bims-engexo 2025-12-28 papers

bims-engexo

Biomed News

on Engineered exosomes

Issue of 2025–12–28
nine papers selected by
Ravindran Jaganathan, Universiti Kuala Lumpur

Therapeutic potential of microRNA let-7a-enriched exosomes in modulating macrophage plasticity (M1) and inhibiting malignant tendencies in breast cancer.
Humanized Extracellular Vesicles for Efficient RNA Delivery.
M2 macrophage-derived exosome nanoplatform for targeted H2S delivery to alleviate hepatic ischemia-reperfusion injury via synergistic anti-oxidative, anti-inflammatory, and anti-ferroptotic effects.
Engineered Extracellular Vesicles for Intervertebral Disc Regeneration: Mechanisms, Strategies, and Translational Potential.
Engineered vesicles enhance oral antibiotic absorption in proximal small intestine and mitigate gut dysbiosis.
The role of exosomes as oligonucleotide delivery system for managing α-synuclein in Parkinson's disease: A systematic review of in vivo studies.
Combined action of suicide gene exosomes from pancreatic cancer-associated fibroblasts and from mesenchymal stem cells as a pancreatic ductal adenocarcinoma treatment approach.
Rapid extracellular vesicle surface decoration with targeting moieties based on a fluorescein binding single chain variable fragment snorkel.
Rat adipose-derived mesenchymal stem cell-derived exosomes loaded with miR-375 promote neurite outgrowth and peripheral nerve regeneration via activation of Akt by targeting EPHA4.

Eur J Pharmacol. 2025 Dec 18. pii: S0014-2999(25)01255-5. [Epub ahead of print]1012 178501

Therapeutic potential of microRNA let-7a-enriched exosomes in modulating macrophage plasticity (M1) and inhibiting malignant tendencies in breast cancer.

Elmira Aghazadeh, Shahla Mohammad Ganji, Fatemeh Ashrafi, Ardeshir Abbasi.

  Tumor-associated macrophages (TAMs) play a pivotal role in cancer progression, and their polarization toward the M2 phenotype contributes to immune suppression and metastasis promotion. One promising immunotherapeutic approach involves reprogramming M2 macrophages into the pro-inflammatory and anti-tumor M1 phenotype within the tumor microenvironment. This study investigated the effect of exosomes derived from 4T1 breast cancer cells, loaded with microRNA let-7a, on macrophage polarization and tumor-related behaviors. Tumor-derived exosomes (TEXs) were isolated and characterized by DLS, Flowcytometry (CD63) and TEM, followed by let-7a loading via electroporation. Peritoneal macrophages from Balb/c mice were extracted, polarized into the immunosuppressive M2 phenotype, and subsequently treated with let-7a -enriched exosomes (TEX + let-7a), leading to their repolarization into M1 macrophages. Anti-cancer assays, including viability, apoptosis, metastasis, angiogenesis, and migration, were then performed. Results demonstrated that these engineered exosomes effectively repolarized M2 macrophages into active, tumor-suppressing M1 macrophages. The induced M1 macrophages significantly reduced cancer cell survival, increased apoptosis through elevated BAX/Bcl-2 ratio, and suppressed the expression of metastasis-associated (MMP2, MMP9) and angiogenesis-related (VEGF) genes. Moreover, cancer cell migration was markedly inhibited. Collectively, these findings indicate that let-7a -enriched exosomes can reprogram macrophage plasticity to effectively suppress malignant behaviors in breast cancer. This novel approach highlights the high therapeutic potential of targeted exosome-based miRNA delivery for refining tumor microenvironment regulation and advancing breast cancer immunotherapy strategies.

Keywords:  Breast cancer; Cancer immunotherapy; Exosomes; Let-7a; Macrophage polarization

DOI:  https://doi.org/10.1016/j.ejphar.2025.178501
bioRxiv. 2025 Dec 17. pii: 2025.12.15.694436. [Epub ahead of print]

Humanized Extracellular Vesicles for Efficient RNA Delivery.

Xiang Ma, Sophia R Zhao, Constance L Cepko.

