R-loops, RNA:DNA hybrids formed during transcription, play critical roles in regulating gene expression and maintaining genome stability. Dysregulation of R-loop formation and resolution has been linked to genomic instability, a hallmark of cancer. Recent advances have highlighted the pivotal role of non-coding RNAs (ncRNAs), particularly long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs), in modulating R-loop dynamics, influencing tumorigenesis, and contributing to therapeutic resistance. These ncRNAs participate in the formation, stabilization, and resolution of R-loops, which in turn regulate critical processes such as transcriptional regulation, DNA repair, and chromatin architecture. For example, circRNAs such as circSMARCA5 and circDMD have been shown to induce R-loop formation, influencing gene expression and sensitizing cancer cells to chemotherapy. Conversely, lncRNAs such as TUG1 and NEAT1 regulate R-loop resolution, maintaining genome stability and enhancing tumor cell survival. The interaction between ncRNAs and R-loops offers promising avenues for targeted therapeutic strategies aimed at restoring R-loop balance to improve cancer treatment outcomes. This review provides an in-depth exploration of the molecular mechanisms by which ncRNAs modulate R-loop dynamics and discusses their potential as biomarkers and therapeutic targets in oncology. Furthermore, we highlight the challenges and future directions in translating these findings into clinical applications.

Atherosclerosis has traditionally been considered a lipid-driven disease. However, emerging evidence highlights the central role of inflammation in the development of atherosclerosis. Multiple cell types, including endothelial cells, macrophages, and other immune cells, interact within lesions to form a chronic inflammatory microenvironment. Unhealthy lifestyle, smoking and metabolic disorders like hyperlipidemia, hyperglycemia, and their derived pro-atherogenic products drive vascular inflammation. Recent evidence reveals that high-sensitivity C-reactive protein surpasses LDL-cholesterol in predicting future cardiovascular risk. Combining lipid-lowering with anti-inflammatory therapies significantly reduces event recurrence, underscoring the need for targeted drugs. Promising results have emerged from trials of anti-inflammatory agents. Statins and other lipid-lowering drugs also exhibit anti-inflammatory properties. However, cholesterol-independent strategies face challenges like infection risk from immunosuppression, requiring extensive safety evaluation. Enhancing intrinsic resilience mechanisms is a promising alternative in combating vascular inflammation and atherosclerosis. High-throughput screening accelerates drug development, while induced pluripotent stem cell-derived vascular organoids better simulate human atherosclerosis pathobiology, improving preclinical predictions. Research on organ interaction networks and trained immunity offers novel therapeutic targets. In this comprehensive review, we provide a state-of-the-art synthesis of the drivers, mechanisms, and potential therapies of atherosclerosis by targeting inflammation, with an aim to reducing residual cardiovascular risk in the post-statin era.

Hypoxia-inducible factors (HIFs) serve as the central signaling hub within the hypoxic tumor microenvironment, coordinating tumor proliferation, metastasis, and therapy resistance. This protein family comprises HIF-1α, HIF-2α, HIF-3α, and HIF-1β, with distinct subcellular localization patterns reflecting their isoform-specific oxygen-sensing mechanisms. Tumor-associated HIF expression is regulated through a multifaceted network involving non-coding RNAs (ncRNAs), mitochondrial metabolites, and post-translational modifications (PTMs). These aberrantly expressed HIFs then orchestrate biologically important processes including metabolic reprogramming, angiogenesis, and pro-tumorigenic inflammation. It is important to note that current evidence regarding HIF-3α function remains limited and requires further validation in vivo. This review first systematically deciphers the mechanisms governing HIF dysregulation in tumors, before elucidating the key biological processes involved. We then synthesize advances in HIF-targeting inhibitors. These pivotal findings provide a solid theoretical foundation and novel insights for both anticancer therapies targeting this critical “molecular switch” and translational research focusing on HIFs as promising therapeutic targets.

Telomeres and telomerase have been extensively implicated in the cellular processes of aging and inflammation. Recent studies have shown that telomere length (TL), the shelterin complex, and telomerase dysfunction are closely related to non-malignant pulmonary diseases, including interstitial lung diseases (ILD), chronic obstructive pulmonary disease (COPD), and asthma. Short telomere defects with or without mutations in telomere maintenance genes and telomerase are relatively common, affecting the progression of non-malignant pulmonary diseases, explaining disease susceptibility, and revealing clinically relevant manifestations. In this review, we examine the biological characteristics and functions of telomeres and telomerase, and investigate the intricate relationship between changes in TL and mutations in telomerase genes in non-malignant pulmonary diseases after a detailed associated literature review. Subsequently, we focus on the clinical features of non-malignant pulmonary diseases related to dysfunctional telomeres/telomerase, as well as the potential molecular mechanisms underlying these associations, along with the current status of therapeutic interventions. Finally, we delineate current knowledge gaps and transformative opportunities in telomere biology research, with a focus on bridging molecular discoveries to clinical innovations for telomere-associated non-malignant pulmonary diseases, aiming to accelerate the development of precision diagnostic tools and mechanism-based therapeutics.

Targeted protein degradation (TPD) offers a revolutionary paradigm to eliminate disease-driving proteins. Given the distinct technical requirements and challenges associated with degrading intracellular versus extracellular proteins, we classify existing TPD strategies based on subcellular localization into two categories: intracellular TPD (iTPD), which targets proteins within the cytoplasm and nucleus, and extracellular TPD (eTPD), which focuses on membrane-bound and secreted proteins. This destination-based framework facilitates precise technology selection and rational design by aligning methods with the biological context of their targets. However, the clinical translation of TPD remains constrained by a significant “delivery gap”. Current nanotechnological approaches are often discussed monolithically, despite the fundamentally distinct delivery requirements between iTPD and eTPD. For iTPD, the primary nanocarrier role is to confer fundamental drug-like properties to overcome systemic pharmacokinetic hurdles. Conversely, for eTPD, the nanoplatform's chief function is to engineer cellular engagement, enhance internalization, and orchestrate correct intracellular trafficking to the lysosome. This review will dissect the distinct challenges inherent to each “geographic” space and detail the tailored nano-playbooks being developed to address them. We will further explore the convergence of these two worlds and the emergence of nanoparticles as intrinsic degraders. Ultimately, we argue that a location-aware design philosophy is essential for unlocking the full therapeutic potential of TPD.

