NLRP3 inflammasome plays a critical role in innate immunity and has been implicated in the pathogenesis of multiple inflammatory diseases. However, pharmacological agents that directly target NLRP3 remain limited. In the present study, we investigated the inhibitory effect of isoalantolactone (IAL), a naturally occurring sesquiterpene lactone, on NLRP3 inflammasome activation. IAL markedly suppressed caspase-1 activation and IL-1β/IL-18 maturation induced by canonical NLRP3 stimuli in macrophages, while showing little effect on affecting AIM2 or NLRC4 inflammasomes, indicating selective inhibition of the NLRP3 inflammasome. Mechanistically, IAL inhibited NLRP3 inflammasome assembly, as evidenced by reduced NLRP3 oligomerization, ASC speck formation, and disruption of the NLRP3-NEK7 interaction. Cellular thermal shift assay (CETSA) and microscale thermophoresis (MST) confirmed direct binding between IAL and NLRP3. Molecular docking, CETSA mutational analyses (E695A, H916A, and the double mutant E695A/H916A), and chemical validation using a reduced, inactive IAL derivative (Re-IAL) collectively demonstrated that residues E695 and H916 are critical for IAL binding and its subsequent inhibitory activity. Furthermore, IAL alleviated monosodium urate-induced gouty inflammation and methionine-choline deficient diet-induced metabolic dysfunction-associated steatotic liver disease (MASLD) in wild-type mice, whereas these protective effects were absent in Nlrp3-deficient mice. Together, these findings identify IAL as a small-molecule inhibitor that directly targets NLRP3 at residues E695/H916 to disrupt inflammasome assembly, highlighting its therapeutic potential for NLRP3-driven inflammatory diseases.

MET fusions represent emerging therapeutic targets in solid tumors; however, functional interpretation of non-canonical variants remains poorly understood, posing a major challenge for precision oncology. We conducted a multicenter, pan-cancer study analyzing 23,299 clinical samples using DNA-based next-generation sequencing (NGS) to profile MET fusions. Transcriptional validation was performed using RNA-based NGS on available samples. Preliminary clinical outcomes were assessed in four patients with advanced malignancies harboring uncommon MET fusions who received MET tyrosine kinase inhibitor therapy. We identified 116 MET fusions (incidence: 0.5

Lynch syndrome (LS) is an inherited disorder caused by germline mutations in mismatch repair (MMR) genes or EPCAM deletions and is primarily associated with an increased risk of gastrointestinal and endometrial cancers. However, LS-associated gliomas are rare, and their clinical and molecular characteristics remain poorly defined. The objective of this study was to systematically characterize the prevalence, genetic landscape, molecular features, and clinical outcomes of LS-associated gliomas. We retrospectively analyzed a multicenter cohort of 5,594 glioma patients who underwent targeted next-generation sequencing to identify pathogenic germline mutations in LS-associated genes. Clinical characteristics, molecular features, treatment histories, and outcomes of LS-associated glioma patients were systematically evaluated and compared with those of LS-associated colorectal cancer (CRC) patients. Among 5,594 glioma patients, 54 individuals (0.97

Ferroptosis is a highly synchronized form of non-apoptotic intracellular iron-dependent cell death which is regulated by multiple cellular metabolic pathways. Extensive studies suggest that ferroptosis could efficiently set the therapy resistant cancer cells on the road to ruin, thus providing new opportunities for cancer therapy. And-1 is an acidic nucleoplasmic DNA binding protein which plays a vital role in DNA replication and repair. Here in this study, we report a novel function of And-1 in regulating erastin-induced ferroptosis in ovarian cancer cells. And-1 overexpression (OE) enhanced erastin-induced ferroptosis by modulating the level of ferroptosis biomarkers such as MDA, GSH, Fe2+ and lipid peroxidation; while knockdown (KD) of And-1 produces opposite results. Moreover And-1(OE) suppressed the expression of NRF2, SLC7A11, and FTH1 and promoted the expression of TFR1 and ATF3 while And-1 (KD) produces opposite results. Further mechanistic study revealed that And-1 inhibits the expression of SLC7A11 by increasing ATF3 expression. Moreover, using pharmacological inhibitors, we have shown that erastin induces ferroptosis through multiple mechanisms. Taken together, our data suggest that And-1 enhanced erastin-induced ferroptosis in OC cells by inhibiting system xc - expression through ATF3 and suppressing NRF2 expression.

Background Herpes simplex encephalitis (HSE), predominantly caused by herpes simplex virus type 1 (HSV-1), is a life-threatening central nervous system disease and the leading cause of sporadic encephalitis globally. Currently, the treatment for HSE mainly relies on nucleoside drugs such as acyclovir (ACV), but the mortality rate remains 10%-25% after treatment, with about half of the survivors suffering from neurological sequelae. Rhoifolin (ROF) is a chromatographic standard component specified in the pharmacopoeia of the traditional Chinese medicine Turpinia leaf. It can be derived from various plants such as rhus succedanea, bitter orange, lemon, grapefruit, tomatoes, lupinus, bananas and grapes. ROF exhibits significant antioxidant, anti-inflammatory and neuroprotective properties, which are intricately linked to the mechanisms underlying neurodamage in HSE. Objective To investigate the therapeutic impact and mechanisms of ROF in treating mice with HSE. Methods We evaluated the efficacy of ROF in combating HSE and its antiviral effects using a lethal HSE mouse model and HSV-1 infected neural cell models. The potential mechanisms were explored through molecular biology and histochemical techniques. Results ROF significantly rescued mice from lethal HSE induced by HSV-1 infection. HSV-1 infection can affect various brain regions, such as the olfactory bulb, cortex, cerebellum and brainstem. Microglia and astrocytes are predominantly infected, whereas neuron infection is rare. ROF exhibited antiviral activity in various neural cell types, with 50% effective concentration (EC50) values ranging from 4.15 to 5.30 μM. It mitigated HSV-1-induced oxidative stress by modulating Sestrin2 (SESN2) to regulate the downstream Kelch-like ECH Associated Protein 1/Nuclear Factor erythroid 2-Related Factor 2 (Keap1/Nrf2) and Protein kinase AMP-activated catalytic subunit alpha 1/ NADPH oxidase (AMPK/Nox) pathway, thus decreasing neural oxidative damage and inflammation. Conclusion ROF treatment successfully saved lethal HSE mice by curbing viral replication and oxidative stress, leading to improved neural damage and reduced neuroinflammation. The underlying mechanism involves enhancing the expression of SESN2 in various types of neural cells and regulating the downstream Keap1/Nrf2 and AMPK/Nox pathways.

