Metabolomics offers a comprehensive and functional snapshot of physiological and pathological states, and its advancement critically depends on high-performance analytical technologies. Laser desorption/ionization mass spectrometry (LDI MS) has attracted increasing attention in metabolomic analysis owing to its minimal sample preparation, rapid analysis speed, and high compatibility with high-throughput workflows. Nevertheless, the analytical reliability of LDI MS is fundamentally dictated by matrix performance. Traditional organic matrices suffer from intrinsic drawbacks in small-molecule analysis, including intense low-mass background interference, inhomogeneous crystallization, and limited reproducibility in complex biological samples. In recent years, inorganic nanomaterials have emerged as a promising class of alternative LDI MS matrices. Benefiting from their tunable composition and structure, inorganic nanomatrices exhibit excellent laser energy absorption, efficient photothermal or plasmonic energy conversion, enhanced charge/electron transfer capability, and superior thermal and chemical stability, thereby enhancing sensitive and selective detection of low-abundance metabolites. This review systematically summarizes the main categories of inorganic nanomatrices, including noble metal nanomaterials, carbon-based nanomaterials, silicon-based nanomaterials, metal oxide, and quantum dots with a particular focus on the relationships between material structure, physicochemical properties, and LDI MS performance. Furthermore, the representative applications of inorganic nanomatrix-assisted LDI MS in metabolomics are discussed, covering targeted metabolic analysis, untargeted metabolic fingerprinting for disease diagnosis, and mass spectrometry imaging. Finally, current challenges and further perspectives are critically evaluated, including matrix standardization, mechanistic understanding of desorption/ionization processes, and the translation of inorganic nanomatrix-based LDI MS platforms toward clinical and precision medicine applications. This review aims to provide a systematic framework to guide material design, methodological optimization, and clinical applications of inorganic nanomatrix-assisted LDI MS, thereby promoting its development in precision medicine and translational research.

The enhanced permeability and retention (EPR) effect, a cornerstone of cancer nanomedicine, has been predominantly interpreted at the macroscopic level, offering limited insight into the microscopic dynamics governing nanocarrier delivery. To address this gap, we directly compared the intratumoral particokinetics of liposomal carriers with that of their drug payload (doxorubicin) through precise tracing and quantitative analysis. Using a second near-infrared (NIR-II) aggregation-caused quenching (ACQ) probe, ACQ5, we achieved accurate in vivo tracking of doxorubicin-loaded liposomes. This approach minimizes artifacts from free labels and ensures superior fluorescence linearity. Our results show substantial sequestration of liposomes by the mononuclear phagocyte system (liver: 16.3% ID/g at 8 h; spleen: 70.5% ID/g at 36 h), with only modest tumor accumulation (∼4% ID/g after 24 h). Importantly, by noninvasively differentiating intravascular from extravascular signals, we established that microdistribution serves as a more meaningful indicator of permeation efficiency than total tumor accumulation. Moreover, we revealed a critical spatial dissociation: although liposomes progressively penetrated the tumor interstitium, doxorubicin readily dissociated and was largely retained in perivascular regions. This study challenges the classical macro-level EPR concept and underscores the necessity of a microscopic reassessment of intratumoral particokinetics to guide the development of next-generation nanocarriers with improved therapeutic efficacy.