Nanoparticle-based delivery systems hold transformative potential for nucleic acid therapeutics. However, the fate of nucleic acid nanodrugs (NANDs) in vivo differs significantly from that observed in vitro, directly impacting their therapeutic efficacy. Upon introduction into biological fluids, NANDs rapidly adsorb proteins onto their surfaces, forming an assembled adsorption layer known as the protein corona (PC). This PC critically influences the physicochemical properties of NANDs and consequently governs their subsequent biological interactions. This review comprehensively introduces the mechanisms underlying PC formation, including dynamic adsorption kinetics and influential physicochemical and environmental factors. We further discuss how the PC modulates key in vivo processes, including penetration of gastrointestinal mucus and epithelial barriers, stability during systemic circulation, biodistribution and cellular tropism, as well as cellular uptake and endolysosome escape of nucleic acid therapeutics. While the PC may obscure engineered ligands and accelerate off-target clearance, it also offers opportunities to harness endogenous proteins for targeting. We therefore highlight emerging design strategies aimed at actively steering PC composition to achieve targeted nucleic acid delivery and enhanced therapeutic outcomes. Finally, we present prospects for translating fundamental knowledge of PC formation and function into the rational design of next-generation engineered nanocarriers for targeted NANDs applications.

Nanocarrier-based delivery of platinum compounds represents the next generation of platinum-based chemotherapy, which is a first-line treatment for many types of tumors. Despite the significant success of Pt(IV) prodrugs as effective antitumor agents, the therapeutic efficacy of most of the reported prodrugs is limited due to rapid biodegradation in the bloodstream and limited accumulation in the tumor. To overcome these limitations, various types of nanomedicines have been developed as drug delivery systems for Pt(IV) prodrugs, with those demonstrating significantly enhanced antitumor effects due to passive and active tumor targeting, stimulus-responsive drug release, effective synergistic therapy, the ability to induce immunogenic cell death, stimulate native and adaptive immunity, and preventing tumor recurrence. Also, the design of Pt(IV)-based theranostic nanoagents opens up photothermal, fluorescent, and photoacoustic imaging modalities for real-time monitoring of drug delivery and therapeutic response. The ability to harness photocontrolled chemotherapy along with immunotherapy, PDT, and PTT holds immense promise for synergistic anticancer effects. In the present review, we highlighted recent advances in the design of Pt(IV)-based NPs reported in 2022-2025 with the focus on the further development of this fast-growing research area.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), a beta-coronavirus, caused the recent global Coronavirus Disease 2019 (COVID-19) pandemic. Among the virus-encoded proteins, the surface spike (S) protein is critical for viral entry, membrane fusion, and pathogenesis, and its receptor-binding domain (RBD) initiates viral entry by binding to a cellular receptor. This makes the S an important therapeutic target for COVID-19. SARS-CoV-2 mutates frequently, giving rise to five major variants of concern, among which the Omicron variant and its subvariants are less sensitive to current therapeutic antibodies. The first part of this review describes the main protein constituents of SARS-CoV-2 and their functions, the S protein-mediated viral entry and fusion processes, and the main SARS-CoV-2 variants. Nanobodies are single-domain antibodies with high target-binding affinity, strong stability, and low production costs, whose small size facilitates their access to protein regions that are inaccessible to conventional antibodies. Thus, in the second part, we comprehensively review SARS-CoV-2-targeting nanobodies, including those that bind specifically to the RBDs of the S proteins, non-RBD S proteins, and non-S proteins of variants and subvariants of SARS-CoV-2, with the hope that this information will be valuable for the generation of novel SARS-CoV-2-targeting nanobodies with improved potency against COVID-19.

Phase change nanodroplets (PCND) consist of a liquid fluorocarbon core, which vaporizes into gas upon energy excitation, and a stabilizing shell. Through this phase-change property, PCND combine nanoparticle-like stability and pharmacokinetics, such as tissue extravasation and uptake by the mononuclear phagocyte system, with distinctive microbubble-like capabilities, namely ultrasound contrast enhancement and energy-triggered therapeutic action. PCND serve both as highly sensitive imaging agents, from molecular imaging to high-resolution vascular mapping, and as versatile therapeutics, used for example for thrombolysis, tissue ablation, biofilm removal, immunomodulation, and drug delivery. This review article provides a focused review of the biological behavior of PCND, including blood half-life, biodistribution, clearance pathways, and tumor accumulation, following a brief overview of the key physicochemical features that shape their in vivo fate. By comparing data across studies, we identify major inconsistencies in reported pharmacokinetics and highlight critical knowledge gaps, particularly regarding safety data. We further summarize advances in PCND-enabled imaging and therapy, and propose strategies to strengthen formulation design, biological evaluation, and stimulation protocols to support future clinical translation. Overall, PCND hold strong potential as a versatile theranostic platform. Continued systematic and clinically focused research is expected to accelerate their path toward clinical translation.

The melanocortin-4 receptor (MC4R) is a key regulator of energy balance and a potential target for weight management. Early drug discovery endeavors were hindered by incomplete understanding of its signal transduction mechanisms and broad activation of Gs pathways. Recent advances indicate that MC4R elicits multiple intracellular responses, with Gq/₁₁ signaling in the paraventricular nucleus (PVN) neurons playing a central role in appetite suppression. High-resolution cryo-electron microscopy structures of MC4R, such as the setmelanotide-MC4R-Gq complex reported here, provide valuable insights into ligand binding, receptor activation, and biased signaling, thereby enabling the design of more selective agonists with improved safety profiles. Combination therapies targeting MC4R alongside glucagon-like peptide-1 mimetics and other agents regulating metabolic pathways have shown promise to enhance weight loss. MC4R modulators are also implicated in treating other disorders, including melanocortin signaling dysfunction. These progresses call for renewed efforts in developing the next-generation MC4R-based therapies against metabolic diseases.

