bims-engexo 2025-11-16 papers

bims-engexo

Biomed News

on Engineered exosomes

Issue of 2025–11–16
ten papers selected by
Ravindran Jaganathan, Universiti Kuala Lumpur

Engineered exosome-mediated delivery of MicroRNA let-7i-5p targets toll-like receptor 4 to modulate inflammation and alleviate acute liver injury in septic mice.
Engineered Exosomes as Programmable Nanocarriers for Targeted Theranostic and Translational Applications.
Production of Genetically Engineered Extracellular Vesicles for Targeted Protein Delivery.
Targeted Delivery of Engineered RVG-BDNF-Exosomes: A Novel Neurobiological Approach for Ameliorating Depression and Regulating Neurogenesis.
Targeted Delivery of Exosomal Bmp2 mRNA via a Bioadhesive Interface Synergistically Enhances Titanium Implant Osseointegration.
TLyp-1-modified exosome-mediated delivery of circRAPGEF5 ASO inhibits lung adenocarcinoma metastasis via regulating the miR-570-3p/SPP1 axis.
Progress of Exosomes in Cancer Immunotherapy.
Exosomes as Nonviral Carrier for Targeted Delivery of CRISPR/Cas12a for Therapeutic HIV-1Proviral DNA Editing.
MicroRNAs as Emerging Therapeutic Targets Modulating the Tumor Microenvironment in Head and Neck Squamous Cell Carcinoma.
Current landscape of cross-line immune checkpoint blockade for cancer treatment.

Int Immunopharmacol. 2025 Nov 10. pii: S1567-5769(25)01815-6. [Epub ahead of print]168(Pt 1): 115827

Engineered exosome-mediated delivery of MicroRNA let-7i-5p targets toll-like receptor 4 to modulate inflammation and alleviate acute liver injury in septic mice.

Hong-Phuc Nguyen Vo, Chao-Yuan Chang, Van Long Le, Hung-Jen Shih, Bing-Hua Su, Van Kim Long Vu, Chun-Jen Huang.

  Engineered exosomes have emerged as a promising platform for the targeted delivery of therapeutic nucleic acids. The microRNA let-7i-5p is recognized for its anti-inflammatory properties, but its molecular targets remain incompletely defined. To elucidate its mechanism of action and facilitate its efficient delivery, we generated let-7i-5p-enriched engineered exosomes (EEXOs) by overexpressing let-7i-5p in genetically modified RAW264.7 cells. Droplet digital polymerase chain reaction confirmed that EEXO treatment significantly increased the intracellular level of let-7i-5p (3-fold increase; p < 0.001). Bioinformatic analyses performed using four microRNA target prediction databases identified toll-like receptor 4 (TLR4) as a potential target. This finding was validated by a luciferase activity assay, which revealed direct binding of let-7i-5p to the 3' untranslated region of TLR4 mRNA, resulting in its suppression (p < 0.001). In lipopolysaccharide (LPS)-stimulated macrophages, EEXO-delivered let-7i-5p downregulated TLR4 and its downstream signaling components, such as myeloid differentiation primary response 88 (MyD88), tumor necrosis factor receptor-associated factor 6 (TRAF6), and nuclear factor-κB (NF-κB), thereby reducing the levels of proinflammatory cytokines such as tumor necrosis factor-α and interleukin-6 (all p < 0.05). In vivo, systemic EEXO administration attenuated LPS-induced liver injury in adult male mice, as indicated by reduced histopathological damage, liver enzyme release, and tissue edema (all p < 0.05). Furthermore, EEXOs suppressed the TLR4/MyD88/TRAF6/NF-κB pathway and proinflammatory cytokine production in liver tissues. In conclusion, engineered exosome-delivered let-7i-5p targets TLR4 and modulates MyD88/TRAF6/NF-κB signaling to alleviate inflammation and protect against LPS-induced liver injury.

Keywords:  Engineered exosomes; Inflammation; Let-7i-5p; Liver injury; TLR4

DOI:  https://doi.org/10.1016/j.intimp.2025.115827
ACS Appl Bio Mater. 2025 Nov 14.

