Journal of Nanobiotechnology最新文献

Characterizing the tumor immune microenvironment (TIME) to explore potential therapeutic targets is fundamental to advancing precision tumor immunotherapy. However, the immunosuppressive nature of "cold" tumors, notably prostate cancer, poses a significant barrier to immunotherapy, demanding new approaches to simultaneously reinvigorate anti-tumor immunity and modulate the molecular drivers of immune evasion. Here, we identified VSIG4 as a key regulator of prostate tumor-resident macrophage fate through single-cell sequencing analysis. Meanwhile, a shikonin (Shik)-mediated downregulation of VSIG4 in macrophages is verified, potentially attenuating its immunosuppressive effects. Building on these findings, cytosine guanine dinucleotide (CpG) oligodeoxynucleotide (ODN)-modified manganese (Mn)-Shik metal-polyphenol network nanodrugs (Mn/Shik@CpG NDs) are designed to reverse the "cold" immune environment of prostate tumor. In this scenario, Mn/Shik@CpG NDs release monomeric components under the stimulation of acidic and glutathione-rich tumor microenvironment (TME), thus exerting their immunomodulatory effects synergistically. Since the released Shik can induce DNA damage by necroptosis promoting reactive oxygen species production, cGAS-STING signaling pathway is initiated, which further activates interferon production in the TME. In addition, the necroptosis of Shik initiates immunogenic cell death, further activating innate immunity and promoting adaptive immune responses. Mn²⁺ is a cGAS-STING sensitizer, which amplifies the intratumoral interferon response. As an immune adjuvant, CpG ODN effectively promotes the maturation of dendritic cells, as well as the helper T cell differentiation and pro-inflammatory cytokine secretion, thus activating both innate and adaptive immunity. In vivo studies suggest that Shik-mediated VSIG4 downregulation, combined with innate and adaptive immune activation, remodels the TIME to evoke a significant anti-tumor response. Furthermore, transcriptomic analysis of rechallenged tumors indicated this durable protection was driven by a genuine immune memory response, revealing a gene signature of T cell activation and immune reprogramming. Collectively, beyond presenting a novel therapeutic candidate for converting immunologically "cold" tumors into "hot" ones, our work validates a data-guided design pipeline, offering a conceptual blueprint to inform the precise engineering of future nanodrugs.

Background: Inducible nitric oxide synthase (iNOS) is a key driver of aberrant angiogenesis in inflammatory conditions and cancer, making it an attractive therapeutic target. Nevertheless, its function can be affected by the complex immune responses and tumor microenvironment (TME). Hence, combinatorial treatment approaches that simultaneously target iNOS and immune-modulatory signaling are strongly recommended for cancer therapy. Moreover, the current iNOS inhibitors are limited by poor pharmacokinetics and a lack of selectivity.

Results: To address these challenges, we developed a glutathione (GSH)-responsive iNOS-inhibiting polymeric prodrug (GRIP) decorated with betamethasone succinate (NP_BeS). These dual-function nanoparticles (NP_BeS) remain stable under physiological conditions but selectively release their payload in response to elevated GSH levels, a hallmark of the TME. Only upon activation by GSH, NP_BeS inhibits iNOS, as evidenced by suppressed lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 macrophages. NP_BeS also normalized vascular endothelial growth factor (VEGF)-mediated tube formation in HUVECs and 3T3-L1 fibroblast cell migration, and angiogenesis in the CAM assay, demonstrating its anti-angiogenic activity. Importantly, GRIP did not impair acetylcholine (ACh)-induced vasodilation in rat aorta, even at elevated concentrations, indicating preservation of eNOS function.

Conclusions: This is the first report of a GSH-responsive polymeric prodrug system that leverages intracellular GSH for both controlled release of anionic therapeutic agents and in situ synthesis of an iNOS antagonist. Through these two complementary pathways, the system enables targeted, sustained anti-angiogenic effects and promotes vascular normalization. This dual-function platform holds strong potential for the treatment of cancer-associated angiogenesis.

