Skip to main content
NCBI home page
Molecular Therapy logoLink to Molecular Therapy
. 2024 Feb 27;32(9):2939–2949. doi: 10.1016/j.ymthe.2024.02.025

Therapeutic potential of RNA-enriched extracellular vesicles: The next generation in RNA delivery via biogenic nanoparticles

Muskan Muskan 1, Pevindu Abeysinghe 2, Riccardo Cecchin 1, Heather Branscome 3, Kevin V Morris 2,∗, Fatah Kashanchi 3,∗∗
PMCID: PMC11403218  PMID: 38414242

Abstract

Exosomes are extracellular vesicles (EVs) (∼50–150 nm) that have emerged as promising vehicles for therapeutic applications and drug delivery. These membrane-bound particles, released by all actively dividing cells, have the ability to transfer effector molecules, including proteins, RNA, and even DNA, from donor cells to recipient cells, thereby modulating cellular responses. RNA-based therapeutics, including microRNAs, messenger RNAs, long non-coding RNAs, and circular RNAs, hold great potential in controlling gene expression and treating a spectrum of medical conditions. RNAs encapsulated in EVs are protected from extracellular degradation, making them attractive for therapeutic applications. Understanding the intricate biology of cargo loading and transfer within EVs is pivotal to unlocking their therapeutic potential. This review discusses the biogenesis and classification of EVs, methods for loading RNA into EVs, their advantages as drug carriers over synthetic-lipid-based systems, and the potential applications in treating neurodegenerative diseases, cancer, and viral infections. Notably, EVs show promise in delivering RNA cargo across the blood-brain barrier and targeting tumor cells, offering a safe and effective approach to RNA-based therapy in these contexts.

Keywords: exosomes, extracellular vesicles, nanoparticles, RNA, non-coding RNA, shRNA, circular RNA

Graphical abstract

graphic file with name fx1.jpg

Open in a new tab

Morris and colleagues describe and compile key studies developing extracellular vesicles (EVs) as therapeutic RNA delivery vehicles. We outline the approaches to engineering EVs and the advantages and disadvantages of using EVs as delivery vehicles as a next-generation gene and cell therapy delivery approach.

Introduction: Nature’s couriers—extracellular vesicles

Extracellular vesicles (EVs) have emerged as versatile and intriguing mediators of intercellular communication, gaining substantial attention in recent years.1 EVs form a diverse group of membrane-bound vesicles that can be classified primarily based on biogenesis, size, content, and function.2 Exosomes, a subcategory of EVs (∼30–150 nm diameter in size), are derived through unique endosomal biogenesis pathways and are released into the intercellular space through the fusion of multivesicular bodies.3 EVs can be further categorized into ectosomes (microvesicles and oncosomes) (∼100–1,000 nm in diameter), which result from direct outward budding from the plasma membrane.4 Apoptotic EVs, another type of EV (∼50–5,000 nm in diameter), are released during the cell death (apoptosis) and have been elucidated as critical downstream mediators of apoptotic cell death.5 These small-membrane-enclosed particles are released by virtually all cell types and play crucial roles in various physiological and pathological processes.6 However, distinguishing between EV subtypes remains challenging after they leave the cell, necessitating further research into EV classification7 or defining more specifically the particular class of EVs of particular interest with an understanding that EVs are native heterogeneous populations. Of particular interest in the context of therapeutic applications are exosomes, small (30–150 nm) lipid-bilayer-enclosed vesicles that serve as intercellular messengers.8 Their unique biogenesis and natural role in cell-to-cell communication have positioned exosomes as ideal candidates for therapeutic cargo delivery.9

EVs harbor an array of bioactive molecules, including proteins, lipids, and nucleic acids, with RNA molecules being the central component, protecting them from degradation in the extracellular environment.10,11,12 This complex molecular cargo is selectively packaged and can reflect the physiological state and pathological conditions of parent cells. Notably, exosomal RNA content encompasses a diverse range of RNA species, such as microRNAs (miRNAs), messenger RNAs (mRNAs), long non-coding RNAs (lncRNAs), and even circular RNAs (circRNAs).13 The versatility and promise of EVs as drug delivery systems, coupled with ongoing efforts to elucidate their precise biogenesis and distinguish between EV subtypes, highlight their potential in the field of therapeutic RNA delivery.

RNA as therapeutic arsenal

RNA molecules have long been recognized as pivotal regulators of cellular processes, and the recent advancements in RNA-based therapeutics have invigorated the field of molecular medicine.14 RNA-based therapeutics include miRNAs, small interfering RNAs (siRNAs), mRNAs, lncRNAs, circRNAs, and short hairpin RNAs. These diverse RNA species hold great promise in regulating gene expression, modulating cellular processes, and, ultimately, offering tailored solutions for a spectrum of medical conditions.15

miRNAs, small non-coding RNA molecules typically 18–24 nt in length, have garnered significant attention for their role in post-transcriptional gene regulation.16 miRNAs can exert potent influence over cellular pathways and are implicated in numerous diseases, making them promising targets for therapeutic intervention. For instance, miR-155, a key regulator of the immune system, has been explored for its therapeutic potential in inflammatory bowel disease.17,18

In addition to miRNAs, mRNA has emerged as a cornerstone in the development of cutting-edge vaccines, including the groundbreaking mRNA vaccines against infectious diseases like COVID-19.19 This paradigm-shifting technology has showcased the remarkable potential of mRNA as a platform for rapid and adaptable therapeutic interventions.

siRNAs are double-stranded RNA molecules designed to target and silence specific genes by mRNA degradation. They hold immense potential for treating genetic diseases and cancer.20 Patisiran (the first marketed siRNA-based drug) has shown consistent benefits in slowing the progression of hereditary transthyretin-mediated amyloidosis, a rapidly progressive disease characterized by neuropathy and cardiomyopathy, and enhancing quality of life in clinical trials.21

lncRNAs and circRNAs represent additional layers of complexity in the RNA landscape. While their precise roles are still being elucidated, they have been linked to various diseases, offering intriguing possibilities for targeted therapies.22,23

Despite their promise, naked RNA molecules face challenges including rapid degradation due to nuclease activity24 and reticuloendothelial clearance,25 limited cellular internalization,26 and potential immune response activation.27 These barriers underscore the need for effective RNA delivery systems. The integration of RNA molecules within the context of EVs expands the therapeutic horizon even further. These RNA-enriched EVs harness the intrinsic intercellular communication machinery of exosomes to deliver their therapeutic payload to specific target cells or tissues.

EVs as a drug delivery system (advantages of EVs as drug carriers)

Synthetic-lipid-based materials, including liposomes and lipid nanoparticles (LNPs), have long been used for drug delivery.28 LNPs are a subset of synthetic particles with sizes similar to small EVs.29 Liposomes and LNPs can encapsulate both hydrophobic and hydrophilic drugs, and their formulation can be modified to enhance their stability and circulation half-life.30 Notable examples of lipid-based drug carriers include Doxil (doxorubicin HCl liposome injection), an anticancer drug, and mRNA-based COVID-19 vaccines by Pfizer-BioNTech and Moderna. However, liposomes and LNPs face challenges related to clearance, uptake by macrophages, and immune responses.31 Modifications, such as incorporation of cholesterol and the use of polymer coatings like polyethylene glycol, have been applied to improve their stability and half-life in circulation.19,32

In contrast to synthetic carriers, EVs such as exosomes are considered to have low immunogenic potential.33 EVs have demonstrated safety in small-animal studies, with minimal systemic effects and no significant deviations in body weight or blood parameters.34,35 Human trials have also shown promising results so far regarding the safety of intravenous injection of EVs from various cell sources.36

Moreover, EVs exhibit unique capabilities in crossing biological barriers, both at the cellular and tissue levels.37 They are efficiently taken up by cells through various pathways, showing enhanced cellular uptake compared to liposomal formulations.38 At the tissue level, EVs can traverse the blood-brain barrier (BBB), which is a challenge for many synthetic particles, thereby opening possibilities for targeted drug delivery to hard-to-reach organs, like the brain.39 Furthermore, EVs can be bioengineered from parent cells to produce therapeutic agents, addressing the complex manufacturing challenges associated with biologics.40

EVs hold great promise as drug delivery vehicles due to their natural characteristics, low immunogenicity, and ability to target specific cells and cross biological barriers. While lipid-based carriers like liposomes and LNPs have their merits, EVs offer unique advantages and are being explored for various therapeutic applications.

Methods of loading RNA in EVs: Loading cargos

The packaging of RNA in EVs is a complex and selective process that involves specific mechanisms and sorting signals and can occur through two main methods: endogenous and exogenous packaging. Understanding the packaging mechanism and RNA sorting in EVs is crucial for unraveling the functional implications of EV-mediated RNA transfer.

Exogenous loading

EVs can be loaded exogenously, after isolation from target cells, through passive incubation with cargo or via active loading methods such as electroporation, sonication, or freeze-thaw cycling (Figure 1).41 Electroporation is considered for drug loading into EVs but might lead to adverse morphological changes, like aggregation.42 Exogenous loading is suitable for situations where the ideal cargo is a small-molecule drug or other molecules that cells cannot produce.43 Santos et al. showed that exogenous loading of miR-195-5p (tumor-suppressor miRNA) into EVs enhanced their antitumoral activity. These modified EVs, when incorporated by bystander tumor cells, reduced proliferation, increased cell death, and improved the response to targeted therapy in melanoma, demonstrating the potential of EV-based strategies to enhance clinical outcomes in B-raf proto-oncogene-mutated patients.44

Figure 1.

