The intratumoral microbiota, as an important component of the tumor microenvironment (TME), impact tumor progression by regulating the arginine-ornithine metabolic axis. It has become a new frontier in tumor research. Arginine is a crucial amino acid in TME, and its metabolites, ornithine and polyamines, directly promote tumor proliferation and induce immunosuppression. Intratumoral microbiota mainly exert their effects through two direct pathways: 1) arginine depletion, such as Streptococcus in gastric cancer. Specific intratumoral microbiota highly express arginine deiminase (ADI) or arginase (Arg) to consume arginine in the TME, leading to T cell dysfunction and enhancing immunosuppressive cells. 2) Ornithine/polyamines supplement, such as fusobacteria in esophageal cancer produce putrescine. The microbiota converts arginine into ornithine, which is then synthesized into polyamines, directly stimulating tumor cell proliferation and reshaping the immunosuppressive TME. Additionally, the metabolic products from the microbiota like short-chain fatty acids (SCFAs) and indole substances, can amplify these effects through signaling pathways including G protein-coupled receptor 43 (GPR43) and aryl hydrocarbon receptors (AHR). The regulation of intratumoral microbiota-arginine metabolism axis has a "double-edged sword" characteristic, relying on the metabolic dependence of the different tumors, which provides a basis for precise treatment. Furthermore, strategies targeting the axis present great potential, including Arg1 inhibitors (CB-1158) in combination with immunotherapy, engineered probiotics to supply arginine and inhibit polyamine synthesis in situ within the TME. These advancements also indicate there is enormous progress from exploring the intratumoral microbiota-metabolism interaction to developing novel tumor microecological therapies.

Breast cancer chemotherapy faces severe limitations due to multidrug resistance and metastasis. Herein, we report a rationally designed ORI-LDH@LP (OLL) nanoplatform that establishes a self-reinforcing vicious cycle among endoplasmic reticulum (ER) stress, mitochondrial calcium overload, and oxidative stress to drive irreversible tumor cell apoptosis and reshape the immunosuppressive tumor microenvironment. This nanoplatform comprises oridonin-loaded Mn2+-based layered double hydroxides encapsulated within folate-targeting liposomes. Upon internalization, released oridonin triggers intense ER stress, activating the PERK-eIF2α-ATF4-CHOP axis and upregulating the IP3R-GRP75-VDAC complex at mitochondria-associated ER membranes to facilitate massive Ca2+ flux into mitochondria. This targeted calcium overload induces mitochondrial permeability transition pore opening, membrane potential collapse, ATP depletion, and reactive oxygen species (ROS) burst. Concurrently, released Mn2+ catalyzes hydroxyl radical generation via Fenton-like reactions, synergistically amplifying oxidative damage. This interplay between ER stress-mediated calcium signaling and Mn2+-catalyzed ROS generation creates a self-amplifying cascade that drives mitochondrial-dependent apoptosis. Beyond direct cytotoxicity, the ROS storm triggers immunogenic cell death, while Mn2+ robustly activates the cGAS-STING pathway, establishing a positive feedback loop that amplifies innate immune signaling. This dual mechanism promotes type I interferon production, driving tumor-associated macrophage repolarization from the protumoral M2 to the antitumoral M1 phenotype and enhancing cytotoxic T lymphocyte infiltration. In 4T1 tumor-bearing mice, OLL achieved remarkable tumor suppression without systemic toxicity, demonstrating potent antitumor efficacy through combined direct killing and systemic immune activation. This study establishes a nanomaterial-driven paradigm for precisely modulating ER-mitochondria crosstalk to overcome drug resistance and ignite antitumor immunity, offering a promising strategy for cancer nanotherapeutics.

