British Journal of Pharmacology最新文献

Background and purpose: Inflammation plays a crucial role in the development and progression of numerous acute and chronic diseases such as myocardial infarction (MI) and heart failure. Targeting ADAM proteases, particularly ADAM10, holds promise for fine-tuning inflammatory responses. However, developing selective inhibitors has proven challenging because of cross-inhibition of other metalloproteases. Protein-based inhibitors, such as the synthetic ADAM10 prodomain, offer potential for improved selectivity and effectiveness.

Experimental approach: A novel ADAM10 inhibitor, VTH144, was designed by modifying the ADAM10 prodomain and evaluated empirically for selectivity, efficacy and toxicity both in vitro and in vivo and by using machine learning-based modelling.

Key results: AlphaFold3 predictions confirm that the ADAM10 prodomain is highly selective for its protease and identify VTH144 as a unique ADAM10-inhibiting peptide. Fluorescence resonance energy transfer (FRET) assays verified VTH144 as highly selective for ADAM10, sparing other metalloproteases. In a MI mouse model, we show that VTH144 applied twice, 4 and 48 h after ligation of the left ventricular descending artery (LAD), is highly effective in preserving cardiac function by reducing scar size. VTH144 significantly attenuates ADAM10-mediated CX₃CL1 shedding, subsequent neutrophil infiltration into the heart and IL-1β-driven systemic inflammation. Importantly, safety pharmacology in naïve mice revealed that ADAM10 inhibition with VTH144 lacks toxic short-term effects in cardiac, pulmonary, hepatic or renal systems.

Conclusions and implications: Our study highlights the potential of the selective ADAM10 inhibitor VTH144 as a promising therapeutic candidate for improving cardiac function and reducing the risk of heart failure following myocardial infarction.

Background and purpose: Mavacamten (MAVA) is a novel small molecule inhibitor of cardiac myosin, mitigating cardiomyocyte hypercontractility in patients with hypertrophic obstructive cardiomyopathy (HOCM). Despite its recent approval for clinical use, the transcriptional and functional impacts of MAVA remain not well understood. In this study we investigate the effects of MAVA across diverse cardiac models, including healthy female porcine cardiomyocytes and myocardial slices, human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), cardiac organoids and living myocardial slices (LMSs) derived from patients with HOCM.

Experimental approach: Long-term LMS culture facilitated continuous force measurements, while SarcTrack and MUSCLEMOTION analyses were used to evaluate contractility in cardiomyocytes and cardiac organoids. Transcriptome profiling of MAVA-treated HOCM hiPSC-CMs and HOCM LMSs allowed in-depth examination of gene expression signatures in response to MAVA treatment.

Key results: Across all models tested, MAVA demonstrated robust force inhibition. In primary disease models, MAVA showed little effect on time to peak or relaxation times and even reduced contraction and relaxation velocities. By contrast, in engineered human HOCM models, MAVA accelerated both contraction and relaxation, suggesting potential model-specific effects. Transcriptome analyses revealed that MAVA treatment not only influenced contraction regulation but also significantly altered cytoskeleton organization, muscle stretch response, and metabolic pathways. Notably, in LMSs derived from three HOCM patients, MAVA treatment up-regulated myosin binding protein H (MyBPH) expression, suggesting that MyBPH may also be involved in contraction regulation.

Conclusion and implications: These data suggest that MAVA not only inhibits force within the sarcomere but also influences transcriptional pathways in model-specific manner.

Background and purpose: The effectiveness of conventional treatments for epithelial ovarian cancer (EOC) is very limited and their side effects are serious. Previous research has demonstrated the inhibitory effects of enterolactone (ENL) on EOC by inhibiting malignant angiogenesis. Gemcitabine (Gem) is a chemotherapeutic agent commonly used for the treatment of EOC with limited efficacy. In this study, we aimed to explore the combined inhibitory effects of ENL and Gem on EOC.