Engineered extracellular vesicles (EVs) are a class of non-viral delivery vectors for RNA-based vaccines and gene therapies. A specialized form of engineered EVs, known as enveloped protein nanocages (EPNs), has been developed to enhance cargo loading and delivery. When EPNs are equipped with a viral fusogen, such as vesicular stomatitis virus glycoprotein (VSV-G), they have been shown to deliver proteins or RNA efficiently into recipient cells. Comparisons across different EPN types and optimization of their different features have been difficult, as assays for their activity have not been reported for single, active units. As we were interested in optimizing EVs, we first developed a biological titration assay inspired by the methods used for infectious viral particles. With this assay, we optimized EVs using a modular platform, creating EVs composed predominantly of human-derived protein components. This system achieved efficient RNA delivery, with functional titers comparable to those of lentiviral vectors. The optimized chimeric proteins comprising the EV particles integrate domains from human epsin 1, human citramalyl-CoA lyase beta-like protein (CLYBL), and human CEP55. The constructs also include a short 21-amino-acid peptide from a non-human source for RNA packaging, resulting in an EV-based RNA delivery system with reduced immunogenicity compared with EPNs and retroviral virus-like particles (VLPs).
Significance Statement: We developed engineered extracellular vesicles (EVs) as RNA delivery vehicles to address limitations of virus-like particles (VLPs) and lipid nanoparticles (LNPs) in gene therapies and vaccines. We first developed an assay for individual active particles, using methods typically employed for viral titrations. This approach allowed iterative optimization of a modular EV platform. Our optimized particles comprise primarily human proteins and reach titers that comparable to those of lentiviral vectors.

DOI: https://doi.org/10.64898/2025.12.15.694436
Acta Biomater. 2025 Dec 18. pii: S1742-7061(25)00948-1. [Epub ahead of print]

M2 macrophage-derived exosome nanoplatform for targeted H2S delivery to alleviate hepatic ischemia-reperfusion injury via synergistic anti-oxidative, anti-inflammatory, and anti-ferroptotic effects.

Tianci Zuo, Chengge Shi, Jiayu Li, Diwei Li, Xiaoming Luo, Mengzhu Xu, Mingli Su, Hongyang Lu, Yanmei Zhang, Xiaowen Hu, Qiang Zhou, Quazi T H Shubhra, Xiaosong He, Xiaojun Cai.

  Hepatic ischemia-reperfusion injury (HIRI) presents a critical challenge in liver surgery, transplantation, and trauma, driven by the interplay of oxidative stress, inflammation, and ferroptosis. Current treatments are limited by poor targeting and insufficient efficacy. Here, we develop a liver-targeted exosomal nanoplatform (DATS@M2-Exos) by encapsulating the H2S donor diallyl trisulfide (DATS) into M2 macrophage-derived exosomes (M2-Exos), enabling liver-specific H2S delivery. In vitro, DATS@M2-Exos exhibit good biocompatibility, efficient cellular uptake, and effective H2S release, resulting in significant suppression of oxidative stress, inflammation, and ferroptosis by restoring GSH levels, enhancing GPX4 expression, and reducing Fe2+ and MDA accumulation. In a murine HIRI model, DATS@M2-Exos demonstrate strong hepatic tropism, significantly decrease serum ALT/AST levels, alleviate histopathological injury, while exhibiting favorable safety. RNA sequencing reveals potent antioxidant and anti-ferroptosis effects of DATS@M2-Exos via redox and lipid metabolic reprogramming, including activation of the GSH metabolic pathway, suppression of iron overload, and enhancement of GPX4 expression. Furthermore, DATS@M2-Exos exert dual immunomodulatory effects by suppressing the TNF-α/IL-1β/MAPK13 axis and promoting M2 macrophage polarization through activation of the PPAR pathway. This study presents an exosome-based nanotherapeutic for the targeted delivery of H2S to coordinately combat oxidative stress, inflammation, and ferroptosis, offering a precise and effective strategy for HIRI treatment. STATEMENT OF SIGNIFICANCE: Hepatic ischemia-reperfusion injury (HIRI) remains a critical challenge in liver surgery and transplantation, driven by oxidative stress, inflammation, and ferroptosis. Current therapies lack targeted delivery and multimodal efficacy. This study develops a liver-targeted exosomal nanoplatform (DATS@M2-Exos) by encapsulating the H₂S donor diallyl trisulfide (DATS) into M2 macrophage-derived exosomes (M2-Exos). DATS@M2-Exos demonstrate exceptional hepatic tropism, biocompatibility, and controlled H2S release, synergistically combating oxidative stress (via GSH/GPX4 axis activation), inflammation (via PPAR-mediated M2 polarization), and ferroptosis (via iron overload suppression). In vivo, DATS@M2-Exos significantly reduce liver damage with no systemic toxicity. This work pioneers an exosome-based gas therapy platform, offering a precise, multifunctional strategy for HIRI and broader oxidative stress-related diseases.