Abstract Background Endometrial cancer (EC) is a common malignancy of the female reproductive tract worldwide. While comprehensive genomic analyses have identified multiple genetic alterations in EC, disease models remain relatively rare, limiting the ability to conduct disease studies. Establishing a hybrid in vitro and in vivo model of EC and comparing it to its in vivo counterpart could greatly advance research into EC mechanisms and treatment strategies. Methods We established genetically engineered EC mice ( Pten flox/+ ; Stk11 flox/flox ; LTF-Cre (PSC)) and constructed a corresponding epithelial cell line from isolated uterine tumor tissues using conditional reprogramming techniques. The cell line was successfully passaged, frozen, and stored in a biobank, providing a valuable resource for future EC research. Results The PSC-derived cell line stably retained the mutant alleles of Pten and Stk11 during passaging, preserving the genetic profile of the original tumor. In addition, we performed various functional assays on isolated mouse cell linesand formed tumors after subcutaneous implantation of cells in immunodeficient mice and C57BL/6 mice with an intact immune system. Hematoxylin and eosin (H&E) staining and immunohistochemical staining were used to characterize the histopathology and biomarkers of the subcutaneously implanted tumors, which were highly similar to those of the parental mouse uterine tumors. Conclusions The isolated mouse cell line provides a powerful platform for basic and translational EC studies, matching well with the corresponding in vivo models. The fidelity of the cell line to human-relevant genetic alterations and tumor biology makes it a valuable tool for investigating EC pathogenesis and evaluating novel therapeutic approaches.

Calcium overload exhibits significant anti-tumor potential by inducing abnormal intracellular Ca2+ accumulation, which disrupts mitochondrial and endoplasmic reticulum (ER) functions, thereby triggering apoptosis. However, its clinical application is currently hindered by challenges such as poor tumor-targeting capabilities, insufficient tumor accumulation, and incomplete mechanistic understanding. This review systematically analyzes the structural and functional coupling between the ER and mitochondria to elucidate the mechanisms of calcium overload-mediated cell death. We highlight how Ca2+ acts as a critical trigger to amplify mitochondria-associated ER stress, fostering a self-amplifying loop of crosstalk that initiates tumor cell death pathways. Furthermore, we summarize recent advances in targeted Ca2+ delivery using calcium-based nanocarriers combined with emerging modalities like sonodynamic therapy (SDT) and photothermal therapy (PTT), highlighting their synergistic antitumor potential. Compared with previous reviews, this work focuses on recent calcium-based nanosystems, sequential ER-mitochondria damage during Ca2+ overload, the MAM-associated IP3R-GRP75-VDAC1-MCU axis, and future strategies for tumor-targeted, TME-responsive, and multimodal synergistic therapy. By summarizing current research, this review aims to provide a prospective outlook for the novel anti-cancer therapies that target the disruption of intracellular Ca2+ homeostasis.

AACOCF3, a cell-permeable arachidonic acid analogue, is widely established as a selective inhibitor of cytosolic phospholipase A2 (cPLA2, PLA2G4A) in studies of metabolic disorders. Although its primary mechanism involves cPLA2 inhibition, emerging evidence indicates that AACOCF3 may target additional protein entities, exemplified by calcium-independent phospholipase A2 (iPLA2, PLA2G6) and fatty acid amide hydrolase (FAAH). Notably, cPLA2 displays a markedly heterogeneous expression profile in non-small cell lung cancer (NSCLC). Our findings establish that AACOCF3 exerts more potent growth inhibition in cPLA2-negative NSCLC cells, with IC50 values of 15.13 μM for H1975 and 15.84 μM for PC9 cells, in contrast to the cPLA2-positive A549 cells (IC50 = 56.23 μM). Mechanistically, AACOCF3 upregulates IFN-α/β signaling-associated genes (e.g., IFNB1, ISG15) specifically in cPLA2-negative NSCLC cells. This aligns with TCGA-LUAD data revealing that PLA2G4A-low tumors predominantly engage immune-activation pathways rather than metabolic programs when compared to PLA2G4A-high counterparts. Through integrated molecular docking and surface plasmon resonance (SPR) analysis, we identified structure-specific recognition protein 1 (SSRP1) as a direct molecular target of AACOCF3 in cPLA2-negative NSCLC, with SPR binding studies confirming a stable interaction (Kd = 25.9 μM). Ectopic SSRP1 expression abrogated AACOCF3-induced phenotypic alterations, concurrently suppressing IFN-α/β signaling. Collectively, these results provide evidence that AACOCF3 exerts its anti-proliferative effect by targeting SSRP1, which leads to the activation of the IFNα/β pathway, thereby underscoring its therapeutic promise for the cPLA2-negative patient subpopulation.

