Theranostics最新文献

Background: Metastasis is the primary cause of cancer-related mortality, and targeting the drivers of this process is a promising strategy to improve patient outcomes. Recent studies have highlighted a role of Carbamoyl Phosphate Synthetase 1 (CPS1), the urea cycle's rate-limiting enzyme, in tumor development. However, its involvement in tumor spreading and metastasis remains unclear. Methods: Transwell assay, wound healing assay and a range of lung cancer metastasis animal models were employed to investigate the impact of genetic knockdown and pharmacological inhibition of CPS1 on lung cancer metastasis both in vitro and in vivo. Quantitative proteomic analysis, RNA sequencing, untargeted metabolomics and targeted metabolomics to urea cycle were conducted to elucidate the underlying mechanisms of CPS1 inhibition. Results: CPS1 was overexpressed in a subset of patients with metastatic lung cancer, and this increased expression correlated with decreased patient survival. Genetic knockdown and pharmacological inhibition of CPS1 significantly reduced the tumor burden and metastasis in mice with the spontaneous (Kras ^G12D/+; p53 ^-/-) and induced metastatic lung cancer. Mechanistically, CPS1 overexpression in metastatic cancer cells resulted in excessive fumarate production, an intermediate metabolite in the urea cycle. Fumarate accumulation inhibited TET2 activity and altered miR200a gene methylation to drive epithelial-to-mesenchymal transition (EMT), thereby enhancing cell migration and invasion. Notably, CPS1 inhibition reduced fumarate accumulation and enhanced TET2 activity, which epigenetically upregulated PD-L1 expression. This activation contributed to impaired CD8⁺ T cell function and ultimately promoted tumor immune evasion. To overcome immune evasion, we investigated a combination therapy. Combining a CPS1 inhibitor with an anti-PD-1 antibody demonstrated a synergistic and potent effect, significantly inhibiting both lung tumor growth and metastasis. Conclusions: These findings define a crucial role for CPS1 in lung cancer metastasis. Targeting CPS1 may offer a valuable therapeutic intervention strategy against metastatic lung cancer.

Background: Berberine, a natural compound with unique bioactivity, has been widely used in the treatment of gastrointestinal inflammatory diseases. Despite its well-documented anti-inflammatory properties, the system-level regulatory network underlying its multifaceted mechanisms remains poorly understood. Methods: In this study, we employed a multi-level analytical approach, integrating single-cell RNA sequencing, targeted metabolomics, 16S rRNA gene sequencing, and drug-target analysis, to elucidate the integrative effects of berberine on gut microbiota-metabolism-immune interactions. Results: Single-cell RNA sequencing revealed that berberine enhances energy metabolism in intestinal cells of DSS-induced mice, thereby maintaining normal physiological functions. Targeted metabolomics analysis of short-chain fatty acids, combined with 16S rRNA gene sequencing, demonstrated that berberine supplementation significantly increases short-chain fatty acid (SCFA) levels in the intestinal environment and selectively enriches the abundance of Akkermansia. Furthermore, single-cell RNA sequencing data indicated that berberine inhibits fibroblast-to-lymphatic transformation and suppresses the expression of interleukin-1β, leading to reduced immune activation in innate immune cells. Drug-target analysis identified shared molecular targets between berberine and various immunotherapeutic agents. Conclusion: This study provides a comprehensive understanding of berberine's multi-target mechanisms and highlights its potential as a therapeutic agent for inflammatory diseases through the modulation of gut microbiota, host metabolism, and immune responses.

