Lipid nanoparticle used in mRNA vaccine promotes tumor metastasis in mouse model via mtDNA-induced neutrophil activation and NETosis

Effect of TU-100 on Colorectal Liver Metastasis in Mouse Model of MASH.

Substituted 1-(benzo[d]thiazol-2-yl)-3-phenylurea derivatives as anti-invasion agents.

Alisol B as a novel inhibitor of TβRs suppresses the growth and metastasis of non-small cell lung cancer by inhibiting TGF-β-induced epithelial-mesenchymal transition.

A pH-responsive layered double hydroxide nanoradiosensitizer for bone metastasis tumor.

Here, we report the development of lectin-directed protein aggregation therapy (LPAT), which combines the strong glycan-targeting capabilities of multivalent lectins with the aggregating propensities of bacterial microcompartment proteins. The design aims to create a system sensitive enough to elicit cell-specific aggregation towards invasive, metastatic tumor cells, while being nontoxic to normal tissues. LPAT agents were screened against a panel of 6 breast cancer cell lines, with the most potent agent showing preferential anti-adhesive and anti-invasive activity against the hypersialylated/MMP9 overexpressing MDA-MB-231cell line. Furthermore, LPAT agents did not exhibit any propensity for hemagglutination, a principal disadvantage of lectin-based targeting systems. Subsequent studies using a metastatic mouse model showed that LPAT agents could prevent the formation of experimental lung metastases caused by the highly metastatic MDA-MB-231-LM2 isoform cell line. Overall, this work has laid the foundation for a potential glycan-targeting therapy aimed at preventing the onset and progression of metastatic tumors in a safe and selective manner.

Atractylodes macrocephala Koidz. has long been used for invigorating Qi and traditionally prescribed for cancer treatment by Chinese medicine practitioners. Atractylenolide I (ATI) is a major active component in A. macrocephala and has inhibitory effects on various malignancies. Triple-negative breast cancer (TNBC)-related deaths are predominantly attributed to metastasis. However, the activity and mechanism of ATI on TNBC metastasis are unclear. Cell viability was detected by CCK-8 assay. Cell migration was determined by wound healing and transwell assays. The invivo experiment was performed in 4 T1 spontaneous metastasis breast cancer nude mice. The mechanism was investigated by RNA-seq analysis, and further verified by bioinformatics, qPCR, western blot, and siRNA transfection analysis. ATI inhibited the growth of 4TI cells. Additionally, ATI repressed the wound healing, invasion, and migration activities of 4TI and MDA-MB-231 cells at a lower concentration. Invivo experiment revealed that while ATI (50 mg/kg) did not significantly inhibit tumor growth but markedly suppressed lung metastasis progression in a 4 T1 spontaneous metastasis mouse model. RNA-seq analysis of lung tissues showed that the extracellular matrix (ECM) signaling pathway was significantly suppressed by ATI. In addition, ATI inhibited the mRNA and protein levels of ECM-related members (SPP1, MMP9, and COL1A1) invitro and invivo. Bioinformatics analysis revealed that the overexpression of SPP1 correlated with the poor prognosis of breast cancer patients. Furthermore, ATI inhibited ECM activator pyrintegrin-induced wound healing. siRNA knockdown of SPP1 abolished ATI's inhibition on wound healing, cell migration, and expression of ECM-related genes/proteins in 4 T1 cells. Molecular docking showed that ATI interacted with Trp-106, Pro-44, and Leu-46 residues in SPP1 (-7.73 kcal/mol). Our findings suggest that ATI could suppress the lung metastasis of TNBC by inhibiting the SPP1-mediated ECM signaling pathway.

