Acta Pharmacologica Sinica最新文献

Heart failure with preserved ejection fraction (HFpEF) is a pervasive clinical syndrome characterized by complex and elusive pathogenic mechanisms. In this study, we identify aberrant cardiomyocyte cell cycle arrest as a previously unrecognized pathological driver of HFpEF that hinders tissue repair. Using in vivo and in vitro models, we further demonstrate that Glabridin (Gla), a bioactive isoflavone, can induce cardiac repair and alleviate myocardial hypertrophy and fibrosis in HFpEF by targeting the cell cycle arrest pathway. Mechanistically, Gla exerts its protective effects by specifically inhibiting the degradation of Cyclin A2 (CCNA2) and mitigating cardiomyocyte oxidative damage. This stabilization of CCNA2 effectively abrogates cell cycle arrest and drives cardiomyocyte cell cycle re-entry, an effect that was successfully recapitulated by CCNA2 gain-of-function experiments. This reparative response is intrinsically coupled with the restoration of metabolic homeostasis and the attenuation of oxidative stress, as evidenced in both cardiac and hepatic cells. Collectively, our findings define cell cycle arrest as a novel target and position Gla as a potent inducer of CCNA2-mediated cardiac repair for HFpEF treatment.

Triggering receptor expressed on myeloid cells 2 (TREM2) is a central regulator of microglial adaptation during Alzheimer's disease (AD), yet the development of small-molecule modulators associated with TREM2 signaling remains limited. Here, through a consensus pharmacophore-guided virtual screening strategy, MD2-IN-1 was identified as a candidate small molecule associated with TREM2 signaling. Biolayer interferometry and cellular thermal shift assays supported cellular target engagement related to TREM2 (K_D = 13.85 μM). In Aβ-stimulated BV2 microglia, MD2-IN-1 increased SYK phosphorylation, promoted Aβ uptake, and shifted microglial responses toward a less proinflammatory phenotype. These effects were markedly attenuated following Trem2 knockdown, indicating the involvement of the TREM2-SYK signaling pathway. Mechanistically, MD2-IN-1 restored lysosomal acidification and increased the expression of the V-ATPase subunits ATP6v0c and ATP6v0a1, accompanied by increased intracellular Aβ degradation. In 5×FAD mice, chronic MD2-IN-1 administration improved cognitive performance without detectable behavioral abnormalities compared with WT animals. Multimodal PET imaging demonstrated reduced amyloid deposition and attenuated neuroinflammatory signals, which were further supported by histological analyses showing decreased plaque burden, reduced inflammatory microglial responses, and preserved neuronal integrity. In addition, MD2-IN-1 reduced P62 accumulation and restored ATP6v0c expression in vivo, whereas these effects were partially attenuated by pharmacological inhibition of TREM2. Collectively, these findings identify MD2-IN-1 as a small-molecule modulator associated with TREM2 signaling that restores microglial lysosomal degradative function and promotes Aβ clearance in AD models, providing pharmacological support for targeting adaptive microglial responses in AD.

Lung adenocarcinoma (LUAD) is a multi-kinase hyperactivation-driven malignancy. Targeted therapies are effective but have limited application, primarily due to a small eligible patient population and the development of drug resistance. Analysis of TCGA and tissue microarray show that Dystonia 8 (DYT8) is specially highly expressed in more than 70% of LUAD patients. Furthermore, increased DYT8 expression is positively associated with tumor malignancy and negatively associated with survival. Using the Dyt8 knock-in (cKras^LSL-G12D/+p53^R172H/+-Dyt8-KI) and knockout (cKras^LSL-G12D/+p53^R172H/+-Dyt8-KO) transgenic mice, we identify DYT8 as key activator that accelerates the initiation and progression of LUAD. Mechanistically, EGFR-mediated phosphorylation of DYT8 at Y120 reinforces its stability by increasing its interaction with PSMC6, a protein that is highly expressed in LUAD and associated with an unfavorable prognosis. DYT8 drives LUAD malignant transformation by activating Src. Although Y419 phosphorylation is a recognized key event in Src-driven LUAD, our study identifies novel phosphorylation sties at S306 and S525 that are activated by DYT8, which these novel sites being more critical for DYT8-driven malignant transformation than Y419 phosphorylation. Furthermore, the knockdown of Dyt8 rescues EGFR-triggered carcinogenesis and resistance to EGFR tyrosine kinase inhibitors (EGFR-TKIs). These findings reveal that DYT8 drives LUAD via a previously unreported Src activation mechanism and identify EGFR-DYT8-PSMC6 axis as the key determinant of high DYT8 level, thereby conferring a novel therapeutic strategy for wider LUAD patients, including those with EGFR-TKIs resistant.