Engineered Exosomes as Programmable Nanocarriers for Targeted Theranostic and Translational Applications.

Jugal Patil, Ankur Singh, Satyam Bhalerao, Syed Mudasir Ahmad, Rakesh M Rawal, Dhiraj Bhatia, Raghu Solanki.

  Exosomes are nanoscale extracellular vesicles secreted by cells that possess molecular and pathological characteristics of their cellular origin. Acting as natural carriers, they efficiently transport a diverse cargo of biomolecules, including proteins, nucleic acids, lipids, metabolites, and small molecules facilitating highly specific intercellular communication. Owing to their inherent biocompatibility, target specificity, and cargo versatility, exosomes have emerged as one of the most promising platforms for diagnostic and therapeutic applications. This review comprehensively elaborates on intricate biogenesis and regulatory pathways governing exosome production, examines their structural composition and cargo loading preferences, and highlights emerging strategies to enhance their functional capabilities. We further explore recent breakthroughs at the intersection of exosome biology and nanotechnology, emphasizing their roles in maintaining cellular homeostasis, advancing disease diagnostics, and enabling targeted therapeutic delivery. Finally, we critically address current challenges and limitations in exosome research, offering insights into innovative solutions and future directions for their clinical translation.

Keywords:  cargo loading; exosome biogenesis; exosomes; surface engineering; therapeutics

DOI:  https://doi.org/10.1021/acsabm.5c01251
Bio Protoc. 2025 Nov 05. 15(21): e5494

Production of Genetically Engineered Extracellular Vesicles for Targeted Protein Delivery.

Leyla A Ovchinnikova, Evgeniy G Evtushenko, Dmitriy V Bagrov, Anton O Goncharov, Alexander G Gabibov, Yakov A Lomakin.

  Extracellular vesicles (EVs) have emerged as promising carriers for the targeted delivery of therapeutic proteins to specific cells. Previously, we demonstrated that genetically engineered EVs can be used for targeted protein delivery. This protocol details the generation of mannose receptor (CD206)-targeted EVs using a modular plasmid system optimized for production in HEK293T cells. Three plasmids enable customizable EV budding, cargo loading, and surface modification for targeting to antigen-presenting cells (APCs). EVs are isolated via differential centrifugation and chromatography, characterized using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA), and validated through functional uptake assays in primary human activated dendritic cells. Our approach combines flexibility in engineering required EVs with robust, reproducible isolation and characterization workflows. Its modularity allows easy adaptation to alternative targets or cargoes, which can be validated immediately through in vitro testing. Key features • First detailed protocol for generating genetically engineered EVs with fusogenic VSV-G protein and CD206-specific targeting. • Enables rapid customization of EVs for diverse therapeutic cargo and cell-targeting applications. • Integrates gold-standard EV isolation with multi-modal characterization to ensure reliability. • A universal platform for customizable cell targeting: swapping VSV-G-linked llama nanobodies with diverse specificities.

Keywords:  Antigen-presenting cells; CD206; EVs; Engineered EVs; NTA; Protein delivery; Target delivery; VSV-G

DOI:  https://doi.org/10.21769/BioProtoc.5494
Research (Wash D C). 2024 ;7 0402

Targeted Delivery of Engineered RVG-BDNF-Exosomes: A Novel Neurobiological Approach for Ameliorating Depression and Regulating Neurogenesis.

Shaobo Liu, Lei Chen, Mei Guo, Yongbiao Li, Qingshan Liu, Yong Cheng.