In the omics era, confident high-throughput analytical tools are crucial for the efficient identification of metabolites. Here, we present DeepHalo, a deep learning-integrated and hierarchically optimized workflow designed for high-throughput exploration of halogenated metabolites from high-resolution mass spectrometry-based metabolomics. DeepHalo leverages deep learning models combined with a comprehensive scoring to enhance the reliability of halogen predictions. It integrates PyOpenMS for fast isotope pattern detection and incorporates a halogen-based dereplication algorithm with GNPS molecular networking to efficiently exploit and annotate halogenates from complex biological matrices. To validate its performance, DeepHalo was applied to explore halogenated metabolites from 1296 microbial culture crudes, leading to the discovery of six families of structurally diverse halogenated molecules. This included a new class of cyclic depsipeptides, aglomycins A‒E, featuring rare 3-chloroanthranilic acid and/or epoxyvaline blocks. Additionally, a plausible biosynthetic pathway of aglomycins was proposed through bioinformatics analyses and targeted gene knockout experiments. Bioassays revealed that aglomycin A exhibits synergistic antibacterial activity with linezolid against vancomycin-resistant Enterococcus faecium (VRE) both in vitro and in vivo. We envision that DeepHalo, a user-friendly standalone executable freely available at https://github.com/xieyying/deephalo/releases/tag/DeepHalo_V1.0.0, will become a powerful tool for accelerating the discovery of halogenated “dark matter”.

Ionizable lipids are a pivotal component for lipid nanoparticles (LNPs) to optimize mRNA expression to fulfill the broad demands of various mRNA therapeutics. Traditionally, the screening of ionizable lipids needs extensive synthetic labor and stringent in vivo efficacy evaluation, which was often time-consuming, costly, and characterized by a lower success rate. Hence, we pioneered a machine-learning framework named Lipid with Artificial Intelligence (LipidAI) to evaluate the novel ionizable lipids rapidly. In this framework, the Methyl Tail Augmentation (MTA) strategy was first developed to triple the data by precisely adjusting the methyl groups on lipid tail chains. This ground-breaking approach compensated for data paucity in ionizable lipids libraries from the previous research and boosts model accuracy. Subsequently, the Ensemble Stacking Learning (ESL) algorithm was exploited to integrate multiple learning algorithms to surpass the predictive accuracy of a single algorithm used in former studies. Finally, we found that the predicted results of LipidAI were highly consistent with the actual data according to the in vivo expression of Luc-mRNA. Overall, this study highlights the remarkable potential of LipidAI in the rapid screening of ionizable lipids, adeptly avoiding the inherent drawbacks of traditional ionizable lipids development and thereby boosting the progress of LNP-based mRNA nano-drugs.

Angiogenesis is a hallmark of lung adenocarcinoma (LUAD) and a leading cause of mortality. Identifying potential therapeutic targets that modulate this process is of critical clinical importance. Here, we established a 12-gene risk-scoring model through bioinformatics screening and systematically delineated the molecular function of H2AC19, a previously poorly understood histone variant within this signature. Analysis of human LUAD specimens revealed elevated H2AC19 expression, which correlated positively with angiogenesis markers and advanced clinical stage and negatively with patient prognosis. Functional validation using CRISPR/Cas9 in patient-derived organoids (PDOs), in vitro cell models and in vivo xenografts demonstrated that H2AC19 promotes angiogenesis and tumor growth. Mechanistically, H2AC19 recruits p300 through its amino acid residues 24-88 to specifically augment H3K27 acetylation at the EGR1 promoter region, thereby triggering EGR1 transcriptional activation and subsequently promoting MMP-1-driven angiogenesis and progression of LUAD. Furthermore, we evaluated the therapeutic potential of lipid nanoparticles (LNP)-encapsulated siRNA targeting H2AC19, which significantly suppressed angiogenesis and LUAD progression. Collectively, our findings establish a critical role for H2AC19 in governing angiogenesis and highlight its potential as a promising therapeutic target for LUAD.

Drug-induced liver injury (DILI), particularly acetaminophen (APAP)-induced acute liver injury (ALI), is the leading cause of acute liver failure. Progranulin (PGRN) is a multifunctional glycoprotein. However, the role of PGRN in APAP-induced ALI remains unknown. Here, we found that PGRN serum concentration increased and correlated with disease severity in the patients with APAP overdose. Mice with PGRN deficiency were protected from APAP-induced ALI or concanavalin A (ConA)-induced ALI. They exhibited substantially increased hepatic recruitment of eosinophils, which depended on up-regulated IL-33 that was primarily released from liver sinusoidal endothelial cells (LSECs). Moreover, treatment of mice with blocking PGRN antibody could prophylactically and therapeutically treat APAP- or ConA-induced ALI, while injection of recombinant PGRN protein enhanced APAP-induced liver damage and worsened survival. Mechanistically, PGRN inhibited APAP-induced IL-33 expression in LSECs by dampening the activation of AMP-activated protein kinase (AMPK)-forkhead box O3 (FOXO3) signaling pathway. Therefore, PGRN should be considered as a new biomarker and potential therapeutic target to treat DILI.

