogenesis, tumor growth, proliferation process, metastasis, and cell migration as well as the epithelial–mesenchymal transition (EMT).³⁸ ExomiRs can also impact the ECM, immune system activation, and recruitment in the area around the tumor.⁴¹

General Roles of ExomiRs in Cancer

ExomiRs are exported by cancer cells to neighboring cancer cells around them. In the tumor microenvironment, they facilitate communication between primary tumor cells and other cells. EXOs produced by healthy cells can influence the behavior of malignant cells, and those produced by virus-infected cells can impact not only their oncology but also that of normal cells.³⁶

ExomiRs and Tumor Immunity

Tumor-derived EXOs aid tumor immune escape by transporting immunosuppressive factors and molecules.⁴² ExomiRs act as information carriers and can modulate the behavior of certain immunological activity factors and immune target cells, such as T lymphocytes, dendritic cells (DCs), and natural killer (NK) cells.⁴³ They are also involved in the biology of T lymphocytes and NK.⁴⁴ Additionally, the process by which ExomiRs affect NK cells’ immunological activity and, consequently, cause tumor immunology resistance involves a multifaceted, multitargeted, and multifactored effect.³⁶

ExomiRs and Tumor Proliferation

Malignant cells can transport genetic information to different cells within the tumor microenvironment through EXOs.³⁶ ExomiRs play a role in the metastasis, angiogenesis process, proliferation, drug resistance, and tumor suppression of cancer cells, as some of them are transferred between donors and recipients.³⁶ Proliferation is a critical factor in the development and spread of cancer, characterized by changes in the expression and function of proteins involved in the cell cycle.³⁶ Additionally, cell proliferation is stimulated by the constitutive activation of several signal transduction pathways.⁴⁵

ExomiRs and Tumor Angiogenesis

The tumor angiogenesis process consists of numerous phases, including the enzymatic breakdown of the vessel's basement membrane and the sprouting, proliferation, branching, migration, and tube creation of endothelial cells. EXOs produced by several cell types, such as stromal, endothelial, and mesenchymal stem cells, have been demonstrated to serve as beneficial mediators in the tumor microenvironment.^46,47 One key variable affecting tumor angiogenesis is hypoxia, which can influence the activities of many different chemicals and encourage the development of ExomiRs.³⁶

ExomiRs and Tumor Metastasis

Numerous signaling molecules participate in intercellular communication. The several stages of the metastatic process require EXOs produced by the tumor.⁴⁸ Research has identified 4 common ways in which ExomiRs distribution occurs when a tumor is developing in the microenvironment.⁴⁹

First, miRNAs released by more invasive tumor cells may be absorbed by less invasive tumor cells through TDEs, which could lead to the worsening of a primary tumor.³⁶

Second, ExomiRs in the tumor microenvironment allow primary tumor cells to communicate with other cells.⁵⁰

Third, EXOs produced by healthy cells or common biological processes serve as a means of intercellular communication and can influence the behavior of tumor cells.⁴⁶

The fourth and final mode focuses on tumors induced by viral infections. Abnormally released ExomiRs from virus-infected cells create precancerous conditions in both healthy cells and the infected cells themselves.⁵¹

ExomiRs as a Cancer Biomarker

Typically used cancer biomarkers suffer from false negatives or false positives and are not specific to tumors.⁵² Therefore, the only accurate diagnostic option currently available is tumor biopsy, which is an invasive and potentially harmful procedure.¹⁰ In contrast, miRNAs are commonly found in circulation and can be released into the bloodstream by extracellular vesicles like EXOs.⁵³ According to mounting evidence, they offer significant advantages over blood-free miRNAs as diagnostic markers for cancer.⁵⁴ ExomiRs show more resistance to degradation than free miRNAs, primarily due to their lipid bilayer structure, which protects the miRNAs from degradation.⁵⁵ They are resistant to freeze-thaw cycles and can remain stable at −20 °C for up to 5 years. Additionally, ExomiRs remain almost unchanged even after 2 weeks at 4 °C.⁵⁶ Consequently, EXOs provide a source of miRNAs that allows for effective preservation and recovery, even under circumstances that would typically cause free miRNAs to degrade.¹⁰ As free miRNAs originate from various cell types, their intrinsic heterogeneity may reduce their sensitivity and specificity as cancer biomarkers. ExomiRs, on the other hand, are more reliable than circulating serum miRNAs, according to many studies.^57,58

Naturally, however, ExomiRs may face some of the same challenges as more conventional tumor biomarkers. For example, they may be released by other cell types, potentially concealing signals that are exclusive to cancer. However, it is anticipated that by profiling a diverse set of ExomiR markers and distinguishing those associated with tumor-specific protein markers, it will be feasible to enhance sensitivity and specificity, addressing the issues associated with existing cancer biomarkers. Through such efforts, ExomiRs hold promising potential for improving cancer diagnostics and contributing to a more accurate and effective approach to cancer detection and management.¹⁰

In the diagnosis of different cancers, false positive cases should also be considered. Some diseases with inflammatory causes report cancer in test results, which should be noted. People with benign bladder diseases such as infection, stones, inflammation, and hematuria can have false-positive findings for bladder cancer.⁵⁹ Glioblastoma has significant levels of exosomal miR-21 expression. However, patients with the Japanese encephalitis virus (JEV) have been found to have a positive expression of the miR-21 gene. Therefore, while diagnosing glioblastoma in virus-infected regions, it is important to pay more attention to the false positive results caused by exosomal miR-21.⁶⁰ The presence of “false” positive serum EXOs in chronic pancreatitis patients is one of the field's limitations. To significantly reduce medical mistakes, it could be required to include another panel that can identify inflammatory markers.⁶¹

Lung Cancer

In a study conducted by Cazzoli et al.⁶² plasma ExomiR expression levels were examined in patients with lung adenocarcinoma (LAC), pulmonary granuloma, and healthy smokers. The researchers discovered that by utilizing ExomiRs miR-200b-5p, miR-379, miR-378a, and miR-139-5p, they could distinguish lung cancer (LC) patients from healthy individuals. Moreover, by using ExomiRs miR-154-3p, miR-629, miR-151a-5p, miR-100, miR-200b-5p, and miR-30a-3p, they could differentiate between patients with LAC and those with lung granulomas. These findings suggest that ExomiRs hold promise as potential biomarkers for diagnosing and differentiating LC from other lung-related conditions, providing a noninvasive and potentially more accurate approach to LC detection and classification.

In the study conducted by Zhou et al.⁶³ 6 ExomiR groups (miR-425-5p, miR-19b-3p, miR-409-3p, miR-221-3p, miR-21-5p, and miR-584-5p) were identified, which could be used to differentiate patients with LAC from healthy individuals. Furthermore, the researchers found that all of the identified miRNAs, except miR-584-5p, were significantly up-regulated in LAC tissues. Indeed, joint diagnoses using several ExomiRs can potentially enhance diagnostic effectiveness. For example, the combination of exosomal markers let-7e-5p, let-7b-5p, miR-21-5p, and miR-24-5p collected from plasma could be valuable in differentiating patients with nonsmall-cell LC (NSCLC) from controls, even in the early stages. Additionally, these ExomiRs can also distinguish between squamous cell carcinoma (SCC) and LAC.⁶⁴

According to Shan et al.⁶⁵ the combination of ExomiRs miR-93-5p, miR-21-5p, miR-181a-5p, and miR-106a-5p could be valuable in detecting SCC. Similarly, Zhang et al.⁶⁶ demonstrated that a combination of 3 ExomiRs, miR-20a-5p, miR-106a-5p, and miR-93-5p, is useful for diagnosing SCC in male patients. They also observed that combinations of these 3 miRNAs were highly effective in distinguishing lung SCC from lung hematoma.

Similarly, Feng et al.⁶⁷ reported that serum EXOs from LAC patients had higher expression levels of miR-140-5p, miR-21-5p, and miR-126-3p compared to healthy controls. In the study by Zhang et al.⁶⁸ ExomiR-17-5p expression was found to be significantly elevated in patients with NSCLC compared to controls. Furthermore, Grimolizzi et al.⁶⁹ found in their study that ExomiRs-126 can be used to differentiate healthy people from patients with early-stage NSCLC.

Wu et al.⁷⁰ recently found that serum levels of miRNAs (miR-486-5p, miR-21-5p, miR-222-3p, and miR-141-3p) and ExomiRs (miR-486-5p and miR-146a-5p) were significantly elevated in the early-stage NSCLC patients. These miRNAs can be combined to aid the early diagnosis of NSCLC. Additionally, Sun et al.⁷¹ demonstrated that serum ExomiR-106b levels were correlated with lymph node metastases and TNM (tumor, nodes, metastases) staging and that they were higher in LC patients than healthy individuals.

Using qRT-PCR, Chen et al.⁷² confirmed that patients with LAC demonstrated elevated levels of ExomiR-7797 in serum and decreased miR-98-3p levels. Additionally, the combination of 2 miRNAs provided more accurate diagnoses.

According to the study by Kim et al.⁷³ patients with LAC showed increased levels of let-7a and ExomiRs-126 in both their tumor tissues and bronchoalveolar lavage (BAL) samples. The work of Roman-Canal et al.⁷⁴ provides additional pertinent evidence. They made it possible to employ ExomiRs from pleural and lavage fluids. Moreover, miR-150-5p, miR-144-5p, and ExomiRs-1-3p were used to diagnose LC specifically (Table 1).

Table 1.

ExomiRs Are Used as LC and CRC Diagnostic Biomarkers.