Traumatic brain injury (TBI)-induced neuroinflammation, driven by inflammatory microglial polarization, continues to pose a significant regenerative and clinical challenge. Small extracellular vesicles (sEVs) have demonstrated great potential in mitigating post-TBI inflammation. Nevertheless, the limited yield and efficacy of sEVs produced via conventional two-dimensional (2D) culture systems (2D-sEVs) substantially hinder their clinical applicability. Moreover, effective strategies for the therapeutic application of sEVs in TBI treatment, along with an understanding of their underlying mechanisms, remain largely unexplored. In this study, we employed a 3D coaxial bioprinting method to encapsulate adipose-derived stem cells (ADSCs) within a hydrogel microfiber, facilitating 3D culturing and large-scale production of 3D-sEVs. Additionally, we utilized GelMA hydrogel for the sustained release of 3D-sEVs and evaluated their effects in LPS-activated microglia as well as in a TBI mouse model. Our results demonstrated that 3D culture significantly enhanced sEV production. GelMA improved sEV stability and prolonged sEV release up to 30 days in vivo. Compared to 2D-sEVs, 3D-sEVs offered superior therapeutic benefits. Specifically, 3D-sEVs substantially reduced neuroinflammation and brain tissue loss while accelerating motor function recovery in TBI mice. Furthermore, 3D-sEVs shifted pro-inflammatory microglia toward an anti-inflammatory polarization state, as evidenced by elevated expression levels of IL-4, IL-10, TGF-β, Arg1, and CD206, alongside reduced expression of IL-6, IL-1β, TNF-α, iNOS, and CD86, both in vitro and in vivo. Additionally, 3D-sEVs attenuated chemotaxis and migration in LPS-activated microglia. Further mechanistic exploration through RNA-seq, proteomic profiling, and GAS6 knockdown in 3D-sEVs, revealed that 3D-sEVs deliver growth arrest-specific protein 6 (GAS6) to modulate the transition of microglia from a pro-inflammatory to an anti-inflammatory state, thereby mitigating neuroinflammation following TBI. Our findings underscore the therapeutic promise of sEVs derived from 3D-cultured ADSCs in treating TBI via modulating microglia polarization.

Cancer continues to be the second leading cause of death globally. Although medical technologies have advanced significantly, cancer treatment still faces major challenges, such as drug resistance and dynamic alterations in tumor microenvironment (TME). Plant-derived exosomes (PDEs) are promising to serve as next-generation anticancer agents to address these issues. In this review, we highlight interdisciplinary progress in the development of PDEs as delivery systems for cancer therapy, focusing on three key advantages: (1) abundant sources and bioactive compounds: PDEs can be extracted from a wide range of plant sources using various methods, providing low immunogenicity and retaining natural pharmacological activity through preserved bioactive compounds. (2) multiple anti-tumor mechanisms: PDEs exert anticancer effects through direct tumor cell killing, modulation of the TME, and metabolic reprogramming. Their ability to engage multiple pathways may help delay or overcome drug resistance. (3) broad applications: due to their strong anti-tumor efficacy and excellent biocompatibility, PDEs have shown great potential in diverse therapeutic contexts. By summarizing cutting-edge research in PDEs, we also propose future directions for optimizing PDE-based delivery systems for clinical applications.

Endothelial cells (ECs) of endothelial-to-mesenchymal transition (EndMT) are drivers of cardiac fibrosis. BRD4 has recently been identified as an epigenetic regulator of EndMT. Proteolysis-targeting chimera (PROTAC) technology has revolutionized targeted protein degradation, offering unprecedented opportunities for BRD4 modulation in diverse pathological contexts. Nevertheless, the non-selective cellular targeting profile of PROTACs poses significant limitations for their therapeutic application in cardiac fibrosis management. To address these limitations, we developed a GSH-responsive nanoscale PROTAC (RGD-PEG-MZ1) that targets activated platelets, leveraging their chemotactic properties to precisely degrade BRD4 in ECs. RGD-PEG-MZ1 exhibits selectivity for ECs and inhibition of EndMT, which can prevent the progression of cardiac fibrosis. The RNA-seq analysis revealed an attenuation of the MAPK signaling pathway following RGD-PEG-MZ1 treatment. The interaction between BRD4 and the MAPK signaling was analyzed through AlphaFold3 and immunoprecipitation assays. The experimental data showed that BRD4 directly interacts with RAF1, a critical effector in MAPK signaling, which suggested that RGD-PEG-MZ1 modulates MAPK signaling by disrupting the BRD4-RAF1 interaction. This innovative GSH-activated PROTAC strategy not only offers a novel therapeutic approach for cardiac fibrosis but also provides insights into the functional role of BRD4 in the disease pathogenesis of cardiac fibrosis.