Sepsis remains a major challenge in global clinical practice due to its persistently high mortality rate. Traditional antibiotics address symptoms rather than root causes, while the problem of drug resistance continues to escalate. Detoxification strategies such as monoclonal antibodies have shown limited efficacy in complex clinical settings, as their single-target specificity and narrow activity spectrum restrict their ability to neutralize diverse bacterial toxins. The therapeutic bottleneck lies in the absence of effective methods capable of simultaneously neutralizing Gram-negative bacterial endotoxins (such as LPS) and Gram-positive bacterial exotoxins (such as Hlα). To overcome this limitation, we drew inspiration from the natural "bait" mechanisms of living organisms to develop a biomimetic nanoscale detoxifier with broad-spectrum toxin adsorption and neutralization capacity. Specifically, chloroquine was used to inhibit autophagy lysosome formation in macrophages, promoting the release of exosomes enriched with toxin receptors CD14 and ADAM10. These exosomes (Exo) were then fused with artificial liposomes (Lip) possessing extensive membrane space to construct exosome membrane hybrid liposomes (Elip). Our results demonstrated that Elip effectively neutralized the hemolytic activity of Hlα and adsorbed LPS in vitro. In a subcutaneous HIα-induced local inflammation model, Elip completely prevented local skin and muscle tissue necrosis. In a systemic inflammation model induced by intravenous HIα, Elip significantly alleviated acute inflammatory damage in the lungs and liver, reducing key pro-inflammatory factor levels to near-normal ranges. In an LPS-induced shock model, all mice in the Elip treatment group survived. This study robustly confirmed that Elip successfully resolves the clinical challenge of simultaneously clearing endotoxins and exotoxins through a "synergistic detoxification" mechanism, offering a novel therapeutic strategy with significant translational potential that transcends traditional antibiotics for conquering sepsis.

Protein complexes are central to cellular function and respond rapidly to pharmacological perturbations. Co-fractionation mass spectrometry (CoFrac-MS) is widely employed to analyze protein complexes by analyzing individual chromatographic fractions, but it is labor-intensive and slow. To address these challenges, we introduce a chromatography-guided strategy enabling rapid identification of drug-perturbed protein complexes. It combines cross-linking enhanced reversed phase liquid chromatography cofractionation (XL-CoFrac) for high-resolution separation with ChromaQuant, a custom tool for precise peak quantification and differential analysis (https://hplcfdu.shinyapps.io/ChromaQuant/). Subsequent targeted MS analyses, guided by ChromaQuant, collectively establish the XL-CoFrac-Q-MS workflow. In proof-of-concept studies, we first adopted XL-CoFrac to MCF7 cells and profiled representative protein complexes. ChromaQuant demonstrated exceptional precision, achieving coefficients of variation below 1% and replicate correlations exceeding 0.98. Furthermore, we analyzed RS4;11 leukemia cells treated with increasing concentrations of the BCL-2 inhibitor ABT-199 using the XL-CoFrac-Q-MS workflow. Seven chromatographic peaks that changed consistently with the drug concentration were selected to be identified by this approach. MS analysis of these peaks revealed cross-linked peptides from the BCL-2 associated protein complex. Specially, cross-linking peptides between BCL-2 and FKBP38 may shed light on the mechanisms underlying resistance to ABT-199. Further pathway enrichment analysis provides new insights into the molecular mechanisms driving ABT-199 induced apoptosis. Collectively, the XL-CoFrac-Q-MS strategy holds significant potential for broad applications, including rapid screening of drug targets and elucidation of protein complex dynamics across various pharmacological and pathological conditions.

Nanovaccines hold significant promise for the prevention and treatment of infectious diseases. However, the efficacy of many nanovaccines is often limited by inadequate stimulation of both innate and adaptive immune responses. Herein, we explore a rational vaccine strategy aimed at modulating innate cell microenvironments within lymph nodes (LNs) to enhance the generation of effective immune responses. Inspired by the structure and natural infection process of viruses, we developed a versatile antigen and adjuvant co-delivery platform, termed virus-mimetic vaccines (VMVs). Specifically, polyarginine-tagged antigens were noncovalently assembled onto nucleic acid nanogels containing cytosine-phosphate-guanine oligodeoxynucleotide via a salt-bridge zipper mechanism, which can activate Toll-like receptor 9. Upon intramuscular immunization, VMVs effectively drained into the LNs, recruiting and activating multiple innate cells, including CD8+ dendritic cells (DCs), CD103+ DCs, macrophages, plasmacytoid DCs, and neutrophils. This activation modulates the innate cell microenvironments and relocates antigen-presenting cells within LNs, optimizing adaptive immune responses. VMVs induced a robust antigen-specific immune response, characterized by high levels of neutralizing antibodies, augmented memory T cell activity, and enhanced development of germinal center B cells. Together, our findings demonstrate that dynamic modulation of innate cell microenvironments by VMVs leads to optimized generation of both humoral and cellular immunity against infectious diseases.