Triple-negative breast cancer (TNBC) remains difficult to treat because of the lack of effective therapeutic targets and limited treatment options. Dual targeting of Focal adhesion kinase 1 (FAK1) and 2 (FAK2) has emerged as a promising strategy, yet current dual-targeted ligands are still limited in structural diversity and binding mode. In this study, a pyrazolo[3,4-b]pyridine scaffold was identified through virtual screening and optimized to generate 16 derivatives. Among them, compound 8a showed the strongest binding affinity for both FAK1 and FAK2, with KD values of 2.9 μM and 2.5 μM, respectively, and exhibited the best selectivity toward TNBC cells. Intracellular target engagement studies indicated that 8a protected specific peptides within the kinase domains of FAK1 and FAK2 in cells. Functionally, 8a induced G2/M-phase arrest and activated DNA damage-related signaling, accompanied by decreased phosphorylation of CDK1. The antiproliferative activity of 8a was markedly attenuated by FAK1/FAK2 knockdown but significantly enhanced in combined treatment with doxorubicin. In addition, 8a inhibited 4 T1 tumor growth in vivo without significant body weight loss. These findings identify pyrazolo[3,4-b]pyridine as a new chemotype for dual FAK1/FAK2 targeting and support 8a as a target-dependent lead compound for anti-TNBC development.

Background:Frailty and systemic inflammation/immune-protein reserve may drive adverse outcomes after endometrial cancer (EC) surgery in older patients, but their incremental predictive value is unclear. Methods:We analyzed a retrospective cohort of women ≥65 years undergoing primary EC surgery between January 2016 and December 2024. Frailty was assessed using mFI‑5. Inflammation-nutrition status (INS) was derived from routine preoperative laboratory tests as exploratory composite. The primary endpoint was 30‑day severe complications (Clavien-Dindo III-V), and the secondary endpoint was 90‑day unplanned readmission to the index hospital. One‑year non-endometrial cancer (non‑EC) death was exploratory with EC death as a competing event. Logistic regression and Fine-Gray models were fitted. Incremental prediction versus a prespecified baseline perioperative clinical model was evaluated with bootstrap optimism‑corrected discrimination, calibration, and decision‑curve analysis. Results:Severe complications occurred in 62 (7.1%) and readmission in 71 (8.1%). One‑year non‑EC and EC deaths were 28 (3.2%) and 19 (2.2%), respectively. mFI‑5 and INS predicted severe complications (OR per 1‑point mFI‑5 1.43 [95% CI 1.22-1.67]; OR per 1‑SD INS 1.34 [1.15-1.55]) and readmission (OR 1.28 [1.10-1.50]; OR 1.22 [1.05-1.41]). For non‑EC death, mFI‑5 (SHR 1.52 [1.23-1.88]) and INS (SHR 1.37 [1.11-1.69]) were significant. Adding mFI‑5+INS improved discrimination (AUC 0.69 to 0.77 for complications; 0.63 to 0.70 for readmission) and the time-dependent C-index for the exploratory competing-risk endpoint (0.66 to 0.73), with acceptable calibration. Decision-curve findings were descriptive rather than definitive. Conclusion:Combining frailty with an objective inflammation-nutrition risk composite modestly improved internal model performance for 30-day severe complications and 90-day readmission in older EC patients, and 1-year non-EC death analysis remains exploratory. These findings should be interpreted as perioperative rather than purely preoperative prediction, and decision thresholds require external validation before protocolized use.