Addressing the urgent need for innovative depression treatments, this study heralds a breakthrough in major depressive disorder (MDD) therapy by intertwining clinical observations with neurobiological advancements. We analyzed brain-derived neurotrophic factor (BDNF) levels in serum exosomes from a diverse group of 60 individuals, including first-episode, drug-free MDD patients, medicated MDD patients, and healthy controls. Our results revealed a significant decrease in BDNF levels within MDD patients' exosomes, which notably increased post-medication, highlighting BDNF's potential as a biomarker for both MDD diagnosis and treatment efficacy. Advancing these clinical findings, we developed RVG-modified exosomes engineered to overexpress BDNF (RVG-BDNF-Exos), designed to directly target neuronal cells. Our findings demonstrate that these engineered exosomes can successfully traverse the blood-brain barrier, targeting neurons in the hippocampus and prefrontal cortex. In our mouse model of depression induced by lipopolysaccharide, RVG-BDNF-Exos treatment led to a significant increase of BDNF in these key brain regions, crucial for mood regulation and neurogenesis. This intervention modulated the BDNF/TrkB/AKT signaling pathway, central to neural plasticity and implicated in depression's pathogenesis. Behavioral assessments exhibited substantial improvements in depressive-like behaviors in mice treated with RVG-BDNF-Exos, including reduced immobility in Tail Suspension and Forced Swim Tests. Additionally, our treatment effectively decreased neuroinflammation, as evidenced by the reduction in microglia and astrocyte numbers. Moreover, RVG-BDNF-Exos treatment enhanced neurogenesis and regulated synaptic plasticity, as indicated by the increased expression of neuronal markers MAP2 and DCX, and synaptic proteins PSD95 and Syn-1. In conclusion, this study not only underscores the clinical potential of serum exosomal BDNF as a diagnostic and therapeutic marker for MDD but also demonstrates the efficacy of RVG-BDNF-Exos in alleviating depressive symptoms. Our findings pave the way for future targeted, personalized psychiatric treatments, offering a promising direction in MDD therapy.

DOI: https://doi.org/10.34133/research.0402
Small. 2025 Nov 09. e10386

Targeted Delivery of Exosomal Bmp2 mRNA via a Bioadhesive Interface Synergistically Enhances Titanium Implant Osseointegration.

Zhujun Yang, Yan Dong, Zhiwei Dong, Pengyu Zhao, Yichen Li, Liang Kong, Yimin Zhao, Zhongshan Wang.

  Rapid osseointegration of titanium implants remains challenging under pathological conditions. This study develops an engineered exosome-based mRNA delivery system for osteogenic regulation. Through co-transfection with BMP2 and Paip2 plasmids, Exosomes (Exoen) loaded with untranslated Bmp2 mRNA are generated, exhibiting an eight-fold increase over naive exosomes and a three-fold enhancement compared to BMP2-only exosomes. These vesicles are functionalized with a cholesterol-anchored E7 peptide for BMSC targeting and immobilized on a titanium implant via a mussel adhesive protein coating (Exoen-E7@TiM). The system enables targeted, sustained delivery of Bmp2 mRNA to BMSCs. Once internalized, the mRNA utilizes an internal ribosome entry site (IRES) to initiate translation, effectively bypassing the original suppression and driving robust BMP2 protein expression. Exoen-E7@TiM significantly enhanced osteogenic differentiation of human bone mesenchymal stem cells (hBMSCs) in vitro, upregulating key markers and accelerating mineralization. In rat tibiae, it substantially improved new bone formation and bone-implant contact vs controls. This integrated strategy combines exosome-mediated nucleic acid delivery, biological targeting, and bioadhesive immobilization to significantly enhance osseointegration, showing considerable promise for clinical applications in compromised bone healing.

Keywords:  Bmp2 mRNA delivery; E7 peptide; engineered exosomes; mussel adhesive protein; osseointegration

DOI:  https://doi.org/10.1002/smll.202510386
Mater Today Bio. 2025 Dec;35 102416

TLyp-1-modified exosome-mediated delivery of circRAPGEF5 ASO inhibits lung adenocarcinoma metastasis via regulating the miR-570-3p/SPP1 axis.

Tianqi Hu, Chenkang Zhou, Yuxin Chen, Jiali Fu, Yingjie Dai, Liqun Ling, Yixin Chen, Yi Lu, Yuyu He, Jie Chen, Yumin Wang.