Figure 1

Open in a new tab

Development of EV-based RNA delivery systems in disease therapy

EVs can be loaded in two ways: one involves using engineered genetic constructs or, alternatively, loading them externally after they have been isolated. This external loading is achieved through methods like electroporation, which disrupts the vesicle membranes. Endogenous loading, on the other hand, utilizes the cell’s internal machinery to load the EVs. Created with BioRender.com.

Endogenous loading

Endogenous loading involves genetic overexpression in parent cells to produce EVs with a desired cargo, leading to a high cytoplasmic RNA concentration and subsequent packaging into EVs (Figure 1).45 This approach allows for more precise control over cargo content and has been used for loading therapeutic proteins, RNAs, and CRISPR-Cas946 complexes into EVs.

Studies have explored various mechanisms for packaging different types of RNA, including miRNA, mRNA fragments, siRNA, and circRNAs, into EVs.

The selective packaging of specific miRNAs into exosomes is influenced by various sorting mechanisms47 involving RNA-binding proteins (such as heterogeneous nuclear ribonucleoprotein A2/B1, argonaute 2,48 Y-box binding protein 1 [YBX-1], and others) and membranous proteins (like caveolin-1 and neutral sphingomyelinase 2) during exosome biogenesis. These mechanisms impact the miRNA content of exosomes, which can be harnessed for various biological effects. For instance, YBX-1 has been linked to the sorting of miR-133 into endothelial progenitor cell-derived exosomes, leading to increased miR-133 levels in cardiac fibroblasts.49 Also, post-transcriptional modifications, such as 3′ end uridylation, contribute to direct miRNA sorting into EVs.50

Valadi et al. was among the first to discover that exosomes naturally carry mRNA.51 However, incorporating foreign mRNA into EVs has been challenging due to ineffective electroporation or chemical methods. Tsai et al. developed an exosome-liposome hybrid for efficient mRNA transfection. In the research, they first treated the purified mRNA with a polycationic lipid coating and then combined it with an equal quantity of purified exosomes. This approach led to the successful development of a multiplexed mRNA COVID-19 vaccine.52 Also, Kojima et al. developed the “EXOtic (EXOsomal Transfer Into Cells)” platform to enhance key properties of EVs, particularly exosomes. They genetically manipulated parent cells to boost exosome production and facilitate efficient mRNA packaging. The system involved overexpressing genes (STEAP3, SDC4, NadB) to increase exosome production and conjugating ribosomal protein L7Ae to cluster of differentiation 63 (CD63), which binds C/D box RNA structures. C/D box motifs were attached to target mRNAs. The overexpression of connexin 43 facilitated cytosolic mRNA delivery.53 These designer exosomes efficiently delivered catalase mRNA to the brains of Parkinsonian mice, attenuating neuroinflammation and demonstrating the potential of genetically engineered EVs for RNA-based therapeutic applications.54

siRNAs, small non-coding RNA molecules that silence gene expression, face challenges in circulation and cell entry due to their hydrophilic nature.55,56 Various methods have also been developed for siRNA loading into EVs, including electroporation, calcium-chloride-mediated transfection, sonication, and the use of hydrophobically modified siRNAs. siRNAs can be easily loaded into EVs due to their small size (20–25 nt).57 Alvarez-Erviti and colleagues were among the first to confirm the concept of delivering siRNA using exosomes by electroporating exogenous siRNA into exosomes, achieving the knockdown of the beta-site APP-cleaving enzyme 1 gene both in vitro and in vivo.58 Several other studies have also employed electroporation to load siRNA into exosomes. Each method has its advantages and limitations in terms of efficiency and potential immunostimulation. Additionally, siRNAs can be preloaded into EVs through cytosolic expression in EV-producing cells, offering a versatile approach for gene silencing.

Uptake of EVs: Unloading cargos

EVs can enter recipient cells through various mechanisms, either via fusion with the plasma membrane or through endocytosis processes, which include clathrin mediated, caveolin mediated, lipid-raft mediated, macropinocytosis, and phagocytosis.59,60 The mechanism of uptake can depend on cell type, its physiological state, and interactions between ligands on EV surfaces and cell receptors. For example, the presence of CD47 on the exosome surface significantly hinders the uptake of EVs by monocytes through the process of phagocytosis.61 Heparin sulfate proteoglycans on recipient cell membranes play a crucial role in EV uptake, and blocking the recipient cell with heparin reduces EV internalization.62,63 Also, membrane fusion can serve as a method of uptake, especially in low-pH tumor environments, and the cytosolic release of EV contents can be enhanced by pH-sensitive fusogenic peptides, allowing content release into the cytosol.64,65

While surface interactions between EVs and cells can trigger signaling without entry, functional effects often depend on EV contents entering the cytoplasm. The mode of entry and effective escape from the endosomal pathway remain complex, but it has been observed that RNA functionality in recipient cells is dependent on successful entry and escape from endosomes.66 Different cell types utilize various mechanisms to either transfer functional cargo or degrade EV content, influenced by cell-specific ligands and receptors.67 Various tracking methods have been explored to monitor EV cargo delivery to recipient cells, including the use of split tag proteins and nanoparticles for insights into these processes.68 Overall, different cell types employ a variety of mechanisms for EV uptake, which may determine whether the cargo is functional or degraded. This complex interplay is influenced by cell-specific ligands and receptors that guide the process. The reliable means of increasing the efficiency of EV cargo escape from the endosomal compartment is crucial for expanding the potential applications of EVs in cargo delivery to recipient cells.

Disease treatment using RNA-enriched EVs

Neurodegenerative disease

Neurological diseases affect the nervous system, which includes the brain, spinal cord, and nerves, and are the second leading cause of death.69 Exosomes hold promise for delivering therapeutic agents to the central nervous system, despite the impermeability of the BBB to 98% of the drugs.70 Exosomes can efficiently carry small RNAs across the BBB, as demonstrated by the studies in zebrafish models71 and other neurological disease models. For instance, exosomes derived from bone marrow mesenchymal stem cells (MSCs) modified with miRNA-29b show promise as therapeutic agents for spinal cord injury (SCI). In a rat model of SCI, intravenous injection of these miRNA-29b exosomes significantly increased miRNA-29b expression in spinal cord tissues, leading to improved hindlimb motor function, enhanced neuronal regeneration, and reduced histopathological damage compared to control groups.72 There are two proposed mechanisms for how exosomes might penetrate the brain.73 One theory is that they go through a process called transcytosis, where they are absorbed by recipient cells and then released to enter other cells, much like some viruses. The other theory suggests direct uptake by endothelial cells through intercellular junctions.73 However, it is important to note that there is no direct experimental evidence supporting either of these mechanisms. Several studies have harnessed exosomes to effectively deliver small RNAs, such as siRNA and miRNA, to the brain to treat various neurological disorders, including glioblastoma, morphine addiction, stroke, and Huntington’s disease (Table 1).

Table 1.

The therapeutic applications of EV-based RNA delivery system

Therapeutic RNA Loading method The source of EVs Type of EVs Disease (cell lines) Therapeutic outcomes Reference
VEGF siRNA Lipofectamine 2000 transfection reagent brain endothelial bEND.3 cells brain endothelial bEND.3 cell-derived exosomes U-87 glioblastoma reduction in the activity of the VEGF gene was observed in brain tumor cells known as glioblastoma-astrocytoma cells; highly effective in preventing the clustering of cancer cells when xenotransplanted into zebrafish Yang et al.125
MOR siRNA Lipofectamine 2000 transfection reagent HEK293T cells RVG exosomes from HEK293T cells morphine relapse precisely targeted the brain for the delivery of siRNA-targeting MOR, resulting in a reduction of MOR mRNA and protein levels Liu et al.126
miR-21a Lipofectamine 2000 transfection reagent neural stem cells (NPCs) NPC-originated exosomes neurological disorders promoted neurogenesis Ma et al.127
miR-133b knockin and knockdown in miR-133b mesenchymal stem cells stem-cell-derived exosome stroke reorganization of neural processes and the restoration of function after a stroke Luarte et al.128
miR-30d transfection of miR-30d adipose-derived stem cells (ADSCs) exosomes secreted from ADSCs acute ischemic stroke polarization of microglia/macrophages and the inhibition of autophagy Jiang et al.129
miR-139-5p mimic Lipofectamine 2000 transfection reagent human umbilical cord mesenchymal stem cells stem-cell-derived exosome bladder cancer (T24, J82, UMUC3, 5637) hindered the ability of cells to grow, move, and invade while also suppressing the expression of PRC1 Wang et al.130
LNA anti-miR-142-3p electroporation bone-marrow-derived mesenchymal stem cells stem-cell-derived exosome breast cancer (4T1) cell proliferation was reduced through the suppression of miR-142-3p and miR-150 expression levels while also increasing the transcription of the regulatory target genes APC and P2X7R Hi et al.131
miR-139 mimic Lipofectamine 2000 transfection reagent cancer-associated fibroblasts cancer-associated fibroblasts gastric cancer (N87, AGS) suppressed both cell proliferation and the spread of cancer, along with a reduction in the levels of MMP11 expression Xu et al.132
SCD-1 siRNA electroporation HEK293 cells – anaplastic thyroid carcinoma (Hth-7) suppressed cell growth and induced cell apoptosis; controlled the metabolism of fatty acids and elevated the level of reactive oxygen species (ROS) Wang et al.133
Survivin siRNA ExoFect exosome transfection reagent HEK293T cells engineering exosome (FA-displaying exosome) human oral epidermal carcinoma (KB) suppressed the expansion of the tumor and downregulated the expression of survivin Lamichhane et al.134
miRNA nucleofection HepG2, human adult liver stem cells (HLSCs) HLSC-derived microvesicles hepatocellular carcinoma suppressed the progression and survival of hepatocellular carcinoma in mice Fansato et al.135
CPT1A siRNA Lipofectamine 2000 transfection reagent HEK293T cell engineered exosomes using iRGD-LAMP-2B plasmid colon cancer model (HCT116-lohp) restored the responsiveness to the chemotherapy drug oxaliplatin by blocking fatty acid oxidation; this was achieved through the interaction between the iRGD molecule located on exosomes and the αvβ3 and Neuropilin-1 receptors found on colon cancer cells
 Lin et al.136
Open in a new tab

A survey of the various therapeutic exosomes strategies being developed and the respective diseases being targeted and outcomes is shown.

