O-linked N-acetylglucosamine (O-GlcNAc) is a monosaccharide modification occurring on serine or threonine residues of most eukaryotic proteins. Only two enzymes, O-GlcNAc transferase and O-GlcNAc hydrolase, regulate the dynamic flux of O-GlcNAc modification, rendering it extremely responsive to nutrition and stress conditions. O-GlcNAcylation stands at the center of epithelial-mesenchymal transition (EMT), sensing nutrient and stress signals to direct the transcriptional and signaling programs that enable phenotypic plasticity, thereby establishing its fundamental role in fibrosis and tumor metastasis. EMT is an essential biological event that confers mesenchymal characteristics to epithelial cells, characterized by the suppression of E-cadherin, a key epithelial adhesion molecule, and the overexpression of N-cadherin, a mesenchymal cadherin that promotes motility, or Vimentin, a mesenchymal intermediate filament protein. This review covers recent insights on the multiple canonical and non-canonical roles of O-GlcNAc, presenting O-GlcNAc cycling as a significant post-translational mechanism involved in various aspects of EMT. Furthermore, we systematically examine the functional connections between O-GlcNAcylation and EMT, focusing on identifying key O-GlcNAcylated proteins that regulate EMT and evaluating the relative contributions of transcriptional and post-translational mechanisms mediated by this modification. A comprehensive understanding of the intricate molecular circuitry governing the interplay between O-GlcNAcylation and EMT will deepen our mechanistic insights into cellular plasticity and offer novel therapeutic avenues for combating metastasis and other EMT-associated pathologies.

Excessive or persistent inflammation intensifies skin tissue damage, leading to abnormal repair processes like scar hyperplasia, and potentially causing chronic wound non-healing or even malignant transformation. The effective enhancement of wound repair and regeneration is impeded by the challenge of precisely suppressing overactivated immune cells, repairing damaged skin cells, and rapidly reconstructing a stable local immune microenvironment. We engineered small extracellular vesicles (sEVs) co-overexpressing PDL1 and Siglec15 (PDL1-Siglec15-sEVs), these PDL1-Siglec15-sEVs were encapsulated within photocurable hydrogels to create a multifunctional hydrogel-based therapeutic system (PDL1-Siglec15-sEVs@NHT). This system continuously releases PDL1-Siglec15-sEVs into the surrounding microenvironment at physiological temperature. In vitro experiments demonstrated that PDL1-Siglec15-sEVs exert an immune reprogramming effect, significantly inhibiting PBMC proliferation and reducing the proportion of CD8+ T cells. Furthermore, they effectively suppressed ROS generation in HaCaT cells and diminished the expression of multiple inflammatory factors. This multifunctional hydrogel composite system accelerated wound closure and promoted re-epithelialization in a rat full-thickness skin defect model by stimulating skin cell proliferation, migration, and extracellular matrix deposition. Transcriptome sequencing revealed that PDL1-Siglec15-sEVs profoundly upregulated Krt32 expression in skin keratinocytes while inhibiting activation of the NF-κB mediated inflammatory signaling pathway. PDL1-Siglec15-sEVs significantly promote skin wound healing through multifaceted mechanisms, synergistically enhancing the immunosuppressive effect mediated by the PD1/PDL1 pathway, reducing keratinocyte inflammatory responses, and fostering the polarization of M2-type macrophages.

Type 2 diabetes mellitus (T2DM) is increasingly viewed as a systemic metabolic disorder rather than as an isolated glycemic defect. It is closely linked to obesity and metabolic dysfunction-associated steatotic liver disease (MASLD), sharing mechanisms such as insulin resistance, chronic inflammation, and interorgan crosstalk among the gut, liver, adipose tissue, pancreas, and central nervous system. Plant-derived nanovesicles (PDNVs) are emerging cross-kingdom nanoplatforms with the potential to modulate metabolic and inflammatory signaling. Evidence from T2DM, obesity, and MASLD models suggests that PDNVs can improve metabolic homeostasis by reprogramming metabolism and reducing inflammation. PDNVs may modulate organ axes, particularly gut-liver and gut-brain axes, and potentially adipose-liver and liver-pancreas axes, partly via context- and tissue-dependent regulation of AMPK-, PI3K-AKT-, and SIRT1-related pathways. Strategies such as structural engineering, surface functionalization, and cargo loading optimization may further enhance the stability, bioavailability, and targeting capacity of PDNVs, thereby supporting their application in metabolic diseases.