Therapeutic challenges in endometrial carcinoma (EC) arise from the limited efficacy and toxicity of current treatments. Although exosome-based RNA interference shows promise, its clinical translation is hindered by inefficient cargo loading, low yields, and poor tumor targeting. We have engineered an exosome platform (cRGD-ExoM) that integrates the following innovations: Firstly, RNA motifs enable the enrichment of shRNA loading by over 80-fold for targeting of ferroptosis regulators (glutathione peroxidase 4/ferroptosis suppressor protein 1/ferritin heavy chain [GPX4/FSP1/FTH]). Secondly, Rab4 silencing amplifies exosome biogenesis via dysregulated endosomal recycling, enhancing tumor cell uptake by impairing endosome maturation-a dual-action mechanism that boosts both production and delivery. Thirdly, cRGD peptides confer αvβ3-integrin-specific targeting. cRGD-ExoM induces potent ferroptosis by increasing lipid peroxidation and downregulating GPX4/FSP1/FTH, significantly suppressing EC tumor growth in vivo without causing systemic toxicity. The platform's modular design allows for spatiotemporal control of loading, production, and targeting, demonstrating its scalability. This study provides new insights into the precision treatment of endometrial cancer by developing engineered, multifunctional, exosome-based therapeutic drugs that combine mechanism precision and translational feasibility in tumor treatment.

The neurokinin-3 receptor (NK3R) has emerged as a promising non-hormonal therapeutic target for menopausal hot flashes, with fezolinetant being the only clinically approved NK3R antagonist to date. To overcome this therapeutic limitation, we designed a series of imidazolepiperazine derivatives (17a-17c, 21, 23a-23u), among which 23i(R) demonstrated superior pharmacological properties including potent NK3R inhibition (IC50 = 65.42 ± 6.54 nM), strong target binding (IC50 = 53.61 ± 3.67 nM), excellent membrane permeability (Papp A-B = 27.3 × 10-6 cm/s; ER = 0.53), and remarkable oral bioavailability (165 %). In ovariectomized rat models, 23i(R) effectively suppressed luteinizing hormone levels while exhibiting favorable pharmacokinetics and tolerability, establishing it as a promising clinical candidate for further development as a next-generation NK3R antagonist.

Uterine leiomyosarcoma (uLMS) is a rare but aggressive malignant mesenchymal tumor, accounting for 2–5

Bacteria-infected macrophages undergo pyroptosis to release inflammatory cytokines, which contributes to host defense. It has been known that activated macrophages involve metabolic reprogramming. However, the metabolic changes and the role of metabolites in pyroptotic macrophages are not fully understood. Here, we revealed that aerobic glycolysis product, lactate, could promote NLRP3 inflammasome activation induced pyroptosis. We found that endogenous lactate facilitates ASC recruitment to NLRP3 cores on the organelle membrane, thus inducing NLRP3 inflammasome complex formation. Mechanistically, we identified NLRP3 as a target protein modified by lactate, which is lactylated by AARS2. We confirmed lactylated sites on NLRP3 by LC-MS/MS analysis and verified that lactylation at K24 and K565 of NLRP3 facilitates inflammasome activation in macrophage. In vivo, inhibition of lactate production alleviates inflammatory responses in polymicrobial sepsis. Overall, our results indicate the role of lactate in regulating macrophage pyroptosis and the crosstalk between metabolism and innate immunity.

Tandem rearrangement-amination of biomass-derived furan alcohols (FAs) with nitrogenous compounds to produce N-cyclopentylanilines is essential for fine chemicals. However, this process is hindered by the in situ poisoning of acidic sites by basic amines over conventional metal-acid bifunctional catalysts. Herein, Ce-doped cobalt metaphosphate catalysts (CoP2/Ce-Co2P4O12), featuring CoP2 nanoparticles and oxygen vacancies (Ov), were fabricated via the precipitation of Ce-doped Prussian blue analogs, followed by oxidation and high-temperature phosphidation. Mechanistic investigation reveals that Ov promotes in situ hydrogen spillover from CoP2 nanoparticles to the Co2P4O12 support, forming H+-H− pairs. These pairs function as both Brønsted acid sites for 1,4-pentanedione aldol condensation and unconventional active sites for C=N bond hydrogenation. Consequently, a one-pot synthesis of N-cyclopentylaniline was achieved for the first time at a low temperature (120 °C), affording an exceptional yield of 86.5%. Furthermore, the catalyst exhibited broad generality for various nitrogen sources (e.g., NH3·H2O, amino compounds, and nitrobenzene) and FAs (e.g., 5-methyl furan alcohol and 2,5-dihydroxymethyl furan) via a similar reaction route. This work demonstrates that Ov-mediated hydrogen spillover can dynamically generate robust bifunctional H+-H− pairs to overcome basic poisoning, offering a new design strategy for cascade catalysis involving basic nitrogen compounds.