The transfer of EVs across the BBB has also been achieved through the use of engineered vesicles that possess a surface protein, which facilitates crossing.74 However, while exosomes hold tremendous potential for RNA-based therapies in neurological diseases, challenges remain, including the need for further investigation into their exact mechanisms of BBB penetration and the discovery of novel targeting ligands for the brain. Nonetheless, these exosome-based RNA delivery systems offer great promise for clinical applications in brain disorder treatment.

Cancer

EVs have garnered significant attention for their potential in cancer diagnosis and therapy.75 These vesicles can serve as effective vehicles for the delivery of therapeutic RNAs to target tumor cells.76 What makes EVs particularly appealing as therapeutic carriers is their intrinsic stability in circulation, homing ability, and inherent capacity for delivering RNAs to specific tumor sites.77 Unlike viral vectors, EVs offer a safer approach to cancer treatment, with minimal risk of causing tumor formation.78 Additionally, their surface can be engineered for improved tumor- or patient-specific targeting.79 A multitude of studies have explored the use of exosomes to deliver small RNAs, such as siRNA and miRNA, with promising outcomes (Table 1). In a research conducted by Zhang and colleagues,80 a method was systematically employed to use exosomes for delivering siRNA-targeting transforming growth factor β1 (TGF-β1), a potential therapeutic target in cancer treatment. The study involved the transfection of TGF-β1 siRNA into mouse fibroblast L929 cells using transfection agents. Subsequently, exosomes containing the TGF-β1 siRNA were isolated from the cell culture supernatant. These exosomes were then assessed for their ability to inhibit the growth and metastasis of murine sarcoma cells both in laboratory settings (in vitro) and in live animals (in vivo). The results of the study indicated that exosomes carrying TGF-β1 siRNA effectively reduced the levels of TGF-β1 in the tumor cells they targeted when compared to the use of free TGF-β1 siRNA. This reduction in TGF-β1 led to a significant decrease in the viability and migration of the implanted mouse sarcoma cells.

This growing body of research highlights the potential of exosomes in RNA-based cancer therapy, paving the way for clinical applications. While these findings are promising, some challenges remain, such as the ability of exosomes to systemically deliver RNA and their potential to escape endosomal degradation.81 EVs also hold significant promise in cancer diagnosis, as they can carry cancer biomarkers and specific RNAs from various cancer types, including prostate, ovarian, and glioblastoma cancers.82 Efforts are underway to leverage EVs for cancer therapy by delivering RNAs and chemotherapeutic drugs, making them a multifaceted player in the fight against cancer.

Viral infections

Viruses and EVs share common strategies in genetic material delivery between cells. EVs, like enveloped viruses, transfer genetic material, proteins, and molecules to recipient cells, influencing their function and gene expression.83 Both EVs and viruses have surface proteins that determine their adhesion to target cells. Some viruses use host cell EVs to enhance their infectivity by transporting viral machinery and proteins to target cells.84 These EVs have been found to play significant roles in various viral infections, including human immunodeficiency virus (HIV) (supporting the idea of “Trojan exosomes”85), human T-lymphotropic virus type 1, coronaviruses, and human adenoviruses.86 For instance, HIV-1 proteins carried by EVs can make cells more susceptible to HIV-1 infection and impair immune responses.87 Additionally, EVs can transport viral receptors, increasing viral binding to cells and enhancing infectivity.84 Studies on cytomegalovirus and herpes simplex virus 1 show that EVs aid viral infectivity by eliminating host proteins critical for antiviral defense.88,89 This complex interplay between EVs and viruses highlights their shared mechanisms in promoting infection and immune evasion. As a result, researchers have explored the use of EV production as a possible means to enhance diagnostics and treatments for viral infections.

EVs can also serve as a defense mechanism against HIV-1 infection. For instance, EVs from breast milk and semen have demonstrated antiviral activity during HIV-1 infection.90,91 Some studies suggest that culturing HIV-1-producing cells in EV-depleted media can limit HIV-1 production.92 Loading exosomes with different types of anti-HIV RNAs can compensate for the limited presence of host-derived anti-HIV-1 RNAs in exosomes.93 Additionally, equipping naturally occurring anti-HIV-1 exosomes with artificial anti-HIV-1 RNAs may enhance their therapeutic effectiveness. To achieve higher antiviral efficacy, it is essential to attach target peptides on the surface of anti-HIV RNA-loaded exosomes. This approach allows for a more specific delivery of therapeutic RNAs to target tissues while reducing the risk of affecting non-target cells. Moreover, assessing the balance between HIV-promoting and HIV-inhibiting exosomes may offer insights into disease progression and serve as potential prognostic biomarkers.

EVs offer distinct advantages in drug delivery as they contain naturally occurring biomolecules. Recent studies have explored their potential for delivering small molecules like mRNA, RNA, peptides, proteins, and other biomolecules to treat various viral diseases, including HIV-associated neurocognitive disorder in the brain and spinal cord. Synthetic EVs, with lower immunogenicity compared to liposomes or lentiviral-based systems, hold promise for delivering RNA therapeutics to block viral infections.

Other diseases

EVs, particularly exosomes, hold promise as innovative therapeutic agents for various other diseases as well. In cardiac diseases like acute myocardial ischemia, miRNA-enriched exosomes derived from MSCs exhibit cardioprotective effects, highlighting their potential for therapeutic interventions.94 However, challenges such as determining optimal delivery methods and addressing potential side effects need to be addressed. In musculoskeletal diseases like osteoarthritis, MSC-derived exosomes offer potential for cartilage regeneration, presenting a promising avenue for therapeutic development.95 However, extensive investigation is necessary to translate these findings into effective clinical applications. The production of clinical-grade EVs following good manufacturing practice standards is crucial for therapeutic development.

Exosomes have emerged as promising tools for treating various diseases, with numerous preclinical studies validating their potential across regenerative medicine and cancer treatments.96,97 Recent years have seen an increase in clinical trials involving two types of exosomes: those derived from human cells and plants.98 These trials reveal that exosomes are primarily investigated for their roles as biomarkers, therapeutic agents, drug delivery systems, and vaccines.

Nanoparticle formulations (like LNPs) have mainly succeeded in reaching the liver in clinical applications so far; expanding delivery beyond the liver can open up more therapeutic possibilities.99 In clinical trials, EVs serve as both biomarkers and active ingredients in pharmaceuticals, leveraging their natural functions and delivery capabilities.100 Clinical trials evaluating the therapeutic potential of EVs in various diseases have been ongoing, but consistent efficacy remains inconclusive.100

EVs and exosomes derived from stem cells have gained immense spotlight as antiaging signals through anti-inflammatory and regenerative function.101 Specifically, exosomes derived from human exfoliated deciduous teeth stem cells have been identified as modulators of histone methylation and nuclear factor κB inhibition, which reverse aged tendon stem/progenitor cells.102 Further, EVs have been identified to slow down kidney and bone degeneration and rejuvenate bone-marrow-derived MSCs in aged mice models.103

Different companies are at various stages of developing EV-based therapies. For example, Codiak Biosciences is working on a versatile platform to produce engineered EVs. Their clinical candidate, CDK-002, is an exosome containing a molecule that activates the stimulator of interferon genes pathway, aimed at treating advanced solid tumors.104 Aruna Bio is focusing on AB126, an unmodified exosome derived from neural stem cells, renowned for its capacity to traverse the BBB.105

In essence, utilizing EVs for delivery is a significant approach in modern medicine, offering promising avenues for therapeutic applications.