Cancer type	Exosome source	MicroRNAs type	Ref.
NSC	Plasma	let-7b-5p, let-7e-5p, miR-24-5p, miR-21-5p	Jin et al.⁶⁴
		miR-342-5p, miR-574-5p, miR-222-3p	Han et al.¹¹⁷
		miR-378a, miR-379, miR-139-5P, miR-200b-5P, miR-151a-5p, miR-629, miR-30a-3p, miR-200b-5p, miR-154-3p, miR-100	Cazzoli et al.⁶²
		miR-126, miR-144	Rodríguez et al.¹¹⁸
		miR-196-3p, miR-21-5P, miR-221-3P, miR-409-3P, miR-425-5P, miR-584-5P	Zhou et al.⁶³
		miR-30a-3p, miR-30e-3p, miR-181-5P, miR-361-5P, miR-15b-5p, miR-320b, miR-10b-5p	Jin et al.⁶⁴
		miR-181-5P, miR-21-5p, miR-106a-5p, miR-93-5p, miR-181a-5p	Shan et al.⁶⁵
		miR-205, miR-19a, miR-19b, miR-30b, miR-20a	Aushev et al.¹¹⁹
		miR-96	Wu et al.¹²⁰
	Serum	miR-5684, miR-125b-5p	Zhang et al.¹²¹
		miR-126	Grimolizzi et al.⁶⁹
		miR-23a	Hsu et al.¹²²
		miR-21-5P, miR-141-3P, miR-222-3p, miR-486-5p, miR-146a-5p, miR-486-5p	Wu et al.⁷⁰
		miR-106b	Sun et al.⁷¹
LC		miR-7797, miR-98-3p	Chen et al.⁷²
		miR-17-5p	Zhang et al.⁶⁸
		miR-21-5p, miR-126-3P, miR-140-5p	Feng et al.¹²³
		miR-20b-5p, miR-3187-5p	Zhang et al.¹²⁴
		miR-620	Tang et al.¹²⁵
		miR-21, miR-210, miR-155, miR-25, miR-486	Liu et al.¹²⁶
		miR-106a-5P, miR-20a-5p, miR-93-5p	Zhang et al.⁶⁶
	BAL	miR-302a, miR-302c	Rodríguez et al.¹¹⁸
	BAL	miR-126	Kim et al.⁷³
	Pleural lavage	miR-1-3P, miR-150-5p, miR-144-5p	Roman-Canal et al.⁷⁴
		miR-205-5p, miR-483-5p, miR-375, miR-200c-3p, miR-429, miR-200b-3p, miR-200a-3p, miR-203a-3p, miR-141-3p	Wang et al.¹²⁷
		miR-205-5p, miR-200b	Lin et al.¹²⁸
		miR-182, miR-210	Tamiya et al.¹²⁹
		miR-200	Hydbring et al.¹³⁰
CRC	Plasma	miR-27a, miR-130a	Liu et al.⁷⁵
		miR-92b	Min et al.⁷⁶
		miR-150-3p, miR-145-3p, miR-139-3p, let-7b-3p	Min et al.⁷⁷
		miR-320c, miR-125a-3p	Wang et al.⁷⁸
		miR-96	Sun et al.¹³¹
		miR-221	Yau et al.¹³²
		miR-451	Phua et al.¹³³
		miR-760, miR-7, miR-93	Wang et al.¹³⁴
		miR-92, miR-92a	Wu et al.¹³⁵
		miR-375	Xu et al.¹³⁶
		miR-106a	Zhang et al.¹³⁷
		miR-183	Yuan et al.¹³⁸
		miR-17-3p	Li et al.¹³⁹
		miR-601	Wang et al.¹⁴⁰
		miR-139-3p, miR-431	Kanaan et al.¹⁴¹
	Serum	miR-99b-5p, miR-150-5p	Zhao et al.¹⁴²
		miR-422a	Zheng et al.¹⁴³
		miR-1290	Toiyama et al.¹⁴⁴
		miR-21	Toiyama et al.¹⁴⁵
		miR-145	Li et al.¹³⁹
		miR-144	Kalimutho et al.¹⁴⁶
		miR-18a	Yau et al.¹³²
		miR-19a	Chen et al.¹⁴⁷
		miR-23a-3p, miR-27a-3p, miR-142-5p, miR-376c-3p	Vychytilova-Faltejskova et al.¹⁴⁸

Abbreviations: ExomirRs, exosomal miRNAs; LC, lung cancer; CRC: colorectal cancer; BAL, bronchoalveolar lavage; miRNA, microRNA; NSCLC: nonsmall cell lung cancer.

Colorectal Cancer

The plasma expression levels of ExomiR-130a and ExomiR-27a are significantly higher in early-stage colorectal cancer (CRC) patients compared to healthy individuals, with area under the curve (AUC) values of 0.742 and 0.773, respectively. Importantly, high expression levels of ExomiR-130a and ExomiR-27a are correlated with poor patient prognoses.⁷⁵ Furthermore, CRC patients exhibit significantly lower levels of plasma ExomiR-92b compared to noncancerous controls (NC), indicating the potential of this EXO as a promising biomarker for early CRC diagnosis, particularly in patients with the TNM stage II (AUC = 0.793). The accuracy of miR-92b reaches 0.867, even in patients of different ages.⁷⁶ Plasma EXOs were found to contain 4 miRNAs (miR-139-3p, miR-145-3p, miR-150-3p, and let-7b-3p), all of which showed enrichment in the early-stage CRC. These miRNAs exhibited good diagnostic efficacy with AUC values of 0.692, 0.679, 0.686, and 0.792, respectively.⁷⁷ miR145-3p combined with miR-139-3p and miR let-7b-3p can diagnose early-stage CRC with an AUC value of 0.927.⁷⁷ In addition, patients with early-stage CRC have significantly higher levels of ExomiR-320c and miR-125a-3p in their plasma than healthy volunteers.⁷⁸ Even though miR-125a-3p on its own has an AUC of 0.685, combined with carcinoembryonic antibody (CEA), its AUC value reaches 0.855⁷⁸ (Table 1).

Hepatocellular Cancer

miRNAs play a critical role in the progression and development of hepatocellular carcinoma (HCC) by functioning as regulators.⁷⁹ They are involved in various regulatory mechanisms such as apoptosis, metastasis, angiogenesis, autophagy, invasion, EMT, drug resistance, and proliferation in HCC. By controlling gene expression in target cells, ExomiRs are also crucial in HCC invasion, metastasis, proliferation, and drug resistance. Additionally, some miRNAs, such as ExomiRs, can be used as possible diagnostic and prognostic markers for HCC⁷⁹ (Tables 2 and 3).

Table 2.

Types of miRNA Functions.

Function	MicroRNA	Ref.
Epithelial–mesenchymal transition (EMT)	miR-320a	Liu et al.¹⁴⁹
Drug resistance	miR-199-3p, miR-744, miR-32-5p	Liu et al.¹⁴⁹
Angiogenesis	miR-21, miR-200b-3p, miR-155, miR-210, miR-182, miR-26a, miR-146a, miR-122	Liu et al.¹⁴⁹
Apoptosis	miR-25, miR-101, miR-337	Xu et al.⁷⁹
Autophagy	miR-7, miR-26, miR-101, miR-142-3p, miR-181a	Xu et al.⁷⁹
Metastasis	miR-320a, miR-1247-3p, miR-92a-2-5p, miR-23a/b, miR-10b, miR-21	Liu et al.¹⁴⁹
Immune reaction	miR-449c-5p, miR-92b, miR-23a	Liu et al.¹⁴⁹
Proliferation	miR-320a, miR-331-3p, miR-320a, miR-365	Xu et al.⁷⁹

Table 3.

ExomiRs Used as HCC Diagnostic Biomarkers.

Cancer type	Exosome source	MicroRNAs type	Ref.
HCC	Serum	miR-21	Wang et al.¹⁵⁰
		miR-665	Qu et al.¹⁵¹
		miR-9-3p	Tang et al.¹⁵²
		miR-122, miR-148a	Wang et al.¹⁵³
		miR-18a, miR-221, miR-222, miR-224, miR-101, miR-106b, miR-122, miR195, miR-18a-5p, miR-215-5p, miR-940, miR-101, miR-106b, miR-93	Sohn et al.¹⁵⁴
		miR-10b-5p, miR-215-5p	Cho et al.¹⁵⁵
		miR-125b, miR-145, miR-192, miR-194, miR-29a, miR-17-5p, miR-106a	Xue et al.¹⁵⁶
		miR-18a	Guan et al.¹⁵⁷
		miR-26a, miR-29c, miR-103, miR-181c, miR-181a	Li et al.¹⁵⁸
		miR-1246	Murakami et al.¹⁵⁹
		miR-10b, miR-200a	Bukong et al.¹⁶⁰
		miR-519d, miR-1228	Lin and Zhang¹⁶¹
		miR-140-3p, miR-30d-5p, miR-29b-3p, miR-130b-3p, miR-330-5p, miR-296-3p	Yu et al.¹⁶²
		miR-638	Shi et al.¹⁶³
		miR-718	Sugimachi et al.¹⁶⁴
		miR-122a	Luo et al.¹⁶⁵
		miR-183	Liang et al.¹⁶⁶
		miR-130b, miR-15b	Liu et al.¹⁶⁷
		miR-618, miR-650	Abdalla and Haj-Ahmad¹⁶⁸
		miR-223,	Xu et al.¹⁶⁹
		miR-16, miR-199a	Qu et al.¹⁷⁰
		miRNA-1247-3p	Fang et al.¹⁷¹
	Cells	miR-335-5p	Wang et al.¹⁷²
	CCS	miR-584, miR-517c, miR-378, miR-520f, miR-142-5p, miR-451, miR-518d, miR-215, miR-376a, miR-133b	Kogure et al.¹⁷³
	CCS	miR-423-5p, miR-21-5, miR-486-5p, miR-10b-5p	Jx et al.¹⁷⁴
	Plasma	miR-139	Li et al.¹⁷⁵
	Plasma	miR-483-5p	Shen et al.¹⁷⁶

Abbreviations: ExomirRs, exosomal miRNAs; HCC, hepatocellular carcinoma; CCS, cell cultural supernatants; miRNA, microRNA.

Gastric Cancer

According to Tang et al.⁸⁰ serum levels of ExomiR-9-5p, let-7g-5p, miR-146b-5p, and miR-92b-3p can be employed as possible markers in patients with early-stage gastric cancer (GC). Similarly, serum levels of ExomiR-590-5p have been observed to be significantly higher in some patients diagnosed with early-stage GC (stage I/II) compared to healthy individuals.⁸¹

Shi et al.⁸² discovered that serum levels of ExomiR-1246 were significantly higher in patients with GC, allowing for the differentiation of healthy individuals from those with early-stage GC. Additionally, GC patients were found to have considerably lower serum levels of ExomiR-92a-3p compared to healthy individuals.⁸³ In a study by Huang et al.⁸⁴ the diagnostic efficacy of 58 circulating miRNAs in the sera of GC patients was investigated in a three-step investigation. They found that the expression of miR-296-5p, miR-20a-3p, miR-10b-5p, and miR-195-5p was noticeably increased in EXOs from serum samples of GC patients. According to Wei et al.⁸⁵ ExomiR-15b-3p expression was seen to be elevated, and it may be used as a prognostic and diagnostic marker in GC (Table 4).

Table 4.

ExomiRs Used as GC, Pancreatic, and Prostate Diagnostic Biomarkers.