ABSTRACTBackgroundCancer‐associated fibroblasts (CAFs) in the bone microenvironment play a key role in breast cancer (BC) osteolytic bone metastases. Differentiated embryo‐chondrocyte expressed gene 1 (DEC1) has been implicated as a potential therapeutic target in CAFs. Here, we investigate the role of DEC1 within the bone microenvironment during BC cell‐induced bone destruction.MethodsIn vivo, a BC bone metastasis mouse model by intratibially injecting 4 T1 cells into DEC1+/+ and DEC1−/− mice was used to explore the effects of DEC1 deficiency in the bone microenvironment on BC osteolytic bone destruction. Bone mesenchymal stromal cells (BMSCs) isolated from DEC1+/+ and DEC1−/− mice were induced to differentiate into CAFs, as in vitro model to explore the roles and underlying mechanisms.ResultsDEC1 deficiency markedly attenuated BC‐induced osteolytic destruction. Notably, DEC1+/+‐4 T1 mice exhibited a higher abundance of CAF biomarker‐positive cells in the bone microenvironment compared to DEC1−/−‐4 T1 mice. Mechanistically, tumor‐conditioned medium (TCM) and IL‐6 were found to promote the differentiation of BMSCs into CAFs through activation of the JAK2/STAT3 pathway, whereas DEC1 deletion suppressed this process by inhibiting JAK2/STAT3 signaling. Furthermore, DEC1 deficiency reduced RANKL secretion, thereby limiting osteoclastogenesis, and decreased PAI‐1 levels, which contributed to remodeling of the tumor microenvironment and suppression of BC cell migration.ConclusionOur findings demonstrate that DEC1 deficiency in the bone microenvironment critically restrains BC cell‐induced bone destruction through inhibition of CAF differentiation.

Metastasis remains the leading cause of cancer-related mortality, yet its early detection is hindered by the subtle and unpredictable nature of the metastatic niche. In this study, we developed a self-assembling peptide-based activatable nanoprobe, 1-CyBr(PBA)-NGR NPs, designed for the early detection of the metastatic niche through circulating tumor cells (CTCs)-mediated hitchhiking and hydrogen peroxide (H2O2)-induced fluorescence activation. The peptide probe spontaneously assembled into stable spherical nanoparticles in aqueous media and undergoes H2O2-triggered fluorescence recovery via oxidation of the phenylboronic acid masking group. Upon intravenous injection, 1-CyBr(PBA)-NGR NPs "hitchhike" CTCs through specific binding via the NGR moiety, utilizing the natural trafficking pathways of CTCs to reach the metastatic lesion. Upon reaching the metastatic site, the nanoparticles are selectively activated by the microenvironment, resulting in fluorescence activation and enabling precise optical imaging of the metastatic site. In vivo imaging in a lung metastatic mouse model revealed distinct fluorescence in the lungs following intravenous administration of 1-CyBr(PBA)-NGR NPs, whereas minimal fluorescence was observed in both the healthy control and antioxidant N-acetylcysteine (NAC)-treated metastatic mice, further confirming the probe's selective activation in response to the elevated H2O2 levels in the metastatic lesion. This study presents a "silent-to-bright" NIR imaging strategy for tumor metastasis monitoring, utilizing CTCs as "hitchhiking" carriers and the metastatic microenvironment as fluorescence activators, offering a promising approach for tumor metastasis diagnosis and intervention.

Immunogenic cell death-primed autophagosome vaccines drive dendritic cell cross-presentation and suppress colon cancer metastasis.

Deoxyribonuclease 1 Like 3 (DNASE1L3) is a member of the deoxyribonuclease I family that is associated with some diseases, including systemic lupus erythematosus and hypocomplementemic urticarial vasculitis. Recently, abnormal DNASE1L3 was preliminarily shown to correlate with tumor pathogenesis. However, its role is still undetermined in nasopharyngeal carcinoma (NPC). Multiple sets of Gene Expression Omnibus (GEO) high-throughput data were utilized to screen the differentially expressed genes. Signal pathway enrichment analysis analyzed the correlation between DNASE1L3 and epithelial-mesenchymal transition (EMT) and cytoskeleton reorganization. An immunohistochemistry assay for analysis of DNASE1L3 expression was used to detect the clinical samples. Woundhealing, migration, invasion assays, and mouse model of lung metastasis were used to assess the role of DNASE1L3 in NPC metastasis. The mechanism of DNASE1L3 inhibition of NPC metastasis by attenuating MYH9/β-catenin/c-Jun/LncRNA-KDM4A-induced E-cadherin ubiquitination degradation was demonstrated by protein stability evaluation, co-immunoprecipitation, immunofluorescence, chromatin immunoprecipitation, dual-luciferase reporter assay, and RNA immunoprecipitation. DNASE1L3 downregulation in patients with NPC was not only negatively related to lymph node metastasis and distant metastasis but was also positively associated with poor prognosis. Overexpression of DNASE1L3 in NPC cells suppresses migration, invasion, and metastasis in vitro and in vivo. Inversely, DNASE1L3 knockdown increased cell migration and invasion abilities. Mechanistically, DNASE1L3 recruited PARK2 to ubiquitinate and degrade MYH9 protein. MYH9 protein activated β-catenin/c-Jun signal and augmented c-Jun-induced LncRNA-KDM4A transcription. In the process of DNASE1L3-induced metastatic suppression, decreased LncRNA-KDM4A attenuated the recruitment of E3 ubiquitin ligase Hakai and thus impeded the degradation of E-cadherin, by which heightened E-cadherin protein stability and finally inactivated the EMT signal. Our data firstly elucidated that DNASE1L3 acts as a metastatic suppressor by attenuating E-cadherin ubiquitination degradation via the MYH9/β-catenin/c-Jun/LncRNA-KDM4A axis in NPC. DNASE1L3 is a potential marker for predicting NPC prognosis.