Exosomes mediate a two-way signaling between gastric cancer (GC) cells and macrophages in the tumor microenvironment (TME) and have significant effects on tumor progression, immune suppression, and treatment outcomes. This paper will bring together existing literature on the vesicular cargo of macrophage-derived and GC-derived exosomes, such as non-coding RNAs (ncRNAs), and proteins, and their contribution to the re-polarization of macrophages, as well as the behavior of GC cells. The cargo contained in macrophage-derived exosomes biases signaling pathways associated with macrophage polarization, tumor-promoting phenotype, and helping GC proliferation, invasion, metastasis, and chemoresistance. Conversely, macrophage exosomal messages may complement anti-tumor immunity through the regulation of PD-L1 and augmentation of T cells. The exosomes of GC also remodel the macrophage phenotype in favor of immunosuppression by delivering cargoes, facilitating metabolic reprogramming, and tumor-associated macrophages (TAMs) polarization. This crosstalk is generalized to more widespread tumor stroma interactions, which involve cancer-associated fibroblasts (CAFs) and angiogenesis. The resulting therapeutic implications are engineered exosomes and cargo-modified vesicles to tip the balance towards anti-tumor immunity and overcome chemoresistance, where a high efficiency of loading and precision of targeting remain problematic, and the translation of preclinical research findings to the clinic.

Abstract Natural killer (NK) cells are potent immune modulators that can quickly lyse tumor cells without prior sensitization or recognition of specific tumor antigens. However, NK cells in tumor microenvironment are frequently dysfunctional. Despite advances in cytokine therapy and NK cell adoptive transfer, tumor-expressed ligands that interact with NK cell-expressed checkpoint receptors can greatly suppress NK cell-mediated tumor lysis, leading to suboptimal therapeutic outcomes. Here we explored a new approach of utilizing cellular nanovesicles (NVs) as artificial controllers of cellular immunity to redirect and activate NK cells for tumor eradication. Through this strategy, we engineered HEK293 cells-derived NVs genetically modified to display two distinct types of surface monoclonal antibodies (mAbs), namely human CD16 and epidermal growth factor receptor 2 (HER2) antibodies, generating dual αCD16/αHER2 NVs that simultaneously target CD16-expressing NK cells and HER2-expressing gastric cancer cells. Following thorough characterization, the αCD16/αHER2 NVs exhibited highly potent and specific anti-tumor activity both in vitro and in vivo. This preclinical work demonstrates the feasibility of using dual-targeting NVs to navigate functional NK cells to recognize and eliminate gastric cancers, presenting αCD16/αHER2 NVs as an effective and versatile platform for cancer immunotherapy.

The search for an efficacious blood substitute has remained a persistent challenge for more than a century. Despite extensive efforts to develop synthetic oxygen carriers, no broadly approved alternative to human blood has yet been realized. In this review, we first provide an overview of the historical development and repeated failures of artificial blood substitutes, while highlighting the continuing clinical demand for transfusion alternatives. We then discuss recent setbacks in blood substitute development, particularly adverse events observed in human trials. Current research efforts with goals to improve biocompatibility are further examined, with a particular focus on oxygen-carrying micro-/nano-particles, emerging fabrication technologies, and unmet clinical needs. Hemoglobin-based cellular and acellular oxygen carriers, as well as novel non-hemoglobin-based systems, are reviewed in detail with emphasis on technical innovations and biological advances. Finally, future development in the field was briefly envisioned to shed light on the rational design and versatile applications of safe and efficient oxygen carriers.