Human embryonic stem cell (hESC)- or induced pluripotent stem cell (iPSC)-derived natural killer (NK) cells serve as promising adoptive cell therapies for cancer patients and as valuable models for studying the regulation of human NK cell development and effector functions. In this study, we used shRNA to knock down the expression of FTO in hESCs and differentiated these cells into NK cells to explore the role of FTO in regulating human NK cell development and effector functions. Our data revealed that the FTO knockdown in hESCs resulted in a comparable effect on haematopoietic progenitor cell generation but significantly impaired CD45⁺CD56⁺ NK cell production, which might be attributed to impaired proliferation, as indicated by reduced Ki-67 expression. Functional assays demonstrated that FTO knockdown in hESC-NK cells effectively increased per-cell cytotoxicity, including the expression of perforin, granzyme B, and IFN-γ, and increased degranulation (as indicated by the CD107a level). Integrated MeRIP-seq and RNA-seq analyses identified ADAMTS1 and ZAP70 as key target genes of FTO, demonstrating that FTO knockdown increased m⁶A methylation near the 3'UTR regions of ADAMTS1 and ZAP70 mRNAs and upregulated their transcript levels. ADAMTS1 is a known proliferation inhibitor, whereas increased ZAP70 expression is accompanied by increased phosphorylation of ERK1/2, a downstream component of ZAP70 signalling cascades, which positively regulates the effector functions of NK cells. Overall, our results demonstrate that FTO maintains human NK cell proliferation but restricts their per-cell effector functions, possibly through regulating the m⁶A methylation of ADAMTS1 and ZAP70 mRNAs. Exploring methods to achieve stage-specific FTO knockdown in mature NK cells may provide a potential strategy to optimize hESC-NK cell-based immunotherapy.

Accurate identification of the most native-like conformation remains a critical bottleneck in protein complex structure prediction, particularly when multiple globally plausible candidate models are generated by deep learning methods such as AlphaFold3 (AF3). While AF3 provides internal confidence metrics such as ipTM and pLDDT, these signals often lack sufficient sensitivity to distinguish subtle but structurally meaningful differences in local interface geometry. Here, we present TINA, a geometry-aware post-hoc scoring framework designed for fine-grained ranking and selection among high-quality protein complex candidates. TINA constructs atom-level interface graphs and explicitly models two-body, three-body, and four-body geometric interactions through a many-body tensor modeling strategy, enabling a more comprehensive characterization of local contact patterns, angular arrangements, and torsional constraints. The model is initially trained using a joint ranking and regression objective on a large decoy set generated by ZDOCK, with additional task-specific fine-tuning for protein-peptide complexes, and is evaluated exclusively on AF3-generated candidate pools across four independent benchmarks covering general protein-protein, antibody-antigen, protein-peptide, and antibody-peptide complexes. TINA consistently achieves superior or competitive performance in terms of top-1 success rate, enrichment factor, and top-1 DockQ quality. Notably, its benefit is most pronounced in low-confidence and antibody-related scenarios, where AF3 internal ranking signals are less effective in discriminating among candidate conformations. Representative case studies demonstrate that TINA successfully identifies near-native conformations that are present in the AF3 decoy pool but overlooked by ipTM. Ablation experiments further confirm the critical contributions of atom-level graphs, many-body geometric modules, and the joint loss design. Overall, TINA demonstrates that independent, interface-focused post-hoc scoring remains a valuable complement to AF3, particularly for challenging complexes where local geometric discrimination is critical for accurate model selection.