Cisplatin remains a cornerstone of treatment for head and neck squamous cell carcinoma (HNSCC), yet its therapeutic efficacy is often undermined by acquired resistance. Here, we identify the PLOD1-PFKP-glycolysis axis as a central driver of cisplatin resistance. PLOD1 is significantly upregulated in cisplatin-resistant tumors and correlates with poor prognosis. Mechanistically, PLOD1 stabilizes the glycolytic enzyme PFKP by promoting its AKT-mediated phosphorylation at serine 386 via the HSP90-AKT complex, thereby enhancing glycolytic flux. This metabolic reprogramming facilitates stem-like properties and sustains epithelial-mesenchymal transition (EMT). Furthermore, increased intracellular acetyl-CoA levels driven by this axis promote histone H3K27 acetylation at the TGFBR2 promoter, thereby activating TGF-β signaling. Genetic depletion or nanoparticle-mediated silencing of PLOD1 reverses EMT and stemness features, restores cisplatin sensitivity, and impairs tumor growth and metastasis in vivo. These findings reveal a PLOD1-PFKP-glycolysis axis as the principal driver of cisplatin resistance, which coordinates metabolic and epigenetic alterations to promote tumor plasticity. Targeting this axis offers a promising strategy to overcome chemoresistance in HNSCC.

Recently, the BRAF inhibitor (Encorafenib) in combination with Cetuximab and mFOLFOX6 has been approved for the treatment of metastatic CRC (mCRC) patients with BRAF V⁶⁰⁰E mutation in the first line. However, intrinsic resistance to BRAFi still limits its therapeutic efficacy, and the clinical response is only ∼5%. One potential strategy to improve mCRC therapy is to combine agents that target key cellular signaling pathways, which may yield synergistic antitumor efficacy and overcome drug resistance. Herein, CDK9 inhibitors (CDK9i) were identified as candidate synergistic agents through kinase library-based high-throughput screening (HTS) and RNA sequencing. CDK9i synergistically sensitizes the therapeutic efficacy of BRAFi (as well as inhibitors of well-known downstream effector of BRAF, MEK and ERK) in multiple intrinsically resistant CRC models. Notably, CDK9i in combination with BRAFi also resulted in an enhanced therapeutic response in chemo-resistant CRC cells and PDOs. Taken together, CDK9 inhibition overcomes intrinsic resistance to BRAFi monotherapy, and targeting CDK9-mediated transcriptional elongation appears to be a promising and tolerable sensitization strategy for BRAFi-based treatment in mCRC, even in chemo-resistant mCRC.

Opioid addiction is driven by maladaptive reward memory, yet its molecular underpinnings remain poorly understood. Circular RNAs (circRNAs) have emerged as key regulators of neuroplasticity, but their roles in addiction-related memory remain unclear. Here, we identify circUnc79, a neuron-enriched and synaptically localized circRNA, as a critical modulator of morphine reward memory in the medial prefrontal cortex (mPFC). In the morphine-induced conditioned place preference (CPP) model, circUnc79 expression is dynamically downregulated during cue-induced memory retrieval. Gain- and loss-of-function experiments demonstrate that circUnc79 bidirectionally regulates the acquisition and persistence of morphine reward memory, without affecting locomotion, anxiety-like behavior, social interaction, or sucrose reward. Mechanistically, circUnc79 acts as a competing endogenous RNA for miR-149-3p, thereby relieving its repression of excitatory amino acid transporter 2 (EAAT2, encoded by Slc1a2). Elevated neuronal EAAT2 enhances presynaptic glutamate reuptake and recycling, supporting reward memory encoding. Manipulation of miR-149-3p produces opposite behavioral and synaptic effects, supporting the functional relevance of this regulatory axis. These findings reveal a circUnc79/miR-149-3p/EAAT2 pathway that regulates synaptic plasticity underlying opioid reward memory and highlight circUnc79 as a potential therapeutic target for opioid use disorders.

Clinical studies have suggested that exendin-4 (Ex-4) exhibits therapeutic potential for anxiety disorders, yet the molecular mechanisms underlying its efficacy remain largely unclear. Our study demonstrates that Ex-4 at specific doses ameliorates anxiety-like behaviors in mice. Mechanistically, Ex-4 could cross the blood-brain barrier and enter the central nervous system, where it exerts anxiolytic effects by activating glucagon-like peptide-1 receptor (GLP-1R) in the basolateral amygdala (BLA). Anterior dorsal bed nucleus of the stria terminalis (adBNST) as a downstream projection target of BLA neurons. Chemogenomic inhibition of the BLA-adBNST circuit induced anxiety-like phenotypes, which were rescued by Ex-4 treatment. Brain-derived neurotrophic factor (BDNF), a critical neurotrophin implicated in neuropsychiatric disorders including anxiety and depression, was functionally interrogated in this pathway. Furthermore, conditional knockdown of Bdnf in the BLA-adBNST circuit abolished the anxiolytic effects of Ex-4, indicating that the activation of GLP-1R in BLA neurons drives BDNF release into adBNST to mitigate anxiety. Taken together, these studies identify a central mechanism whereby Ex-4 attenuates anxiety-like behaviors by promoting BDNF release in the BLA-adBNST circuit.

Loss of TET2 protein leads to reduced DNA 5-hydroxymethylation (5-hmC), a key epigenetic alteration in melanoma, yet the regulatory mechanism governing TET2 stability remains unclear. Here, TET2 sulfhydration was analyzed in clinical melanoma samples, with sulfhydration sites identified by mass spectrometry, and the effects of active sulfur on TET2 stability, catalytic activity, and global 5-hmC assessed in melanoma cells. TET2 sulfhydration was significantly depleted in melanoma and further reduced in advanced stages. The cysteine-rich zinc finger domain at C1186/1202 was identified as the key sulfhydration site, which is essential for maintaining TET2 stability and enzymatic activity. Active sulfur restored TET2 sulfhydration, up-regulated TET2 protein, and reprogrammed global DNA hydroxymethylation. Notably, active sulfur synergized with anti-PD-1 therapy by enhancing IFN-γ-induced Th1-type chemokine and MHC-I expression in melanoma cells, thereby boosting CD8⁺ T-cell-mediated immune responses. These findings demonstrate that TET2 sulfhydration at C1186/1202 is a crucial post-translational modification for TET2 stability and function, playing a key role in epigenetic regulation and antitumor immunity, and highlight the immunoadjuvant potential of reactive sulfur species in melanoma immunotherapy.