Abstract Background Endometrial cancer (EC) is a gynecological malignancy that originates from the endometrial epithelium and has a poor prognosis when advanced, recurrent, or metastatic. The limited therapeutic efficacy and severe adverse effects of conventional chemotherapy in advanced EC highlight the urgent need to develop more effective therapeutic drugs. Accumulating clinical evidence has revealed that natural compounds possess pharmacological advantages, including low toxicity and multi-target mechanisms. Erianin is a natural, small-molecule compound isolated from Dendrobium chrysotoxum Lindl that has multiple pharmacological effects. However, the effects of erianin on EC have not been confirmed and its anticancer mechanisms remain unclear. Methods Erianin was identified as a potent natural compound against EC through compound library screening. CCK-8 assays, colony formation assays, Edu experiments, and Live/Dead cell staining assays were used to analyze the anti-proliferative activity of erianin. Morphological characteristics, transmission electron microscopy, lactate dehydrogenase release assays, and western blot assays were used to evaluate the activation of pyroptosis. A transcriptome sequencing analysis was conducted to identify the potential mechanism of erianin. Biotin–erianin was synthesized and 20-k human proteome microarray was used to identify its direct targets. Molecular docking and cellular thermal shift assays (CETSA) were used to investigate whether erianin would bind to YTH domain family proteins (YTHDF1). To evaluate the in vivo therapeutic potential of erianin, an EC xenograft model was established and mechanistic investigations incorporating hematoxylin and eosin and immunohistochemical (IHC) staining, and western blot assays were conducted. Results Erianin inhibited the cell proliferation of EC cells and promoted pyroptosis through the caspase-3/gasdermin E (GSDME) pathway. Mechanistically, a crucial role for FOXM1/RRM2-mediated DNA damage in erianin-induced pyroptosis was established. Protein microarrays indicated that erianin–biotin directly targeted the m6A reader, YTHDF1. Erianin was confirmed to bind to YTHDF1 using molecular docking and CETSA. Molecular studies indicated that erianin inhibited YTHDF1, recognized m6A-modified FOXM1, and promoted FOXM1 mRNA degradation, which led to DNA damage and caspase-3-mediated GSDME cleavage. Erianin also substantially inhibited EC tumor growth in EC models. Conclusion Erianin directly targeted YTHDF1 to suppress the FOXM1/RRM2 axis and consequently promoted caspase-3/GSDME-dependent pyroptosis in EC cells. Our findings provide a new strategy for further clinical exploration of EC. Graphical Abstract

Although strategies combining multidrug resistance (MDR) reversal agents with chemotherapeutic drugs have demonstrated efficacy in clinical practice, their widespread application remains limited by potential toxicity. Natural thiazole-containing macrocycles have displayed promising MDR reversal potential. However, the 18-membered macrocycles bistratamides, despite their structural similarity to the known MDR reversal agent dendroamide A, remain unexplored regarding potential roles as MDR reversal agents and their molecular targets. In this study, we report the synthesis and initial discovery of bistratamides as potential MDR reversal agents. Structure-activity relationship analysis revealed that the oxazoline or oxazole motifs embedded within the macrocyclic framework are essential for MDR reversal activity in bistratamides. The oxazoline-containing bistratamide derivative 15 demonstrated the greatest synergistic anticancer effect among the examined compounds, exhibiting > three-fold greater activity than verapamil (VRP) with no observable toxicity in KBV cells and normal human podocytes. The MDR reversal effect of 15 in combination with paclitaxel was further validated through combination assays and flow cytometry. Subsequent rhodamine 123 (Rh123) accumulation assays and western blotting confirmed that 15 exerts its MDR reversal activity by inhibiting P-gp efflux function rather than affecting its expression. Our findings suggest that 15 represents a promising starting point for the development of novel chemosensitizing agents.