Solid tumors pose a spatial "delivery-at-depth" bottleneck: therapeutics that reach tumors often remain sequestered near vessels and fail to distribute uniformly into tumor cores. This limitation arises from heterogeneous perfusion, elevated interstitial fluid pressure, and dense extracellular matrix, which together restrict convection-diffusion balance and amplify binding-site barriers. We organize transformable and bioinspired nanomedicines using a barrier-centric lens and summarize five strategy families to deepen and homogenize intratumoral transport: (i) stimuli-responsive size/charge switching, (ii) microenvironment remodeling to restore perfusion and decompress stroma, (iii) ligand-guided transcytosis and CendR pathway engagement, (iv) cell-based and biomimetic vectors leveraging homing and immune evasion, and (v) multistage designs that sequence priming, switching, and payload activation. We compare representative systems by trigger specificity, activation timing, affinity tuning, and corona susceptibility, and highlight recurring failure modes including stimulus heterogeneity, premature/off-target activation, and escalating chemistry-manufacturing-controls burdens with added components. We conclude with translational priorities: couple barrier priming with a single well-characterized switching event, favor moderated or activatable affinity to avoid perivascular trapping, and validate spatial gains using standardized intratumoral distribution metrics linked to therapeutic endpoints.

Multidrug-resistant bacterial infections pose a severe threat to sepsis, where uncontrolled bacterial proliferation and the accompanying inflammatory response lead to multiorgan dysfunction. Recent clinical isolation of a super-resistant Enterobacter asburiae (E. asburiae) strain co-harboring mcr-10 and bla_NDM-1 underscores the urgent need for more effective antimicrobial strategies. Herein, we develop a synergistic strategy for the chemodynamic and photothermal therapy of super-resistant E. asburiae using a multifunctional mesoporous nanocomposite. This nanocomposite is composed of mesoporous silica nanoparticle loaded with polymyxin B, polydopamine, and palladium nanoparticles, possessing properties including photothermal therapy, chemodynamic therapy, and drug cargo. This synergistic approach achieves bactericidal effects through localized hyperthermia, enhanced generation of reactive oxygen species, and membrane-disrupting antibiotic action at a low dose of 42.1 µg mL^-1. In a murine peritonitis-sepsis model, treatment with this multifunctional mesoporous nanocomposite reduces the bacterial burden by 100%, decreases serum levels of inflammatory cytokines to normal levels, and improves the 3-day survival rate to 100%. These findings highlight a promising therapeutic strategy that leverages nanomaterial-mediated synergy to combat super-resistant pathogens.

Background: Acute kidney injury (AKI) remains a major clinical challenge resulting from the intertwined processes of oxidative stress and macrophage-driven inflammation, both converging on mitochondrial dysfunction. We developed a glucosylated albumin nanoplatform (Glc⁶-AD¹¹-Alb) designed to exploit glucose transporter 1-mediated uptake in inflammatory M1 macrophages while preserving the intrinsic antioxidant properties of albumin.

Results: Physicochemical characterization confirmed reproducible synthesis with defined degrees of functionalization and stable physicochemical properties. In vitro, Glc⁶-AD¹¹-Alb demonstrated selective uptake in M1 macrophages and significantly reduced intracellular reactive oxygen species, validating its dual role in immune modulation and redox regulation. Positron emission tomography imaging with Cu-64 radiolabeling revealed preferential renal accumulation in ischemia-reperfusion injury (IRI) models, supporting macrophage-targeted delivery. In vivo, administration of Glc⁶-AD¹¹-Alb attenuated renal dysfunction, suppressed pro-inflammatory and oxidative markers, and promoted tubular regeneration in pre- and post-treatment settings. Importantly, Glc⁶-AD¹¹-Alb directly protected renal tubular epithelial cells by restoring mitochondrial membrane potential under oxidative and hypoxic stress. Seahorse metabolic flux analysis further confirmed enhanced oxidative phosphorylation, reduced glycolytic dependency, and improved coupling efficiency, indicating recovery of mitochondrial bioenergetics. Transmission electron microscopy demonstrated preservation of mitochondrial ultrastructure, including intact cristae and elongated morphology, consistent with improved ATP synthesis capacity.