Challenges and future perspectives

The use of EVs in therapeutic applications presents both substantial potential and a set of challenges. EVs derived from different sources of MSCs, including bone marrow, adipose, and umbilical cord, have demonstrated especially high therapeutic potential. These EVs naturally retain the biological cargo and regenerative properties of their parental cell stem cells. In addition to the numerous preclinical studies that have evaluated their reparative functions, there are also several ongoing clinical trials that are using MSC EVs as a therapeutic agent against various pathological conditions.106 With respect to EV-associated RNAs, studies have found that MSC EVs are packaged with certain reparative miRNAs (i.e., miR-23a, miR-21, miR-125b, miR-145) in abundance relative to their parental cells.107,108 Others have also found that MSC EVs carry certain lncRNAs, such as MALAT1, that may be attributed to their reparative functions.109 In addition to their natural biological cargo, MSC EVs can be further enriched with various molecules, including RNAs. For example, parental MSCs have been genetically engineered to overexpress different miRNAs (i.e., miRNA-133b, miR-17-92 cluster, miRNA-138-5p, miRNA-223-3p, miR21), and EVs isolated from these cells have demonstrated increased reparative properties in vivo.110,111,112,113,114 Interestingly, Son et al. showed that MSC EVs isolated from a 3D culture platform overexpressed miR-132-3p and miR-27a-3p, which are associated with brain recovery, relative to MSC EVs isolated from 2D cultures.115 It has also been found that MSCs cultured under hypoxic conditions release EVs with boosted therapeutic properties, and this has been attributed to the modification of EV-associated RNA cargo.116,117 MSC EVs can also be exogenously loaded with therapeutic cargo using the methods already described above.

It is important to note that MSC EVs are generally regarded as hypoimmunogenic and thus are regarded as a potentially safer alternative to cell-based therapies. There is also evidence to suggest that MSC EVs possess natural homing abilities, which allow them to migrate to damaged or diseased tissues where they can deliver their therapeutic payload.118,119 For these reasons, MSC EVs may represent a superior choice when considering a future EV-based therapeutic.

While EVs hold promise for clinical use due to their exceptional biocompatibility and double-layered lipid structures, their application in clinical settings faces obstacles like limited yield and a need for specific isolation techniques to remove impurities as well as challenges related to loading efficiency.120 Engineered EVs carrying therapeutic molecules hold promise as alternative therapies. However, the translation of exosomes into clinical therapies is impeded by the complexity of precisely targeting specific cells while minimizing off-target distribution and the presence of naturally integrated genetic materials from cells, which could potentially trigger immune responses.59,121,122 Additionally, the isolation of exosomes may inadvertently result in the loss or masking of certain proteins and lipids, and typical isolation protocols are not specific for exosomes, which may influence their targeting behavior.123 Traditional methods of EV isolation, involving ultracentrifugation and filtration, may yield heterogeneous EV populations and pose translational difficulties.124

The potential of EV-based therapies for RNA delivery is promising, but it faces several challenges that need to be overcome, including efficient isolation techniques, scalable production, loading methods that maintain EV integrity, and a deeper understanding of molecular mechanisms. Despite these challenges, the potential of EVs as unique delivery vehicles is clear, changing the landscape of gene and cell therapeutics and offering new possibilities for treating a variety of diseases. Nevertheless, addressing methodological limitations, developing standardized production protocols, and optimizing EV characterization criteria are essential for realizing the full clinical potential of EV-based therapies.

Acknowledgments

This work is supported by NIH R01 1R01MH134389-01 NIMH to F.K. and K.V.M.

Author contributions

M.M. wrote the entire manuscript with P.A. and H.B. R.C. and M.M. generated the figure, and K.V.M. and F.K. edited the draft and coordinated the submission process.

Declaration of interests

K.V.M. is the chief scientific officer for The Gene Company, which is actively developing RNA exosome therapeutics to treat various human diseases. K.V.M. has submitted a provisional patent (202290305) on some of those technologies reported here.

Contributor Information

Kevin V. Morris, Email: morrisk5@qut.edu.au.

Fatah Kashanchi, Email: fkashanc@gmu.edu.