Abstract Extracellular vesicles (EVs) have emerged as promising nanotherapeutics for kidney diseases due to their innate biocompatibility, barrier penetration, and regenerative cargo delivery. However, native EVs face critical limitations including low drug‐loading efficiency, poor renal targeting, and batch heterogeneity, hampering their clinical utility. Recent advances in EV bioengineering—encompassing cargo loading (electroporation, transfection), membrane modification (ligand conjugation, biomimetic vesicles), and donor cell preconditioning (hypoxia, pharmacological priming)—have significantly enhanced therapeutic precision and efficacy. Integration with biomaterials (e.g., responsive hydrogels, scaffolds) further enables sustained release and targeted delivery, improving outcomes in kidney injury models. This review systematically analyzes these innovative strategies, highlighting mechanistic insights, comparative advantages, and unresolved challenges. We critically evaluate ongoing clinical trials and propose scalable manufacturing solutions and regulatory frameworks to accelerate translation. Engineered EVs represent a next‐generation platform for personalized renal nanomedicine, poised to bridge the regenerative potential with clinical reality.

ABSTRACT Small extracellular vesicles (sEVs)‐derived circular RNA (circRNA) serves as a crucial biomarker for diagnosing gastric cancer and as a key regulator of tumor progression, orchestrating intercellular crosstalk within the tumor microenvironment (TME). In gastric cancer (GC), tissue‐derived mesenchymal stem cells (GC‐MSCs) critically drive tumor progression; however, the interplay between sEVs and circRNA in GC‐MSCs remains incompletely understood. We identified sEVs‐hsa_circ_0006718 (circ6718) as significantly upregulated in gastric cancer patients. Elevated levels of sEVs‐circ6718 correlated with clinical stage, distant metastasis and poor prognosis, confirming its utility as both an early diagnostic and prognostic biomarker in GC. Mechanistically, circ6718 functions as a competing endogenous RNA (ceRNA) by sequestering hsa‐miR‐561‐3p, thereby derepressing the expression of SAAL1 (Serum amyloid A‐like 1). SAAL1 enhances the transcriptional activity of PRRX1 (Paired related homeobox 1), which directly activates the TGFβ1 promoter. Consequently, the TGFβ1/Smad2/3 signaling pathway drives the transdifferentiation of GC‐MSCs into cancer‐associated fibroblasts (CAFs)—promoting stromal remodeling and tumor aggressiveness. Our findings unveil a novel sEVs‐circRNA‐mediated axis in GC progression, revealing dual utility in diagnostics and targeted therapy.

Osteoporotic bone defects remain a major clinical challenge due to impaired osteogenesis, disrupted angiogenesis, and poor scaffold integration. To overcome these limitations, we developed hierarchical micro-/nanostructured hydroxyapatite (nwHA) scaffolds by integrating morphology-specific nanohydroxyapatite (nHA) onto whisker-reinforced hydroxyapatite (wHA) scaffolds. This modular strategy decouples mechanical strength from interfacial bioactivity, enabling programmable topographical control. Five distinct nHA morphologies were used to functionalize wHA scaffolds, which were systematically evaluated both in vitro and in osteoporotic rat models. Among them, nanofiber-coated scaffolds (nwHA1) significantly enhanced bone volume fraction, mineral apposition rate, mechanical strength, and neovascularization. Histological analysis identified three distinct ossification patterns—type I (wall-penetrating), type II (surface-appositional), and a hybrid endochondral–intramembranous mode—whose distribution varied with nHA morphology and the local microenvironment. Mechanistically, nwHA1 activated canonical Hedgehog signaling and upregulated HIF-1α in both MSCs and HUVECs, thereby promoting coordinated osteogenic and angiogenic responses. Pharmacological inhibition with cyclopamine, as well as siRNA-mediated knockdown of GLI1 or HIF-1α, significantly attenuated these pro-osteoangiogenic markers, confirming functional crosstalk between Hedgehog and hypoxia signaling pathways in response to scaffold-induced topographic cues. These findings establish nHA morphology as a critical topographical regulator of bone regeneration and provide a versatile platform for designing adaptive bioceramics tailored to osteoporotic bone repair.