Cancer type	Exosome source	MicroRNAs type	Ref.
Gastric	Serum	miR-92a-3p	Isobe et al.⁸³
		let-7g-5p, miR-92b-3p, miR-146b-5p, miR-9-5p	Tang et al.⁸⁰
		miR-590-5p	Zheng et al.⁸¹
		miR-1246	Shi et al.⁸²
		miR-15b-3p	Wei et al.⁸⁵
		miR-10b-5p, miR-132-3p, miR-185-5p, miR-195-5p, miR-20a-3p, miR-296-5p	Huang et al.⁸⁴
Pancreatic cancer	Serum	miR-21	Pu et al.⁸⁶ Goto et al.⁸⁹ Lai et al.¹⁷⁷ and Que et al.¹⁷⁸
		miR-451a, miR-191	Goto et al.⁸⁹
		miR-17-5p	Que et al.¹⁷⁸
		miR-4644, miR-1246, miR-3976, miR-4306	Madhavan et al.¹⁷⁹
		miR-10b, miR-30c, miR-181a, miR-let7a	Lai et al.¹⁷⁷
	Pancreatic juice	miR-21, miR-155	Nakamura et al.⁹¹
	Plasma	miR-196a, miR-1246	Xu et al.⁸⁷ and Xu et al.¹⁸⁰
		miR-451a	Takahasi et al.⁸⁸
		miR-483-3p	Abue et al.¹⁸¹
	Blood	miR-196a, miRNA-16a	Engle et al.⁹³ and Kondo et al.¹⁸²
	Saliva	miR-1246, miR-4644	Machida et al.⁹²
Prostate	Serum	miR-375, miR-141	Bryant et al.⁹⁶
		miR-1246	Bhagirath et al.¹⁸³
		miR-103, miR-451, miR-24, miR-26b, miR-30c, miR-93, miR-106a, miR-223, miR-874, miR-146a, miR-125b, miR-100, miR-107, miR-130b	Mihelich et al.¹⁸⁴
		miR-200a, miR-200c, miR-210	Cheng et al.¹⁸⁵
		miR-298	Selth et al.¹⁸⁶
		miR-378	Nguyen et al.¹⁸⁷
		miR-433, miR-605, miR-135a	Alhasan et al.¹⁸⁸
	Plasma	miR-1290	Huang et al.¹⁸⁹
		miR-200c-3p	Endzeliņš et al.¹⁹⁰
		miR-622, miR−1285	Chen et al.¹⁹¹
		miR-106a/miR-130b, miR-106a/miR-223	Sharova et al.¹⁹²
		miR-16, miR-148a, miR-195	Al-Qatati et al.¹⁹³
	Urine	miR-196a-5p, miR-501-3p, miR-92a-1-5p, miR-143-3p	Rodríguez et al.¹⁹⁴
		miR-34a-5p, miR-141-5p, miR-574-3p, miR-21-5p, miR-574	Samsonov et al.¹⁹⁵
		miR-2909	Wani et al.¹⁹⁶
		miR-21	Foj et al.¹⁹⁷
		miR-19b	Bryzgunova et al.¹⁹⁸
		miR-145	Xu et al.¹⁹⁹
	Cell-free urine samples	miR-222-3p, miR-24-3p/miR-30c-5p, miR-125b-5p	Fredsøe et al.²⁰⁰
	Urinary pellets and urinary exosomes	miR-21, miR-214	Foj et al.¹⁹⁷

Abbreviations: ExomirRs, exosomal miRNAs; GC, gastric cancer.

Pancreatic Cancer

Early-stage pancreatic cancer patients exhibit significantly elevated serum levels of ExomiR-21 when compared to healthy individuals.⁸⁶ Importantly, clinical risk factors like blood type, gender, smoking history, drinking history, age, body mass index, and diabetes mellitus have no significant impact on ExomiR-21 levels. Researchers have discovered that in stages I and IIa PDAC patients, intraductal papillary mucinous neoplasms (IPMN) have significantly elevated plasma ExomiR-196a and ExomiR-1246 levels compared to healthy people.⁸⁷ In addition, the plasma EXOs of stages I and II patients show a significant increase in miR-451a compared to healthy individuals, implying that ExomiR-451a could be used for the early detection of PDAC.⁸⁸

Likewise, Goto et al.⁸⁹ discovered that early-stage pancreatic cancers can be diagnosed using serum ExomiR-191, ExomiR-451a, and ExomiR-21. The receiver operating characteristic (ROC) analysis of these EXOs is more accurate than that of carcinoembryonic antigen (CEA) (AUC of 0.754, 0.935, 0.741, and 0.601, respectively). Additionally, a high ExomiR-21 expression level is an independent predictor of overall survival. With a 0.9% positive prediction rate, ExomiRs are more effective than CA19-9 in making an early diagnosis of pancreatic cancer.⁹⁰

ExomiRs have been also discovered in other body fluids, such as pancreatic juice (miR-21 and miR155),⁹¹ saliva (miR-1246 and miR-4644),⁹² and blood (miR-196a and miR-16a)^93,94 (Table 4).

Prostate Cancer

It has been demonstrated that EXOs, which include proteins, DNAs, and RNAs (miRNAs), play a crucial role in tumor development and represent a rich source of potential biomarkers, especially for miRNA content profiling⁹⁵ (Table 4). ExomiRs such as miR-375 and miR-141 have been studied in plasma for prostate cancer.⁹⁶

Brain Cancer

Santangelo et al.⁹⁷ reported an AUC of 0.87 for a glioblastoma multiforme (GBM) serum panel made up of ExomiR-21, miR-222, and miR-124-3p.

In patients with advanced gliomas, the expression of these miRNAs dropped dramatically after tumor excision. In another study, miR-320 and miR-574-3p levels were shown to be significantly higher in EXOs extracted from the sera of 75 individuals with GBM, and they were associated with the diagnosis of this disease.⁹⁸

Serum ExomiR-301a levels are considerably higher in glioma patients than in controls, corresponding with advancing pathological stages. It was also discovered that serum ExomiR-301a levels are dramatically decreased following surgical removal of tumors but then rise again after disease recurrence.⁹⁹ It has been shown that miR-1246 collected from cerebrospinal fluid (CSF) can be used as a possible diagnostic biomarker in gliomas and that miR-1246 targeting therapy may damage the immunosuppressive tumor microenvironment and provide insight into anticancer immunotherapy.⁹⁵ According to Shao et al.¹⁰⁰ ExomiR-454-3p has a sensitivity and specificity of 79.17% and 91.67%, respectively, showing it to potentially be a glioma diagnostic biomarker (Table 5).

Table 5.

ExomiRs Are Used as Brain Cancer, OC, Breast Cancer, OSCC, and Bladder Cancer Diagnostic Biomarkers.

Cancer type	Exosome source	MicroRNAs type	Ref.
Brain cancer	Serum	miR-21, miR-222, miR-124-3p	Santangelo et al.⁹⁷
		miR-320, miR-574-3p	Manterola et al.⁹⁸
		miR-301a	Lan et al.⁹⁹
	CSF	miR-1246	Qian et al.⁹⁵
	Plasma	miR-454-3p	Shao et al.¹⁰⁰
Ovarian	Serum	miR-200a-3p, miR-766-3p, miR-26a-5p, miR-142-3p, let-7d-5p, miR-328-3p	Yokoi et al.¹⁰¹
		miR-373, miR-200a, miR-200b, miR-200c	Meng et al.²⁰¹
		miR-1290	Kobayashi et al.¹⁰² and Jeon et al.²⁰²
		miR-222-3p	Ying et al.²⁰³
		miR-145, miR-200c, miR-93	Kim et al.²⁰⁴
		miR-34a	Maeda et al.²⁰⁵
	Peritoneal effusions	miR-21	Cappellesso et al.¹⁰³
	Urine	miR-92a	Zavesky et al.¹⁰⁴
	Urine	miR-30a-5p	Zhou et al.¹⁰⁵
	Plasma	miR-205	Zhu et al.¹⁰⁶
		miR-320d, miR-4479, miR-6763-5p	Wang et al.²⁰⁶
		miR-4732-5p	Liu et al.¹⁰⁷
Breast cancer	Plasma	miR-223-3p	Yoshikawa et al.¹¹²
		miR-93	Ni et al.¹¹³
		miR-1246	Hannafon et al.²⁰⁷
		miR-155, miR-301	Stevic et al.²⁰⁸
	Cell culture	miR-21	Hannafon et al.²⁰⁷
		miR-16	Ni et al.¹¹³
		miR-24	Jang et al.²⁰⁹
		miR-423-5p	Zhong et al.²¹⁰
	Serum	miR-19b-3p, miR-20b-5p, miR-106a-363 cluster	Li et al.¹¹¹
		miR-340-5p, miR-17-5p, miR-130a-3p, miR-93-5p	Sueta et al.²¹¹
		miR-372, miR-373, miR-101	Eichelser et al.¹⁰⁸
		miR-206, miR-1246	Jang et al.²⁰⁹
		miR-93-5p	Ni et al.¹¹³
		miR-18a-3p	Zhang et al.¹⁰⁹
OSCC	Plasma	miR-223	Tachibana et al.²¹²
Bladder	Urine	miR-1285-3p, miR-142-3p, miR-16-1-3p, miR-195-3p, miR-196b-5p, miR-23b-3p, miR-28-5p, miR-299-3p, miR-3155a, miR-3162-5p, miR-3678-3p, miR-4283, miR-4295, miR-4311, miR-4531, miR-492, miR-5096, miR-513b-5p, miR-5187-5p, miR-92a-2-5p, miR-601, miR-619-5p	Yasui et al.¹¹⁶
		miR-375, miR-146a	Andreu et al.²¹³
		miR-155-5p miR-15a-5p, miR-21-5p, miR-132-3p, miR-31-5p	Matsuzaki et al.²¹⁴
		miR-21, miR-93, miR-940	De Long et al.²¹⁵
		miR-30a-5p, miR-486-5p	Pardini et al.²¹⁶
		miR-652, miR-7-5p, miR-22-3p, miR-29a-3p, miR-126-5p, miR-200a-3p, miR-375, miR-423-5p	Du et al.²¹⁷
		miR-199a-3p, miR-200a, miR-222, miR-429, miR-143, miR-106b, miR-200c, miR-491-5p, miR-146b-5p, miR-191, miR-141, miR-140-3p, miR-99b, miR-223, miR-766, miR-96, miR-224, miR-30a, miR-1305, miR-142-5p, miR-93, miR-140-5p	Urquidi et al.²¹⁸
		miR-6124/miR-4511	Piao et al.²¹⁹
		miR-99a, miR-125b	Zhang et al.²²⁰
		miR-22-3p, miR-200a-3p	Du et al.²¹⁷
		miR16, miR34a, miR221, miR21, miR205, miR200c	Sapre et al.²²¹
		miR-214	Kim et al.²²²
	Serum	miR-30a-5p, miR-152, miR-27a-3p, miR-15b-5p, miR-3187-3p, miR-148b-3p	Jiang et al.²²³
		miR-152	Jiang et al.²²⁴
		miR-422a-3p, miR-27a-3p, miR-103a-3p, miR-486-3p	Jiang et al.²²⁵
	Cells	miR-30a-3p, miR-205-5p, miR-141-3p, miR-137-3p, miR-99a-5p	Baumgart et al.²²⁶
	Cells	miR-21-5p, miR-Let-7i-3p	Fanous et al.²²⁷

Abbreviations: ExomirRs, exosomal miRNAs; OSCC, oral squamous cell carcinoma; OC: ovarian cancer; CSF, cerebrospinal fluid.