Liver metastasis is the primary cause of mortality in colorectal cancer (CRC) patients. To decipher the underlying mechanisms, we performed single-cell RNA sequencing (scRNA-seq) on paired primary colorectal tumors, adjacent tissues and liver metastases from three CRC liver metastasis (CRLM) patients, alongside colorectal tumors and adjacent tissues from three non-metastatic CRC patients. Our analysis revealed a significant enrichment of Enolase 2-expressing (ENO2⁺) cancer cells in CRLM patients compared to their non-metastatic counterparts. Functional characterization, supported by bioinformatics and murine models, demonstrated that ENO2⁺ cancer cells exhibit enhanced epithelial-mesenchymal transition (EMT) and are critical drivers of CRLM. Mechanistically, the ENO2 protein directly binds to macrophage migration inhibitory factor (MIF) within cancer cells, stabilizing MIF by inhibiting its C-terminus of Hsc70-Interacting Protein (CHIP)-mediated ubiquitination and degradation. This ENO2-MIF interaction activates MIF signaling, fostering robust tumor cell-macrophage crosstalk that promotes M2 macrophage polarization, which is validated by spatial transcriptomics showing the colocalization of ENO2⁺ cancer cells and M2 macrophages. Crucially, both organoid and in vivo models confirmed that ENO2 in CRC cells is essential for inducing M2 macrophage polarization via the MIF pathway, thereby facilitating liver metastasis. Knockout of ENO2 significantly suppressed tumor growth and liver metastasis in mouse models. An inhibitor of the ENO2-MIF interaction, pyrithioxin, can effectively reduce the burden of liver metastasis in mice. Collectively, our findings identify ENO2 as a key driver of CRLM by stabilizing MIF to orchestrate M2 macrophage polarization, highlighting the ENO2-MIF axis as a promising therapeutic strategy for CRLM.