Background: ²²⁴Ra, an alpha-emitting radionuclide with a half-life of 3.63 d, holds significant promise in cancer therapy. However, like many other medical alpha-emitters, the development of ²²⁴Ra radiopharmaceuticals has long been impeded by dosimetry limitation caused by the off-target toxicity, which is tightly related to the secondary radioactivity biodistribution. Methods: In this work, we propose leveraging radionuclide trap preorganized in nanoscale barium-based metal-organic framework (AEMOF-6) to overcome the off-target effects of ²²⁴Ra therapy. Functional side chains with high binding affinity towards ²²⁴Ra and its decay daughters were preinstalled inside the cavity of nanoscale AEMOF-6, constructing radionuclide trap capable of inhibiting the radioactivity leaking effectively. Results: The ²²⁴Ra-labeled radiopharmaceutical ²²⁴Ra-AEMOF-6@CS demonstrates effective in vivo radioactivity localization ability, significant antitumor efficacy, and favorable biosafety. It was obtained with a radiochemical yield of 92.87% and a radiochemical purity of 94.75%, maintaining over 87% in vitro stability throughout the observation period. Integrated micro-PET/CT and micro-SPECT/CT imaging, complemented by biodistribution analyses, validated the robust stability and radioactivity localization capability of the AEMOF-6@CS nanocarrier in vivo. A dose-dependent antitumor effect accompanied by excellent biosafety was observed, achieving complete tumor eradication in 20%, 40%, and 60% of mice at 36 d after injection of 18.5, 37.0, and 55.5 kBq of ²²⁴Ra-AEMOF-6@CS, respectively. Conclusion: This discovery provides a potential approach to address the challenges of radioactivity migration of ²²⁴Ra radiopharmaceuticals via radionuclide trap preorganized in nanoscale MOFs, which can also be beneficial to other alpha-emitting radiopharmaceuticals.

Background: Chronic myocarditis (CMYO) progresses to fibrosis and heart failure, yet no therapies effectively target fibrosis. Fibroblast activation protein (FAP) marks pathogenic myofibroblasts, but its therapeutic potential remains unexplored in inflammatory settings. Methods: Using bulk/scRNA-seq of human myocarditis samples, we identified FAP as a fibrosis-specific marker. We engineered FAP-targeted CAR-T (FAP.CAR-T) cells and tested their efficacy in autoimmune (EAM) and viral (CVB3) myocarditis models. Human cardiac organoids (hCOs) treated with IL-17A modeled inflammatory fibrosis. Results: FAP expression correlated with fibrosis severity in patients (r = 0.96, P = 0.0028). In EAM and CVB3 models, FAP.CAR-T cells reduced fibrosis by 65% and 55%, respectively (P < 0.001), restored ejection fraction to higher than 65%. hCOs treated with FAP.CAR-T cells showed 55% less fibrosis (P < 0.05). No toxicity was observed in healthy mice. Conclusions: FAP.CAR-T cells eliminate fibrosis-driving myofibroblasts, reversing cardiac dysfunction in chronic myocarditis. This strategy, validated in human organoids, offers translatable immunotherapy for fibrosis-driven heart disease.

Background: Perineural invasion (PNI) is a key biological feature underpinning the high malignancy and poor prognosis of pancreatic ductal adenocarcinoma (PDAC). Lysine lactylation (Kla), a metabolite-stress-induced post-translational modification, plays crucial regulatory roles in diverse biological processes. The RNA methyltransferase NSUN2 is essential for cancer invasion and metastasis. However, the mechanisms by which NSUN2 contributes to lactylation-driven PNI in PDAC remain to be elucidated. Methods: We assessed tumor lactate / pan-lactylation, NSUN2 lactylation, and PNI in human PDAC cohorts with survival follow-up. Functional studies used PDAC cell lines for migration/invasion assays, dorsal-root-ganglion (DRG) co-culture, and neurite-outgrowth assays under lactate or enzymatic perturbations. Mechanistic interrogation combined NSUN2 knockout, CRISPR knock-in mutants at K692 (K692R/E), co-immunoprecipitation, RIP-seq, MeRIP-qPCR, and actinomycin-D chase to test mRNA binding, m5C modification, and stability of CDCP1/STC1. In vivo validation employed a sciatic nerve invasion model and a KPC genetically engineered mouse model to assess tumor-nerve infiltration and disease progression. Results: Lactylated NSUN2 is markedly upregulated in mice and human PDAC with more severe PNI, and is significantly associated with poorer prognosis. Functionally, inhibiting lactylation or blocking NSUN2 markedly attenuated tumor-nerve interactions and neural invasion. Mechanistically, lactate accumulation leads to the lactylation of NSUN2 at lysine 692 (K692), subsequently inhibiting its ubiquitination and degradation. lactylation of NSUN2 mediated m5C modification on CDCP1 and STC1 mRNA, enhanced their mRNA stability. Conclusions: This study identifies lactate-driven NSUN2 K692 lactylation as a key driver of perineural invasion in PDAC. We define a lactate-NSUN2-m5C-CDCP1/STC1 axis that links metabolic stress-induced lysine lactylation to mRNA methylation-dependent stabilization of pro-invasive transcripts, highlighting actionable therapeutic targets to restrain neural invasion and improve patient outcomes.