Ovarian Cancer

According to Yokoi et al.¹⁰¹ ExomiRs miR-766-3p, miR-200a-3p, miR-26a-5p, miR-374a-5p, miR-142-3p, miR-328-3p, and let-7d-5p show higher expression levels in the serum of early-stage ovarian cancer (OC) patients (n = 15) compared to healthy individuals. These findings suggest that these miRNAs have the potential to distinguish between early-stage OC patients and healthy individuals. Other researchers Kobayashi et al.¹⁰² have analyzed the combination of miR-200a, miR-200b, and miR-200c and found that this EXO compound could differentiate benign ovarian disorders from EOC patients with a specificity of 90% and a sensitivity of 88%.

According to Cappellesso et al.¹⁰³ miR-21 contributes to the oncogenesis of ovarian serous carcinoma (OSC). ExomiR-21 has the potential to stimulate neoplastic transformation in target cells and might be employed as a diagnostic tool. Urine ExomiRs are easily accessed and have lately been more extensively studied, especially in gynecological and urological illnesses. Zavesky et al.¹⁰⁴ demonstrated a significant increase in miR-92a levels in the urine of OC patients, suggesting that it could serve as a diagnostic tool.

According to miRNA microarray data, miR-30a-5p shows higher levels in urine samples from ovarian serous adenocarcinoma (OSA) patients compared to healthy individuals. Conversely, decreased concentrations of miR-30a-5p were observed in the urine of individuals with colon cancer and GC, indicating that urinary levels of miR-30a-5p might be specific to OC. These findings suggest that exosomal urinary miR-30a-5p could potentially serve as a specific diagnostic biomarker for OC.¹⁰⁵ The expression of miR-205 was considerably greater in the plasma EXOs of OC patients than in the benign tumor group and healthy individuals, and throughout stages III and IV of OC and lymph node metastases, miR-205 levels were raised. Thus, miR-205 concentration in plasma EXOs is a promising tumor biomarker to help diagnose OC.¹⁰⁶

ExomiR-4732-5p extracted from plasma has a specificity of 82.4% and a sensitivity of 85.7% for distinguishing EOC patients from healthy individuals. Thus, it could act as a possible marker for tracking the development of EOC from early to late stages. It may also be a potential new biomarker for detecting EOC¹⁰⁷ (Table 5).

Breast Cancer

miR-372,¹⁰⁸ miR-18a-3p,¹⁰⁹ miR-101, miR-423-5p,¹¹⁰ and 8 miRNAs of the miR-106a-363 cluster¹¹¹ can differentiate breast cancer patients from healthy individuals and are associated with cancer proliferation, cell properties, and migration. Triple-negative patients have higher levels of other miRNAs, like miR-373, than healthy controls or even luminal cancer patients; miR-223-3p¹¹² levels are higher in invasive ductal carcinoma patients than in those preoperatively diagnosed with ductal carcinoma in situ (DCIS); miR-93¹¹³ is also upregulated in DCIS.

Breast cancer patients have higher levels of plasma ExomiR-223-3p, which can differentiate them from the general population with early breast cancer. Moreover, ExomiR-223-3p expression is significantly increased in biopsy-proven invasive DCIS (n = 13) compared to early ductal carcinoma, suggesting that ExomiR-223-3p can be used to predict the risk of invasive lesions in DCIS patients.¹¹² Additionally, DCIS patients have significantly higher levels of plasma ExomiR-93 expression than healthy individuals (n = 80)¹¹³ (Table 5).

Oral and Oropharyngeal Cancer

Several ExomiRs related to oral SCC (OSCC) have been identified, such as miR-342-3p¹¹⁴ and miR-138¹¹⁵ (Table 5).

Bladder Cancer

miR-1285-3p, miR-142-3p, miR-16-1-3p, miR-195-3p, miR-196b-5p, miR-23b-3p, miR-28-5p, and miR-299-3p¹¹⁶ are identified as ExomiRs that can distinguish bladder cancer patients from healthy individuals. These miRNAs are associated with cancer proliferation, cell properties, and migration (Table 5).

Conclusion

EXOs serve as efficient and dependable carriers of miRNAs found in various body secretions. Moreover, the expression patterns of ExomiRs in tumor cells differ significantly from those in normal cells. As a result, ExomiRs offer a promising noninvasive approach to cancer detection. This innovative approach may lead to improved cancer therapy. Various industries are currently exploring the applications of EXOs in biotechnology. EXOs Diagnostics, now a part of Bio-Techne, has been at the forefront of developing molecular diagnostics using biological fluid samples. Their precision EXO technology has enabled liquid biopsy for the detection of lung and prostate malignancies. However, there are still challenges to overcome before this technology can be widely applied in clinical settings. One of the major hurdles is the standardization of EXO extraction methods from different body fluids. Future applications should focus on more efficient techniques that require minimal biofluid volume while ensuring high purity and yield of EXOs.

Footnotes

Abbreviations

Declaration of Conflicting Interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Funding for this work was provided by Kermanshah University of Medical Sciences, Kermanshah, Iran. This study was carried out under the approval code IR.KUMS.REC.1402.039 at Kermanshah University of Medical Sciences, Kermanshah, Iran.

ORCID iD

Leila Rezakhani

References

Rezakhani

Fekri

Rostaminasab

Rahmati

. Exosomes: special nano-therapeutic carrier for cancers, overview on anticancer drugs. Med Oncol. 2023;40(1):31.

Khazaei

Rezakhani

Alizadeh

Mahdavian

Khazaei

. Exosomes and exosome-loaded scaffolds: characterization and application in modern regenerative medicine. Tissue Cell. 2022;80:102007.

Rashidi

Bijari

Khazaei

Shojaei-Ghahrizjani

Rezakhani

. The role of milk-derived exosomes in the treatment of diseases. Front Genet. 2022;13:1-14.

Mattick

Makunin

. Small regulatory RNAs in mammals. Hum Mol Genet. 2005;14(suppl_1):R121-R132.

Abels

Breakefield

. Introduction to extracellular vesicles: biogenesis, RNA cargo selection, content, release, and uptake. Cell Mol Neurobiol. 2016;36(3):301-312.

Gilligan

Dwyer

. Engineering exosomes for cancer therapy. Int J Mol Sci. 2017;18(6):1122.

Anakor

Le Gall

Dumonceaux

Duddy

Duguez

. Exosomes in ageing and motor neurone disease: biogenesis, uptake mechanisms, modifications in disease and uses in the development of biomarkers and therapeutics. Cells. 2021;10(11):2930.

Mao

Gong

, et al. Serum exosomes contain ECRG 4 mRNA that suppresses tumor growth via inhibition of genes involved in inflammation, cell proliferation, and angiogenesis. Cancer Gene Ther. 2018;25(9-10):248-259.

Xie

Dang

Zhang

, et al. The role of exosomal noncoding RNAs in cancer. Mol Cancer. 2019;18(37):1-10.

10.

Thind

Wilson

. Exosomal miRNAs as cancer biomarkers and therapeutic targets. J Extracell Vesicles. 2016;5(1):31292.

11.

Sun

Shi

Yang

, et al. Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer. 2018;17(147):1-19.

12.

Whiteside

. Tumor-derived exosomes and their role in cancer progression. Adv Clin Chem. 2016;74:103-141.

13.

Rahmati

Khazaei

Nadi

Alizadeh

Rezakhani

. Exosome-loaded scaffolds for regenerative medicine in hard tissues. Tissue Cell. 2023;82:102102.

14.

Gurunathan

Kang

M-H

Jeyaraj

Qasim

Kim

J-H

. Review of the isolation, characterization, biological function, and multifarious therapeutic approaches of exosomes. Cells. 2019;8(4):307.

15.

Dai

Zhong

, et al. Exosomes: key players in cancer and potential therapeutic strategy. Signal Transduction Targeted Ther. 2020;5(1):1-10.

16.

Rezakhani

Alizadeh

Sharifi

Soleimannejad

Alizadeh

. Isolation and characterization of crab haemolymph exosomes and its effects on breast cancer cells (4T1). Cell Journal (Yakhteh). 2021;23(6):658.

17.

Colletti

Ceglie

Di Giannatale

Nazio

. Autophagy and exosomes relationship in cancer: friends or foes? Front Cell Dev Biol. 2021;8:614178.

18.

Hessvik

Llorente

. Current knowledge on exosome biogenesis and release. Cell Mol Life Sci. 2018;75(2):193-208.

19.

Mashouri

Yousefi

Aref

Molaei

Alahari

. Exosomes: Composition, biogenesis, and mechanisms in cancer metastasis and drug resistance. Mol Cancer. 2019;18(1):1-14.

20.

Roucourt

Meeussen

Bao

Zimmermann

David

. Heparanase activates the syndecan-syntenin-ALIX exosome pathway. Cell Res. 2015;25(4):412-428.

21.

Shi

Stewart

, et al. Reduced oligodendrocyte exosome secretion in multiple system atrophy involves SNARE dysfunction. Brain. 2020;143(6):1780-1797.

22.

Staubach

Wenzel

Beck

Rinschen

Müller

Hanisch

. Autosomal tubulointerstitial kidney disease—MUC1 type: differential proteomics suggests that mutated MUC1 (insC) affects vesicular transport in renal epithelial cells. Proteomics. 2018;18(7):1700456.

23.

Ostrowski

Carmo

Krumeich

, et al. Rab27a and Rab27b control different steps of the exosome secretion pathway. Nat Cell Biol. 2010;12(1):19-30.

24.

Gardiner

Vizio

Sahoo

, et al. Techniques used for the isolation and characterization of extracellular vesicles: results of a worldwide survey. J Extracell Vesicles. 2016;5(1):32945.

25.

Konoshenko

Lekchnov

Vlassov

Laktionov

. Isolation of extracellular vesicles: General methodologies and latest trends. BioMed Res Int. 2018;2018:1-27.