  Exosome circRAPGEF5 promotes lung adenocarcinoma (LUAD) metastasis and may serve as an intervention target, but its mechanism of action is unclear. Consequently, using engineered exosomal preparations of circular RNAs for delivery into tumor cells represents a safe and effective strategy for cancer treatment. The results confirmed the successful construction of tLyp-1-exosome (EXO)-circRAPGEF5 antisense oligonucleotides (ASO). tLyp-1 modified exosomes efficiently targeted LUAD tumors in vivo and in vitro. The tLyp-1-EXO-circRAPGEF5 ASO significantly reduced the proliferation, invasion, migration, and levels of N-cadherin and vimentin in A549 and HCC827 cells. circRAPGEF5 acted as an miR-570-3p sponge, facilitating LUAD progression, and miR-570-3p directly regulated secreted phosphoprotein 1(SPP1) expression. The knockdown of circRAPGEF5 inhibited LUAD progression via the miR-570-3p/SPP1 axis. The tLyp-1-EXO-circRAPGEF5 ASO did not damage BEAS-2B (Human Normal Lung Epithelial) cells. Nude mouse model experiments using hematoxylin and eosin staining and biochemical indexes demonstrated that tLyp-1-EXO-circRAPGEF5 ASO significantly inhibited LUAD metastasis and did not cause biological damage to vital organs. This study clarified the mechanism of tLyp-1-EXO-circRAPGEF5 ASO in inhibiting LUAD metastasis, indicating that circRAPGEF5 may serve as a new intervention target for LUAD metastasis.

Keywords:  Engineered exosomes; Lung adenocarcinoma (LUAD); Metastasis; circRAPGEF5; miR-570-3p

DOI:  https://doi.org/10.1016/j.mtbio.2025.102416
Cell Biochem Funct. 2025 Nov;43(11): e70140

Progress of Exosomes in Cancer Immunotherapy.

Yueyou Wang, Jingyuan Wang, Han Li, Siyun Deng, Yanli Li.

  Cancer immunotherapy, focusing on breaking tumor microenvironment (TME) immunosuppression, is limited by heterogeneity and drug resistance. Exosomes, 30-150 nm extracellular vesicles(EVs) carrying proteins, lipids, and noncoding RNAs, mediate intercellular communication and play dual roles in tumors. This review explores their multifaceted functions in cancer immunotherapy: in TME, tumor-derived exosomes (TDEs) drive immunosuppression, cancer-associated fibroblasts(CAFs) activation, and angiogenesis to promote progression and immune checkpoint inhibitors (ICIs) resistance; diagnostically, exosomal biomolecules (e.g., urinary miR-424/423/660/let-7i, serum LINC01125) serve as sensitive liquid biopsy markers for early detection and monitoring; therapeutically, engineered exosomes (e.g., DC-derived antigen-loaded ones) activate antitumor immunity and reverse ICIs resistance. These findings highlight exosomes' potential as diagnostic and therapeutic tools, laying a foundation for personalized cancer treatment.

Keywords:  cancer immunotherapy; exosomes; noncoding RNAs

DOI:  https://doi.org/10.1002/cbf.70140
Mol Ther. 2025 Nov 11. pii: S1525-0016(25)00949-9. [Epub ahead of print]

Exosomes as Nonviral Carrier for Targeted Delivery of CRISPR/Cas12a for Therapeutic HIV-1Proviral DNA Editing.

Yan Ma, Wanrong Wei, Zihao Yang, Ying Zhou, Ting Dong, Tao Wang, Xuejiao Xia, Yige Ma, Mengying Zhou, Yang Gao, Boyu Yu, Changjun Wang, Lianguo Ruan, Ke Hong, Chaojiang Gu.

Current strategies to treat HIV infection including traditional cART and immunotherapy can effectively suppress viral replication but unable to eliminate the latent viral reservoir, particularly within circulating immune cells. Although genome editing by CRISPR-Cas provides a promising cure for HIV-1, gene delivery efficiency in vivo remains an obstacle to overcome. Here, we developed an exosome-mediated targeted CRISPR-Cas12a delivery system (EMT-Cas12a), an engineered exosome system enabling targeted delivery of mRNA of Cas12a and crRNAs to CD4+ T cells. The EMT-Cas12a system uniquely optimizes cell-specific targeting, CRISPR-Cas12a expression, crRNAs maturation, nuclear entry efficiency, accuracy cleavage with major Delins and achieving dramatically HIV suppression in both cellular and humanized mouse models. Compared to single-crRNA approaches, the multiple crRNA arrays strategy demonstrates enhanced antiviral efficacy in HIV -infected mouse model, ex vivo expanded PBMCs from HIV+ subjects and especially in vitro cell line without detectable HIV DNA. Critically, the system exhibits no detectable off-target effects and restores CD4+ T cell counts in vivo and ex vivo PBMCs, indicating its dual therapeutic potential for viral clearance and immune reconstitution. Altogether, in vitro and in vivo excision of HIV-1 proviral DNA can be achieved via EMT-Cas12a delivery, which could advance efforts toward human clinical trials.