DNA-damaging agents combined with agonists of the cGAS-STING pathway can effectively suppress colorectal cancer (CRC) by inducing cancer cell death and eliciting an antitumor immune response. In this study, we demonstrate that the natural compound Bruceine A (BA) inhibits CRC progression through a dual mechanism involving nuclear-to-cytoplasmic translocation of Ku70. Cytoplasmic Ku70 loses its canonical DNA repair function while simultaneously enhancing its interaction with cGAS, leading to increased cGAS oligomerization and elevated levels of double-stranded DNA (dsDNA), both of which amplify cGAS-STING signaling. Furthermore, Bruceine A-mediated Ku70 translocation exacerbates DNA damage accumulation, further enhancing tumor immunogenicity. In the murine CRC model, Bruceine A significantly inhibited tumor growth and enhanced tumor sensitivity to chemotherapy, radiotherapy, and anti-PD-1 treatment. Notably, genetic ablation of STING and CD8⁺ T cells in mice substantially abolished the antitumor effects of Bruceine A, confirming its reliance on cGAS-STING activation and adaptive immunity. Our findings establish Ku70 as a novel therapeutic target in CRC, where its subcellular redistribution disrupts genomic stability and bridges innate immune activation, synergistically promoting tumor cell death and antitumor immunity. Modulating Ku70 localization thus represents a promising strategy to enhance CRC treatment.

Through various murine models and 5 R-16 S sequencing, we identified Bacteroides thetaiotaomicron (B.t) as a sensitizer for the combined treatment of Apatinib and anti-PD-1 therapy. Clinically, B.t enrichment was associated with improved neoadjuvant treatment outcomes and a favorable prognosis in HCC patients. Mechanistically, B.t produces formic acid in tumor cells, inhibits aryl hydrocarbon receptor (AhR) nuclear translocation by methylation, and suppresses downstream pathways, thereby mitigating pro-angiogenic effects and immunosuppression. The addition of formate or AhR inhibitors with combined treatment significantly enhanced therapeutic efficacy in preclinical models.

RNA-binding proteins (RBPs) are emerging as crucial regulators in cancer, but the development of therapeutic strategies targeting RBPs remains limited. Here, through chemical proteomics approaches, we identify DIQ01 as a novel small-molecule inhibitor of KH-type splicing regulatory protein (KHSRP), an RBP that is aberrantly hyper-expressed in human tumors and plays an essential role in proliferation, metastasis, and tumor progression. DIQ01 specifically binds to the KH3 and KH4 domains (Phe358 as the key residue) of KHSRP, inhibiting its interaction with mRNAs. Furthermore, the binding of DIQ01 diminishes the PRMT5-mediated arginine methylation of KHSRP, a post-translational modification required for its oncogenic activity. Mechanistically, transcriptomic and proteomic profiling suggested that DIQ01 functionally inactivates KHSRP, triggering destabilization and downregulation of its target PLK1 mRNA. This leads to S-phase arrest, DNA damage, and apoptosis in HCT116 cells. Both in vitro and in vivo studies, including CRC xenograft and patient-derived organoid models, demonstrate that DIQ01 exhibits potent antitumor efficacy with minimal systemic toxicity. Our findings underscore the innovative approach of concurrently targeting the RNA-binding function of an RBP and its post-translational modification, highlighting DIQ01's unique mechanism and significant translational potential as a targeted cancer therapy.

Myeloproliferative neoplasms (MPNs) are a group of hematologic malignancies for which current treatment options remain limited, underscoring the urgent need to explore novel therapeutic targets and intervention strategies. Through a high-throughput screen of an epigenetic compound library, we identified the SIRT6 allosteric agonist MDL-800 as a potent suppressor of neutrophil hyperplasia. We established an endogenous sirt6-mutant zebrafish model that develops a myeloproliferative neoplasm (MPN)-like phenotype, with a 64% incidence in adult zebrafish. Mechanistically, Sirt6 was found to regulate neutrophil proliferation in vivo and in vitro by deacetylating histone H3K9 at the c-myb promoter. Sirt6 deficiency led to aberrant proliferation of neutrophils and hematopoietic stem/progenitor cells, whereas Sirt6 overexpression significantly alleviated neutrophil hyperplasia and MPN-related symptoms. Furthermore, the SIRT6 activator MDL-800 enhanced the efficacy of imatinib and reduced neutrophil proliferation in a zebrafish leukemia model. In xenograft mouse models, the combination of MDL-800 and imatinib significantly inhibited leukemia progression and restored drug sensitivity in imatinib-resistant cases. This study establishes the Sirt6-c-Myb axis as a core epigenetic pathway for myeloid homeostasis, providing a novel strategy for simultaneously suppressing neutrophil hyperplasia and enhancing chemotherapeutic efficacy in hematologic malignancies.