Probiotics play a crucial role in colon cancer treatment by metabolizing prebiotics to generate short-chain fatty acids (SCFAs). Colon cancer patients are frequently propositioned to supplement with probiotics to enhance the conversion and utilization of prebiotics. Nevertheless, the delivery and colonization of probiotics is hindered by the harsh conditions of gastrointestinal tract (GIT). Here, we devised a straightforward yet potent modified prebiotic-based “shield” (Gelatin-Inulin, GI), employing dietary inulin and natural polymer gelatin crosslinked via hydrogen bonding for enveloping Lactobacillus reuteri (Lr) to formulate synbiotic hydrogel capsules (Lr@Gl). The GI “shield” serves as a dynamic barrier, augmenting the resistance of Lr to gastric acid and facilitating its bioactivity and adherence in the GIT, synergizing with Lr to elicit an anti-tumor effect. Simultaneously, Lr@GI demonstrates anti-tumor effects by depleting glutathione to release reactive oxygen species, accompanied by the activation of NLRP3 (NOD-like receptor family pyrin domain containing 3), and the induction M1 macrophage polarization. Furthermore, Lr@GI can not only promote the recovery of intestinal barrier but also regulate intestinal flora, promoting the production of SCFAs and further exerting anti-tumor effect. Crucially, Lr@GI also potentiates the anti-tumor effect of 5-Fluorouracil. The construction and synergistic anti-tumor mechanism of synbiotic hydrogel capsules system provide valuable insights for gut microbial tumor therapy.

The high heterogeneity of basal-like breast cancer (BLBC) and the absence of effective therapeutic targets pose ongoing treatment challenges. Pyroptosis, a type of cell death characterized by cell swelling and membrane perforation, offers a promising therapeutic weathervane for BLBC, particularly when induced by physical therapies. In this study, a high-intensity focused ultrasound (HIFU)-driven targeted pyroptosis strategy is developed for BLBC therapy. Through integration of HIFU-driven gene regulation analysis and bioinformatics analysis of pyroptosis-related genes from the TCGA dataset, 20 potential pyroptosis inducers are identified to work synergistically with HIFU. Mitoxantrone, a promising inducer, is encapsulated in platelet membrane-hybridized liposomes to enhance targeted delivery and therapeutic efficacy. Importantly, the combination of HIFU and Plp enhanced tumor delivery of liposomes by 2.46 fold and dramatically inhibited tumor growth, with 50% of female BALB/c mice remaining tumor-free compared to liposome-only treatment. Mechanistically, HIFU significantly downregulated the expression of histone deacetylases 4 and 9, while promoting cathepsin-L (CTSL) gene transcription. Simultaneously, Plp and HIFU synergistically suppressed BCL-2 via CTSL, increasing ROS production. This activated Caspase8 and the NLRP3 inflammasome, leading to GSDMC cleavage and initiating pyroptosis. Collectively, this study provides an innovative pyroptosis therapy strategy combining physical treatment and chemotherapy for BLBC and other refractory diseases.

Efficient protection and precise delivery of biomolecules are of critical importance in the intervention and therapy of various diseases. Although diverse specific marker-functionalized drug carriers have been developed rapidly, current approaches still encounter substantial challenges, including strong immunogenicity, limited target availability, and potential side effects. Herein, we developed a biomimetic exosome-sheathed magnetic mesoporous anchor modified with glucose oxidase (MNPs@mSiO2-GOx@EM) to address these challenges and achieve synergistic targeting and starving of tumor cells. The MNPs@mSiO2-GOx@EM anchor integrated the unique characteristics of different components. An external decoration of exosome membrane (EM) with high biocompatibility contributed to increased phagocytosis prevention, prolonged circulation, and enhanced recognition and cellular uptake of loaded particles. An internal coated magnetic mesoporous core with rapid responsiveness by the magnetic field guidance and large surface area facilitated the enrichment of nanoparticles at the specific site and provided enough space for modification of glucose oxidase (GOx). The inclusion of GOx in the middle layer accelerated the energy-depletion process within cells, ultimately leading to the starvation and death of target cells with minimal side effects. With these merits, in vitro study manifested that our nanoplatform not only demonstrated an excellent targeting capability of 94.37% ± 1.3% toward homotypic cells but also revealed a remarkably high catalytical ability and cytotoxicity on tumor cells. Assisted by the magnetic guidance, the utilization of our anchor obviously inhibits the tumor growth in vivo. Together, our study is promising to serve as a versatile method for the highly efficient delivery of various target biomolecules to intended locations due to the fungibility of exosome membranes and provide a potential route for the recognition and starvation of tumor cells.