Renal interstitial fibrosis is a critical feature of renal injury in diabetic kidney disease (DKD), and the degree of fibrosis progressively worsens as the disease advances. Therefore, understanding the complexity of the renal fibrosis microenvironment is vital for improving anti-fibrotic therapeutic outcomes. In this study, we found that CD68+VEGF+TGF-β1+ macrophages promote the overexpression of NUAK family SNF1-like kinase 1 (NUAK1) in tubular epithelial cells, as revealed by single-cell transcriptome sequencing. Mechanistically, increased NUAK1 directly binds to Yes1-associated protein (YAP), disrupts its interaction with large tumor suppressor kinase 1 (LATS1), and promotes YAP nuclear translocation, thereby accelerating the progression of DKD fibrosis. In addition, umbilical cord mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) attenuate renal interstitial fibrosis in DKD by inhibiting the NUAK1/YAP pathway. Analysis of conditional Nedd4 loss-of-function transgenic mice showed that MSC-EVs exert anti-fibrotic effects by delivering the E3 ubiquitin ligase Nedd4, which mediates NUAK1 degradation. Furthermore, we constructed dual-targeted engineered MSC-EVs modified with superparamagnetic nanoparticles and loaded with Nedd4 (SPION-EVs-Nedd4), which improved their targeted anti-fibrotic therapeutic effects. Together, these findings provide a new strategy for the prevention and treatment of DKD interstitial fibrosis, with significant scientific value and clinical translational potential.

Liver fibrosis (LF) is the final common pathological outcome of various chronic liver diseases. Advanced LF can progress to severe complications, such as cirrhosis, liver failure, and hepatocellular carcinoma (HCC). Currently, liver transplantation remains the main clinical treatment for advanced LF, but its application is limited by donor availability and unavoidable complications. Extracellular vesicles (EVs), nanoscale particles actively released by hepatic cells, including hepatocytes, hepatic stellate cells (HSCs), and macrophages), circulate in bodily fluids carrying cell-specific cargoes (e.g., RNAs, proteins). EVs mediate intercellular communication via their specific cargo profiles and contribute to the progression in LF. Increasing evidence indicates that tracking changes in the quantity and composition of EVs in LF can aid in disease diagnosis and prognosis prediction. This review discusses the pathological role of EVs in LF development and their potential as biomarkers and therapeutic targets, and provides new perspectives for future research and treatment advances.

Triple-negative breast cancer (TNBC) is a highly heterogeneous subtype with a poor prognosis and limited therapeutics, for which metastasis is a primary driver. In this study, we explore the function of EGFR in breast cancer progression and the mechanistic basis of EGFR-targeted therapies, focusing on kinase inhibitors and protein degraders. In a murine breast cancer model, EGFR knockout exhibited minimal impact on primary tumor growth but significantly suppressed metastasis, supporting the clinical association between high EGFR expression and poor prognosis. At high doses, EGFR degraders exert their antitumor effect through kinase inhibition, not degradation, whereas low-dose EGFR inhibition has limited anti-proliferative activity. Gene-drug interaction screening identified gemcitabine (GEM) and decitabine (DAC) as agents interacting with EGFR, a finding confirmed by the observed resistance to these drugs in TNBC cell lines with high EGFR expression. Both low-dose EGFR degraders and inhibitors chemosensitize cells to GEM/DAC in vitro, with the EGFR degrader plus GEM combination exhibiting an enhanced inhibitory effect in a lung colonization model. Mechanistically, both low-dose EGFR degraders and inhibitors elevate ROS levels and induce lipid peroxidation, thereby sensitizing cancer cells to GEM/DAC-triggered ferroptosis. In summary, this study demonstrates that low-dose EGFR targeting creates a susceptible state for ferroptosis in tumor cells and, consequently, defines a combination therapy strategy of EGFR-targeted agents with chemotherapy for TNBC. Our results provide a foundation for the clinical translation of low-dose EGFR-targeting strategies and their rational combination with other agents.

Accumulating evidence indicates that environmental exposures, particularly to nitrites, play a critical role in the initiation and progression of gastric cancer (GC). During carcinogenesis, exosomes act as key mediators of intercellular communication. Exosomes derived from N‑methyl-N'‑nitro‑N‑nitrosoguanidine (MNNG)‑induced malignantly transformed GES‑1 cells (TGES‑1), as well as serum exosomes from gastric cancer patients with a history of high nitrite exposure, were found to influence normal cells and promote GC initiation. The present study established a malignant transformation model and applied bioinformatics analyses to screen and validate candidate circRNAs. A series of functional and mechanistic experiments were performed to elucidate the regulatory role of exosomes in GC progression. Circ0000549 was markedly upregulated in MNNG‑exposed GES‑1 cells, their derived exosomes and serum exosomes from patients with GC. Further investigations revealed that circ0000549 overexpression enhanced GES‑1 cell malignant features, while also modulating epithelial‑mesenchymal transition and stemness‑related properties. Nude mouse experiments demonstrated that circ0000549, carried by malignantly transformed exosomes, plays a crucial role in MNNG‑induced gastric carcinogenesis. Mechanistically, miR‑15b‑5p was identified as a potential target of circ0000549. Circ0000549 functioned as a sponge for miR‑15b‑5p, leading to increased KIF1B expression and subsequent activation of the PI3K/AKT signaling pathway. Collectively, these findings reveal that exosomal circ0000549 promotes malignant transformation of GES‑1 cells through the miR‑15b‑5p/KIF1B/PI3K/AKT axis. Exosomal circ0000549 may serve as a promising biomarker for GC diagnosis and prognosis, highlighting its potential as a target for future therapeutic investigation.