Metastatic castration-resistant prostate cancer (mCRPC) represents the terminal and most challenging stage of prostate cancer progression, characterized by limited therapeutic options and poor prognosis. Although AKT inhibitors have been explored as a potential treatment strategy for mCRPC, their efficacy as monotherapy is often constrained by the rapid development of resistance. Emerging evidence indicates that this acquired resistance is frequently associated with the elevated expression of the c-MYC oncogene. Bromodomain and extraterminal (BET) inhibitors, such as those targeting BRD4, can disrupt the interaction between BRD4 and chromatin, thereby attenuating c-Myc-driven oncogenic signaling. To address this dual-pathway challenge, we designed and synthesized a novel series of dual BRD4/AKT inhibitors. Through structure-activity relationship analyses, compound 21d was identified as a potent dual inhibitor, exhibiting activity against both BRD4 and AKT1 with IC50 values of 66.12 ± 7.69 nM and 143.81 ± 12.21 nM, respectively. In vitro, 21d effectively modulated key downstream effectors of both AKT1 and BRD4, leading to significant suppression of mCRPC cell proliferation, migration, and colony formation. Mechanistically, 21d induced G0/G1 phase cell-cycle arrest by downregulating phosphorylated RB (p-RB), cyclin E1, and CDK2. Concurrently, 21d elevated the protein levels of the autophagy marker LC3B, promoting apoptosis in 22Rv1 cells. In vivo, 21d demonstrated a favorable pharmacokinetic profile and markedly inhibited tumor growth in an mCRPC xenograft mouse model, achieving a tumor growth inhibition (TGI) rate of 62.0%. Collectively, our findings establish 21d as the first-in-class dual BRD4/AKT inhibitor, offering a promising therapeutic strategy to overcome c-Myc-associated resistance in mCRPC.

Rationale: Given the crucial role of the Nrf2 pathway in cellular adaptability to stress, targeting small-molecule activation of Nrf2 represents a promising therapeutic strategy for acute ischemic stroke (AIS). However, the clinical translation of existing Nrf2 activators is hindered by adverse effects, such as liver damage, and none are currently approved for AIS. Therefore, we aimed to develop a novel Nrf2 activator that specifically activates neuronal Nrf2 while mitigating adverse effects, with the goal of providing a lead compound for AIS. Methods: We validated the anti-AIS effects and mitochondrial protective functions of the novel Nrf2 activator Cpd.51 through multiple in vivo and in vitro experiments. Mechanistic studies involving surface plasmon resonance, cellular thermal shift assay, co-immunoprecipitation, chromatin immunoprecipitation, GST pull-down, and RNA sequencing were used to determine how Cpd.51 activates Nrf2. A comparative toxicological evaluation was conducted to demonstrate its superior safety profile over parent compound (Omaveloxolone). Results: Cpd.51 exhibited favorable blood-brain barrier permeability, improved safety profile, enhanced mitochondrial function protection and significant neuroprotective effect through the specific activation of neuronal Nrf2. Mechanistically, Cpd.51 interacted with Cys151 and Gly148 in the BTB domain of Keap1, inhibiting Nrf2 degradation, consequently suppressing the transcription of its downstream target DHRS3, a member of the short-chain dehydrogenase/reductase family. Furthermore, Cpd.51 exerted additional Nrf2-activating activity by disrupting protein-protein interactions between Nrf2 and DHRS3. Conclusions: Our work identified Cpd.51 as a novel and safe Nrf2 activator and unveils a unique feedback mechanism involving Nrf2-DHRS3 interaction, providing a new therapeutic avenue for AIS.

While type I interferon is critical for viral clearance in host defense, its production needs to be tightly regulated in both physiological and infectious contexts. Herein, we reported that an Iroquois homeobox protein IRX2 negatively regulates the activation of type I interferon signaling during influenza infection. We found that IRX2 binds to the IRF3 and negatively regulates IRF3 activation and IRF3-mediated interferon signaling. Knockdown of IRX2 upregulated the phosphorylation of IRF3 and augmented the production of type I interferon cytokines and antiviral immunity. We further showed that genetic abrogation of IRX2 protected mice from lethal influenza infection. Our results reveal a previously unrecognized non-transcriptional function of IRX2, and suggest a potential therapeutic target in viral infections.