BACKGROUND:Colorectal cancer (CRC) incidence is increasing in recent years due to intestinal flora imbalance, making oral probiotics a hotspot for research. However, numerous studies related to intestinal flora regulation ignore its internal mechanisms without in-depth research.RESULTS:Here, we developed a probiotic microgel delivery system (L.r@(SA-CS)2) through the layer-by-layer encapsulation technology of alginate (SA) and chitosan (CS) to improve gut microbiota dysbiosis and enhance anti-tumor therapeutic effect. Short chain fatty acids (SCFAs) produced by L.r have direct anti-tumor effects. Additionally, it reduces harmful bacteria such as Proteobacteria and Fusobacteriota, and through bacteria mutualophy increases beneficial bacteria such as Bacteroidota and Firmicutes which produce butyric acid. By binding to the G protein-coupled receptor 109A (GPR109A) on the surface of colonic epithelial cells, butyric acid can induce apoptosis in abnormal cells. Due to the low expression of GPR109A in colon cancer cells, MK-6892 (MK) can be used to stimulate GPR109A. With increased production of butyrate, activated GPR109A is able to bind more butyrate, which further promotes apoptosis of cancer cells and triggers an antitumor response.CONCLUSION:It appears that the oral administration of L.r@(SA-CS)2 microgels may provide a treatment option for CRC by modifying the gut microbiota.

Due to low success rates and long cycles of traditional drug development, the clinical tendency is to apply omics techniques to reveal patient-level disease characteristics and individualized responses to treatment. However, the heterogeneous form of data and uneven distribution of targets make drug discovery and precision medicine a non-trivial task. This study takes pyroptosis therapy for triple-negative breast cancer (TNBC) as a paradigm and uses data mining of a large TNBC cohort and drug databases to establish a biofactor-regulated neural network for rapidly screening and optimizing compound pyroptosis drug pairs. Subsequently, biomimetic nanococrystals are prepared using the preferred combination of mitoxantrone and gambogic acid for rational drug delivery. The unique mechanism of obtained nanococrystals regulating pyroptosis genes through ribosomal stress and triggering pyroptosis cascade immune effects are revealed in TNBC models. In this work, a target omics-based intelligent compound drug discovery framework explores an innovative drug development paradigm, which repurposes existing drugs and enables precise treatment of refractory diseases.

Transforming growth factor β (TGF-β), a versatile immunosuppressive cytokine, has gained increasing attention as a potential target for cancer immunotherapy. However, current strategies are constrained by tumor heterogeneity and drug resistance. Therapeutic probiotics, such as Escherichia coli Nissle1917 (EcN), not only regulate the gut microbiota to increase beneficial bacteria with anti-tumor effects, but also modulate immune factors within the body, thereby enhancing immunity. In this study, we developed an oral microgel delivery system of EcN@(CS-SA)2 by electrostatic interaction between chitosan (CS) and sodium alginate (SA), aiming to enhance its bioavailability in the gastrointestinal tract (GIT). Notably, EcN@(CS-SA)2 microgel showed a synergistic enhancement of the anti-tumor efficacy of Galunisertib (Gal, a TGF-β inhibitor) by inducing apoptosis and immunogenic cell death (ICD) in tumor cells, as well as promoting increased infiltration of CD8+ T cells into the tumor microenvironment (TME).