Rationale: Nuclear factor erythroid 2-like 1 (Nrf1/NFE2L1) is a crucial redox-sensitive factor essential for mitochondrial homeostasis. However, its function in controlling macrophage-associated liver inflammation and fibrosis remains to be fully understood. Herein, this study was conducted to elucidate the roles of macrophage Nrf1 in regulating liver fibrosis. Methods: Expression levels were analyzed in human liver tissues collected from individuals diagnosed with or without liver fibrosis. High-fat diet feeding, carbon tetrachloride injection or bile duct ligation was performed respectively to established three mouse models of liver fibrosis. Myeloid-specific Nrf1-knockout (Nrf1^M-KO ) mice were developed to investigate the role and underlying mechanisms of macrophage Nrf1 in vivo and in vitro. Results: Macrophage Nrf1 expression was markedly reduced in liver samples from both humans and mice with liver fibrosis. The deletion of myeloid Nrf1 remarkably accelerated liver inflammation and fibrosis. Macrophages from Nrf1^M-KO mice exhibited enhanced M1 polarization and mitochondrial dysfunction. Mechanistically, Nrf1 directly binds to Foxo1 and inhibits its transcriptional activity. The target gene KLF16, regulated by the Nrf1-Foxo1 complex, is crucial for modulating mitochondrial function and immune response. Conclusions: Our study highlights the functional properties of macrophage Nrf1-Foxo1 axis in controlling mitochondrial reprogramming and liver fibrosis progression.

Rationale: High-intensity psychological and physiological stress contributed greatly to development of cardiac disorders in contemporary society. However, the underlying molecular mechanisms remain largely unknown. Synaptotagmin-7 (Syt7), a Ca²⁺ sensor with high affinity, has been associated with synaptic transmission and tumor progression, but its role in cardiac stress responses remains poorly defined. Methods: Corticosterone (CORT) was used to induce stress injury in vivo and in vitro. The expression of Syt7 was modulated by genetic knockout, injection of adenoviral siRNA or injection of adeno-associated virus serotype 9 (AAV9) shRNA. Cardiac function and remodeling were evaluated by echocardiography, electrocardiography, and histological staining. Necroptosis was analyzed by propidium iodide (PI) staining, lactate dehydrogenase (LDH) release detection, and necroptosis marker levels. Ca²⁺ overload, ROS production, mitochondrial permeability transition pore (mPTP) opening, and bioenergetic profiling were assessed to evaluate mitochondrial function. Co-IP assay was performed to detect protein interaction, and ChIP- qPCR was performed to assess transcriptional regulation. Results: Syt7 expression was significantly upregulated in both cardiomyocytes and heart tissues exposed to CORT. Both genetic knockout and cardiomyocyte-specific knockdown of Syt7 significantly preserved cardiac function and rhythm, and alleviated myocardial hypertrophy and fibrosis in CORT-treated mice. Mechanistically, Syt7 regulated necroptosis by promoting calcium overload, ROS production, mitochondrial ΔΨm dissipation, and mPTP prolonged opening. Notably, Syt7 interacted with transcription factor p53 and enhanced p53- mediated transcription of Bcl-2 homologous antagonist/killer (Bak). Syt7, p53 and Bak constitute a novel signaling axis to regulate mitochondrial dysfunction and necroptosis. Therapeutically, cardiac delivery of Syt7-targeting siRNA via adenoviral vectors significantly alleviated structural remodeling, electrophysiological instability, and myocardial necrosis in CORT-challenged mice. Conclusions: The study identified Syt7 as a novel upstream regulator involved in cardiomyocyte necroptosis triggered by stress stimuli through a p53-Bak-mPTP pathway. Therapeutic targeting of Syt7 offers a promising strategy for protecting the heart against psychological or neuroendocrine stress-related injury.