26.

Ludwig

Razzo

Yerneni

Whiteside

. Optimization of cell culture conditions for exosome isolation using mini-size exclusion chromatography (mini-SEC). Exp Cell Res. 2019;378(2):149-157.

27.

Rekker

Saare

Roost

, et al. Comparison of serum exosome isolation methods for microRNA profiling. Clin Biochem. 2014;47(1-2):135-138.

28.

Tang

Y-T

Huang

Y-Y

Zheng

, et al. Comparison of isolation methods of exosomes and exosomal RNA from cell culture medium and serum. Int J Mol Med. 2017;40(3):834-844.

29.

Serrano-Pertierra

Oliveira-Rodríguez

Rivas

, et al. Characterization of plasma-derived extracellular vesicles isolated by different methods: a comparison study. Bioengineering. 2019;6(1):8.

30.

Sharma

Ludwig

Muller

, et al. Immunoaffinity-based isolation of melanoma cell-derived exosomes from plasma of patients with melanoma. J Extracell Vesicles. 2018;7(1):1435138.

31.

O’Brien

Hayder

Zayed

Peng

. Overview of microRNA biogenesis, mechanisms of actions, and circulation. Front Endocrinol (Lausanne). 2018;9:402.

32.

Cech

Steitz

. The noncoding RNA revolution—trashing old rules to forge new ones. Cell. 2014;157(1):77-94.

33.

Martellucci

Orefice

Angelucci

Luce

Caraglia

Zappavigna

. Extracellular vesicles: new endogenous shuttles for miRNAs in cancer diagnosis and therapy? Int J Mol Sci. 2020;21(18):6486.

34.

Acunzo

Romano

Wernicke

Croce

. MicroRNA and cancer—a brief overview. Adv Biol Regul. 2015;57:1-9.

35.

Melo

Esteller

. Disruption of microRNA nuclear transport in human cancer. Paper presented at: Seminars in cancer biology, 2014.

36.

Sun

Shi

Yang

, et al. Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer. 2018;17(1):1-19.

37.

Akers

Gonda

Kim

Carter

Chen

. Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies. J Neuro-Oncol. 2013;113(1):1-11.

38.

Mao

Gong

, et al. Serum exosomes contain ECRG4 mRNA that suppresses tumor growth via inhibition of genes involved in inflammation, cell proliferation, and angiogenesis. Cancer Gene Ther. 2018;25(9):248-259.

39.

Chiarugi

Cirri

. Metabolic exchanges within tumor microenvironment. Cancer Lett. 2016;380(1):272-280.

40.

Tkach

Théry

. Communication by extracellular vesicles: where we are and where we need to go. Cell. 2016;164(6):1226-1232.

41.

Wang

, et al. Systematic characterization of A-to-I RNA editing hotspots in microRNAs across human cancers. Genome Res. 2017;27(7):1112-1125.

42.

Greening

Gopal

Simpson

Chen

. Exosomes and their roles in immune regulation and cancer. Paper presented at: Seminars in cell & developmental biology, 2015.

43.

Que

R-s

Lin

Ding

G-p

Z-r

Cao

L-p

. Increasing the immune activity of exosomes: the effect of miRNA-depleted exosome proteins on activating dendritic cell/cytokine-induced killer cells against pancreatic cancer. J Zhejiang Univ-Sci B. 2016;17(5):352-360.

44.

Berzins

Ritchie

. Natural killer T cells: drivers or passengers in preventing human disease? Nat Rev Immunol. 2014;14(9):640-646.

45.

Feitelson

Arzumanyan

Kulathinal

, et al. Sustained proliferation in cancer: Mechanisms and novel therapeutic targets. Paper presented at: Seminars in cancer biology, 2015.

46.

Zhang

Yao

, et al. Microenvironment-induced PTEN loss by exosomal microRNA primes brain metastasis outgrowth. Nature. 2015;527(7576):100-104.

47.

Liang

Zhang

Wang

Han

Zhao

. Exosomes secreted by mesenchymal stem cells promote endothelial cell angiogenesis by transferring miR-125a. J Cell Sci. 2016;129(11):2182-2189.

48.

Gomes

Nedel

Alves

Nör

Tarquinio

SBC

. Tumor angiogenesis and lymphangiogenesis: tumor/endothelial crosstalk and cellular/microenvironmental signaling mechanisms. Life Sci. 2013;92(2):101-107.

49.

Cao

Shen

Feng

. Tumor-derived exosomes in cancer progression and treatment failure. Oncotarget. 2015;6(35):37151-37168.

50.

Zhou

Fong

Min

, et al. Cancer-secreted miR-105 destroys vascular endothelial barriers to promote metastasis. Cancer Cell. 2014;25(4):501-515.

51.

Nanbo

Katano

Kataoka

, et al. Infection of Epstein–Barr virus in type III latency modulates biogenesis of exosomes and the expression profile of exosomal miRNAs in the Burkitt lymphoma Mutu cell lines. Cancers (Basel). 2018;10(7):237.

52.

Perkins

Slater

Sanders

Prichard

. Serum tumor markers. Am Fam Physician. 2003;68(6):1075-1082.

53.

Avgeris

Panoutsopoulou

Papadimitriou

M-A

Scorilas

. Circulating exosomal miRNAs: clinical significance in human cancers. Expert Rev Mol Diagn. 2019;19(11):979-995.

54.

Zhu

Zhao

Wang

, et al. Circulating exosomal miRNAs and cancer early diagnosis. Clin Transl Oncol. 2022;24(3):393-406.

55.

Zhou

Bai

Xie

. miRNA in plasma exosome is stable under different storage conditions. Molecules. 2014;19(2):1568-1575.

56.

Weber

Baxter

Zhang

, et al. The microRNA spectrum in 12 body fluids. Clin Chem. 2010;56(11):1733-1741.

57.

Lin

, et al. Combination of serum miRNAs with serum exosomal miRNAs in early diagnosis for non-small-cell lung cancer. Cancer Manag Res. 2020;12:485.

58.

Zhang

Z-J

Song

X-G

Xie

, et al. Circulating serum exosomal miR-20b-5p and miR-3187-5p as efficient diagnostic biomarkers for early-stage non-small cell lung cancer. Exp Biol Med. 2020;245(16):1428-1436.

59.

Stenzl

Sharma

Vasdev

. Urinary biomarkers in bladder cancer: a review of the current landscape and future directions. Paper presented at: Urologic Oncology: Seminars and Original Investigations, 2021.

60.

Shi

. Considering exosomal miR-21 as a biomarker for cancer. J Clin Med. 2016;5(4):42.

61.

Erb

Zöller

. Progress and potential of exosome analysis for early pancreatic cancer detection. Expert Rev Mol Diagn. 2016;16(7):757-767.

62.

Cazzoli

Buttitta

Di Nicola

, et al. microRNAs derived from circulating exosomes as noninvasive biomarkers for screening and diagnosing lung cancer. J Thorac Oncol. 2013;8(9):1156-1162.

63.

Zhou

Wen

Shan

, et al. A six-microRNA panel in plasma was identified as a potential biomarker for lung adenocarcinoma diagnosis. Oncotarget. 2017;8(4):6513-6525.

64.

Jin

Chen

, et al. Evaluation of tumor-derived exosomal miRNA as potential diagnostic biomarkers for early-stage non-small cell lung cancer using next-generation sequencing exosomal miRNA as early diagnostic biomarkers for NSCLC. Clin Cancer Res. 2017;23(17):5311-5319.

65.

Shan

Zhang

, et al. Identification of four plasma microRNAs as potential biomarkers in the diagnosis of male lung squamous cell carcinoma patients in China. Cancer Med. 2018;7(6):2370-2381.

66.

Zhang

Shan

Wang

, et al. A three-microRNA signature for lung squamous cell carcinoma diagnosis in Chinese male patients. Oncotarget. 2017;8(49):86897-86907.

67.

Feng

Zhao

Wang

Liu

. Upregulated expression of serum exosomal microRNAs as diagnostic biomarkers of lung adenocarcinoma. Annals of Clinical & Laboratory Science. 2018;48(6):712-718.

68.

Zhang

Yin

. Detection of circulating exosomal miR-17-5p serves as a novel non-invasive diagnostic marker for non-small cell lung cancer patients. Pathol Res Pract. 2019;215(8):152466.

69.

Grimolizzi

Monaco

Leoni

, et al. Exosomal miR-126 as a circulating biomarker in non-small-cell lung cancer regulating cancer progression. Sci Rep. 2017;7(1):1-12.

70.

Lin

, et al. Combination of serum miRNAs with serum exosomal miRNAs in early diagnosis for non-small-cell lung cancer. Cancer Manag Res. 2020;12:485-495.

71.

Sun

Chen

, et al. Exosomal miR-106b serves as a novel marker for lung cancer and promotes cancer metastasis via targeting PTEN. Life Sci. 2020;244(117297):1-7.

72.

Chen

Cao

Huang

Chen

. Serum exosomal miR-7977 as a novel biomarker for lung adenocarcinoma. J Cell Biochem. 2020;121(5-6):3382-3391.

73.

Kim

Eom

Kim

, et al. Diagnostic value of microRNAs derived from exosomes in bronchoalveolar lavage fluid of early-stage lung adenocarcinoma: A pilot study. Thorac Cancer. 2018;9(8):911-915.

74.

Roman-Canal

Moiola

Gatius

, et al. EV-associated miRNAs from pleural lavage as potential diagnostic biomarkers in lung cancer. Sci Rep. 2019;9(1):15057.

75.

Liu

Pan

Sun

, et al. Circulating exosomal miR-27a and miR-130a act as novel diagnostic and prognostic biomarkers of colorectal CancerDiagnostic circulating exosomal miRNAs in colorectal cancer. Cancer Epidemiol Biomarkers Prev. 2018;27(7):746-754.

76.

Min

Chen

Liu

, et al. Loss of circulating exosomal miR-92b is a novel biomarker of colorectal cancer at early stage. Int J Med Sci. 2019;16(9):1231.

77.

Min

Zhu

Chen

, et 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(1):1643670.

78.

Wang

Yan

Zhao

, et al. Circulating exosomal miR-125a-3p as a novel biomarker for early-stage colon cancer. Sci Rep. 2017;7(1):1-9.

79.

Tao

Shan

, et al. The role of microRNAs in hepatocellular carcinoma. J Cancer. 2018;9(19):3557.

80.

Tang

Cheng

Yao

, et al. Combination of four serum exosomal miRNAs as novel diagnostic biomarkers for early-stage gastric cancer. Front Genet. 2020;11:237.

81.