Surgical removal together with chemotherapy and radiotherapy has used to be the pillars of cancer treatment. Although these traditional methods are still considered as the first-line or standard treatments, non-operative situation, systemic toxicity or resistance severely weakened the therapeutic effect. More recently, synthetic biological nanocarriers elicited substantial interest and exhibited promising potential for combating cancer. In particular, bacteria and their derivatives are omnipotent to realize intrinsic tumor targeting and inhibit tumor growth with anti-cancer agents secreted and immune response. They are frequently employed in synergistic bacteria-mediated anticancer treatments to strengthen the effectiveness of anti-cancer treatment. In this review, we elaborate on the development, mechanism and advantage of bacterial therapy against cancer and then systematically introduce the bacteria-based nanoprobes against cancer and the recent achievements in synergistic treatment strategies and clinical trials. We also discuss the advantages as well as the limitations of these bacteria-based nanoprobes, especially the questions that hinder their application in human, exhibiting this novel anti-cancer endeavor comprehensively.

The scarcity and dynamic nature of phosphotyrosine (pTyr)-modified proteins pose a challenge for researching protein complexes with pTyr modification, which are assembled through multiple protein-protein interactions. We developed an integrated complex-centric platform for large-scale quantitative profiling of pTyr signaling complexes based on cofractionation/mass spectrometry (CoFrac-MS) and a complex-centric algorithm. We initially constructed a trifunctional probe based on pTyr superbinder (SH2-S) for specifically binding and isolation of intact pTyr protein complexes. Then, the CoFrac-MS strategy was employed for the identification of pTyr protein complexes by integrating ion exchange chromatography in conjunction with data independent acquisition mass spectrometry. Furthermore, we developed a novel complex-centric algorithm for quantifying protein complexes based on the protein complex elution curve. Utilizing this algorithm, we effectively quantified 216 putative protein complexes. We further screened 21 regulated pTyr protein complexes related to the epidermal growth factor signal. Our study engenders a comprehensive framework for the intricate examination of pTyr protein complexes and presents, for the foremost occasion, a quantitative landscape delineating the composition of pTyr protein complexes in HeLa cells.

As an essential branch of targeted drug delivery, oral targeted delivery is attracting growing attention in recent years. In addition to site-specific delivery for the treatment of locoregional diseases in the gastrointestinal tract (GIT), oral targeted delivery to remote sites beyond the GIT emerges as a cutting-edge research topic. This review aims to provide an overview of the fundamental concepts and most recent advances in this field. Owing to the physiological barriers existing in the GIT, carrier systems should be transported across the enteric epithelia to target remote sites. Recently, pioneer investigations have validated the transport of intact micro- or nanocarriers across gastrointestinal barriers and subsequently to various distal organs and tissues. The microfold (M) cell pathway is the leading mechanism underlying the oral absorption of particulates, but the contribution of the transcellular and paracellular pathways should not be neglected either. In addition to well-acknowledged physicochemical and biological factors, the formation of a protein corona may also influence the biological fate of carrier systems. Although in an early stage of conceptualization, oral targeted delivery to remote diseases has demonstrated promising potential for the treatment of inflammation, tumors, and diseases inflicting the lymphatic and mononuclear phagocytosis systems.

Bacteria-mediated antitumor therapy has gained widespread attention for its innate tumor-targeting capability and excellent immune activation properties. Nevertheless, the clinical approval of bacterial therapies remains elusive primarily due to the formidable challenge of balancing safety with enhancing in vivo efficacy. In this study, leveraging the probiotic Escherichia coli Nissle1917 (EcN) emerges as a promising approach for colon cancer therapy, offering a high level of safety attributed to its lack of virulence factors and its tumor-targeting potential owing to its obligate anaerobic nature. Specifically, we delineate the erythrocyte (RBC) membrane-camouflaged EcN, termed as Trojan horse EcN@RBC, which triggers apoptosis in tumor cells by mitigating mitochondrial membrane potential (MMP) and subsequently activating the PINK1/Parkin pathway associated with mitophagy. Concurrently, the decline in MMP induced by mitophagy disrupts the mitochondrial permeability transition pore (MPTP), leading to the release of Cytochrome C and subsequent apoptosis induction. Moreover, synergistic effects were observed through the combination of the autophagy activator rapamycin, bolstering the antitumor efficacy in vivo. These findings offer novel insights into probiotic-mediated antitumor mechanisms and underscore the therapeutic potential of EcN@RBC for colon cancer patients.