Keywords:  Ferroptosis inhibition; Hepatic ischemia–reperfusion injury; Immunomodulation; M2 macrophage-derived exosomes; Targeted H(2)S delivery

DOI:  https://doi.org/10.1016/j.actbio.2025.12.041
Cureus. 2025 Nov;17(11): e97119

Engineered Extracellular Vesicles for Intervertebral Disc Regeneration: Mechanisms, Strategies, and Translational Potential.

Jiliang Wang, Bing Ran, Jun Wei.

  Intervertebral disc degeneration (IVDD) is a primary contributor to chronic low back pain and disability, yet current treatment strategies remain largely palliative, with limited regenerative potential. Engineered extracellular vesicles (EVs) have emerged as a promising class of bio-nanocarriers, offering low immunogenicity, high biocompatibility, and precise therapeutic cargo delivery. This review systematically summarizes recent advancements in EV engineering approaches, including cargo loading techniques, surface functionalization, and integration with stimuli-responsive biomaterials, and delineates their mechanistic roles in modulating inflammation, cellular senescence, and extracellular matrix remodeling in the degenerated disc microenvironment. Furthermore, we structure the discussion around a "carrier-mechanism-application" axis, providing a conceptual and technical framework to guide the development of EV-based regenerative therapies. Finally, we address current translational challenges and propose future directions to bridge the gap between bench and bedside in the context of IVDD treatment.

Keywords:  biomaterials based scaffolds; extracellular vesicles; intervertebral disc degeneration (ivdd); tissue engineering and regenerative medicine; translational potential

DOI:  https://doi.org/10.7759/cureus.97119
Nat Commun. 2025 Dec 25.

Engineered vesicles enhance oral antibiotic absorption in proximal small intestine and mitigate gut dysbiosis.

Yinglan Yu, Yongfeng Xu, Zehui Yu, Xinrui Liu, Jiayue Guo, Min Xiang, Junmeng Chen, Riyanto Teguh Widodo, Lei Luo.

Oral antibiotics are a mainstay for treating bacterial infections, but unabsorbed portions can reach the caecum and colon, leading to gut dysbiosis. Herein, we engineer a biohybrid delivery system through the integration of milk extracellular vesicles with liposomes. The hybrid vesicles employ targeting mechanisms via neonatal Fc receptor and peptide transporter 1, facilitating antibiotic transport across the proximal small intestine. These vesicles exhibit superior drug encapsulation efficiency, stable release behavior, efficient mucus traversal, higher endocytosis, increased basolateral exocytosis, and improved oral absorption, achieving a 3.24-fold increase in oral bioavailability compared to free antibiotics. In lung bacterial infections and bacteremia models, hybrid vesicle-encapsulated cefdinir outperforms free antibiotics in eliminating infections. Notably, this approach also mitigates adverse effects on the intestinal microbiota, safeguarding the animals from dysbiosis-associated metabolic syndromes and opportunistic pathogen infections. This innovative hybrid vesicle system holds promise for the oral delivery of other drugs that suffer from limited absorption or cause gut dysbiosis.

DOI: https://doi.org/10.1038/s41467-025-68082-9
Mol Ther Nucleic Acids. 2025 Dec 09. 36(4): 102776

The role of exosomes as oligonucleotide delivery system for managing α-synuclein in Parkinson's disease: A systematic review of in vivo studies.

Soodeh Salarpour, Fatemeh Dourandish, Arsalan Salajegheh, Sara Hashemi.

  Although Parkinson's disease (PD) is primarily idiopathic, genetic mutations-accounting for approximately 5%-15% of cases with regional variability-have prompted the development of gene expression modulators, such as oligonucleotides, to target and reduce alpha-synuclein (α-syn) accumulation. However, challenges in delivering these agents to the brain have limited their therapeutic potential. This study systematically reviews the use of exosomes as delivery systems for oligonucleotides aimed at reducing α-syn aggregation in PD. A comprehensive literature search was conducted using Scopus, Embase, OVID, and ISI Web of Science databases up to January 2022, targeting in vivo studies relevant to the subject. Of 904 initial records, five eligible studies were selected. Three utilized transgenic mouse models and two used induced models to simulate PD. All reported a reduction in α-syn aggregation in the midbrain-particularly in the substantia nigra-following treatment with exosome-delivered oligonucleotides. This reduction was associated with decreased neuronal death and improved motor function. No significant toxicity or immune response was reported. Exosome-mediated oligonucleotide delivery appears to be a promising approach to reduce α-syn aggregation, protect dopaminergic neurons, and improve motor symptoms in animal models of PD.