To evaluate the clinical applicability of deep learning (DL) models based on automatic segmentation in preoperatively predicting tumor spread through air spaces (STAS) in peripheral stage I lung adenocarcinoma (LUAD). This retrospective study analyzed data from patients who underwent surgical treatment for lung tumors from January 2022 to December 2023. An external validation set was introduced to assess the model's generalizability. The study utilized conventional radiomic features and DL models for comparison. ROI segmentation was performed using the VNet architecture, and DL models were developed with transfer learning and optimization techniques. We assessed the diagnostic accuracy of our models via calibration curves, decision curve analysis, and ROC curves. The DL model based on automatic segmentation achieved an AUC of 0.880 (95 www.researchregistry.com ).

Alzheimer’s disease is an increasingly prevalent neurodegenerative disorder characterized by the accumulation of misfolded Aβ aggregates. The efficacy of upconversion nanomaterials in photodynamic therapy has been proven, yet their applicability is constrained by their inherent toxicity. In this study, exosomes derived from stem cells, combined with targeting molecules, were utilized to encapsulate upconversion nanoparticles for targeted removal of pathogenic amyloid aggregates in Alzheimer’s disease. Evidence from AFM and TEM imaging, fluorescence spectroscopy, and cell-based experiments demonstrates that the clearance efficiency of amyloid aggregates is enhanced, and the cytotoxicity induced by misfolded Aβ aggregates is significantly reduced in nerve cells. Notably, the viability of neural PC12 cells is partially restored even in the presence of toxic amyloid aggregates.

BACKGROUND:Hesperetin, a flavonoid predominantly present in citrus fruits, exhibits significant intervention effects on both the initiation and progression of gastric cancer. However, the specific mechanisms underlying this effect remain unclear. AIM:To investigate the interventional role of hesperetin on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-induced exosomes in inducing gastric carcinogenesis. METHODS:Bioinformatics technology was used to identify the critical molecular components underlying hesperetin-mediated inhibition of MNNG induced gastric carcinogenesis through exosomal circular RNA. Biological experiments were conducted to validate these findings. RESULTS:Exosomes derived from TGES-1 cells (TGES-1-EX) significantly enhanced the proliferation, migration, invasion, epithelial-mesenchymal transition (EMT), and stemness of GES-1 cells. The oncogenic potential of TGES-1-EX was significantly diminished following hesperetin pretreatment. TGES-1-EX with overexpressed or knocked down circ0008274 was extracted and GES-1 cells were treated in combination with hesperetin or alone. Our investigation revealed that hesperetin exerted significant inhibitory effects on MNNG-induced gastric carcinogenesis by exosomal circ0008274. Bioinformatics prediction identified microRNA (miR)-526b-5p as a potential miRNA binding to circ0008274. Functional experiments demonstrated that hesperetin may mediate its intervention in MNNG-induced gastric cancer initiation by targeting miR-526b-5p through exosomal circ0008274. TGES-1-EX circ0008274 promoted the proliferation, EMT, and cancer stem cell-like characteristics in GES-1 cells through miR-526b-5p-mediated regulatory mechanisms. CONCLUSION:Hesperetin exerted an interventional effect on the gastric carcinogenesis process, particularly through the modulation of exosomal circ0008274 and its interaction with miR-526b-5p.