Zheng

G-D

Z-Y

, et al. Exosomal miR-590-5p in serum as a biomarker for the diagnosis and prognosis of gastric cancer. Front Mol Biosci. 2021;8:636566.

82.

Shi

Wang

Zhu

, et al. Exosomal miR-1246 in serum as a potential biomarker for early diagnosis of gastric cancer. Int J Clin Oncol. 2020;25(1):89-99.

83.

Isobe

Hisamori

Hogan

, et al. miR-142 regulates the tumorigenicity of human breast cancer stem cells through the canonical WNT signaling pathway. Elife. 2014;3(1):e01977.

84.

Huang

Zhu

, et al. Six serum-based miRNAs as potential diagnostic biomarkers for gastric cancer. Cancer Epidemiol Biomarkers Prev. 2017;26(2):188-196.

85.

Wei

Peng

Yang

, et al. Exosomal transfer of miR-15b-3p enhances tumorigenesis and malignant transformation through the DYNLT1/caspase-3/caspase-9 signaling pathway in gastric cancer. J Exp Clin Cancer Res. 2020;39(1):32.

86.

Ding

Zhou

Jia

Cao

. Elevated expression of exosomal microRNA-21 as a potential biomarker for the early diagnosis of pancreatic cancer using a tethered cationic lipoplex nanoparticle biochip. Oncol Lett. 2020;19(3):2062-2070.

87.

Y-F

Hannafon

Zhao

Postier

Ding

W-Q

. Plasma exosome miR-196a and miR-1246 are potential indicators of localized pancreatic cancer. Oncotarget. 2017;8(44):77028.

88.

Takahasi

Iinuma

Wada

, et al. Usefulness of exosome-encapsulated microRNA-451a as a minimally invasive biomarker for prediction of recurrence and prognosis in pancreatic ductal adenocarcinoma. J Hepato-Biliary-Pancreatic Sci. 2018;25(2):155-161.

89.

Goto

Fujiya

Konishi

, et al. An elevated expression of serum exosomal microRNA-191, -21, -451a of pancreatic neoplasm is considered to be efficient diagnostic marker. BMC Cancer. 2018;18(1):1-11.

90.

Lee KT

KIMJE

Lee

Paik

Rhee

Choi

. Clinical usefulness of carbohydrate antigen 19-9 as a screening test for pancreatic cancer in an asymptomatic population. J Gastroenterol Hepatol. 2004;19(2):182-186.

91.

Nakamura

Sadakari

Ohtsuka

, et al. Pancreatic juice exosomal microRNAs as biomarkers for detection of pancreatic ductal adenocarcinoma. Ann Surg Oncol. 2019;26(7):2104-2111.

92.

Machida

Tomofuji

Maruyama

, et al. Mir-1246 and miR-4644 in salivary exosome as potential biomarkers for pancreaticobiliary tract cancer. Oncol Rep. 2016;36(4):2375-2381.

93.

Engle

Tiriac

Rivera

, et al. The glycan CA19-9 promotes pancreatitis and pancreatic cancer in mice. Science. 2019;364(6446):1156-1162.

94.

Kondo

Murakami

Uemura

, et al. Prognostic impact of perioperative serum CA 19-9 levels in patients with resectable pancreatic cancer. Ann Surg Oncol. 2010;17(12):2321-2329.

95.

Qian

Wang

Guo

, et al. Hypoxic glioma-derived exosomes deliver microRNA-1246 to induce M2 macrophage polarization by targeting TERF2IP via the STAT3 and NF-κB pathways. Oncogene. 2020;39(2):428-442.

96.

Bryant

Pawlowski

Catto

, et al. Changes in circulating microRNA levels associated with prostate cancer. Br J Cancer. 2012;106(4):768-774.

97.

Santangelo

Imbrucè

Gardenghi

, et al. A microRNA signature from serum exosomes of patients with glioma as complementary diagnostic biomarker. J Neuro-Oncol. 2018;136(1):51-62.

98.

Manterola

Guruceaga

Pérez-Larraya

, et al. A small noncoding RNA signature found in exosomes of GBM patient serum as a diagnostic tool. Neuro Oncol. 2014;16(4):520-527.

99.

Lan

Qing

Pan

Yue

. Serum exosomal miR-301a as a potential diagnostic and prognostic biomarker for human glioma. Cell Oncol. 2018;41(1):25-33.

100.

Shao

Xue

Wang

Luo

Zhi

Lan

. miR-454-3p is an exosomal biomarker and functions as a tumor suppressor in Glioma. Mol Cancer Ther. 2019;18(2):459-469.

101.

Yokoi

Yoshioka

Hirakawa

, et al. A combination of circulating miRNAs for the early detection of ovarian cancer. Oncotarget. 2017;8(52):89811-89823.

102.

Kobayashi

Sawada

Nakamura

, et al. Exosomal miR-1290 is a potential biomarker of high-grade serous ovarian carcinoma and can discriminate patients from those with malignancies of other histological types. J Ovarian Res. 2018;11(1):81.

103.

Cappellesso

Tinazzi

Giurici

, et al. Programmed cell death 4 and microRNA 21 inverse expression is maintained in cells and exosomes from ovarian serous carcinoma effusions. Cancer Cytopathol. 2014;122(9):685-693.

104.

Zavesky

Jandakova

Turyna

Langmeierova

Weinberger

Minar

. Supernatant versus exosomal urinary microRNAs. Two fractions with different outcomes in gynaecological cancers. Neoplasma. 2016;63(1):121-132.

105.

Zhou

Gong

Tan

, et al. Urinary microRNA-30a-5p is a potential biomarker for ovarian serous adenocarcinoma. Oncol Rep. 2015;33(6):2915-2923.

106.

Zhu

Chen

Wang

, et al. Detection of plasma exosomal miRNA-205 as a biomarker for early diagnosis and an adjuvant indicator of ovarian cancer staging. J Ovarian Res. 2022;15(1):27.

107.

Liu

Yoo

, et al. Plasma-derived exosomal miR-4732-5p is a promising noninvasive diagnostic biomarker for epithelial ovarian cancer. J Ovarian Res. 2021;14(1):1-14.

108.

Eichelser

Stückrath

Müller

, et al. Increased serum levels of circulating exosomal microRNA-373 in receptor-negative breast cancer patients. Oncotarget. 2014;5(20):9650.

109.

Zhang

Nguyen

Hickey

, et al. Immunomagnetic sequential ultrafiltration (iSUF) platform for enrichment and purification of extracellular vesicles from biofluids. Sci Rep. 2021;11(1):1-17.

110.

Liu

Shi

, et al. Cross-platform genomic identification and clinical validation of breast cancer diagnostic biomarkers. Aging (Albany NY). 2021;13(3):4258.

111.

Zhou

Xia

, et al. Circulating microRNAs from the miR-106a-363 cluster on chromosome X as novel diagnostic biomarkers for breast cancer. Breast Cancer Res Treat. 2018;170(2):257-270.

112.

Yoshikawa

Iinuma

Umemoto

Yanagisawa

Matsumoto

Jinno

. Exosome-encapsulated microRNA-223-3p as a minimally invasive biomarker for the early detection of invasive breast cancer. Oncol Lett. 2018;15(6):9584-9592.

113.

Stevic

Pan

, et al. Different signatures of miR-16, miR-30b and miR-93 in exosomes from breast cancer and DCIS patients. Sci Rep. 2018;8(1):1-10.

114.

Sakha

Muramatsu

Ueda

Inazawa

. Exosomal microRNA miR-1246 induces cell motility and invasion through the regulation of DENND2D in oral squamous cell carcinoma. Sci Rep. 2016;6(1):1-11.

115.

Xie

L-Y

Guo

Cheng

. Exosomes derived from microRNA-101-3p-overexpressing human bone marrow mesenchymal stem cells suppress oral cancer cell proliferation, invasion, and migration. Mol Cell Biochem. 2019;458(1-2):11-26.

116.

Yasui

Yanagida

Ito

, et al. Unveiling massive numbers of cancer-related urinary-microRNA candidates via nanowires. Sci Adv. 2017;3(12):e1701133.

117.

Han

Zhang

, et al. Tumor-derived circulating exosomal miR-342-5p and miR-574-5p as promising diagnostic biomarkers for early-stage lung adenocarcinoma. Int J Med Sci. 2020;17(10):1428-1438.

118.

Rodríguez

Silva

López-Alfonso

, et al. Different exosome cargo from plasma/bronchoalveolar lavage in non-small-cell lung cancer. Genes Chromosomes Cancer. 2014;53(9):713-724.

119.

Aushev

Zborovskaya

Laktionov

, et al. Comparisons of microRNA patterns in plasma before and after tumor removal reveal new biomarkers of lung squamous cell carcinoma. PLoS One. 2013;8(10):e78649.

120.

Zhou

Mei

, et al. Circulating exosomal microRNA-96 promotes cell proliferation, migration and drug resistance by targeting LMO7. J Cell Mol Med. 2017;21(6):1228-1236.

121.

Zhang

Tang

Song

Xie

Zhao

Song

. Tumor-derived exosomal miRNAs as diagnostic biomarkers in non-small cell lung cancer. Front Oncol. 2020;10:560025.

122.

Hsu

Hung

Chang

, et al. Hypoxic lung cancer-secreted exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1. Oncogene. 2017;36(34):4929-4942.

123.

Feng

Zhao

Wang

Liu

. Upregulated expression of Serum exosomal microRNAs as diagnostic biomarkers of lung adenocarcinoma. Ann Clin Lab Sci. 2018;48(6):712-718.

124.

Zhang

Song

Xie

, et al. Circulating serum exosomal miR-20b-5p and miR-3187-5p as efficient diagnostic biomarkers for early-stage non-small cell lung cancer. Exp Biol Med (Maywood). 2020;245(16):1428-1436.

125.

Tang

Zhang

Song

, et al. Tumor-derived exosomal miR-620 as a diagnostic biomarker in non-small-cell lung cancer. J Oncol. 2020;2020:6691211.

126.

Liu

Kannisto

, et al. Non-invasive detection of exosomal microRNAs via tethered cationic lipoplex nanoparticles (tCLN) biochip for lung cancer early detection. Front Genet. 2020;11:258.

127.

Wang

Zou

, et al. Identification of differential expressed PE exosomal miRNA in lung adenocarcinoma, tuberculosis, and other benign lesions. Medicine (Baltimore). 2017;96(44):e8361.

128.

Lin

Wang

Zou

, et al. Differential miRNA expression in pleural effusions derived from extracellular vesicles of patients with lung cancer, pulmonary tuberculosis, or pneumonia. Tumour Biol. 2016;37(10):15835-15845.