Keywords:  MT; Oligonucleotides; Parkinson’s disease; Therapies and Applications; alpha-synuclein; antisense oligonucleotides; blood–brain barrier targeting; exosome; extracellular vesicles; gene therapy; neurodegeneration; oligonucleotide delivery

DOI:  https://doi.org/10.1016/j.omtn.2025.102776
Cancer Cell Int. 2025 Dec 20.

Combined action of suicide gene exosomes from pancreatic cancer-associated fibroblasts and from mesenchymal stem cells as a pancreatic ductal adenocarcinoma treatment approach.

Jana Jakubechova, Ursula Altanerova, Michal Andrezal, Dajana Vanova, Tatiana Zeleznikova, Peter Bartek, Marina Cihova, Bozena Smolkova, Verona Buocikova, Peter Makovicky, Kristina Jakic, Monika Burikova, Maria Urbanova, Veronika Repaska, Benjamin Spanik, Miroslav Tomas, Peter Dubovan, Georgina Kolnikova, Zbynek Zdrahal, Vaclav Pustka, David Potesil, Cestmir Altaner.

   BACKGROUND: The pancreatic cancer-associated fibroblasts (pCAFs) are among the most active components of the pancreatic ductal adenocarcinoma (PDAC). The pCAFs being of mesenchymal stem/stromal cell origin, interact directly with tumor stromal elements, modulate tumor development, and are involved in the formation of pre-metastatic niches that result in unsatisfactory PDAC treatment outcomes. This study aimed to develop an innovative approach for the treatment of desmoplastic pancreatic carcinoma via intracellularly targeted exosomes derived from pCAFs and from mesenchymal stem cells (MSCs) transduced with the suicide gene - yeast cytosine deaminase::uracil phosphoribosyl transferase (yCD::UPRT).
METHODS: pCAFs were isolated from four PDAC tumor specimens and MSCs from various tissues. Their transduction with yCD::UPRT gene produce homogenous gene transduced cell populations capable of secreting suicide gene exosomes. Both gene- transduced and naive cells and their exosomes underwent characterization by biophysical, biochemical, microscopic, and LC-MS/MS proteomic methods. Tumor cell-killing functionality was assessed using three pancreatic cancer cell lines. The killing efficacy of combined suicide gene exosomes of MSCs and pCAFs was measured in a mixture of pCAFs and MIA PaCa-2 cells as a simulated desmoplastic pancreatic tumor in vitro.
RESULTS: MSCs and pCAFs suicide gene exosomes act as cancer cell-targeted drugs, effectively killing pancreatic carcinoma cells. Exosomes intracellular convert the non-toxic prodrug 5-fluorocytosine into cytotoxic 5-fluorouracil and its metabolites in a dose-dependent manner. In experiments simulating the desmoplastic microenvironment of PDAC, we have found that the suicide gene exosomes from both cells conjugated with prodrug effectively target and inhibit the growth of simulated PDAC.
CONCLUSION: Exosomes containing the yCD::UPRT gene from pCAFs and MSCs function as "Trojan horse" therapies, efficiently and dose-dependently eliminating pancreatic cancer cells. PDAC environment-targeted yCD::UPRT-gene exosomes from MSCs and pCAFs show promise for a novel PDAC treatment.

Keywords:  Gene-directed enzyme prodrug therapy; Mesenchymal stem cells; Pancreatic cancer intracellular treatment; Pancreatic cancer-associated fibroblasts; Suicide gene exosomes

DOI:  https://doi.org/10.1186/s12935-025-04130-0
J Control Release. 2025 Dec 18. pii: S0168-3659(25)01172-1. [Epub ahead of print] 114558

Rapid extracellular vesicle surface decoration with targeting moieties based on a fluorescein binding single chain variable fragment snorkel.

Marieke Theodora Roefs, Johanna Gamauf, Barbara Kroenigsberger, Alessia Brancolini, Michael W Traxlmayr, Elsa Arcalis, Jacak Jaroslaw, Marcelle van Mechelen, Jean-Paul Prieels, Regina Grillari-Voglauer, Johannes Grillari, Madhusudhan Reddy Bobbili.