References

  • 1.Couch Y., Buzàs E.I., Di Vizio D., Gho Y.S., Harrison P., Hill A.F., Lötvall J., Raposo G., Stahl P.D., Théry C., et al. A brief history of nearly EV-erything–The rise and rise of extracellular vesicles. J. Extracell. Vesicles. 2021;10 doi: 10.1002/jev2.12144. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Doyle L.M., Wang M.Z. Overview of Extracellular Vesicles, Their Origin, Composition, Purpose, and Methods for Exosome Isolation and Analysis. Cells. 2019;8 doi: 10.3390/cells8070727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Dimik M., Abeysinghe P., Logan J., Mitchell M. The exosome: a review of current therapeutic roles and capabilities in human reproduction. Drug Deliv. Transl. Res. 2023;13:473–502. doi: 10.1007/s13346-022-01225-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Möller A., Lobb R.J. The evolving translational potential of small extracellular vesicles in cancer. Nat. Rev. Cancer. 2020;20:697–709. doi: 10.1038/s41568-020-00299-w. [DOI] [PubMed] [Google Scholar]
  • 5.Santavanond J.P., Rutter S.F., Atkin-Smith G.K., Poon I.K.H. Apoptotic Bodies: Mechanism of Formation, Isolation and Functional Relevance. Subcell. Biochem. 2021;97:61–88. doi: 10.1007/978-3-030-67171-6_4. [DOI] [PubMed] [Google Scholar]
  • 6.Lee Y., Kim J.-H. The emerging roles of extracellular vesicles as intercellular messengers in liver physiology and pathology. Clin. Mol. Hepatol. 2022;28:706–724. doi: 10.3350/cmh.2021.0390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Cecchin R., Troyer Z., Witwer K., Morris K.V. Extracellular vesicles: The next generation in gene therapy delivery. Mol. Ther. 2023;31:1225–1230. doi: 10.1016/j.ymthe.2023.01.021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Li X., Corbett A.L., Taatizadeh E., Tasnim N., Little J.P., Garnis C., Daugaard M., Guns E., Hoorfar M., Li I.T.S. Challenges and opportunities in exosome research-Perspectives from biology, engineering, and cancer therapy. APL Bioeng. 2019;3 doi: 10.1063/1.5087122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.van Niel G., D'Angelo G., Raposo G. Shedding light on the cell biology of extracellular vesicles. Nat. Rev. Mol. Cell Biol. 2018;19:213–228. doi: 10.1038/nrm.2017.125. [DOI] [PubMed] [Google Scholar]
  • 10.Kim K.M., Abdelmohsen K., Mustapic M., Kapogiannis D., Gorospe M. Vol. 8. Wiley Interdiscip Rev RNA; 2017. RNA in Extracellular Vesicles. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11.Arroyo J.D., Chevillet J.R., Kroh E.M., Ruf I.K., Pritchard C.C., Gibson D.F., Mitchell P.S., Bennett C.F., Pogosova-Agadjanyan E.L., Stirewalt D.L., et al. Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc. Natl. Acad. Sci. USA. 2011;108:5003–5008. doi: 10.1073/pnas.1019055108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Pua H.H., Happ H.C., Gray C.J., Mar D.J., Chiou N.T., Hesse L.E., Ansel K.M. Increased Hematopoietic Extracellular RNAs and Vesicles in the Lung during Allergic Airway Responses. Cell Rep. 2019;26:933–944.e4. doi: 10.1016/j.celrep.2019.01.002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Narang P., Shah M., Beljanski V. Exosomal RNAs in diagnosis and therapies. Noncoding. RNA Res. 2022;7:7–15. doi: 10.1016/j.ncrna.2022.01.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Damase T.R., Sukhovershin R., Boada C., Taraballi F., Pettigrew R.I., Cooke J.P. The Limitless Future of RNA Therapeutics. Front. Bioeng. Biotechnol. 2021;9 doi: 10.3389/fbioe.2021.628137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 15.Sasso J.M., Ambrose B.J.B., Tenchov R., Datta R.S., Basel M.T., DeLong R.K., Zhou Q.A. The Progress and Promise of RNA Medicine—An Arsenal of Targeted Treatments. J. Med. Chem. 2022;65:6975–7015. doi: 10.1021/acs.jmedchem.2c00024. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Ratti M., Lampis A., Ghidini M., Salati M., Mirchev M.B., Valeri N., Hahne J.C. MicroRNAs (miRNAs) and Long Non-Coding RNAs (lncRNAs) as New Tools for Cancer Therapy: First Steps from Bench to Bedside. Target. Oncol. 2020;15:261–278. doi: 10.1007/s11523-020-00717-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Aggeletopoulou I., Mouzaki A., Thomopoulos K., Triantos C. miRNA Molecules-Late Breaking Treatment for Inflammatory Bowel Diseases? Int. J. Mol. Sci. 2023;24 doi: 10.3390/ijms24032233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Shao X., Li J., Xu F., Chen D., Liu K. MiR-155-Mediated Deregulation of GPER1 Plays an Important Role in the Gender Differences Related to Inflammatory Bowel Disease. Can J. Infect Dis. Med. Microbiol. 2020;2020 doi: 10.1155/2020/8811477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Kiaie S.H., Majidi Zolbanin N., Ahmadi A., Bagherifar R., Valizadeh H., Kashanchi F., Jafari R. Recent advances in mRNA-LNP therapeutics: immunological and pharmacological aspects. J. Nanobiotechnology. 2022;20:276. doi: 10.1186/s12951-022-01478-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Dana H., Chalbatani G.M., Mahmoodzadeh H., Karimloo R., Rezaiean O., Moradzadeh A., Mehmandoost N., Moazzen F., Mazraeh A., Marmari V., et al. Molecular Mechanisms and Biological Functions of siRNA. Int. J. Biomed. Sci. 2017;13:48–57. [PMC free article] [PubMed] [Google Scholar]
  • 21.Kristen A.V., Ajroud-Driss S., Conceição I., Gorevic P., Kyriakides T., Obici L. Patisiran, an RNAi therapeutic for the treatment of hereditary transthyretin-mediated amyloidosis. Neurodegener. Dis. Manag. 2019;9:5–23. doi: 10.2217/nmt-2018-0033. [DOI] [PubMed] [Google Scholar]
  • 22.Winkle M., El-Daly S.M., Fabbri M., Calin G.A. Noncoding RNA therapeutics — challenges and potential solutions. Nat. Rev. Drug Discov. 2021;20:629–651. doi: 10.1038/s41573-021-00219-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Liu X., Zhang Y., Zhou S., Dain L., Mei L., Zhu G. Circular RNA: An emerging frontier in RNA therapeutic targets, RNA therapeutics, and mRNA vaccines. J. Control Release. 2022;348:84–94. doi: 10.1016/j.jconrel.2022.05.043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Aagaard L., Rossi J.J. RNAi therapeutics: principles, prospects and challenges. Adv. Drug Deliv. Rev. 2007;59:75–86. doi: 10.1016/j.addr.2007.03.005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25.Orlandini von Niessen A.G., Poleganov M.A., Rechner C., Plaschke A., Kranz L.M., Fesser S., Diken M., Löwer M., Vallazza B., Beissert T., et al. Improving mRNA-Based Therapeutic Gene Delivery by Expression-Augmenting 3' UTRs Identified by Cellular Library Screening. Mol. Ther. 2019;27:824–836. doi: 10.1016/j.ymthe.2018.12.011. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Wadhwa A., Aljabbari A., Lokras A., Foged C., Thakur A. Opportunities and Challenges in the Delivery of mRNA-based Vaccines. Pharmaceutics. 2020;12 doi: 10.3390/pharmaceutics12020102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Park K.S., Sun X., Aikins M.E., Moon J.J. Non-viral COVID-19 vaccine delivery systems. Adv. Drug Deliv. Rev. 2021;169:137–151. doi: 10.1016/j.addr.2020.12.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28.Mehta M., Bui T.A., Yang X., Aksoy Y., Goldys E.M., Deng W. Lipid-Based Nanoparticles for Drug/Gene Delivery: An Overview of the Production Techniques and Difficulties Encountered in Their Industrial Development. ACS Mater. Au. 2023;3:600–619. doi: 10.1021/acsmaterialsau.3c00032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29.Wang B.Z., Luo L.J., Vunjak-Novakovic G. RNA and Protein Delivery by Cell-Secreted and Bioengineered Extracellular Vesicles. Adv. Healthc. Mater. 2022;11 doi: 10.1002/adhm.202101557. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Tenchov R., Bird R., Curtze A.E., Zhou Q. Lipid Nanoparticles—From Liposomes to mRNA Vaccine Delivery, a Landscape of Research Diversity and Advancement. ACS Nano. 2021;15:16982–17015. doi: 10.1021/acsnano.1c04996. [DOI] [PubMed] [Google Scholar]
  • 31.Zahednezhad F., Saadat M., Valizadeh H., Zakeri-Milani P., Baradaran B. Liposome and immune system interplay: Challenges and potentials. J. Control Release. 2019;305:194–209. doi: 10.1016/j.jconrel.2019.05.030. [DOI] [PubMed] [Google Scholar]
  • 32.Hou X., Zaks T., Langer R., Dong Y. Lipid nanoparticles for mRNA delivery. Nat. Rev. Mater. 2021;6:1078–1094. doi: 10.1038/s41578-021-00358-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Liu S., Wu X., Chandra S., Lyon C., Ning B., Jiang L., Fan J., Hu T.Y. Extracellular vesicles: Emerging tools as therapeutic agent carriers. Acta Pharm. Sin. B. 2022;12:3822–3842. doi: 10.1016/j.apsb.2022.05.002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34.Al-Masawa M.E., Alshawsh M.A., Ng C.Y., Ng A.M.H., Foo J.B., Vijakumaran U., Subramaniam R., Ghani N.A.A., Witwer K.W., Law J.X. Efficacy and safety of small extracellular vesicle interventions in wound healing and skin regeneration: A systematic review and meta-analysis of animal studies. Theranostics. 2022;12:6455–6508. doi: 10.7150/thno.73436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 35.Zhu X., Badawi M., Pomeroy S., Sutaria D.S., Xie Z., Baek A., Jiang J., Elgamal O.A., Mo X., Perle K.L., et al. Comprehensive toxicity and immunogenicity studies reveal minimal effects in mice following sustained dosing of extracellular vesicles derived from HEK293T cells. J. Extracell. Vesicles. 2017;6 doi: 10.1080/20013078.2017.1324730. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Sanz-Ros J., Mas-Bargues C., Romero-García N., Huete-Acevedo J., Dromant M., Borrás C. Extracellular Vesicles as Therapeutic Resources in the Clinical Environment. Int. J. Mol. Sci. 2023;24 doi: 10.3390/ijms24032344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Elsharkasy O.M., Nordin J.Z., Hagey D.W., de Jong O.G., Schiffelers R.M., Andaloussi S.E., Vader P. Extracellular vesicles as drug delivery systems: Why and how? Adv. Drug Deliv. Rev. 2020;159:332–343. doi: 10.1016/j.addr.2020.04.004. [DOI] [PubMed] [Google Scholar]