Parkinson's disease (PD), the second most prevalent neurodegenerative disorder, is characterized by progressive loss of dopaminergic neurons in the substantia nigra. Although the molecular mechanisms of PD remain incompletely understood, mitochondrial dysfunction has emerged as a central pathological driver, highlighting the urgent need for therapies targeting mitochondrial homeostasis. In this study, we demonstrate that rhynchophylline (Rhy), a bioactive alkaloid from Uncaria species, exerts neuroprotective effects by restoring mitochondrial dynamics. Thermal proteome profiling identified dihydrolipoamide acetyltransferase (DLAT) as a direct target of Rhy. Genetic ablation of DLAT induced mitochondrial fragmentation and abolished Rhy-mediated beneficial effects on mitochondrial structure and function. Mechanically, Rhy binds to the N-terminal lipoyl domain of DLAT, allosterically disrupting its interaction with sirtuin 4 (SIRT4) and subsequently enhancing DLAT lipoylation, a critical post-translational modification for mitochondrial energy metabolism. In vivo, Rhy administration ameliorated motor deficits and dopaminergic neurodegeneration in both the 6-OHDA-induced and A53T α-synuclein transgenic PD mouse models. Single-nucleus RNA sequencing further highlighted the clinical relevance of DLAT dysregulation in PD. Collectively, our findings establish Rhy as a promising PD therapeutic candidate and delineate DLAT as a pivotal node in therapeutic targets by promoting mitochondrial fusion and bioenergetics, offering a novel mechanistic avenue for neuroprotection.

In recent years, immunotherapy has shown obvious advantages in treating cancers. The close interaction between cancer cells and immune cells in the tumor microenvironment (TME) underlies the progression of glioblastoma multiforme (GBM). However, there are no effective immune-related targets against GBM. Here, in silico analyses and experimental data showed that Interferon Gamma Inducible Protein 30 (IFI30), modulated by histone modifications both H3K4me3 and H3K27ac, was up-regulated in GBM and had a potential role in the antitumor immune responses. In vitro and in vivo experiments further revealed that IFI30 modulated the infiltration of tumor-associated macrophages (TAMs) and reduced the proportion of CD8⁺ T cells. Mechanistically, IFI30 induced PGE2 expression in GBM cells via the MAFF/PTGS2 pathway, and PGE2 bound to macrophage EP2/EP4, activating the downstream ERK1/2 and KLF4/STAT6 pathways, stimulating the infiltration of TAMs. Taken together, we characterized the role and mechanisms of IFI30 in the malignant progression of GBM by regulating TAMs, highlighting that IFI30 may benefit GBM patients as a therapeutic target.

Irregular, curved, and uneven shapes of tumor vessels contribute to a malignant microenvironment, promoting metastasis. In this study, using patient-derived xenograft models, we observed that tumors derived from metastatic colorectal cancer (CRC) tissues exhibited increased vascular density, hypoxia, and permeability, but reduced perfusion and pericyte coverage, compared with tumors derived from non-metastatic CRC tissues. We conducted a high-throughput microarray analysis to determine the molecular mechanisms underlying compromised vessel structures. Dentin sialophosphoprotein (DSPP) was identified as the most upregulated gene in metastatic CRC with abnormal vasculature. Clinically, high DSPP expression is strongly correlated with poor prognosis and advanced CRC stages. DSPP stimulates tumor vessel abnormalization and CRC metastasis in vivo, and acts as a novel ligand of alpha(v)beta (3) integrin (αvβ3), which is predominantly expressed in tumor vessels. DSPP could directly bind to the peptide segment (amino acids 368-411) of αvβ3 on the cytoplasmic membrane of endothelial cells, activating the mitogen-activated protein kinase signaling pathway. Subsequently, therapeutic targeting of DSPP with human DSPP antibody or TFA, a selective inhibitor of the αvβ3, was investigated to effectively induce tumor vessel normalization and suppress tumor metastasis in mice. Additionally, interleukin-17 F (IL-17F) was identified as an upstream regulator of DSPP expression, which promotes DSPP transcription via p65 binding to its promoter. Therefore, targeting the DSPP/αvβ3 axis to promote tumor vessel normalization and inhibit tumor metastasis represents a new strategy for the clinical implementation of combination targeted therapies in patients with advanced CRC.

Renal fibrosis induces irreversible renal failure and lacks therapies. The tubular epithelial cells (TEC) largely depend on exosomes to create a pro-fibrotic microenvironment, initiating fibroblast activation. However, how exosome secretion of TEC is over-activated during renal fibrogenesis remains unknown. Herein, in vitro high-throughout screen uncovered acetyltransferase NAT10 as promising target to interfere with TEC-fibroblast communication. Further experiments showed that NAT10 was upregulated mainly in the TEC of fibrotic kidneys, and its conditional depletion in TEC alleviated renal fibrosis in vivo. Mechanistically, nuclear NAT10 coordinated with cytoplasmic NAT10 to promote exosome secretion of TEC to induce fibroblast activation via combining mRNA ac4C modification and lysine acetylation (Kac). Moreover, NAT10 upregulated the abundance of exosomal Gli in TEC, which were vital for fibroblast activation. In summary, NAT10-mediated orchestration of ac4C and Kac modifications is the mechanism of over-activated exosome secretion of TEC during renal fibrogenesis. And targeting NAT10 with Remodelin is a promising therapy for renal.

Effective therapy for relapsed or refractory central nervous system lymphoma (r/r CNSL) remains an unmet medical need. Meanwhile, developing antitumor drugs for CNS malignancies faces the dual challenge of achieving effective blood‒brain barrier (BBB) penetration and potent tumor cell killing. To address these challenges, a comprehensive predictive system was established to support decision-making during the discovery of HZ-A-018, a potent and BBB-permeable Bruton tyrosine kinase (BTK) inhibitor. This study further presents key preclinical results for HZ-A-018, as well as efficacy/safety data from a multicenter Phase 1 trial in r/r CNSL patients. HZ-A-018 demonstrated manageable safety, with only 19.2% of patients experienced grade 3 or higher adverse events according to the Common Terminology Criteria for Adverse Events version 5.0. Treatment with HZ-A-018 at the recommended phase II dose (RP2D) of 600 mg achieved an overall response rate (ORR) of 72.7% (95% CI, 39.0-94.0) and a 12-month survival rate of 90.5%. The Center for Drug Evaluation in China has authorized the initiation of this single-arm Phase II study as a pivotal registrational clinical trial for accelerated approval of HZ-A-018 for monotherapy in patients with r/r PCNSL. This trial has been registered under the identifiers ChiCTR2400091821 at www.chictr.org.cn and CTR20210181 at www.chinadrugtrials.org.cn.