Experimental approach: We detected the proliferation ability of EOC cells after ENL/Gem/ENL + Gem by CCK8, crystal violet assays, migration and invasion ability by wound healing and transwell assays, in vivo evaluation of the anti-neovascularisation efficacy of zebrafish and in vitro tube formation assays to detect angiogenesis, network pharmacology, Western-blot and immunohistochemistry to analyse molecular pathways, and in vivo animal experiments on tumour progression.

Key results: Our results demonstrated that the ENL and Gem combination synergistically inhibited the proliferation, migration and invasion of EOC. Tube formation and zebrafish neovascularization assays showed potent anti-angiogenic activity of the ENL + Gem combination. In animal experiments, the combined use of ENL and Gem also synergistically inhibited tumour growth and in the meantime markedly reduced the side effects of Gem. ENL ameliorated gut dysbacteriosis of ovarian cancer animals, which significantly enhanced the synergistic anti-cancer effect of ENL and Gem.

Conclusions and implications: ENL and Gem synergistically inhibit the proliferation, migration, invasion, and angiogenesis of EOC by modulating the Akt-Bax and Akt-MMP9-VEGFR-2 pathways and ameliorating gut dysbacteriosis.

Background and purpose: ATP-dependent activation of P2X receptors has been implicated in chronic cough. The mechanisms of ATP release in cough and how P2X-receptor engagement produces cough hypersensitivity are not clear. It is also unclear whether P2X receptor blockade could compromise the essential defensive functions of cough regulated by the vagal Aδ-fibres terminating in the extrapulmonary airways.

Experimental approach: We measured ATP release from isolated human and guinea pig airways. P2X receptor-dependent regulation of vagal afferent nerve discharge in vitro and respiratory reflexes in anaesthetised guinea pigs also were studied using the P2X blockers AF-353 and AF-454.

Key results: ATP was released continually and metabolised by human and guinea-pig airways. ATP release was augmented by mild mechanical stimulation (blocked by carbenoxolone) and hypotonic challenges (blocked by ruthenium red), dwarfing the ATP release evoked by acid or by TRPV4 activation. Epithelium removal prevented ATP release evoked by either mechanical stimulation of the airways or by hypotonic challenges. AF-454 inhibited α, β-methylene ATP evoked action potential discharge in bronchopulmonary nodose C-fibres, but was without effect on extrapulmonary Aδ-fibre excitability and did not prevent acid or mechanically induced cough responses evoked in anaesthetised guinea pigs. AF-454 and AF-353 did, however, inhibit α, β-methylene ATP evoked increases in airway cholinergic tone and changes in respiratory patterns, respectively.

Conclusions and implications: These results predict a desirable safety profile for P2X receptor blockade and identify multiple potential biomarkers that may differentiate responders from non-responders in studies evaluating P2X blockers in chronic cough patients.

Background and purpose: Cellular senescence, a stress-induced cell cycle arrest state, plays a dual role in cancer. Circular RNAs (circRNAs), endogenous noncoding RNAs, regulate physiological processes and are linked to diseases including cancer. This study investigates the role and mechanism of circPTK2 in non-small cell lung cancer (NSCLC).

Experimental approach: Lentivirus-mediated knockdown or overexpression of circPTK2 were used to assess effects on proliferation and senescence. circPTK2 expression in NSCLC tissues was examined using tissue microarrays and in situ hybridization. RNA-seq identified circPTK2-regulated signalling pathways and downstream targets. Transcriptional regulation of FOXM1 by NFYA was investigated using chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays. Co-immunoprecipitation (Co-IP) was conducted to assess the interaction between NFYA and MDM2, as well as NFYA ubiquitination.

Key results: circPTK2 was highly expressed in NSCLC tissues and was associated with poor prognosis. Knockdown of circPTK2 significantly inhibited cancer cell growth and enhanced cellular senescence. RNA-seq analysis identified FOXM1 as a downstream target of circPTK2. Database predictions suggested that NFYA could transcriptionally regulate FOXM1. Knockdown of circPTK2 induced NFYA degradation. circPTK2 was found to bind to the E3 ubiquitin ligase MDM2 in the cytoplasm, thereby preventing MDM2 from interacting with the transcription factor NFYA. This interaction blocked MDM2-mediated ubiquitylation and degradation of NFYA. Additionally, NFYA bound to the FOXM1 gene promoter, ultimately leading to the up-regulation of FOXM1 and suppression of cellular senescence.

