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 (cKrasLSL-G12D/+p53R172H/+-Dyt8-KI) and knockout (cKrasLSL-G12D/+p53R172H/+-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.
Ischemic stroke triggers profound neuroinflammation and autophagic stress, driven predominantly by microglial overactivation. However, effective therapies targeting this pathogenesis remain elusive. Here, we report the use of 3-hydroxydehydroleucodin (3-Hyd), the primary active component from Kudiezi injection used in China for ischemic stroke patients, as a potent neuroprotective agent that markedly reduces the cerebral infarct area and improves neurological deficits in a mouse model of transient middle cerebral artery occlusion (tMCAO). Integrated bulk RNA sequencing and in vitro assays revealed that 3-Hyd significantly suppressed the microglial proinflammatory phenotype and excessive autophagic flux. Mechanistically, through chemical biology approaches, we discovered that 3-Hyd directly binds to the V51 and R128 residues of peroxiredoxin 1 (PRDX1), which physically disrupts the pathological binding of PRDX1 to its E3 ubiquitin ligase TRIM21, thus preventing the polyubiquitination of PRDX1 at the K109 residue and its subsequent proteasome degradation. Consequently, stabilized PRDX1 impedes TRAF6 ubiquitination, effectively blocking the downstream NF-κB signaling cascade. Strikingly, the anti-neuroinflammatory and cerebroprotective effects of 3-Hyd were largely abolished in microglia-specific Prdx1 conditional knockdown (Cx3cr1Cre/ERT2) mice. Together, our findings elucidate a novel TRIM21-PRDX1-TRAF6 signaling axis that governs microglial homeostasis and highlight the pharmacological stabilization of PRDX1 by 3-Hyd to block the binding of TRIM21 with PRDX1 as a promising therapeutic strategy for ischemic stroke.