129.

Tamiya

Mitani

Saito

, et al. Exosomal microRNA expression profiling in patients with lung adenocarcinoma-associated malignant pleural effusion. Anticancer Res. 2018;38(12):6707-6714.

130.

Hydbring

De Petris

Zhang

, et al. Exosomal RNA-profiling of pleural effusions identifies adenocarcinoma patients through elevated miR-200 and LCN2 expression. Lung Cancer. 2018;124:45-52.

131.

Sun

Liu

Cogdell

, et al. Examining plasma microRNA markers for colorectal cancer at different stages. Oncotarget. 2016;7(10):11434.

132.

Yau

Dong

, et al. microRNA-221 and microRNA-18a identification in stool as potential biomarkers for the non-invasive diagnosis of colorectal carcinoma. Br J Cancer. 2014;111(9):1765-1771.

133.

Phua

Chue

Koh

Cheah

Chan

ECY

. Global fecal microRNA profiling in the identification of biomarkers for colorectal cancer screening among Asians. Oncol Rep. 2014;32(1):97-104.

134.

Wang

Xiang

, et al. A plasma microRNA panel for early detection of colorectal cancer. Int J Cancer. 2015;136(1):152-161.

135.

Dong

, et al. Detection of miR-92a and miR-21 in stool samples as potential screening biomarkers for colorectal cancer and polyps. Gut. 2012;61(5):739-745.

136.

Wang

Yan

Feng

. The expression of microRNA-375 in plasma and tissue is matched in human colorectal cancer. BMC Cancer. 2014;14(1):1-11.

137.

Zhang

Meng

Fan

Liu

Pei

Zhao

. Expression of plasma miR-106a in colorectal cancer and its clinical significance. J South Med Univ. 2014;34(3):354-357.

138.

Yuan

Zhu

Yan

Dang

. Plasma miR-183 predicts recurrence and prognosis in patients with colorectal cancer. Cancer Biol Ther. 2015;16(2):268-275.

139.

Liu

Wang

, et al. Serum miRNA expression profile as a prognostic biomarker of stage II/III colorectal adenocarcinoma. Sci Rep. 2015;5(1):1-13.

140.

Wang

Huang

, et al. Plasma miR-601 and miR-760 are novel biomarkers for the early detection of colorectal cancer. 2012.

141.

Kanaan

Roberts

Eichenberger

, et al. A plasma microRNA panel for detection of colorectal adenomas: a step toward more precise screening for colorectal cancer. Ann Surg. 2013;258(3):400-408.

142.

Zhao

Song

Niu

Tang

Song

Xie

. Circulating exosomal miR-150-5p and miR-99b-5p as diagnostic biomarkers for colorectal cancer. Front Oncol. 2019;9:1129.

143.

Zheng

Yang

, et al. Serum microRNA panel as biomarkers for early diagnosis of colorectal adenocarcinoma. Br J Cancer. 2014;111(10):1985-1992.

144.

Toiyama

Imaoka

Fujikawa

, et al. Circulating microRNA-1290 as a novel diagnostic and prognostic biomarker in human colorectal cancer. Cancer Res. 2016;76(14_Supplement):968-968.

145.

Toiyama

Takahashi

Hur

, et al. Serum miR-21 as a diagnostic and prognostic biomarker in colorectal cancer. JNCI: J National Cancer Inst. 2013;105(12):849-859.

146.

Kalimutho

Del Vecchio Blanco

Di Cecilia

, et al. Differential expression of miR-144* as a novel fecal-based diagnostic marker for colorectal cancer. J Gastroenterol. 2011;46(12):1391-1402.

147.

Chen

Xia

H-W

X-J

Zhang

Y-C

Tang

Q-L

. Serum miR-19a predicts resistance to FOLFOX chemotherapy in advanced colorectal cancer cases. Asian Pac J Cancer Prev. 2013;14(12):7421-7426.

148.

Vychytilova-Faltejskova

Radova

Sachlova

, et al. Serum-based microRNA signatures in early diagnosis and prognosis prediction of colon cancer. Carcinogenesis. 2016;37(10):941-950.

149.

Liu

Mao

Chen

. Exosomal microRNAs in hepatocellular carcinoma. Cancer Cell Int. 2021;21(1):1-12.

150.

Wang

Hou

Duan

Gao

Song

. Expression of serum exosomal microRNA-21 in human hepatocellular carcinoma. BioMed Res Int. 2014;2014:1-5.

151.

, et al. Exosomal miR-665 as a novel minimally invasive biomarker for hepatocellular carcinoma diagnosis and prognosis. Oncotarget. 2017;8(46):80666.

152.

Tang

Liu

, et al. Exosomal miR-9-3p suppresses HBGF-5 expression and is a functional biomarker in hepatocellular carcinoma. Minerva Med. 2018;109(1):15-23.

153.

Wang

Zhang

, et al. Serum exosomal micro RNAs combined with alpha-fetoprotein as diagnostic markers of hepatocellular carcinoma. Cancer Med. 2018;7(5):1670-1679.

154.

Sohn

Kim

Kang

, et al. Serum exosomal microRNAs as novel biomarkers for hepatocellular carcinoma. Exp Mol Med. 2015;47(9):e184-e184.

155.

Cho

Eun

Baek

, et al. Serum exosomal microRNA, miR-10b-5p, as a potential diagnostic biomarker for early-stage hepatocellular carcinoma. J Clin Med. 2020;9(1):281.

156.

Xue

Zhao

Wang

Qin

. Development and validation of serum exosomal microRNAs as diagnostic and prognostic biomarkers for hepatocellular carcinoma. J Cell Biochem. 2019;120(1):135-142.

157.

Guan

Yang

, et al. Clinical significance of microRNA-155 expression in hepatocellular carcinoma. Oncol Lett. 2016;11(2):1574-1580.

158.

Zhang

Liu

Xiang

Jiang

. Identification of endogenous controls for analyzing serum exosomal miRNA in patients with hepatitis B or hepatocellular carcinoma. Dis Markers. 2015;2015:893594.

159.

Murakami

Toyoda

Tanahashi

, et al. Comprehensive miRNA expression analysis in peripheral blood can diagnose liver disease. PloS One. 2012;7(10):e48366.

160.

Bukong

Momen-Heravi

Kodys

Bala

Szabo

. Exosomes from hepatitis C infected patients transmit HCV infection and contain replication competent viral RNA in complex with AgO2-miR122-HSP90. PLoS Pathog. 2014;10(10):e1004424.

161.

Lin

Zhang

. Diagnostic value of a microRNA signature panel in exosomes for patients with hepatocellular carcinoma. Int J Clin Exp Pathol. 2019;12(4):1478.

162.

L-X

Zhang

B-L

Yang

, et al. Exosomal microRNAs as potential biomarkers for cancer cell migration and prognosis in hepatocellular carcinoma patient-derived cell models. Oncol Rep. 2019;41(1):257-269.

163.

Shi

Jiang

Yang

Yan

Wang

. Decreased levels of serum exosomal miR-638 predict poor prognosis in hepatocellular carcinoma. J Cell Biochem. 2018;119(6):4711-4716.

164.

Sugimachi

Matsumura

Hirata

, et al. Identification of a bona fide microRNA biomarker in serum exosomes that predicts hepatocellular carcinoma recurrence after liver transplantation. Br J Cancer. 2015;112(3):532-538.

165.

Luo

Chen

Huang

, et al. Circulating microRNA-122a as a diagnostic marker for hepatocellular carcinoma. Onco Targets Ther. 2013;6:577-583.

166.

Liang

Gao

Shi

, et al. Expression and significance of microRNA-183 in hepatocellular carcinoma. Sci World J. 2013;2013:381874.

167.

Liu

Yao

Wang

, et al. Circulating miR-15b and miR-130b in serum as potential markers for detecting hepatocellular carcinoma: a retrospective cohort study. BMJ Open. 2012;2(2):e000825.

168.

Abdalla

Haj-Ahmad

. Promising candidate urinary microRNA biomarkers for the early detection of hepatocellular carcinoma among high-risk hepatitis C virus Egyptian patients. J Cancer. 2012;3:19-31.

169.

Che

, et al. Circulating microRNAs, miR-21, miR-122, and miR-223, in patients with hepatocellular carcinoma or chronic hepatitis. Mol Carcinog. 2011;50(2):136-142.

170.

Zhang

Afdhal

Albitar

. Circulating microRNAs as biomarkers for hepatocellular carcinoma. J Clin Gastroenterol. 2011;45(4):355-360.

171.

Fang

, et al. Tumor-derived exosomal miR-1247-3p induces cancer-associated fibroblast activation to foster lung metastasis of liver cancer. Nat Commun. 2018;9(1):191.

172.

Wang

Piontek

Sakaguchi

Selaru

. Exosome miR-335 as a novel therapeutic strategy in hepatocellular carcinoma. Hepatology. 2018;67(3):940-954.

173.

Kogure

Lin

Yan

Braconi

Patel

. Intercellular nanovesicle-mediated microRNA transfer: a mechanism of environmental modulation of hepatocellular cancer cell growth. Hepatology. 2011;54(4):1237-1248.

174.

, et al. Vps4A functions as a tumor suppressor by regulating the secretion and uptake of exosomal microRNAs in human hepatoma cells. Hepatology. 2015;61(4):1284-1294.

175.

Yin

Yuan

, et al. Downregulation of microRNA-139 is associated with hepatocellular carcinoma risk and short-term survival. Oncol Rep. 2014;31(4):1699-1706.

176.

Shen

Wang

, et al. Exploration of genome-wide circulating microRNA in hepatocellular carcinoma: MiR-483-5p as a potential BiomarkerCirculating miRNAs in differentiation of hepatocellular carcinoma from control. Cancer Epidemiol Biomarkers Prev. 2013;22(12):2364-2373.

177.

Lai

Wang

McElyea

Sherman

House

Korc

. A microRNA signature in circulating exosomes is superior to exosomal glypican-1 levels for diagnosing pancreatic cancer. Cancer Lett. 2017;393:86-93.

178.

Que

Ding

Chen

Cao

. Analysis of serum exosomal microRNAs and clinicopathologic features of patients with pancreatic adenocarcinoma. World J Surg Oncol. 2013;11(1):1-9.

179.

Madhavan

Yue

Galli

, et al. Combined evaluation of a panel of protein and miRNA serum-exosome biomarkers for pancreatic cancer diagnosis increases sensitivity and specificity. Int J Cancer. 2015;136(11):2616-2627.

180.

Y-F

Hannafon

Khatri

Gin

Ding

W-Q

. The origin of exosomal miR-1246 in human cancer cells. RNA Biol. 2019;16(6):770-784.