Conclusions and implications: Elevated circPTK2 expression correlates with poorer survival, representing a potential therapeutic target in NSCLC.

Background and purpose: Myocardial fibrosis (MF), a hallmark of structural cardiac remodelling, drives disease progression across most forms of heart failure and plays a central role in heart failure with preserved ejection fraction (HFpEF). Despite its clinical relevance, effective treatments remain scarce. In preclinical models, current methods for quantifying MF fail to capture its regional heterogeneity, limiting reliable assessment of novel anti-fibrotic compounds. This study aimed to develop a whole-heart imaging and deep learning (DL)-based quantification pipeline for MF, and to validate its utility by evaluating the efficacy of a glucagon-like peptide-1 receptor (GLP-1R) agonist in a mouse model of HFpEF.

Experimental approach: By utilising fluorescent collagen-labelling dye, tissue clearing and three-dimensional light sheet fluorescence microscopy (3D LSFM), we developed a high-throughput imaging platform for MF. We established a DL framework to quantify interstitial, perivascular and replacement fibrosis, as well as hypertrophy, in 17 left ventricular (LV) segments. The antifibrotic efficacy of the GLP-1R agonist semaglutide was evaluated in the db/db UNx-ReninAAV mouse model, which exhibits diabetes, kidney failure, obesity and hypertension.

Key results: Whole-heart 3D LSFM, combined with DL, enabled micrometre-resolution mapping of MF in rodents. Using this approach, we observed that interstitial collagen content increases proportionally with cardiac hypertrophy. Chronic treatment with semaglutide reduced LV hypertrophy and perivascular fibrosis but did not affect the extent of replacement fibrosis.

Conclusions and implications: The established 3D imaging and quantification approach provides a powerful tool for evaluating the therapeutic efficacy of antifibrotic compounds and studying the pathological mechanisms underlying cardiovascular diseases.

Background and purpose: Metabolic dysfunction associated fatty liver disease (MAFLD) is a leading cause of irreversible liver fibrosis and finally liver cancer. Imperatorin was reported to be effective for liver injury and fibrosis, but its effects and mechanisms on MAFLD remain unidentified.

Experimental approach: The high fat diet (HFD)-induced MAFLD model in the mice and palmitic acid-induced primary mouse hepatocytes were introduced to investigate the effects of imperatorin on lipid metabolism and inflammation. Molecular docking technique, cellular thermal shift assay and surface plasmon resonance (SPR) were performed to confirm the targeting of imperatorin on suppressor of variegation 3-9 homologue 1 (Suv39h1). RNA-sequencing was used to screen the downstream genes affected by Suv39h1 overexpression. The functional relationship between Suv39h1 and its downstream genes of fatty acid-binding proteins (Fabps) was elucidated by CHIP, DNA Pull Down and dual-luciferase reporter assays. Point mutation and knockdown of Set domain of Suv39h1 in palmitic acid-stimulated AML12 cells and HFD mice were adopted to confirm the function of Set domain in MAFLD.

Key results: Imperatorin could delay MAFLD by affecting lipid metabolism, inflammation and insulin resistance. Knockdown of Suv39h1, the target of imperatorin, weakened the therapeutic effects of imperatorin on MAFLD. Knockdown of  Fabps and choline/ethanolamine phosphotransferase 1 (Cept1), the downstream of Suv39h1, could alleviate the disorder of lipid metabolism. Histone methylation modification of Fabp promoters was mediated by Suv39h1. Knockdown of Set domain of Suv39h1 could increase the protein expression of Fabps and aggravate the progression of MAFLD.