DOI: https://doi.org/10.1016/j.ymthe.2025.11.012
Int J Mol Sci. 2025 Nov 06. pii: 10794. [Epub ahead of print]26(21):

MicroRNAs as Emerging Therapeutic Targets Modulating the Tumor Microenvironment in Head and Neck Squamous Cell Carcinoma.

Roxana Daniela Brata, Lavinia Marcut, Alina Cristina Barb, Alexia Manole, Alexandru Ciolofan, Cristina Stefania Dumitru, Flavia Zara, Raul Patrascu.

  Head and neck squamous cell carcinoma (HNSCC) remains one of the most aggressive solid tumors, characterized by marked molecular heterogeneity and a complex tumor microenvironment (TME). Recent evidence highlights the pivotal role of microRNAs (miRNAs) in regulating tumor progression, immune evasion, angiogenesis, and stromal remodeling. This review synthesizes current insights into miRNA-mediated molecular pathways that modulate the TME in HNSCC and discusses emerging therapeutic strategies, including nanocarrier- and exosome-based miRNA delivery systems, targeting these molecules. Key miRNAs, including miR-21, miR-146a, and miR-221, orchestrate bidirectional signaling between cancer cells, fibroblasts, and immune infiltrates, thereby shaping tumor aggressiveness and therapy resistance. Advances in nanotechnology have facilitated the development of miRNA-based therapeutics-such as mimics, antagomiRs, and exosome-mediated systems-capable of restoring physiological expression patterns and reprogramming the TME toward an anti-tumor state. However, clinical translation remains hindered by challenges in targeted delivery, molecular stability, and tumor heterogeneity. By integrating molecular and translational perspectives, this review underscores how miRNA-targeting strategies may evolve into a new generation of precision therapies, bridging the gap between molecular oncology and personalized treatment of head and neck cancer.

Keywords:  exosomes; head and neck cancer; immune modulation; microRNA; nanotherapy; therapeutic targets; tumor microenvironment

DOI:  https://doi.org/10.3390/ijms262110794
Cancer Lett. 2025 Nov 12. pii: S0304-3835(25)00707-4. [Epub ahead of print] 218135

Current landscape of cross-line immune checkpoint blockade for cancer treatment.

Pinchao Fan, Jing Wu, Chengjun Zhu, Xiaoxiang Guan.

  Substantial advances from bench to bedside have arisen in the realm of cancer immunotherapy, endowing unprecedented survival benefits and even curing a minor subset of patients with advanced cancer. However, the clinical application of immune checkpoint blockade (ICB) is impeded by several critical challenges, and it tends to be dismissed after disease progression or severe immune-related adverse events following previous ICB attempt. Pursuing further augmentation of ICB efficacy and optimization of overall prognosis, cross-line immunotherapy has emerged as a promising rechallenging tactic for patients who are refractory, relapsed or intolerant to prior ICB. By far, cross-line immunotherapy remains immature and controversial due to the absence of a constructed framework, comprehensive profiling and convincing evidence. In this review, we delve into this burgeoning field from conventional ICB to cross-line immunotherapy, elucidating the current landscape, latest explorations, practical hurdles, viable counteractions and future perspectives. Characterizing the rationales would update our knowledge and unveil the latent therapeutic potential of cross-line immunotherapy, thereby benefiting post-ICB patients to the utmost.

Keywords:  Advanced cancer; Cross-line immunotherapy; Immune checkpoint blockade; Immunotherapy rechallenge; Tumor immune microenvironment

DOI:  https://doi.org/10.1016/j.canlet.2025.218135