Colon cancer (CC) is a highly prevalent malignant tumor with a high mortality rate worldwide. Despite recent advancements in diagnosis and treatment, the overall prognosis for patients remains poor, especially for those with metastasis. Exploring key genes associated with the prognosis of patients with colon cancer, establishing effective molecular models, and validating their functions are necessary to optimize patient management and develop novel therapeutic strategies. This study aimed to reveal the role of the key gene SERPINE1 in the progression of colon cancer and its potential clinical application value through bioinformatics analysis and experimental validation. Multimodal data from colon cancer patients were obtained from The Cancer Genome Atlas (TCGA) database. Differentially expressed genes (DEGs) with a fold change ≥ 2 and an adjusted P value less than 0.05 were screened. A protein‒protein interaction network was constructed using the STRING database, and 40 core genes were identified using Cytoscape software. Genes closely related to the prognosis of colon cancer patients were further selected using LASSO regression and Cox proportional hazards regression models to construct a risk scoring model for assessing patients' survival risk. The model performance was evaluated in combination with immune cell infiltration analysis and ROC curve assessment, and the associations between the target genes and the tumor immune microenvironment were explored. The expression and function of SERPINE1 were subsequently investigated. The expression level of SERPINE1 in colon cancer tissues was analyzed by Western blotting. The effects of SERPINE1 knockdown on the migration and invasion abilities of colon cancer cells were assessed by scratch wound healing and Transwell assays. The impact of SERPINE1 knockdown on tumor formation and metastasis in colon cancer cells was verified through mouse tumor and metastasis models. The bioinformatics analysis identified seven target genes related to the prognosis of colon cancer, namely, CAMK2B, KIF1A, OPCML, SCN5A, SERPINE1, TH, and UCHL1, based on which a risk scoring model was constructed. Further analysis revealed that the risk score was not only associated with patients' survival prognosis but also significantly correlated with the infiltration of immune cells such as T cells and macrophages in the tumor immune microenvironment. Among these seven genes, SERPINE1 was highly expressed in colon cancer and was significantly positively correlated with a poor prognosis. In vitro experiments revealed that SERPINE1 knockdown inhibited the proliferation, migration, and invasion of colon cancer cells. Western blot experiments indicated that SERPINE1 knockdown upregulated the expression of epithelial cadherin (E-cadherin, E-CAD) and downregulated the expression of neural cadherin (N-cadherin, N-CAD) and vimentin (VIM), suggesting that SERPINE1 knockdown may inhibit the epithelial‒mesenchymal transition (EMT). In a mouse subcutaneous tumor experiment, SERPINE1 knockdown significantly inhibited tumor growth. The liver metastasis model revealed that inhibiting SERPINE1 expression significantly reduced the number of liver metastases, further verifying the effect of high SERPINE1 expression on promoting the progression of colon cancer. A risk scoring model constructed based on the results of a bioinformatics analysis can effectively predict the survival prognosis of patients with colon cancer and reveals that SERPINE1 promotes the proliferation of colon cancer cells and accelerates the distant metastasis of tumor cells by inducing the EMT. These results provide an important reference for the prognostic assessment of colon cancer patients and pave the way for the development of therapeutic strategies targeting SERPINE1.

The metastatic progression of breast cancer involves complex interactions between tumor cells and immune cells, including T cells that exert cytotoxic pressure to limit metastasis. Tumor cells reprogram their metabolism to evade immune surveillance, a critical step to achieving metastatic outgrowth. Using an unbiased CRISPR screen targeting metabolism-related genes and a clinically relevant spontaneous metastasis mouse model, we identified CPT1A, the rate-limiting enzyme in fatty acid β-oxidation, as a suppressor of immune-dependent metastasis. Loss of CPT1A enhances lung metastasis in immunocompetent mice, but not Rag1 KO mice that lack mature lymphocytes. Loss of CPT1A triggers cytosolic mitochondrial DNA (mtDNA) release via the mPTP pore. Cytosolic mtDNA release triggers a STING-dependent inflammatory response, creating an environment that impairs CD8+ T cell function, promoting metastatic outgrowth. Among breast cancer patients, low CPT1A expression correlates with poor survival when CD8+ T cell infiltration is high. These findings reveal an extrinsic role for CPT1A in immune-tumor dynamics and suggest therapeutic opportunities targeting inflammation in metastatic breast cancer.

Cancer-associated hyperfibrinolysis are potentially fatal outcomes of advanced cancer. Unfortunately, this knowledge has not yet led to any breakthrough in cancer therapy. Plasmin is the key enzyme of fibrinolytic system. The identification of a direct link between plasmin and the tumor progression remains unknown. Here, we demonstrated that plasmin expression was significantly upregulated in HCC tissue, especially tumor tissues from patients who had received chemotherapy. Through in vitro and in vivo models, we showed that plasmin enhanced HCC cell invasion and migration. Pharmacological inhibition of plasmin with tranexamic acid (TXA) suppressed tumor invasion and metastasis. Mechanistically, we identified plasmin could directly bind to CXCR4, which potentiated the CXCL12/CXCR4 interaction and activated the PI3K/AKT/mTOR signaling pathway. Moreover, CDDP-resistant cells had higher expression of plasmin and stronger invasion and metastasis ability, which were effectively suppressed by plasmin inhibitor, and combining TXA with CDDP synergistically inhibited tumor progression and metastasis in mouse model and human HCC organoids. Our findings reveal a novel plasmin-CXCR4 axis driving HCC dissemination and suggest that targeting plasmin may enhance the efficacy of conventional chemotherapy.