Acute myeloid leukemia (AML) is a heterogeneous and devastating hematologic malignancy characterized by differentiation blockage and immature progenitor accumulation, positioning differentiation therapy as a promising therapeutic strategy. However, clinical success is largely confined to acute promyelocytic leukemia (APL) and isocitrate dehydrogenase (IDH)-mutated AML, leaving most AML subtypes with unmet needs. Herein, novel noscapine derivative ES428 is discovered that induces AML differentiation and exhibits potent anti-AML efficacy across diverse AML cell lines, primary patient samples, as well as cell line- and patient-derived xenograft models. Target deconvolution with combinatorial strategies identifies dihydroorotate dehydrogenase (DHODH), a rate-limiting enzyme in de novo pyrimidine synthesis, as the direct functional target. Integration of molecular dynamics simulations and comprehensive structure-activity relationship studies elucidates ES428’s unique mechanism via simultaneous engagement with DHODH and mitochondrial membrane lipids. This dual-engagement underpins ES428’s enhanced target engagement, efficacy, and selectivity in physiologically relevant mitochondrial membrane environment, potentially through stabilizing ES428-DHODH interaction in situ and facilitating ES428’s selective mitochondrial localization. Furthermore, ES428 triggers a mechanistic cascade linking decreased pyrimidine synthesis, reduced O-linked N-acetylglycosylation (O-GlcNAcylation), EP300/CREBBP catalytic inhibition, and transcriptional reprogramming. Our findings identify promising lead candidates, establish a novel DHODH-targeting strategy, and provide important mechanistic insights to advance differentiation therapies for myeloid malignancies.

Dual FLT3/HDAC inhibition represents a promising synergistic strategy to address tumor heterogeneity. Building upon our prior lead 25h, we developed novel 6-ethylpyrazine-2-carboxamide derivatives via systematic structural optimization to enhance pharmacokinetic properties and target selectivity. The optimized compound, CF-2-17, demonstrated potent dual inhibition of FLT3 (IC₅₀ = 1.1 nmol/L) and HDACs (IC₅₀ = 9.6 nmol/L), and exhibited a 27-fold selectivity for HDAC1 over HDAC6. Its improved physicochemical properties, including enhanced solubility and metabolic stability, translated into favorable plasma exposure in vivo. In the MOLM-13 (FLT3-ITD) xenograft model, oral administration of CF-2-17 showed antitumor efficacy comparable to combination therapy, without observable toxicity. CF-2-17 also exhibited antiproliferative activity against non-FLT3-ITD hematological malignancies and solid tumors, outperforming single-target agents. Furthermore, CF-2-17 effectively remodeled the tumor immune microenvironment through CD4⁺ T cell activation and IFN-γ elevation, achieving 87% tumor growth inhibition in LLC syngeneic models. Mechanistically, CF-2-17 reversed FLT3 blockade-induced DC dysfunction via activation of the NF-κB pathway, thereby reinstating DC-mediated antitumor immunity. This dual FLT3/HDAC inhibitor demonstrates synergistic epigenetic-immune modulation, offering a promising approach for heterogeneous malignancies.

Mismatch repair-proficient (pMMR)/microsatellite-stable (MSS) tumors, representing the majority of solid cancers, remain largely unresponsive to immune checkpoint inhibitors, highlighting the urgent need for effective therapeutic strategies. Tumor cell vaccines, serving as delivery platforms for a broad repertoire of tumor antigens, hold potential for eliciting systemic antitumor immunity, yet their clinical efficacy has remained limited. Here, we developed Tβ1KO/PolyIC-Vac, an off-the-shelf therapeutic whole-tumor cell vaccine generated by CRISPR-Cas9-mediated TGF-β1 knockout and formulated with the TLR3 agonist Poly(I:C). In therapeutic vaccination models, subcutaneous vaccination significantly suppressed distal tumor growth, reduced lung metastases, and delayed spontaneous colorectal tumor progression. It also synergized with anti-PD-1 blockade, enhancing complete response rates and establishing baseline antitumor immunity. Mechanistically, knockout of tumor-derived TGF-β1 reprogrammed immunity by reducing MDSC accumulation to relieve local immunosuppression and activate dendritic cells in draining lymph nodes, while intrinsically triggering the tumor STAT1-IL-15 axis to enhance vaccine immunogenicity. Poly(I:C) further enhanced cross-presentation and adaptive immunity, conferring long-term cross-protection against heterologous tumors. Together, these findings establish Tβ1KO/PolyIC-Vac as a universal tumor vaccine platform capable of efficiently delivering a comprehensive repertoire of tumor antigens and eliciting robust, durable, and cross-protective tumor-specific immunity in immunotherapy-resistant pMMR/MSS tumors.

The approval of radionuclide therapy strategies in nuclear medicine has revolutionized the treatment landscape for patients with advanced malignancies. Due to significant HER2 overexpression in some solid tumors, radionuclide therapy targeting HER2 is a viable strategy. The traditional monoclonal antibody (mAb) direct radiolabelling system may lead to off-target radiation exposure. To address this limitation, we employed the established inverse-electron demand Diels-Alder (IEDDA)-based pretargeting strategy and designed a novel tetrazine probe. First, we identified the optimal targeting molecule (Pertuzumab) and optimal metabolic time (48 h) through micro-positron emission tomography/computed tomography (PET/CT) scans and biodistribution studies of [⁸⁹Zr]Zr-DFO-Per, [⁸⁹Zr]Zr-DFO-Per-F(ab')₂ and [⁸⁹Zr]Zr-DFO-Per-Fab. Next, TCO-Pertuzumab (TCO-Per) was administered to HER2-overexpressing SKOV3 tumor-bearing mice models, and after 48 h of circulation and clearance, a novel radiolabelled small molecule Tz ([¹³¹I]I-Tyr-d-peptide-PEG₁₁-Tz) was introduced. Through a series of in vivo studies, we observed prolonged retention of [¹³¹I]I-Tyr-d-peptide-PEG₁₁-Tz in SKOV3 tumors with rapid renal clearance, along with promising therapeutic efficacy. Our study demonstrates that the novel tetrazine probe within a pretargeting delivery system overcomes limitations of traditional strategies and shows promise for clinical translation.