Background: Regulating the morphology structure of sonosensitizers and nanozymes is crucial to improve sonodynamic and enzyme-mimic activities. Methods: We report for the first time the utilization of Cu₂O nanospheres as the sacrificial templates for the synthesis of hollow RuO₂ nanospheres (H-RuO₂) for high-efficiency sonodynamic and chemodynamic therapy (SDT/CDT). We then utilized NIR phosphorescence carbon dots (CDs) as the auxiliary sonosensitizers to sensitize H-RuO₂ for the construction of CD@H-RuO₂ heterojunctions. Results: Compared with solid nanoparticles, nanosheets, and other structures, the hollow RuO₂ (H-RuO₂) nanostructures are expected to exhibit stronger catalytic activity due to their larger specific surface area and more catalytic active sites. The improved electron-hole separation kinetics enable CD@H-RuO₂ nanozymes with significantly enhanced sonodynamic and multienzyme-mimic activities. CD@H-RuO₂-triggered cascade amplification of antitumor immune response was realized by the heterojunction construction, GSH depletion, and relief of hypoxia co-augmented ROS yield, which significantly induced a robust ICD. Conclusion: CD@H-RuO₂-mediated SDT and CDT co-amplified immunotherapy have shown significant antitumor effects, resulting in the eradication of primary tumors and the inhibition of distant tumor growth. This study offers hopeful insights into the fabrication of heterojunctions for sonodynamic/chemodynamic-activated immunotherapy.

Background: Although autologous whole tumour cells provide broad-spectrum antigens for personalised cancer vaccines, their weak immunogenicity necessitates adjuvant co-delivery systems. Methods: We developed a conjugate adjuvant (G-PL) by coupling modified yeast β-glucan with poly-D-lysine. Electron microscopy confirmed its binding to GL261 cell membranes. The adjuvant-cell complex (ICC@G-PL) was constructed by coating irradiated tumour cells with G-PL. We evaluated the recruitment/activation of dendritic cells (DCs), lymph node priming, tumour-specific immunity, and therapeutic efficacy in glioblastoma, colon cancer, and melanoma models. Dectin-1-mediated Th17 induction was analysed via Western blotting and flow cytometry. Results: G-PL (≤ 500 μg/mL) rapidly adhered to cell membranes without cytotoxicity. In vitro, it enhanced DC uptake of tumour components, maturation, and non-pathogenic Th17 differentiation. In vivo, ICC@G-PL recruited DCs at injection sites, activated draining lymph nodes, and elevated plasma levels of IL-12, TNF-α, and IFN-γ. The vaccine prolonged survival in both therapeutic and preventive models, increasing intratumoral CD8⁺/CD4⁺ T cell ratios, M1 macrophages, and neutrophils. Dectin-1 downregulation in DCs correlated with Th17-driven anti-tumour responses. Conclusions: G-PL, a novel β-glucan-based adjuvant, enables rapid construction of autologous whole-cell vaccines. This strategy enhances tumour-specific immunity and reprogrammes the tumour microenvironment, offering a universal platform for personalised cancer immunotherapy.

Rationale: Ferroptosis-induced tumor cell death and immune activation represent promising strategies for overcoming therapeutic resistance in triple-negative breast cancer (TNBC). However, clinical application remains limited by poor oral absorption, transient immune activation, and systemic toxicity. Methods: We developed an orally administrable nanoplatform (MCT-NE#9) co-delivering docetaxel (DTX) and atorvastatin (ATV), designed to enhance intestinal uptake via bile acid and vitamin transporters. Pharmacokinetic, in vitro, and in vivo studies were conducted to evaluate drug absorption, sustained ferroptosis, and immune modulation. Results: MCT-NE#9 markedly improved oral bioavailability (659% for ATV, 851% for DTX) and sustained intratumoral drug levels under a low-dose metronomic regimen. Mechanistically, it induced sustained ferroptosis by promoting iron accumulation, lipid peroxidation, and GPX4 suppression, while remodeling the tumor immune microenvironment. Treatment increased M1 macrophages and antigen-presenting cells and reduced TGFβ1, regulatory T cells, and M2 macrophages. In vivo, oral MCT-NE#9 suppressed tumor growth by 50.4%, with enhanced efficacy (70.3% inhibition) when combined with anti-CD47 therapy. Conclusion: MCT-NE#9 enables a synergistic, low-toxicity chemo-immunotherapeutic strategy by sustaining ferroptosis and reprogramming the immune microenvironment via transporter-targeted oral delivery. This ligand-directed nanoplatform offers a clinically translatable approach for effective TNBC treatment.