Conclusions: Glc⁶-AD¹¹-Alb acts through complementary mechanisms of macrophage-targeted immune modulation and mitochondrial protection, thereby disrupting the vicious cycle of inflammation and oxidative stress in AKI. This nanoplatform represents a clinically translatable therapeutic strategy with potential to improve outcomes in patients with ischemic kidney injury.

PANoptosis, a distinct type of inflammation-associated programmed cell death that integrates features of apoptosis, necroptosis and pyroptosis, is closely related to the pathogenesis and progression of articular cartilage degeneration in osteoarthritis (OA). Meanwhile, excessive reactive oxygen species (ROS), pro-inflammatory pathways, and inflammatory cytokines in the OA microenvironment mutually reinforce one another, exacerbating synovial inflammation. To simultaneously target these interconnected pathological drivers, we developed an injectable nanocomposite thermosensitive hydrogel (DHA-PRO@NMs@TSH). This hierarchical system was constructed by integrating a ROS-responsive dihydroartemisinin prodrug (DHA-PRO) into Soluplus/TPGS-based nanomicelles (NMs), which were subsequently embedded within a Poloxamer-based thermosensitive hydrogel (TSH). This hierarchical and smart drug delivery system enables stable dihydroartemisinin (DHA) delivery and demonstrates a graded, ROS-triggered release profile, achieving sustained DHA retention in OA joints. The formulated system demonstrated excellent injectability, thermosensitivity, and physicomechanical stability, achieving sustained drug retention in rat joints for over 7 days, as confirmed by in vivo imaging. In vitro studies demonstrated that DHA-PRO@NMs significantly suppressed PANoptosis in chondrocytes by downregulating key markers (e.g., Bax/Bcl-2, RIPK3/p-RIPK3, NLRP3/Caspase-1/GSDMD) and inhibited inflammation by blocking the NF-κB pathway and subsequent cytokine production (e.g., TNF-α, IL-1β). In a rat OA model, treatment with DHA-PRO@NMs@TSH robustly attenuated disease progression, as evidenced by near-complete restoration of gait function, marked inhibition of cartilage degradation, a remarkable nearly 90% reduction in the Mankin score, and a comprehensive suppression of PANoptosis and inflammation pathological markers. This work not only presents a promising translational strategy for OA but also pioneers a therapeutic paradigm of leveraging advanced drug delivery system to dual-targeting PANoptosis and inflammation.

The functional exhaustion of CD8⁺ T cells in the tumor microenvironment (TME) severely limits anti-tumor immunity in gastric cardia adenocarcinoma (GCA). Here, we developed CD8a antibody-functionalized biomimetic red blood cell membrane ectosomes (CD8a-NVEs) encapsulating the p300 inhibitor C646 to selectively target and reprogram exhausted CD8⁺ T cells. Single-cell RNA sequencing of human GCA tissues revealed lactate-driven epigenetic remodeling, characterized by elevated H3K18 lactylation (H3K18la) at the PDCD1 promoter, which correlated with impaired CD8⁺ T cell function. In vitro, C646 effectively reduced H3K18la, suppressed PDCD1 transcription, and restored effector molecule expression, including IFN-γ and GZMB. CD8a-NVEs@C646 exhibited superior targeting specificity, biocompatibility, and functional efficacy, markedly enhancing CD8⁺ T cell proliferation and cytotoxicity compared with free C646. In a humanized orthotopic GCA model, CD8a-NVEs@C646 significantly inhibited tumor growth, and its combination with anti-PD-1 therapy further enhanced T cell infiltration and tumor apoptosis. This biomimetic nanoplatform enables precise epigenetic reprogramming of tumor-infiltrating CD8⁺ T cells, overcoming lactate-induced histone modifications and reversing exhaustion. Collectively, these findings present a translational nanobiotechnology-based strategy to potentiate immunotherapy efficacy in GCA and potentially other malignancies driven by T cell dysfunction.