Hematopoietic stem and progenitor cells (HSPCs) rely on tightly regulated proteostasis to preserve their self-renewal and differentiation potential throughout life. However, how proteostasis stress induced by translational dysregulation affects HSPC homeostasis remains poorly understood. Here, we found that knockout of the cytoplasmic threonyl-tRNA synthetase (Tars), which is a component of the translation machinery, caused HSPC exhaustion and definitive, but not primitive, hematopoiesis defects in zebrafish. Similarly, TARS functional deficiency resulted in human CD34⁺ HSC exhaustion and a decrease in colony-forming unit-granulocyte, macrophage (CFU-GM) and colony-forming unit-macrophage (CFU-M) numbers while sparing burst-forming unit-erythroid (BFU-E) numbers in vitro. Functional rescue experiments demonstrated that the enzymatic activity of Tars is indispensable for HSPC maintenance and definitive hematopoiesis. Interestingly, these phenotypes occurred prior to the apparent decrease in global protein translation and were independent of global translation inhibition, hinting that these defective HSPCs were eliminated by an unknown mechanism. Transcriptomic and proteomic profiling revealed that Tars deletion caused widespread proteomic reprogramming and mTORC1 signaling activation, whereas pharmacological or genetic inhibition of mTORC1 partially rescued HSPC exhaustion in tars^-/- embryos. These findings uncover a link between translational machinery integrity, mTORC1 signaling, and HSPC fate determination, providing critical insight into how proteostasis stress is sensed and resolved to maintain hematopoietic homeostasis.

Under conditions of chronic pressure overload, cardiac hypertrophy can progressively develop into arrhythmias and heart failure, and in severe cases, may lead to sudden death. Previous studies have indicated that protein kinases play significant roles in cardiovascular diseases. This study aims to elucidate the function of the WEE1 G₂ checkpoint kinase (WEE1) in the progression of myocardial hypertrophy. We evaluated the expression of WEE1 in the hearts of mice with transverse aortic constriction (TAC). The functional role of WEE1 was investigated using siWEE1 in an angiotensin II (Ang II)-induced cardiomyocyte hypertrophy model. Furthermore, the contribution of WEE1 to cardiac hypertrophy was assessed in TAC mice via intraperitoneal administration of Adavosertib (WEE1-specific inhibitor). RNA-sequencing (RNA-seq) was employed to identify the downstream effector molecules of WEE1 in cardiomyocytes. The expression of both phosphorylated WEE1 and WEE1 was upregulated in hearts subjected to transverse aortic constriction and in cardiac cells treated with angiotensin II. Knockdown of WEE1 effectively alleviated Ang II-induced cardiac cell hypertrophy. Furthermore, application of the specific inhibitor Adavosertib significantly attenuated cardiac cell hypertrophy and improved cardiac function. Mechanistic studies revealed that WEE1 promotes the translocation of the transcription factor P65 from the cytoplasm to the nucleus, thereby activating the nuclear factor κB (NF-κB) signaling pathway, which ultimately upregulates the transcriptional level of Lipocalin2 (LCN2). LCN2 mediates the process of ferroptosis in cardiac cells by binding to the ferroptosis receptor solute carrier family 22 member 17 (SLC22A17), thereby inducing cardiac hypertrophy. Our findings indicate that WEE1 exacerbates cardiac hypertrophy by activating the NF-κB signaling pathway, which upregulates LCN2 expression and subsequently aggravates ferroptosis in cardiac cells. Inhibition of WEE1 significantly improves cardiac function, highlighting its potential value as a therapeutic target of cardiac hypertrophy.