Chronic polystyrene microplastics exposure promotes lung adenocarcinoma metastasis through EREG-regulated phosphorylation-dependent NF-κB activation

Melanoma is a highly aggressive cancer with significant treatment challenges due to its immunosuppressive tumor microenvironment. Among its subtypes, uveal melanoma (UM), as a distinct subtype arising in the immune-privileged ocular site, currently lacks effective therapies to improve overall survival. We developed a glutathione (GSH)-responsive poly(β-amino ester) (PBAE) platform for intracellular IL-12 mRNA delivery. By optimizing the ratio of GSH-responsive diacrylate monomers and amine structures, we synthesized and screened a library of candidate polymers for transfection efficiency. Nanoparticle size, stability, GSH-triggered disassembly, and mRNA release were characterized. Cellular uptake, protein expression, endo/lysosomal escape, cytotoxicity, and systemic safety were assessed in vitro and in vivo. Efficacy was evaluated in subcutaneous, intraocular, and lung metastasis mouse models using the B16F10 melanoma cell line. The optimized polymer PSS0.5-A2B2 formed stable mRNA-loaded nanoparticles (approximately 100 nm) and exhibited time-dependent disassembly under GSH-rich conditions, enabling efficient mRNA release and enhanced IL-12 expression. In vitro, it enhanced cellular uptake, IL-12 expression, and endo/lysosomal escape. In vivo, no significant systemic toxicity was observed. IL-12 mRNA delivery significantly suppressed tumor growth, promoted apoptosis, and reduced metastasis. Combined intravitreal and intravenous administration showed superior antitumor efficacy. This GSH-responsive PBAE platform enables biocompatible intracellular IL-12 mRNA delivery, offering a promising local-systemic combinatorial immunotherapy strategy for melanoma, holding potential promise for UM.

Accurate early detection of tumor metastasis remains a formidable clinical challenge owing to the lack of imaging tools that can simultaneously respond sensitively to early metastatic signals and maintain enduring intracellular functionality. Here, we report a programmable morphing DNA nanodevice (PMDN) that integrates catalytic hairpin assembly (CHA) with a hybrid network amplification mechanism to achieve dual-stage intracellular assembly and ultrasensitive detection of metastatic biomarkers. The acidic lysosomal milieu induces i-motif-mediated conformational folding into a compact and nuclease-resistant structure. Following lysosomal escape, cytoplasmic miR-221 triggers a secondary-stage CHA cascade, driving large-scale crosslinking of DNA monomers into a stable nanonetwork. This dynamic bottom-up assembly ensures prolonged intracellular structural integrity of PMDN and concomitantly enables triple-stage signal amplification. Compared with conventional CHA systems, PMDN achieves more than a 200-fold improvement in detection sensitivity, exhibits remarkably persistent fluorescence in MDA-MB-231 cells, and maintains durable tumor localization in vivo for over 10 days. In a metastatic mouse model, PMDN enables early visualization of pulmonary micrometastases through miR-221-activated signal amplification. These results establish environment-adaptive morphing DNA architectures as a powerful platform for real-time monitoring of early metastasis and long-term molecular imaging in complex biological environments.

Mammary tumor cells reshape the bone marrow niche inducing B cell lossBone marrow B cell development is impaired in mammary tumor metastasisExperimental depletion of B cells promotes bone metastasisG-CSF mediates B cell loss in mammary tumor metastasis.

Metastasis and cancer-induced cachexia significantly reduce survivorship and quality of life for cancer patients. GDF10 (BMP3b) is a TGF-ß superfamily ligand with little knowledge of its role in cancer progression. Some studies have shown that GDF10 exerts tumor-suppressive effects in a range of cancer types and also plays a protective role against muscle wasting. Basal transcription of GDF10 was described previously to be downregulated in both primary tumors and cachectic muscle. Here, we set out to investigate the therapeutic potential of GDF10 in the 4T1.2 mouse model of breast cancer metastasis and in the C-26 mouse model of cancer cachexia, hypothesizing that GDF10 would ameliorate both metastatic and cachectic disease pathology. Systemic rAAV6:GDF10 administration to mice did not alter primary tumor growth; however, metastatic burden was increased in the mice bearing 4T1.2 tumors. Similarly, increased intramuscular rAAV6:GDF10 expression exacerbated skeletal muscle wasting in C-26 tumor-bearing mice. These results contradicted our initial hypothesis and highlight the complexity of signaling mechanisms utilized by BMP family ligands. Our data point to the need for more research to understand how to target GDF10 in anti-cancer therapy.