  • 38.Gandek T.B., van der Koog L., Nagelkerke A. A Comparison of Cellular Uptake Mechanisms, Delivery Efficacy, and Intracellular Fate between Liposomes and Extracellular Vesicles. Adv. Healthc. Mater. 2023;12 doi: 10.1002/adhm.202300319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 39.Terstappen G.C., Meyer A.H., Bell R.D., Zhang W. Strategies for delivering therapeutics across the blood-brain barrier. Nat. Rev. Drug Discov. 2021;20:362–383. doi: 10.1038/s41573-021-00139-y. [DOI] [PubMed] [Google Scholar]
  • 40.de Almeida Fuzeta M., Gonçalves P.P., Fernandes-Platzgummer A., Cabral J.M.S., Bernardes N., da Silva C.L. From Promise to Reality: Bioengineering Strategies to Enhance the Therapeutic Potential of Extracellular Vesicles. Bioengineering (Basel) 2022;9 doi: 10.3390/bioengineering9110675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 41.Han Y., Jones T.W., Dutta S., Zhu Y., Wang X., Narayanan S.P., Fagan S.C., Zhang D. Overview and Update on Methods for Cargo Loading into Extracellular Vesicles. Processes (Basel) 2021;9 doi: 10.3390/pr9020356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 42.Johnsen K.B., Gudbergsson J.M., Skov M.N., Christiansen G., Gurevich L., Moos T., Duroux M. Evaluation of electroporation-induced adverse effects on adipose-derived stem cell exosomes. Cytotechnology. 2016;68:2125–2138. doi: 10.1007/s10616-016-9952-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 43.Zeng H., Guo S., Ren X., Wu Z., Liu S., Yao X. Current Strategies for Exosome Cargo Loading and Targeting Delivery. Cells. 2023;12 doi: 10.3390/cells12101416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 44.Santos N.L., Bustos S.O., Reis P.P., Chammas R., Andrade L.N.S. Extracellular Vesicle-Packaged miR-195-5p Sensitizes Melanoma to Targeted Therapy with Kinase Inhibitors. Cells. 2023;12 doi: 10.3390/cells12091317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 45.Wiklander O.P.B., Brennan M.Á., Lötvall J., Breakefield X.O., El Andaloussi S. Advances in therapeutic applications of extracellular vesicles. Sci. Transl. Med. 2019;11 doi: 10.1126/scitranslmed.aav8521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 46.Gee P., Lung M.S.Y., Okuzaki Y., Sasakawa N., Iguchi T., Makita Y., Hozumi H., Miura Y., Yang L.F., Iwasaki M., et al. Extracellular nanovesicles for packaging of CRISPR-Cas9 protein and sgRNA to induce therapeutic exon skipping. Nat. Commun. 2020;11:1334. doi: 10.1038/s41467-020-14957-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 47.Groot M., Lee H. Sorting Mechanisms for MicroRNAs into Extracellular Vesicles and Their Associated Diseases. Cells. 2020;9 doi: 10.3390/cells9041044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 48.McKenzie A.J., Hoshino D., Hong N.H., Cha D.J., Franklin J.L., Coffey R.J., Patton J.G., Weaver A.M. KRAS-MEK Signaling Controls Ago2 Sorting into Exosomes. Cell Rep. 2016;15:978–987. doi: 10.1016/j.celrep.2016.03.085. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 49.Lin F., Zeng Z., Song Y., Li L., Wu Z., Zhang X., Li Z., Ke X., Hu X. YBX-1 mediated sorting of miR-133 into hypoxia/reoxygenation-induced EPC-derived exosomes to increase fibroblast angiogenesis and MEndoT. Stem Cell Res. Ther. 2019;10:263. doi: 10.1186/s13287-019-1377-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 50.Koppers-Lalic D., Hackenberg M., Bijnsdorp I.V., van Eijndhoven M.A.J., Sadek P., Sie D., Zini N., Middeldorp J.M., Ylstra B., de Menezes R.X., et al. Nontemplated nucleotide additions distinguish the small RNA composition in cells from exosomes. Cell Rep. 2014;8:1649–1658. doi: 10.1016/j.celrep.2014.08.027. [DOI] [PubMed] [Google Scholar]
  • 51.Valadi H., Ekström K., Bossios A., Sjöstrand M., Lee J.J., Lötvall J.O. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat. Cell Biol. 2007;9:654–659. doi: 10.1038/ncb1596. [DOI] [PubMed] [Google Scholar]
  • 52.Guo C., Tsai S.J., Ai Y., Li M., Pekosz A., Cox A., Atai N., Gould S.J. The D614G Mutation Enhances the Lysosomal Trafficking of SARS-CoV-2 Spike. bioRxiv. 2020 doi: 10.1101/2020.12.08.417022. Preprint at. [DOI] [Google Scholar]
  • 53.Villamizar O., Waters S.A., Scott T., Grepo N., Jaffe A., Morris K.V. Mesenchymal Stem Cell exosome delivered Zinc Finger Protein activation of cystic fibrosis transmembrane conductance regulator. J. Extracell. Vesicles. 2021;10 doi: 10.1002/jev2.12053. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 54.Kojima R., Bojar D., Rizzi G., Hamri G.C.-E., El-Baba M.D., Saxena P., Ausländer S., Tan K.R., Fussenegger M. Designer exosomes produced by implanted cells intracerebrally deliver therapeutic cargo for Parkinson’s disease treatment. Nat. Commun. 2018;9:1305. doi: 10.1038/s41467-018-03733-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 55.Ozpolat B., Sood A.K., Lopez-Berestein G. Liposomal siRNA nanocarriers for cancer therapy. Adv. Drug Deliv. Rev. 2014;66:110–116. doi: 10.1016/j.addr.2013.12.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 56.Ali Zaidi S.S., Fatima F., Ali Zaidi S.A., Zhou D., Deng W., Liu S. Engineering siRNA therapeutics: challenges and strategies. J. Nanobiotechnology. 2023;21:381. doi: 10.1186/s12951-023-02147-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 57.Joshi B.S., Ortiz D., Zuhorn I.S. Converting extracellular vesicles into nanomedicine: loading and unloading of cargo. Mater. Today Nano. 2021;16 doi: 10.1016/j.mtnano.2021.100148. [DOI] [Google Scholar]
  • 58.Alvarez-Erviti L., Seow Y., Yin H., Betts C., Lakhal S., Wood M.J.A. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nat. Biotechnol. 2011;29:341–345. doi: 10.1038/nbt.1807. [DOI] [PubMed] [Google Scholar]
  • 59.Gurung S., Perocheau D., Touramanidou L., Baruteau J. The exosome journey: from biogenesis to uptake and intracellular signalling. Cell Commun. Signal. 2021;19:47. doi: 10.1186/s12964-021-00730-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 60.Mulcahy L.A., Pink R.C., Carter D.R.F. Routes and mechanisms of extracellular vesicle uptake. J. Extracell. Vesicles. 2014;3 doi: 10.3402/jev.v3.24641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 61.Kamerkar S., LeBleu V.S., Sugimoto H., Yang S., Ruivo C.F., Melo S.A., Lee J.J., Kalluri R. Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer. Nature. 2017;546:498–503. doi: 10.1038/nature22341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 62.Atai N.A., Balaj L., van Veen H., Breakefield X.O., Jarzyna P.A., Van Noorden C.J.F., Skog J., Maguire C.A. Heparin blocks transfer of extracellular vesicles between donor and recipient cells. J. Neurooncol. 2013;115:343–351. doi: 10.1007/s11060-013-1235-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 63.Christianson H.C., Svensson K.J., van Kuppevelt T.H., Li J.P., Belting M. Cancer cell exosomes depend on cell-surface heparan sulfate proteoglycans for their internalization and functional activity. Proc. Natl. Acad. Sci. USA. 2013;110:17380–17385. doi: 10.1073/pnas.1304266110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 64.Parolini I., Federici C., Raggi C., Lugini L., Palleschi S., De Milito A., Coscia C., Iessi E., Logozzi M., Molinari A., et al. Microenvironmental pH is a key factor for exosome traffic in tumor cells. J. Biol. Chem. 2009;284:34211–34222. doi: 10.1074/jbc.M109.041152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 65.Nakase I., Futaki S. Combined treatment with a pH-sensitive fusogenic peptide and cationic lipids achieves enhanced cytosolic delivery of exosomes. Sci. Rep. 2015;5 doi: 10.1038/srep10112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 66.Hung M.E., Leonard J.N. A platform for actively loading cargo RNA to elucidate limiting steps in EV-mediated delivery. J. Extracell. Vesicles. 2016;5 doi: 10.3402/jev.v5.31027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 67.Kooijmans S.A.A., de Jong O.G., Schiffelers R.M. Exploring interactions between extracellular vesicles and cells for innovative drug delivery system design. Adv. Drug Deliv. Rev. 2021;173:252–278. doi: 10.1016/j.addr.2021.03.017. [DOI] [PubMed] [Google Scholar]
  • 68.Toribio V., Morales S., López-Martín S., Cardeñes B., Cabañas C., Yáñez-Mó M. Development of a quantitative method to measure EV uptake. Sci. Rep. 2019;9 doi: 10.1038/s41598-019-47023-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 69.Dumurgier J., Tzourio C. Epidemiology of neurological diseases in older adults. Rev. Neurol. 2020;176:642–648. doi: 10.1016/j.neurol.2020.01.356. [DOI] [PubMed] [Google Scholar]
  • 70.Smyth T., Kullberg M., Malik N., Smith-Jones P., Graner M.W., Anchordoquy T.J. Biodistribution and delivery efficiency of unmodified tumor-derived exosomes. J. Control Release. 2015;199:145–155. doi: 10.1016/j.jconrel.2014.12.013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 71.Yang T., Martin P., Fogarty B., Brown A., Schurman K., Phipps R., Yin V.P., Lockman P., Bai S. Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio. Pharm. Res. 2015;32:2003–2014. doi: 10.1007/s11095-014-1593-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 72.Yu T., Zhao C., Hou S., Zhou W., Wang B., Chen Y. Exosomes secreted from miRNA-29b-modified mesenchymal stem cells repaired spinal cord injury in rats. Braz. J. Med. Biol. Res. 2019;52 doi: 10.1590/1414-431x20198735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 73.Jarmalavičiūtė A., Pivoriūnas A. Exosomes as a potential novel therapeutic tools against neurodegenerative diseases. Pharmacol. Res. 2016;113:816–822. doi: 10.1016/j.phrs.2016.02.002. [DOI] [PubMed] [Google Scholar]