Myocardial remodeling, an essential pathological development in cardiovascular disease, is largely determined by disruptions in redox homeostasis and mitochondrial bioenergetic function. Iristectorigenin A (ITG), a naturally occurring isoflavone, exhibits a variety of pharmacological effects, including antioxidant and anti-inflammatory activities. While so many protective effects, its specific effect on cardiac remodeling has not been reported previously. In this study, we aims to investigate the role of ITG in pressure overload-induced cardiac remodeling with a particular focus on mitochondrial function and oxidative stress. We revealed that ITG mitigated TAC-induced cardiac remodeling and improved cardiac function, with the 20 mg/kg dose showing optimal efficacy; further escalation to 40 mg/kg provided no additional benefit. ITG attenuated cardiomyocyte hypertrophy and inhibited activation of fibroblasts in vitro. Additionally, ITG treatment effectively suppressed oxidative stress and inflammation in the hearts of TAC mice, while improving mitochondrial morphology and the integrity of inner membrane structure. Notably, surface plasmon resonance (SPR) analysis indicated a direct interaction between SIRT6 and ITG with micromolar affinity. Besides, sustained TAC- or PE-stimulation impaired cardiac sirtuin 6 (SIRT6) protein levels, which were partially reversed by ITG. More importantly, silencing SIRT6 and SIRT6 knockdown separately inhibited the protective effects of ITG in vitro and in vivo. Mechanistically, ITG promotes SIRT6 levels by inhibiting its ubiquitination degradation. Furthermore, the cardioprotective effect of ITG was concomitant with SIRT6 upregulation, deacetylation and dephosphorylation of forkhead box protein O1 (FOXO1) and the subsequent increased transcription of antioxidant genes. Collectively, we firstly discover that ITG inhibits oxidative stress and safeguards mitochondrial morphofunctional homeostasis in response to pathological stress through activating SIRT6/FOXO1 axis, suggesting the therapeutic potential of ITG in the treatment of cardiac hypertrophy and remodeling, providing a promising avenue for future research and potential clinical application. Schematic illustration of ITG's potential protective mechanism against cardiac hypertrophy.

Chronic pain has emerged as the third leading health issue, following cardiovascular and cerebrovascular diseases and cancer. Clinically, patients with chronic pain often exhibit comorbid depression-like behaviors. However, the underlying mechanisms of chronic pain-related depression remain unclear. In this study, a chronic pain-related depression model was established in C57BL/6J, Vgat-Cre, and TH-Cre mice through sciatic nerve ligation (SNI) surgery. Viral-mediated DNA methyltransferase 1 (DNMT1) knockdown, optogenetic/chemogenetic modulation, electrophysiological recordings and molecular biology techniques were used to investigate the role of DNMT1 in regulating the projection from central amygdala GABAergic neurons (CeA^GABA) to locus coeruleus noradrenergic neurons (LC^NA) and its impact on chronic pain-related depression-like behaviors. Viral tracing identified LC as a major downstream target of the CeA. Immunohistochemical staining and electrophysiological recordings confirmed the functional synaptic connection between CeA-GABAergic neurons and LC-NAergic neurons. Behavioral assays showed the bidirectional modulation of pain-related depression phenotypes through LC-NAergic system: activation of LC-NAergic neurons induced depression-like behaviors, whereas their inhibition improved SNI-induced depression-like behaviors. Notably, CeA-GABAergic neurons were found to specifically innervate LC-NAergic neurons in mediating chronic pain-related depression-like behaviors. Furthermore, inhibition of the CeA^GABA → LC^NA circuit abolished the improvement of depressive behaviors in DNMT1 knockdown SNI mice, establishing DNMT1 as a key epigenetic regulator of this pathway. Taken together, our findings suggest that DNMT1 may suppress the GABAergic projections from the CeA to LC-NAergic neurons, thereby contributing to chronic pain-related depression-like behaviors. These findings provide novel insight into the circuit-related epigenetic basis of pain-depression comorbidity.