ABSTRACT Background Colorectal cancer (CRC) incidence has risen significantly in China, potentially linked to dietary Westernization and increased consumption of high‐fat diets (HFD). Methods This study examines the differential effects of various fatty acids—saturated (palmitic acid, PA), monounsaturated (oleic acid, OA), and polyunsaturated (arachidonic acid, AA; docosahexaenoic acid, DHA)—on CRC progression, focusing on the IDO1/AhR signaling pathway. Results Clinical data indicate that CRC patients exhibit elevated serum lipid levels, with PA promoting cell proliferation, migration, and invasion more strongly than unsaturated fatty acids. The pro‐tumorigenic effects of PA are enhanced in the presence of lipopolysaccharide (LPS), suggesting an interaction between diet and gut microbiota. In vivo experiments corroborate that a high‐PA diet significantly elevates tumor growth and IDO1 expression compared to DHA. Mechanistic analyses reveal that PA and LPS co‐stimulation activates the IDO1‐AhR‐PI3K/Akt‐NF‐κB pathway, which is implicated in CRC progression. Conclusions These findings suggest that saturated fatty acids, particularly PA, may exacerbate CRC development risk through metabolic dysregulation, highlighting the potential of IDO1 as a biomarker for CRC associated with HFD. Reducing dietary saturated fat intake may thus be a viable strategy for CRC prevention.

E3 ligases are crucial to PROTAC technology, and identifying novel E3 ligase ligands could accelerate the advancement of PROTACs. DCAF11 has shown considerable potential for PROTAC applications. However, the ligands targeting DCAF11 remain limited, highlighting the need for the development of novel ligands for this E3 ligase. In this study, leveraging previous research on DCAF11 ligands, we designed a class of arylidene-thiazoldione scaffolds and applied it to develop PROTACs, resulting in the identification of a potent BRD4 degrader, LGF308. Degradation activity and mechanistic studies demonstrated that the compound LGF308 efficiently induces BRD4 degradation through the proteasomal pathway and via recruitment of DCAF11. This scaffold represents a reliable ligand, capable of facilitating the degradation of various proteins, including CDK4/6, BTK, and FKBP12. Therefore, this study introduces the arylidene-thiazoldione scaffold as a novel DCAF11 ligand and validates its application in PROTAC design, providing strong support for the development of DCAF11-based PROTACs.

BACKGROUND:Obesity is a serious multifactorial disease that involves epigenetic mechanisms like DNA methylation. Tyrosine kinase inhibitors (TKIs), originally synthesized and approved for cancer therapy, have recently been linked to the regulation of obesity. We evaluated whether LPM4870108, a small-molecule TKI with antitumor efficacy, contributes to obesity via DNA methylation. METHODS:LPM4870108 (TKI) was administered orally to rats at 0, 1.25, 2.5, or 5.0 mg/kg (for 28 days, twice daily). Body weights (BWs) and food intake were recorded daily. After 28 days of administration, ventromedial hypothalamus (VMH) tissues were collected for whole-genome transcriptomic and methylation sequencing to identify candidate genes. The mRNA expression and promoter methylation of candidate genes were analyzed by real-time RT-PCR and pyrosequencing, respectively. Protein levels of DNA methyltransferases (DNMTs) were determined by Western blot. RESULTS:LPM4870108 treatment substantially increased food intake and BW. Whole-genome transcriptomic and methylation profiling identified 415 differentially expressed genes (DEGs) and 124,935 differentially methylated regions (DMRs) within the VMH of LPM4870108-treated rats. The transcriptomic results were combined with whole-genome methylation sequencing data, followed by further verification via RT-PCR and pyrosequencing, through which the obesity-related candidate gene arginine vasopressin (AVP) was identified. Among all DEGs, AVP showed the most prominent change, with a negative association between its promoter methylation and mRNA expression level. Reduced protein expression of DNA methyltransferase 3A (DNMT3A) in the VMH was also detected in LPM4870108-treated rats. CONCLUSIONS:These findings indicate that LPM4870108-stimulated hyperphagia and weight gain were associated with DNA hypomethylation and concomitant upregulation of the AVP gene. LPM4870108, tyrosine kinase inhibitors; VMH, ventromedial hypothalamus; RRBS, reduced representation bisulfite sequencing; AVP, arginine vasopressin; DNMT, DNA methyltransferases.