A delayed healing process in diabetic wounds is intractable. In this study, a high-glucose condition was found to be responsible for skin structure destruction, inflammatory infiltration, and vital cell dysfunction. Extracellular vesicles, particularly exosomes secreted by hucMSCs, contribute to improved diabetic wound healing, largely by promoting tissue repair and re-establishing normal function in affected cells. Small RNA-sequencing revealed that hucMSC-derived exosomes (hucMSC-Ex) were highly enriched in NC_000019.10_13474 (miR-13474), which was predicted to be an miRNA with an undiscovered function. miR-13474 showed a reduced expression level in high-glucose-treated skin cells as well as diabetic foot ulcer (DFU) rats. Moreover, there is also a significant expression difference between the wound area and the wound edge in DFU patients, indicating the potential clinical value of miR-13474. Blocking miR-13474 in hucMSC-Ex obviously diminished the therapeutic effects. Furthermore, exosomal miR-13474 was found to target the CPEB2/TWIST1 axis to improve the impaired function of skin cells. On this basis, hucMSC-Ex were used as a vehicle for the delivery of therapeutic miR-13474 to optimize the repairing effect. The study has revealed the role of hucMSC-derived exosomes and the underlying molecular mechanism in diabetic wound healing and proposes a cell-free-based modification strategy for refractory wound management.

Cells produce metabolic intermediates through catalytic reactions, mainly via post-translational modifications. The modification of proteins by O-linked N-acetylglucosamine, known as O-GlcNAcylation, is one of the most common post-translational modifications. As O-GlcNAcylation and phosphorylation can occur at serine or threonine residues, it is crucial that the interplay between these two modifications is vital to bioenergetic and biosynthetic demand. Although emerging recognition linking O-GlcNAc modification and phosphorylation to protein functions has been obtained, the issue of how altered O-GlcNAcylation or phosphorylation regulates each other in the metabolic system remains uncertain. The combination of cell biological and proteomic approaches over the recent few years has not only highlighted the interactions between O-GlcNAcylation and phosphorylation in protein function but also prompted us to elucidate the underlying mechanisms behind this crosstalk controlling metabolic homeostasis. The purpose of this review is to summarize recent advances in the O-GlcNAcylation/phosphorylation regulation of the metabolic process. An extensive exploration of this interplay has significant implications for metabolic control systems, including glucose, lipid, and nucleotide metabolism, where dysregulation in O-GlcNAcylation and phosphorylation of metabolic syndrome is essential.

Background Alcohol-associated liver disease (ALD) is a global health problem without an effective treatment. Mallory-Denk body (MDB) is a protein aggregate commonly found in alcohol-associated hepatitis (AH). MDB primarily contains ubiquitinated proteins, cytokeratin 8 and sequestosome 1 (SQSTM1)/p62. Stress granule (SG) is a cytosolic, membrane-less aggregate composed of various RNA-binding proteins and untranslated mRNA. However, the role and mechanisms of MDB and SG induced by alcohol and their implications in the pathogenesis of ALD remain largely unknown.Methods SQSTM1/p62 whole body knockout and matched wild-type mice were subjected to the Gao-binge alcohol model or fed a 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet alongside Gao-binge alcohol model. ALD mouse liver tissues and human AH liver tissues underwent immunohistochemistry (IHC) staining and western blot analysis for SG and MDB markers.Results We found that the livers of patients with AH had higher levels of SQSTM1/p62 (MDB marker) and Ras-GTPase-activating protein-binding protein 1 (an SG marker) using IHC staining, and these increased protein levels were enriched in detergent-insoluble fractions compared with healthy individuals. We further discovered that Gao-binge alcohol feeding increased insoluble SG markers, such as phosphorylated eukaryotic initiation factor 2 in mouse livers. Mice fed a DDC diet with Gao-binge alcohol had greater hepatic MDB formation and liver injury than those fed either diet alone. Loss of SQSTM1/p62 led to reduced protein aggregation involved in SGs and MDBs but increased liver injury in DDC plus Gao-binge alcohol-fed mice, indicating that SQSTM1/p62 is required for MDB formation and protects against alcohol-induced liver injury.Conclusion Chronic plus binge alcohol exposure increases hepatic MDBs and moderate levels of SGs. p62/SQSTM1 is critical for the formation but is not essential for the clearance of MDBs, a process that may act as an adaptive protective mechanism against ALD.