  • 74.Hill A.F. Extracellular Vesicles and Neurodegenerative Diseases. J. Neurosci. 2019;39:9269–9273. doi: 10.1523/jneurosci.0147-18.2019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 75.Tan M., Ge Y., Wang X., Wang Y., Liu Y., He F., Teng H. Extracellular Vesicles (EVs) in Tumor Diagnosis and Therapy. Technol. Cancer Res. Treat. 2023;22 doi: 10.1177/15330338231171463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 76.Du R., Wang C., Zhu L., Yang Y. Extracellular Vesicles as Delivery Vehicles for Therapeutic Nucleic Acids in Cancer Gene Therapy: Progress and Challenges. Pharmaceutics. 2022;14 doi: 10.3390/pharmaceutics14102236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 77.Hernandez-Oller L., Seras-Franzoso J., Andrade F., Rafael D., Abasolo I., Gener P., Schwartz S., Jr. Extracellular Vesicles as Drug Delivery Systems in Cancer. Pharmaceutics. 2020;12 doi: 10.3390/pharmaceutics12121146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 78.Zhang F., Guo J., Zhang Z., Duan M., Wang G., Qian Y., Zhao H., Yang Z., Jiang X. Application of engineered extracellular vesicles for targeted tumor therapy. J. Biomed. Sci. 2022;29:14. doi: 10.1186/s12929-022-00798-y. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 79.Johnson V., Vasu S., Kumar U.S., Kumar M. Surface-Engineered Extracellular Vesicles in Cancer Immunotherapy. Cancers (Basel) 2023;15 doi: 10.3390/cancers15102838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 80.Zhang Y., Li L., Yu J., Zhu D., Zhang Y., Li X., Gu H., Zhang C.-Y., Zen K. Microvesicle-mediated delivery of transforming growth factor β1 siRNA for the suppression of tumor growth in mice. Biomaterials. 2014;35:4390–4400. doi: 10.1016/j.biomaterials.2014.02.003. [DOI] [PubMed] [Google Scholar]
  • 81.Koh H.B., Kim H.J., Kang S.W., Yoo T.H. Exosome-Based Drug Delivery: Translation from Bench to Clinic. Pharmaceutics. 2023;15 doi: 10.3390/pharmaceutics15082042. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 82.Mathew M., Zade M., Mezghani N., Patel R., Wang Y., Momen-Heravi F. Extracellular Vesicles as Biomarkers in Cancer Immunotherapy. Cancers (Basel) 2020;12 doi: 10.3390/cancers12102825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 83.Nolte-'t Hoen E., Cremer T., Gallo R.C., Margolis L.B. Extracellular vesicles and viruses: Are they close relatives? Proc. Natl. Acad. Sci. USA. 2016;113:9155–9161. doi: 10.1073/pnas.1605146113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 84.Moulin C., Crupi M.J.F., Ilkow C.S., Bell J.C., Boulton S. Extracellular Vesicles and Viruses: Two Intertwined Entities. Int. J. Mol. Sci. 2023;24 doi: 10.3390/ijms24021036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 85.Gould S.J., Booth A.M., Hildreth J.E.K. The Trojan exosome hypothesis. Proc. Natl. Acad. Sci. USA. 2003;100:10592–10597. doi: 10.1073/pnas.1831413100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 86.Saad M.H., Badierah R., Redwan E.M., El-Fakharany E.M. A Comprehensive Insight into the Role of Exosomes in Viral Infection: Dual Faces Bearing Different Functions. Pharmaceutics. 2021;13 doi: 10.3390/pharmaceutics13091405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 87.Lee J.H. The complex role of extracellular vesicles in HIV infection. BMB Rep. 2023;56:335–340. doi: 10.5483/BMBRep.2023-0073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 88.Kutchy N.A., Peeples E.S., Sil S., Liao K., Chivero E.T., Hu G., Buch S. Extracellular Vesicles in Viral Infections of the Nervous System. Viruses. 2020;12 doi: 10.3390/v12070700. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 89.Streck N.T., Zhao Y., Sundstrom J.M., Buchkovich N.J. Human Cytomegalovirus Utilizes Extracellular Vesicles To Enhance Virus Spread. J. Virol. 2020;94 doi: 10.1128/jvi.00609-20. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 90.Madison M.N., Roller R.J., Okeoma C.M. Human semen contains exosomes with potent anti-HIV-1 activity. Retrovirology. 2014;11:102. doi: 10.1186/s12977-014-0102-z. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 91.Näslund T.I., Paquin-Proulx D., Paredes P.T., Vallhov H., Sandberg J.K., Gabrielsson S. Exosomes from breast milk inhibit HIV-1 infection of dendritic cells and subsequent viral transfer to CD4+ T cells. Aids. 2014;28:171–180. doi: 10.1097/qad.0000000000000159. [DOI] [PubMed] [Google Scholar]
  • 92.Liao Z., Muth D.C., Eitan E., Travers M., Learman L.N., Lehrmann E., Witwer K.W. Serum extracellular vesicle depletion processes affect release and infectivity of HIV-1 in culture. Sci. Rep. 2017;7:2558. doi: 10.1038/s41598-017-02908-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 93.Yong -.Q., Jing -.M., Yi -.Z. Vol. 31. - Biomedical and Environmental Sciences -; 2018. Therapeutic Potential of Anti-HIV RNA-Loaded Exosomes; p. 215. [DOI] [PubMed] [Google Scholar]
  • 94.Luo Q., Guo D., Liu G., Chen G., Hang M., Jin M. Exosomes from MiR-126-Overexpressing Adscs Are Therapeutic in Relieving Acute Myocardial Ischaemic Injury. Cell. Physiol. Biochem. 2017;44:2105–2116. doi: 10.1159/000485949. [DOI] [PubMed] [Google Scholar]
  • 95.Mao G., Zhang Z., Hu S., Zhang Z., Chang Z., Huang Z., Liao W., Kang Y. Exosomes derived from miR-92a-3p-overexpressing human mesenchymal stem cells enhance chondrogenesis and suppress cartilage degradation via targeting WNT5A. Stem Cell Res. Ther. 2018;9:247. doi: 10.1186/s13287-018-1004-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 96.Mendt M., Rezvani K., Shpall E. Mesenchymal stem cell-derived exosomes for clinical use. Bone Marrow Transpl. 2019;54:789–792. doi: 10.1038/s41409-019-0616-z. [DOI] [PubMed] [Google Scholar]
  • 97.Kalluri R. The biology and function of exosomes in cancer. J. Clin. Invest. 2016;126:1208–1215. doi: 10.1172/jci81135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 98.Rezaie J., Feghhi M., Etemadi T. A review on exosomes application in clinical trials: perspective, questions, and challenges. Cell Commun. Signal. 2022;20:145. doi: 10.1186/s12964-022-00959-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 99.Loughrey D., Dahlman J.E. Non-liver mRNA Delivery. Acc. Chem. Res. 2022;55:13–23. doi: 10.1021/acs.accounts.1c00601. [DOI] [PubMed] [Google Scholar]
  • 100.Suzuki Y., Katsurada Y., Hyodo K. Differences and Similarities of the Intravenously Administered Lipid Nanoparticles in Three Clinical Trials: Potential Linkage between Lipid Nanoparticles and Extracellular Vesicles. Mol. Pharm. 2023;20:4883–4892. doi: 10.1021/acs.molpharmaceut.3c00547. [DOI] [PubMed] [Google Scholar]
  • 101.Ha D.H., Kim H.-k., Lee J., Kwon H.H., Park G.-H., Yang S.H., Jung J.Y., Choi H., Lee J.H., Sung S., et al. Mesenchymal Stem/Stromal Cell-Derived Exosomes for Immunomodulatory Therapeutics and Skin Regeneration. Cells. 2020;9:1157. doi: 10.3390/cells9051157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 102.Jin S., Wang Y., Wu X., Li Z., Zhu L., Niu Y., Zhou Y., Liu Y. Young Exosome Bio-Nanoparticles Restore Aging-Impaired Tendon Stem/Progenitor Cell Function and Reparative Capacity. Adv. Mater. 2023;35 doi: 10.1002/adma.202211602. [DOI] [PubMed] [Google Scholar]
  • 103.Lei Q., Gao F., Liu T., Ren W., Chen L., Cao Y., Chen W., Guo S., Zhang Q., Chen W., Wang H., et al. Extracellular vesicles deposit PCNA to rejuvenate aged bone marrow–derived mesenchymal stem cells and slow age-related degeneration. Sci. Translational Med. 2021;13:eaaz8697. doi: 10.1126/scitranslmed.aaz8697. [DOI] [PubMed] [Google Scholar]
  • 104.Jang S.C., Economides K.D., Moniz R.J., Sia C.L., Lewis N., McCoy C., Zi T., Zhang K., Harrison R.A., Lim J., et al. ExoSTING, an extracellular vesicle loaded with STING agonists, promotes tumor immune surveillance. Commun. Biol. 2021;4:497. doi: 10.1038/s42003-021-02004-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 105.Sun M.K., Passaro A.P., Latchoumane C.F., Spellicy S.E., Bowler M., Goeden M., Martin W.J., Holmes P.V., Stice S.L., Karumbaiah L. Extracellular Vesicles Mediate Neuroprotection and Functional Recovery after Traumatic Brain Injury. J. Neurotrauma. 2020;37:1358–1369. doi: 10.1089/neu.2019.6443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 106.Lee B.C., Kang I., Yu K.R. Therapeutic Features and Updated Clinical Trials of Mesenchymal Stem Cell (MSC)-Derived Exosomes. J. Clin. Med. 2021;10 doi: 10.3390/jcm10040711. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 107.Fang S., Xu C., Zhang Y., Xue C., Yang C., Bi H., Qian X., Wu M., Ji K., Zhao Y., et al. Umbilical Cord-Derived Mesenchymal Stem Cell-Derived Exosomal MicroRNAs Suppress Myofibroblast Differentiation by Inhibiting the Transforming Growth Factor-β/SMAD2 Pathway During Wound Healing. Stem Cells Transl. Med. 2016;5:1425–1439. doi: 10.5966/sctm.2015-0367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 108.Mitchell R., Mellows B., Sheard J., Antonioli M., Kretz O., Chambers D., Zeuner M.T., Tomkins J.E., Denecke B., Musante L., et al. Secretome of adipose-derived mesenchymal stem cells promotes skeletal muscle regeneration through synergistic action of extracellular vesicle cargo and soluble proteins. Stem Cell Res. Ther. 2019;10:116. doi: 10.1186/s13287-019-1213-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 109.Cooper D.R., Wang C., Patel R., Trujillo A., Patel N.A., Prather J., Gould L.J., Wu M.H. Human Adipose-Derived Stem Cell Conditioned Media and Exosomes Containing MALAT1 Promote Human Dermal Fibroblast Migration and Ischemic Wound Healing. Adv. Wound Care. 2018;7:299–308. doi: 10.1089/wound.2017.0775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 110.Xin H., Wang F., Li Y., Lu Q.E., Cheung W.L., Zhang Y., Zhang Z.G., Chopp M. Secondary Release of Exosomes From Astrocytes Contributes to the Increase in Neural Plasticity and Improvement of Functional Recovery After Stroke in Rats Treated With Exosomes Harvested From MicroRNA 133b-Overexpressing Multipotent Mesenchymal Stromal Cells. Cell Transpl. 2017;26:243–257. doi: 10.3727/096368916x693031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 111.Xin H., Katakowski M., Wang F., Qian J.Y., Liu X.S., Ali M.M., Buller B., Zhang Z.G., Chopp M. MicroRNA cluster miR-17-92 Cluster in Exosomes Enhance Neuroplasticity and Functional Recovery After Stroke in Rats. Stroke. 2017;48:747–753. doi: 10.1161/strokeaha.116.015204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 112.Deng Y., Chen D., Gao F., Lv H., Zhang G., Sun X., Liu L., Mo D., Ma N., Song L., et al. Exosomes derived from microRNA-138-5p-overexpressing bone marrow-derived mesenchymal stem cells confer neuroprotection to astrocytes following ischemic stroke via inhibition of LCN2. J. Biol. Eng. 2019;13:71. doi: 10.1186/s13036-019-0193-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 113.Lu F.B., Chen D.Z., Chen L., Hu E.D., Wu J.L., Li H., Gong Y.W., Lin Z., Wang X.D., Li J., et al. Attenuation of Experimental Autoimmune Hepatitis in Mice with Bone Mesenchymal Stem Cell-Derived Exosomes Carrying MicroRNA-223-3p. Mol. Cells. 2019;42:906–918. doi: 10.14348/molcells.2019.2283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 114.Cheng X., Zhang G., Zhang L., Hu Y., Zhang K., Sun X., Zhao C., Li H., Li Y.M., Zhao J. Mesenchymal stem cells deliver exogenous miR-21 via exosomes to inhibit nucleus pulposus cell apoptosis and reduce intervertebral disc degeneration. J. Cell. Mol. Med. 2018;22:261–276. doi: 10.1111/jcmm.13316. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 115.Son J.P., Kim E.H., Shin E.K., Kim D.H., Sung J.H., Oh M.J., Cha J.M., Chopp M., Bang O.Y. Mesenchymal Stem Cell-Extracellular Vesicle Therapy for Stroke: Scalable Production and Imaging Biomarker Studies. Stem Cells Transl. Med. 2023;12:459–473. doi: 10.1093/stcltm/szad034. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 116.Shao M., Jin M., Xu S., Zheng C., Zhu W., Ma X., Lv F. Exosomes from Long Noncoding RNA-Gm37494-ADSCs Repair Spinal Cord Injury via Shifting Microglial M1/M2 Polarization. Inflammation. 2020;43:1536–1547. doi: 10.1007/s10753-020-01230-z. [DOI] [PubMed] [Google Scholar]
  • 117.Zhu D., Johnson T.K., Wang Y., Thomas M., Huynh K., Yang Q., Bond V.C., Chen Y.E., Liu D. Macrophage M2 polarization induced by exosomes from adipose-derived stem cells contributes to the exosomal proangiogenic effect on mouse ischemic hindlimb. Stem Cell Res. Ther. 2020;11:162. doi: 10.1186/s13287-020-01669-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 118.Zhang F., Guo J., Zhang Z., Qian Y., Wang G., Duan M., Zhao H., Yang Z., Jiang X. Mesenchymal stem cell-derived exosome: A tumor regulator and carrier for targeted tumor therapy. Cancer Lett. 2022;526:29–40. doi: 10.1016/j.canlet.2021.11.015. [DOI] [PubMed] [Google Scholar]
  • 119.Harrell C.R., Jovicic N., Djonov V., Arsenijevic N., Volarevic V. Mesenchymal Stem Cell-Derived Exosomes and Other Extracellular Vesicles as New Remedies in the Therapy of Inflammatory Diseases. Cells. 2019;8 doi: 10.3390/cells8121605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 120.Tran P.H.L., Xiang D., Nguyen T.N.G., Tran T.T.D., Chen Q., Yin W., Zhang Y., Kong L., Duan A., Chen K., et al. Aptamer-guided extracellular vesicle theranostics in oncology. Theranostics. 2020;10:3849–3866. doi: 10.7150/thno.39706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 121.Ha D., Yang N., Nadithe V. Exosomes as therapeutic drug carriers and delivery vehicles across biological membranes: current perspectives and future challenges. Acta Pharm. Sin. B. 2016;6:287–296. doi: 10.1016/j.apsb.2016.02.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 122.Bunggulawa E.J., Wang W., Yin T., Wang N., Durkan C., Wang Y., Wang G. Recent advancements in the use of exosomes as drug delivery systems. J. Nanobiotechnology. 2018;16:81. doi: 10.1186/s12951-018-0403-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 123.Kooijmans S.A.A., Schiffelers R.M., Zarovni N., Vago R. Modulation of tissue tropism and biological activity of exosomes and other extracellular vesicles: New nanotools for cancer treatment. Pharmacol. Res. 2016;111:487–500. doi: 10.1016/j.phrs.2016.07.006. [DOI] [PubMed] [Google Scholar]
  • 124.De Sousa K.P., Rossi I., Abdullahi M., Ramirez M.I., Stratton D., Inal J.M. Vol. 15. Wiley Interdiscip Rev Nanomed Nanobiotechnol; 2023. Isolation and Characterization of Extracellular Vesicles and Future Directions in Diagnosis and Therapy. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 125.Yang T., Fogarty B., LaForge B., Aziz S., Pham T., Lai L., Bai S. Delivery of Small Interfering RNA to Inhibit Vascular Endothelial Growth Factor in Zebrafish Using Natural Brain Endothelia Cell-Secreted Exosome Nanovesicles for the Treatment of Brain Cancer. AAPS J. 2017;19:475–486. doi: 10.1208/s12248-016-0015-y. [DOI] [PubMed] [Google Scholar]
  • 126.Liu Y., Li D., Liu Z., Zhou Y., Chu D., Li X., Jiang X., Hou D., Chen X., Chen Y., et al. Targeted exosome-mediated delivery of opioid receptor Mu siRNA for the treatment of morphine relapse. Sci. Rep. 2015;5 doi: 10.1038/srep17543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 127.Ma Y., Li C., Huang Y., Wang Y., Xia X., Zheng J.C. Exosomes released from neural progenitor cells and induced neural progenitor cells regulate neurogenesis through miR-21a. Cell Commun. Signal. 2019;17:96. doi: 10.1186/s12964-019-0418-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 128.Luarte A., Henzi R., Fernández A., Gaete D., Cisternas P., Pizarro M., Batiz L.F., Villalobos I., Masalleras M., Vergara R., et al. Astrocyte-Derived Small Extracellular Vesicles Regulate Dendritic Complexity through miR-26a-5p Activity. Cells. 2020;9:930. doi: 10.3390/cells9040930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 129.Jiang M., Wang H., Jin M., Yang X., Ji H., Jiang Y., Zhang H., Wu F., Wu G., Lai X., et al. Exosomes from MiR-30d-5p-ADSCs Reverse Acute Ischemic Stroke-Induced, Autophagy-Mediated Brain Injury by Promoting M2 Microglial/Macrophage Polarization. Cell. Physiol. Biochem. 2018;47:864–878. doi: 10.1159/000490078. [DOI] [PubMed] [Google Scholar]
  • 130.Wang X., Qin X., Yan M., Shi J., Xu Q., Li Z., Yang W., Zhang J., Chen W. Loss of exosomal miR-3188 in cancer-associated fibroblasts contributes to HNC progression. J. Exp. Clin. Cancer Res. 2019;38:151. doi: 10.1186/s13046-019-1144-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 131.He Z., Li W., Zheng T., Liu D., Zhao S. Human umbilical cord mesenchymal stem cells-derived exosomes deliver microRNA-375 to downregulate ENAH and thus retard esophageal squamous cell carcinoma progression. J. Exp. Clin. Cancer Res. 2020;39:140. doi: 10.1186/s13046-020-01631-w. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 132.Xu G., Zhang B., Ye J., Cao S., Shi J., Zhao Y., Wang Y., Sang J., Yao Y., Guan W., et al. Exosomal miRNA-139 in cancer-associated fibroblasts inhibits gastric cancer progression by repressing MMP11 expression. Int. J. Biol. Sci. 2019;15:2320–2329. doi: 10.7150/ijbs.33750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 133.Wang M.H., Ye Y., Zhang M., Zhou B.R., Wang J.N., Song Y.N., Xia W. Exosome-mediated delivery of SCD-1 siRNA promoted the death of anaplastic thyroid carcinoma cells via regulating ROS level. Clin. Transl. Oncol. 2022;24:288–296. doi: 10.1007/s12094-021-02682-x. [DOI] [PubMed] [Google Scholar]
  • 134.Lamichhane T.N., Jeyaram A., Patel D.B., Parajuli B., Livingston N.K., Arumugasaamy N., Schardt J.S., Jay S.M. Oncogene Knockdown via Active Loading of Small RNAs into Extracellular Vesicles by Sonication. Cell. Mol. Bioeng. 2016;9:315–324. doi: 10.1007/s12195-016-0457-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 135.Fonsato V., Collino F., Herrera M.B., Cavallari C., Deregibus M.C., Cisterna B., Bruno S., Romagnoli R., Salizzoni M., Tetta C., Camussi G. Human Liver Stem Cell-Derived Microvesicles Inhibit Hepatoma Growth in SCID Mice by Delivering Antitumor MicroRNAs. Stem Cells. 2012;30:1985–1998. doi: 10.1002/stem.1161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 136.Lin D., Zhang H., Liu R., Deng T., Ning T., Bai M., Yang Y., Zhu K., Wang J., Duan J., et al. iRGD-modified exosomes effectively deliver CPT1A siRNA to colon cancer cells, reversing oxaliplatin resistance by regulating fatty acid oxidation. Mol. Oncol. 2021;15:3430–3446. doi: 10.1002/1878-0261.13052. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular Therapy are provided here courtesy of The American Society of Gene & Cell Therapy

RESOURCES