Xuefu Wang

Aberrant activation of the NLRP3 inflammasome contributes to the development of multiple inflammatory disorders, but clinically approved inhibitors targeting this pathway are still lacking. We investigated whether niflumic acid (NFA), a clinically used nonsteroidal anti-inflammatory drug, regulates NLRP3 inflammasome activation and explored the underlying mechanism. NLRP3 inflammasome activation was induced in murine bone marrow-derived macrophages and PMA-differentiated THP-1 cells using nigericin, ATP, and monosodium urate crystals. Inflammasome activation, chloride efflux, mitochondrial injury, and mitochondrial ROS production were evaluated by immunoblotting, ELISA, flow cytometry, confocal microscopy, co-immunoprecipitation, and ASC speck analysis. TMEM16F was examined by molecular docking, cellular thermal shift assay, siRNA-mediated silencing, and pharmacological inhibition. The anti-inflammatory effects of NFA were further assessed in mouse models of LPS-induced endotoxemia, MSU-induced peritonitis, and acetaminophen-induced liver injury. NFA preferentially inhibited NLRP3 inflammasome activation, as evidenced by reduced caspase-1 cleavage and decreased IL-1β and IL-18 maturation and secretion, without significantly affecting TNF-α production or the activation of the AIM2 and NLRC4 inflammasomes. Mechanistically, NFA suppressed ASC speck formation and disrupted NLRP3-ASC interaction, indicating impaired inflammasome assembly. NFA also blocked chloride efflux, alleviated mitochondrial damage, and reduced mitochondrial reactive oxygen species (mtROS) generation, while having minimal effects on K⁺ efflux and Ca²⁺ influx. Moreover, TMEM16F was found to contribute to NFA-mediated inhibition of NLRP3 inflammasome activation, and genetic depletion of TMEM16F or pharmacological inhibition of Ca²⁺-activated chloride channel activity mimicked the effects of NFA on chloride efflux, mtROS production, and NLRP3 inflammasome activation. In vivo, NFA reduced inflammatory injury and inflammasome-associated readouts in multiple inflammatory models. NFA suppresses NLRP3 inflammasome activation through a mechanism associated, at least in part, with TMEM16F-related chloride efflux and mtROS production. These findings suggest an anti-inflammatory mechanism of NFA that is not fully explained by canonical COX inhibition and support further investigation of NFA as a potential modulator of inflammasome-associated inflammation.

Background: Chimeric antigen receptor (CAR) T-cell therapy is a promising immunotherapy for patients with refractory or relapsed lymphoma or leukemia. However, inefficient activity and insufficient persistence of CAR T cells impede the durability of its therapeutic efficacy. Methods: Krüppel-like factor 4 (KLF4)-overexpressing CAR T cells were generated and assessed for function, exhaustion, and transcriptional changes using in vitro stimulation models and RNA sequencing, with antitumor efficacy validated in mouse models. Results: We report that KLF4 overexpression enhances the durability of CAR T-cell antitumor efficacy. KLF4 overexpression enhanced CAR T-cell activity and promoted the differentiation into effector CAR T cells. Furthermore, KLF4 overexpression prevented CAR T-cell exhaustion induced by continuous tumor antigen stimulation. RNA sequencing data confirmed that KLF4 overexpression led to the upregulation of genes involved in effector functions and the downregulation of genes associated with exhaustion in CAR T cells. Notably, KLF4-overexpressing CAR T cells delayed tumor growth, prolonged the survival of mice with hematological malignancies, and exhibited reduced exhaustion and enhanced effector functions in vivo relative to normal CAR T cells. Conclusions: Our findings highlight the potential of KLF4 overexpression as a therapeutic approach to counteract CAR T-cell exhaustion and enhance antitumor activity, underscoring its therapeutic promise in clinical immunotherapy.

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory lung disease with progressive airflow limitation. However, the key pathogenic immune

Progenitor exhausted CD8 ⁺ T (Tpex) cells have recently been identified as a stem‐like T cell subset that mediates durable anti‐tumor immune responses and represents a pivotal population responsive to immunotherapies. Here, it is demonstrated that diisopropylamine dichloroacetate (DADA) facilitates CD8 ⁺ T cell‐mediated anti‐tumor immunity and promotes Tpex cells accumulation in the tumor microenvironment. Mechanistically, DADA promotes the conversion from pyruvate to Acetyl‐CoA by inhibiting pyruvate dehydrogenase kinase. This process leads to increased oxidative phosphorylation (OXPHOS) and mitochondrial fitness, thereby enhancing CD8 ⁺ T cells stemness. Treatment of mice with DADA improves the efficacy of PD‐1 blockade. Furthermore, the in vitro expansion of chimeric antigen receptor (CAR)‐T cells supplemented with DADA confers them with stemness characteristics, contributing to improved anti‐tumor efficacy. Collectively, this study illustrates how DADA‐mediated metabolic reprogramming in CD8 ⁺ T cell enhances their stemness, underscoring its potential for anti‐tumor therapy.