181.

Abue

Yokoyama

Shibuya

, et al. Circulating miR-483-3p and miR-21 is highly expressed in plasma of pancreatic cancer. Int J Oncol. 2015;46(2):539-547.

182.

Kondo

Murakami

Uemura

, et al. Prognostic impact of perioperative serum CA 19-9 levels in patients with resectable pancreatic cancer. Ann Surg Oncol. 2010;17(9):2321-2329.

183.

Bhagirath

Yang

Bucay

, et al. microRNA-1246 is an exosomal biomarker for aggressive prostate cancer. Cancer Res. 2018;78(7):1833-1844.

184.

Mihelich

Maranville

Nolley

Peehl

Nonn

. Elevated serum microRNA levels associate with absence of high-grade prostate cancer in a retrospective cohort. PLoS One. 2015;10(4):e0124245.

185.

Cheng

Mitchell

Kroh

, et al. Circulating microRNA profiling identifies a subset of metastatic prostate cancer patients with evidence of cancer-associated hypoxia. PLoS One. 2013;8(7):e69239.

186.

Selth

Townley

Gillis

, et al. Discovery of circulating microRNAs associated with human prostate cancer using a mouse model of disease. Int J Cancer. 2012;131(3):652-661.

187.

Nguyen

Xie

Yang

, et al. Expression differences of circulating microRNAs in metastatic castration resistant prostate cancer and low-risk, localized prostate cancer. Prostate. 2013;73(4):346-354.

188.

Alhasan

Scott

, et al. Circulating microRNA signature for the diagnosis of very high-risk prostate cancer. Proc Natl Acad Sci U S A. 2016;113(38):10655-10660.

189.

Huang

Yuan

Liang

, et al. Exosomal miR-1290 and miR-375 as prognostic markers in castration-resistant prostate cancer. Eur Urol. 2015;67(1):33-41.

190.

Endzeliņš

Melne

Kalniņa

, et al. Diagnostic, prognostic and predictive value of cell-free miRNAs in prostate cancer: a systematic review. Mol Cancer. 2016;15(1):41.

191.

Chen

Zhang

, et al. A panel of five circulating microRNAs as potential biomarkers for prostate cancer. Prostate. 2012;72(13):1443-1452.

192.

Sharova

Grassi

Marcer

, et al. A circulating miRNA assay as a first-line test for prostate cancer screening. Br J Cancer. 2016;114(12):1362-1366.

193.

Al-Qatati

Akrong

Stevic

, et al. Plasma microRNA signature is associated with risk stratification in prostate cancer patients. Int J Cancer. 2017;141(6):1231-1239.

194.

Rodríguez

Bajo-Santos

Hessvik

, et al. Identification of non-invasive miRNAs biomarkers for prostate cancer by deep sequencing analysis of urinary exosomes. Mol Cancer. 2017;16(156):1-6.

195.

Samsonov

Shtam

Burdakov

, et al. Lectin-induced agglutination method of urinary exosomes isolation followed by mi-RNA analysis: application for prostate cancer diagnostic. Prostate. 2016;76(1):68-79.

196.

Wani

Kaul

Mavuduru

Kakkar

Bhatia

. Urinary-exosomal miR-2909: a novel pathognomonic trait of prostate cancer severity. J Biotechnol. 2017;259:135-139.

197.

Foj

Ferrer

Serra

, et al. Exosomal and non-exosomal urinary miRNAs in prostate cancer detection and prognosis. Prostate. 2017;77(6):573-583.

198.

Bryzgunova

Zaripov

Skvortsova

, et al. Comparative study of extracellular vesicles from the urine of healthy individuals and prostate cancer patients. PLoS One. 2016;11(6):e0157566.

199.

Qin

Tang

Huang

Zheng

. Mir-145 detection in urinary extracellular vesicles increase diagnostic efficiency of prostate cancer based on hydrostatic filtration dialysis method. Prostate. 2017;77(10):1167-1175.

200.

Fredsøe

Rasmussen

Thomsen

, et al. Diagnostic and prognostic microRNA biomarkers for prostate cancer in cell-free urine. Eur Urol Focus. 2018;4(6):825-833.

201.

Meng

Müller

Milde-Langosch

Trillsch

Pantel

Schwarzenbach

. Diagnostic and prognostic relevance of circulating exosomal miR-373, miR-200a, miR-200b and miR-200c in patients with epithelial ovarian cancer. Oncotarget. 2016;7(13):16923-16935.

202.

Jeon

Seo

Kim

, et al. Circulating exosomal miR-1290 for diagnosis of epithelial ovarian cancer. Curr Issues Mol Biol. 2022;44(1):288-300.

203.

Ying

, et al. Epithelial ovarian cancer-secreted exosomal miR-222-3p induces polarization of tumor-associated macrophages. Oncotarget. 2016;7(28):43076-43087.

204.

Kim

Choi

Jeong

, et al. Serum exosomal miRNA-145 and miRNA-200c as promising biomarkers for preoperative diagnosis of ovarian carcinomas. J Cancer. 2019;10(9):1958-1967.

205.

Maeda

Sasaki

Ueda

, et al. Serum exosomal microRNA-34a as a potential biomarker in epithelial ovarian cancer. J Ovarian Res. 2020;13(1):1-9.

206.

Wang

Song

Wang

, et al. Plasma exosomal miR-320d, miR-4479, and miR-6763-5p as diagnostic biomarkers in epithelial ovarian cancer. Front Oncol. 2022;12:986343.

207.

Hannafon

Trigoso

Calloway

, et al. Plasma exosome microRNAs are indicative of breast cancer. Breast Cancer Res. 2016;18(1):1-14.

208.

Stevic

Müller

Weber

, et al. Specific microRNA signatures in exosomes of triple-negative and HER2-positive breast cancer patients undergoing neoadjuvant therapy within the GeparSixto trial. BMC Med. 2018;16(1):1-16.

209.

Jang

Kim

Kang

Kim

Park

Kim

. Multiple microRNAs as biomarkers for early breast cancer diagnosis. Mol Clin Oncol. 2021;14(2):1-1.

210.

Zhong

Chen

Wang

, et al. MicroRNA expression profiles of drug-resistance breast cancer cells and their exosomes. Oncotarget. 2016;7(15):19601.

211.

Sueta

Yamamoto

Tomiguchi

Takeshita

Yamamoto-Ibusuki

Iwase

. Differential expression of exosomal miRNAs between breast cancer patients with and without recurrence. Oncotarget. 2017;8(41):69934-69944.

212.

Tachibana

Sho

Takeda

, et al. Circulating miR-223 in oral cancer: its potential as a novel diagnostic biomarker and therapeutic target. PloS One. 2016;11(7):e0159693.

213.

Andreu

Oshiro

Redruello

, et al. Extracellular vesicles as a source for non-invasive biomarkers in bladder cancer progression. Eur J Pharm Sci. 2017;98:70-79.

214.

Matsuzaki

Fujita

Jingushi

, et al. MiR-21-5p in urinary extracellular vesicles is a novel biomarker of urothelial carcinoma. Oncotarget. 2017;8(15):24668-24678.

215.

De Long

Sullivan

Humphrey

, et al. A non-invasive miRNA based assay to detect bladder cancer in cell-free urine. Am J Transl Res. 2015;7(11):2500.

216.

Pardini

Cordero

Naccarati

, et al. microRNA profiles in urine by next-generation sequencing can stratify bladder cancer subtypes. Oncotarget. 2018;9(29):20658.

217.

Jiang

Duan

, et al. Cell-free microRNA expression signatures in urine serve as novel noninvasive biomarkers for diagnosis and recurrence prediction of bladder cancer. Oncotarget. 2017;8(25):40832-40842.

218.

Urquidi

Netherton

Gomes-Giacoia

, et al. A microRNA biomarker panel for the non-invasive detection of bladder cancer. Oncotarget. 2016;7(52):86290-86299.

219.

Piao

Jeong

Kim

, et al. Urinary cell-free microRNA biomarker could discriminate bladder cancer from benign hematuria. Int J Cancer. 2019;144(2):380-388.

220.

Zhang

Lau

Chan

, et al. Cell-free urinary microRNA-99a and microRNA-125b are diagnostic markers for the non-invasive screening of bladder cancer. PLoS One. 2014;9(7):e100793.

221.

Sapre

Macintyre

Clarkson

, et al. A urinary microRNA signature can predict the presence of bladder urothelial carcinoma in patients undergoing surveillance. Br J Cancer. 2016;114(4):454-462.

222.

Kim

Kang

Kim

, et al. Cell-free microRNA-214 from urine as a biomarker for non-muscle-invasive bladder cancer. Korean J Urol. 2013;54(11):791-796.

223.

Jiang

Wang

, et al. Serum microRNA expression signatures identified from genome-wide microRNA profiling serve as novel noninvasive biomarkers for diagnosis and recurrence of bladder cancer. Int J Cancer. 2015;136(4):854-862.

224.

Jiang

Wang

225.

Jiang

Duan

, et al. Serum microRNA expression signatures as novel noninvasive biomarkers for prediction and prognosis of muscle-invasive bladder cancer. Oncotarget. 2016;7(24):36733-36742.

226.

Baumgart

Meschkat

Edelmann

, et al. MP78-05 invasion-associated MIRNAS S as possible diagnostic biomarkers of muscle invasive bladder cancer in tumor tissues and urinary exosomes. J Urol. 2018;199(4S):e1038-e1038.

227.

Fanous

Sullivan

Rieger-Christ

. MP88-15 Distinct exosomal miRNA profiles in chemoresistant bladder carcinoma cell lines. J Urol. 2017;197(4S):e1179-e1180.

The Emerging Role of Exosomal miRNAs as Biomarkers for Early Cancer Detection: A Comprehensive Literature Review

Abstract

Keywords

Introduction

Definition and Biogenesis of EXOs

Current Challenges in EXO Isolation

Definition and Biogenesis of miRNAs

Role of ExomiRs in Cancer

General Roles of ExomiRs in Cancer

ExomiRs and Tumor Immunity

ExomiRs and Tumor Proliferation

ExomiRs and Tumor Angiogenesis

ExomiRs and Tumor Metastasis

ExomiRs as a Cancer Biomarker

Lung Cancer

Colorectal Cancer

Hepatocellular Cancer

Gastric Cancer

Pancreatic Cancer

Prostate Cancer

Brain Cancer

Ovarian Cancer

Breast Cancer

Oral and Oropharyngeal Cancer

Bladder Cancer

Conclusion

Footnotes

Abbreviations

Declaration of Conflicting Interests

Funding

ORCID iD

References