Targeted therapy and immunotherapy have brought hopes for precision cancer treatment. However, complex physiological barriers and tumor immunosuppression result in poor efficacy, side effects, and resistance to antitumor therapies. Bacteria-mediated antitumor therapy provides new options to address these challenges. Thanks to their special characteristics, bacteria have excellent ability to destroy tumor cells from the inside and induce innate and adaptive antitumor immune responses. Furthermore, bacterial components, including bacterial vesicles, spores, toxins, metabolites, and other active substances, similarly inherit their unique targeting properties and antitumor capabilities. Bacteria and their accessory products can even be reprogrammed to produce and deliver antitumor agents according to clinical needs. This review first discusses the role of different bacteria in the development of tumorigenesis and the latest advances in bacteria-based delivery platforms and the existing obstacles for application. Moreover, the prospect and challenges of clinical transformation of engineered bacteria are also summarized.

The facultative intracellular bacterium Listeria monocytogenes (Lmo) has great potential for development as a cancer vaccine platform given its properties. However, the clinical application of Lmo has been severely restricted due to its rapid clearance, compromised immune response in tumors, and inevitable side effects such as severe systemic inflammation after intravenous administration. Herein, an immunotherapy system was developed on the basis of natural red blood cell (RBC) membranes encapsulated Lmo with selective deletion of virulence factors (Lmo@RBC). The biomimetic Lmo@RBC not only generated a low systemic inflammatory response but also enhanced the accumulation in tumors due to the long blood circulation and tumor hypoxic microenvironment favoring anaerobic Lmo colonization. After genome screening of tumors treated with intravenous PBS, Lmo, or Lmo@RBC, it was first found that Lmo@RBC induced extensive pore-forming protein gasdermin C (GSDMC)-dependent pyroptosis, which reversed immunosuppressive tumor microenvironment and promoted a systemic strong and durable anti-tumor immune response, resulting in an excellent therapeutic effect on solid tumors and tumor metastasis. Overall, Lmo@RBC, as an intravenous living bacterial therapy for the selective initiation of tumor pyrolysis, provided a proof-of-concept of live bacteria vaccine potentiating tumor immune therapy.

Although low-temperature photothermal therapy (PTT) can sensitize tumors to immune checkpoint inhibition, its efficacy is still restricted in the deep and internal tumors without enough oxygen and lymphocytic infiltration. Non-oxygen-dependent alkyl radicals have been demonstrated to synergistically enhance PTT through up-regulating lipid peroxidation and reactive oxygen species (ROS). Herein, an innovative strategy based on alkyl radicals to augment immunogenetic cell death (ICD) caused by mild PTT was proposed to improve poor efficacy of immunotherapy, which composed of a photothermal material of Chinse ink, an azo-initiator of 2,2-azobis[2-(2-imidazoline-2-acyl)propane]dihydrochloride (AIPH) and a PD-L1 inhibitor of HY19991 (HY). Upon near-infrared-II laser irradiation, low-temperature (<45℃) stimulation induced a high expression of immune checkpoint receptor (PD-L1) in tumors and triggered a large amount alkyl radicals generated by AIPH. Significantly, the alkyl radicals augmented the ICD and increased the recruitment of tumor-infiltrating lymphocytes against tumors after transformation of the immunologically cold tumor microenvironment into hot by mild PTT. The released HY further enhanced the immunotherapy effect by blocking the binding of activated T lymphocytes and PD-L1. In vivo studies exhibited that the all-in-one hydrogel with synergistic mechanisms had an extraordinary ability to reverse the immunosuppressive microenvironment, stimulate innate and adaptive immune responses to eliminate tumors and prevent metastasis.