Background: The KRAS ^G12C mutation is one of the special mutation types in patients with colorectal cancer liver metastasis (CRLM). Although several small molecule inhibitors specifically targeting KRAS ^G12C mutation have been developed, they have only shown limited clinical benefits for CRLM patients. Thus, alternative approaches are still needed. Methods: We screened out the differentially expressed gene Aldo–keto reductase family 1 member B10 (AKR1B10) between the KRAS ^G12C mutation and wild-type CRLM through RNA sequencing, and characterized the tumor microenvironment (TME) changes of the KRAS ^G12C mutation CRLM using multi-omics analysis. The role of AKR1B10 in the TME and its progression of KRAS ^G12C mutation CRLM was confirmed by in vitro and in vivo experiments, and the molecular mechanism of lactate on neutrophils reprogramming was detected by immunofluorescence, western blot and Chip-qPCR. Results: AKR1B10 was highly expressed in the KRAS ^G12C mutation CRLM and was associated with a poor prognosis. Mechanistically, AKR1B10 promotes the recruitment of neutrophils in the TME by CXCL8/CXCR2 pathway. Meanwhile, AKR1B10 could promote the production of lactate by regulating crucial glycolytic enzymes. The increased lactate accumulation in the TME promoted histone lactylation of neutrophils, which induced PD-L1 transcription and prompted the reprogramming of neutrophils to an immunosuppressive phenotype. Conclusion: AKR1B10 facilitated immune evasion of KRAS ^G12C mutation CRLM by recruiting and reprogramming neutrophils to remodel the immunosuppressive TME, providing a potential therapeutic target for KRAS ^G12C mutation CRLM patients. Graphical

Acetaminophen (APAP) overdose induces acute liver injury and represents the most frequent cause of drug-induced liver injury worldwide. Macrophage-mediated inflammation plays detrimental roles during the early stage of liver injury. However, the potential targets regulating inflammation to improve drug-induced liver injury remains undefined. In this study, we reported that G protein-coupled receptor 35 (GPR35) improves drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cyclic AMP-protein kinase A (Gαs-cAMP-PKA) pathway. The ablation of GPR35 exacerbates APAP-induced liver injury, characterized by higher levels of alanine aminotransferase and aspartate aminotransferase in sera, larger damaged areas, and increased levels of pro-inflammatory cytokines. More hepatic macrophages appeared in the inflamed liver of mice with GPR35 deficiency. In contrast, the agonists of GPR35 alleviated APAP-induced liver injury. The depletion of macrophages abolished GPR35-mediated protection. Mechanistically, GPR35 ablation facilitated the activation of pro-inflammatory AKT, MAPK, and NF-κB signaling pathways at the downstream of Toll-like receptors in macrophages. GPR35 agonists activated Gαs-cAMP-PKA signaling to inhibit the activation of these pro-inflammatory signaling pathways and then suppress the inflammatory response in macrophages. Thus, our findings demonstrate that GPR35 prevents drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cAMP-PKA pathway, indicating that GPR35 is a potential target for the development of novel medicines that control drug-induced liver injury.

Acetaminophen (APAP) overdose induces acute liver injury and represents the most frequent cause of drug-induced liver injury worldwide. Macrophage-mediated inflammation plays detrimental roles during the early stage of liver injury. However, the potential targets regulating inflammation to improve drug-induced liver injury remains undefined. In this study, we reported that G protein-coupled receptor 35 (GPR35) improves drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cyclic AMP-protein kinase A (Gαs-cAMP-PKA) pathway. The ablation of GPR35 exacerbates APAP-induced liver injury, characterized by higher levels of alanine aminotransferase and aspartate aminotransferase in sera, larger damaged areas, and increased levels of pro-inflammatory cytokines. More hepatic macrophages appeared in the inflamed liver of mice with GPR35 deficiency. In contrast, the agonists of GPR35 alleviated APAP-induced liver injury. The depletion of macrophages abolished GPR35-mediated protection. Mechanistically, GPR35 ablation facilitated the activation of pro-inflammatory AKT, MAPK, and NF-κB signaling pathways at the downstream of Toll-like receptors in macrophages. GPR35 agonists activated Gαs-cAMP-PKA signaling to inhibit the activation of these pro-inflammatory signaling pathways and then suppress the inflammatory response in macrophages. Thus, our findings demonstrate that GPR35 prevents drug-induced liver injury by blocking macrophage-mediated inflammation via the Gαs-cAMP-PKA pathway, indicating that GPR35 is a potential target for the development of novel medicines that control drug-induced liver injury.