Conclusions and implications: Imperatorin ameliorates MAFLD through modulating Suv39h1/Fabps/Cept1 signalling pathway.

The cannabinoid receptor 2 (CB2), which is encoded by the Cnr2 gene, is a G-protein-coupled receptor that controls immune responses and has recently emerged as a regulator of renal injury and repair. Over the past 15 years, numerous pharmacological and genetics studies have explored the role of CB2 in acute kidney injury (AKI) and chronic kidney disease (CKD). Although the precise localisation of CB2 within renal compartments remains under debate, pharmacologically mediated CB2 agonism, across a wide array of chemical, metabolic, ischaemia and obstructive mouse models, has consistently preserved tubular epithelial cell integrity, reduced inflammation, and limited tubulointerstitial fibrosis. These protective effects of CB2 activation are further supported by the opposing outcomes in Cnr2 knockout mice, which showed worsened injury. More selective CB2 ligands with defined pharmacokinetic and pharmacodynamic profiles in preclinical animal models along with late-stage clinical studies have further substantiated the safety and efficacy of this type of therapeutic approach. Nevertheless, a few studies have reported conflicting results, suggesting a CB2-dependent contribution towards tubular damage, although these findings are complicated by differences in ligand selectivity, possible off-target activity, and experimental design. With this in mind, the prevailing evidence supports CB2 activation as beneficial in both AKI and CKD and warrants continued translational progress to develop CB2 agonists for therapeutic applications in kidney disease.

Immune checkpoint inhibitors (ICIs) have been a breakthrough in cancer therapy, inducing durable remissions in responding patients. However, they are associated with variable outcomes, spanning from disease hyperprogression to complete responses with the onset of immune-related adverse events. The consequences of checkpoint inhibition on Foxp3⁺ regulatory T (T_reg) cells remain unclear but could provide key insights into these variable outcomes. In this review, we first cover the mechanisms that underlie the development of hot and cold tumour microenvironments, which determine the efficacy of immunotherapy. We then outline how differences in tumour-intrinsic immunogenicity, T-cell trafficking, local metabolic environments and inhibitory checkpoint signalling differentially impair CD8⁺ T-cell function in tumour microenvironments, all the while promoting T_reg-cell suppressive activity. Finally, we focus on the mechanisms that enable the induction of polyfunctional CD8⁺ T-cells upon checkpoint blockade and discuss the role of ICI-induced T_reg-cell reactivation in acquired resistance to treatment. LINKED ARTICLES: This article is part of a themed issue Immunotherapy in Cancer. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v183.6/issuetoc.

Background and purpose: Osteoarthritis (OA) is the most common form of arthritis worldwide. Here, we have sought to clarify the regulatory mechanisms by which the cellular retinoic acid-binding protein 2 (Crabp2) modulated OA occurrence and to elucidate the role of the insular cortex in regulating OA and anxiety.

Experimental approach: A model of OA was established following intra-articular injection of monosodium iodoacetate in mice, and a series of assessments and behavioural experiments were conducted to investigate the pathological features of OA and anxiety-related behaviours.

Key results: RNA-sequencing analysis revealed an increased Crabp2 expression in the articular cartilage of OA mice. Using the adeno-associated virus (AAV) strategy, Crabp2 overexpression in articular cartilage was shown to exacerbate progression of OA and anxiety-related behaviours. Neural linkages from the insular cortex to the knee joints were identified using a retrograde transneuronal viral tracing technique. Hyperexcitability of glutamatergic neurons in the insular cortex of OA mice was assessed by monitoring expression of FosB and Ca²⁺-sensitive fibre photometry recordings. Activation of glutamatergic neurons in the insular cortex promoted the development of OA and anxiety-related behaviours, whereas inhibiting these neurons attenuated the pathological features of OA and anxiety phenotypes.

Conclusion and implications: Our results emphasized the role of Crabp2 in the regulation of OA and related anxiety disorders by interacting with the insular cortex. This brain area may function as a pathogenetic gene and serve as a therapeutic target in the treatment of OA and its related anxiety disorders.