Hypothyroidism, caused by insufficient thyroid hormone production or impaired responsiveness, is typically managed with lifelong daily oral levothyroxine sodium (LT₄). However, oral administration often suffers from variable gastrointestinal absorption and poor adherence. Here, we report a mussel-inspired biphasic microneedle (MN) patch integrating LT₄ microcrystals and a bioadhesive PDA-PAM hydrogel backing for sustained and patient-friendly hypothyroidism management. The biphasic MNs, composed of rapidly dissolving PVA/sucrose tips and a robust polystyrene base, efficiently deposit LT₄ microcrystals intradermally upon insertion, enabling gradual dissolution and sustained release. The hydrogel backing offered strong and flexible adhesion, ensuring reliable skin attachment during movement. In vitro, the patches exhibited first-order LT₄ release for 9 days. In hypothyroid rats, a single patch application maintained therapeutic plasma LT₄ concentrations for ∼6 days, with bioavailability comparable to intravenous injection. In a preliminary study in healthy volunteers (n= 13), placebo MN patches demonstrated high insertion efficiency, good short-term tolerability, and favorable user acceptability. While further studies are required to establish long-term safety and efficacy in patients with hypothyroidism, these findings support the feasibility of a minimally invasive, extended-interval LT₄ delivery strategy.

Peritoneal metastatic colorectal cancer (PMC) is highly aggressive and resistant to anti-angiogenic monotherapy due to the angiogenesis-immunosuppression vicious cycle. This study develops dual-ligand modified nanoparticles (Reg/DMX@BPF NPs), co-loaded with the angiogenesis inhibitor regorafenib (Reg) and the stimulator of interferon genes (STING) agonist DMXAA (DMX). Reg prevents DMX aggregation as a molecular scaffold via π-π stacking. The folic acid (FA) and phenylboronic acid (PBA)-functionalized BSA (BPF) facilitates active tumor targeting and metastatic site enrichment. Upon internalization into lysosomes, the acidic pH triggers boronate ester bond formation between PBA and glycoproteins, inducing lysosomal disruption and efficient cytosolic release. In addition to STING activation, the BPF potently activates toll-like receptor 4 signaling, synergistically inducing M1 tumor-associated macrophages polarization and dendritic cells maturation. In vivo results demonstrate that Reg/DMX@BPF NPs synergistically inhibit tumor proliferation, normalize pathological vasculature, and reprogram the immunosuppressive microenvironment, which leads to reduced tumor burden and ascites. Collectively, the targeted lysosome-escape nanoparticles provide a novel strategy to overcome the poor efficacy of anti-angiogenic therapy against PMC.

Drug-induced liver injury (DILI) is a predominant cause of acute liver failure, intricately associated with excessive reactive oxygen species (ROS) production and the activation of inflammatory cascades. In this study, a metal-drug coordinated nanozyme (M-dCNs) is rationally designed to simultaneously scavenge ROS and polarize Kupffer cells to alleviate DILI. Specifically, the coordination between ferric ions (Fe³⁺) and the complementary therapeutic agents of hesperetin (HST) and KPLH (KPLH1130) facilitates the formation of M-dCNs with uniform particle distribution and enhances structural stability. Notably, M-dCNs exhibits intrinsic antioxidant enzyme-mimetic activity, efficiently neutralizing intracellular ROS in damaged hepatocytes and thereby disrupting apoptosis-related signaling pathways. In parallel, M-dCNs synergistically reprograms hepatic macrophages toward an anti-inflammatory phenotype, leading to a marked reduction in pro-inflammatory cytokines such as tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β), and ultimately suppressing inflammatory cascades. In vivo studies demonstrate the hepatoprotective potential and therapeutic efficacy of M-dCNs in attenuating oxidative stress, reducing inflammatory cell infiltration, and restoring liver function. Collectively, this work presents a promising strategy for the treatment of DILI via the concurrent modulation of redox homeostasis and inflammatory microenvironment.

Terpenoids are key specialized metabolites in Cannabis sativa, shaping cultivar-specific aromas and potentially modulating cannabinoid effects. This study provided a comprehensive analysis of the terpene synthase (TPS) gene family in C. sativa, integrating haplotype-resolved genomic data, volatile terpene profiling, transcriptomics, and functional assays. The comprehensive volatile terpene profiling across 28 spatiotemporal samples spanning weekly developmental intervals and distinct maturity stages from six cultivars identified 227 cannabis volatile terpenes, including 88 monoterpenes and 139 sesquiterpenes, exhibiting distinct tissue, developmental stage, and cultivar-specific patterns. Comparative expression and co-expression network analysis revealed coordinated regulation between MEP/MVA pathways, TPS, and cannabinoid genes, underscoring a shared metabolic foundation. Genome annotation identified 41 full-length CsTPSDK genes, exhibiting extensive expansion and subfamily-specific clusters with structural divergence between haplotypes. Transcript profiling across developmental stages and cultivars distinguished a core set of highly expressed inflorescence-associated CsTPSs from genes exhibiting cultivar-specific regulation. Functional characterization of six previously unreported CsTPSDKs uncovered diverse mono- and sesquiterpene synthase activities, including unexpected substrate promiscuity across subfamilies. These findings deliver the most comprehensive functional annotation of the C. sativa TPS repertoire to date, elucidating the genetic and biochemical bases of terpene diversity and providing a foundation for targeted metabolic engineering and cultivar improvement.