  Extracellular vesicles (EVs) are cell-derived nanovesicles with promising potential for drug delivery due to their low toxicity and immunogenicity. However, their clinical application is limited by poor targeting to sites of interest. Existing strategies to engineer targeted EVs often require genetic donor cell modification for each specific target, making the process time-consuming and costly. To overcome this, we developed a versatile targeting platform using the fluorescein-specific single-chain variable fragment (scFv) 4 M5.3, integrated into a CD81-based Snorkel-tag construct for surface display on EVs. A C-terminal HA-tag, separated by a PreScission protease (PS) site, allows selective purification of targeted EVs and removal of unbound targeting moieties. This design enables functionalization of EVs with any fluorescein-conjugated targeting molecule. We tested various construct modifications (cMyc, FLAG, PS-HA), which showed differing expression levels and FITC-antibody binding by HEK293 cells and their EVs. As proof of concept, we generated EVs targeting human HER2 and mouse CCR2 by capturing FITC-labeled antibodies, which bound specifically to HER2+ NCI-N87 and CCR2+ RAW264.7 cells. The technology was also successfully applied to transmembrane protein CD9 and WJ-MSC/TERT273-derived EVs. In summary, we present a robust, adaptable method for generating EVs with customizable targeting, enabling high-throughput target screening and accelerating the development of EV-based therapeutics.

Keywords:  Engineering; Extracellular vesicles (EVs); HER2; Targeting; WJ-MSC/TERT273, snorkel-tag

DOI:  https://doi.org/10.1016/j.jconrel.2025.114558
Regen Ther. 2025 Dec;30 491-502

Rat adipose-derived mesenchymal stem cell-derived exosomes loaded with miR-375 promote neurite outgrowth and peripheral nerve regeneration via activation of Akt by targeting EPHA4.

Zhiwei Liu, Yuanyuan Han, Chunjie Song.

   Background: MicroRNAs (miRNAs) carried by mesenchymal stem cells (MSCs)-derived exosomes participate in peripheral nerve regeneration. Our study intended to determine the role of miR-375-loaded exosomes secreted by adipose-derived MSCs (ADMSCs) in dorsal root ganglion (DRG) neurons in vitro and in rat models of sciatic nerve injury as well as the underlying mechanisms.
Methods: After the isolation of primary rat ADMSCs and DRG neurons, the characteristics of ADMSC-derived exosomes were identified by western blotting and nanoparticle tracking analysis. MiR-375 mimics or NC mimics were transfected into ADMSCs to prepare exo-miR-375 or exo-NC. Then, DRG neurons were co-cultured with exo-miR-375 or exo-NC to analyze the influence of exosomes loaded with miR-375 on axon extension by neurofilament immunofluorescence staining and neurotrophic factor production by RT-qPCR. A walking track analysis was conducted to assess the effects of exo-miR-375 or exo-NC injection on the recovery of rat sciatic nerve functions. Axon and myelinated fiber regeneration in injured nerves was observed through toluidine blue staining, transmission electron microscopy (TEM), and neurofilament immunofluorescence staining. TargetScan and miRDB databases were used to screen for miR-375 downstream target genes. The miR-375 and EPHA4 interaction relationship was validated through dual luciferase reporter assay. The phosphorylation of Akt in sciatic nerve tissues was determined via western blotting.
Results: ADMSCs-derived exosomes with overexpressed miR-375 stimulated axon extension and enhanced neurotrophic factor expression in DRG neurons as well as improved limb function recovery, facilitated axon myelinated fiber regeneration, and alleviated gastrocnemius muscle atrophy in rats after sciatic nerve injury. EPHA4 targeted by miR-375. Overexpressing EPHA4 reversed the promotion of exo-miR-375 on neurite outgrowth in vitro. Additionally, exo-miR-375 significantly reduced EPHA4 levels but elevated phosphorylated-Akt levels in rat sciatic nerves.
Conclusion: ADMSCs-derived exosomes with overexpressed miR-375 promoted neurite outgrowth and peripheral nerve regeneration by activating Akt through downregulating EPHA4.

Keywords:  Adipose-derived mesenchymal stem cells; Akt; EPHA4; Exosomes; Peripheral nerve injury; miR-375

DOI:  https://doi.org/10.1016/j.reth.2025.06.010