Sarcomas are rare, heterogeneous mesenchymal malignancies with notably high misdiagnosis rates. Despite sarcoma patients in China representing about one-quarter of the global disease burden, large-scale NGS-based diagnostic studies remain scarce, with limited sample sizes failing to capture the extensive subtype complexity of sarcomas. To address diagnostic gaps, we conducted the largest multicenter study in China involving 788 patients with soft tissue or bone sarcomas. All samples underwent targeted RNA sequencing (Fusioncapture) alongside standard histopathology, immunohistochemistry, and DNA-based next-generation sequencing (NGS). Compared with DNA-NGS, RNA-based profiling clarified ambiguous fusion calls and uncovered numerous additional and clinically relevant events, including 281 fusions not captured by the DNA panel. Notably, 114 recurrent alterations were strongly subtype-associated, and 20 newly identified receptor tyrosine kinase fusions had therapeutic significance, expanding targetable cases from 3.3% to 6.5%. Furthermore, integrated RNA data led to subtype reclassification in 11.9% of patients, including 22% of those initially diagnosed as "not otherwise specified". These findings confirm the utility of targeted RNA sequencing for detecting transcriptionally active fusions, refining pathological classifications, and identifying actionable variants in Chinese sarcoma patients. Despite retrospective design and limited orthogonal validation of some fusions, our results strongly support incorporating RNA-based assays into routine clinical workflows. Ultimately, this integrated approach can improve diagnostic precision, guide personalized treatment strategies, and enhance outcomes for sarcoma patients.

Identifying robust diagnostic biomarkers for gastric cancer (GC) remains a significant challenge. Emerging studies highlight extracellular vesicle (EV)-derived RNAs in cancer biology, but the diagnostic potential of circulating EV-derived small non-coding RNAs (sncRNAs) in GC is poorly understood. Using panoramic RNA display by overcoming RNA modification aborted sequencing (PANDORA-seq), we mapped non-canonical sncRNAs—specifically ribosomal RNA-derived small RNAs (rsRNAs) and transfer RNA-derived small RNAs (tsRNAs)—in plasma EVs. We identified a three-rs/tsRNA signature that discriminates GC patients from healthy individuals with high sensitivity (80.42

Phosphoglycerate kinase 1 (PGK1) is traditionally recognized for its pivotal role in glycolysis. Our findings reveal that PGK1 also functions as a protein kinase phosphorylating valosin-containing protein (VCP) at S746, which subsequently reduces Beclin 1 deubiquitination and impairs autophagy. Inhibition of PGK1 initiates autophagy in T315I-mutant chronic myeloid leukemia (CML) cells, thereby enhancing their sensitivity to first-generation Tyrosine Kinase Inhibitor (TKI) imatinib and third-generation TKI ponatinib. Despite the significant clinical implications, few PGK1-targeting inhibitors have been approved for clinical use to date. Through a comprehensive high-throughput screening of ∼20,000 natural compounds, we identified flavonoid as potent inhibitors of the enzymatic activity of PGK1. Subsequent structural optimization of these flavonoid derivatives led to the development of CPU-216, a compound that binds to the GLU344 and PHE292 residues of PGK1, effectively inhibiting its enzymatic and kinase activity. Notably, CPU-216 induces autophagy via VCP and Beclin 1 in CML-T315I cells, enhancing their responsiveness to TKIs. These discoveries propose a novel therapeutic strategy for T315I-mutant CML, underscoring the potential to develop targeted treatments that leverage the kinase functions of PGK1.

BACKGROUND:Despite the increasing approval and ongoing clinical trials of FGFR-targeted therapies, accurately detecting FGFR fusions remains a challenge due to limited research, low incidence rates, complex fusion partner distribution, and unique kinase domain distribution. METHODS:We conducted a multicenter study to comprehensively profile FGFR fusions in the largest Chinese pan-cancer cohort to date, comprising 118 FGFR fusions from 114 individuals. Both DNA- and RNA-based sequencing approaches were utilized to reveal novel and fundamental features of FGFR fusion. RESULTS:Our research reveals an incidence rate of 0.96% for FGFR rearrangements within this Chinese cohort, including a high incidence rate of FGFR fusions (40%) in parotid gland carcinoma. However, this is based on a small sample size of 5 tumors and should be interpreted cautiously pending validation in larger cohorts. We also uncovered distinct breakpoint distribution patterns across various FGFR rearrangements. For example, a primary breakpoint in intron17 of FGFR2 was predominant (21/22), while FGFR1/3 breakpoints displayed substantial diversity. For the first time, we identified "hot" breakpoints in FGFR1 intron17, exon18, and FGFR3's 3' untranslated region. These findings underline the importance of incorporating these regions in targeted sequencing to ensure comprehensive detection of FGFR1/3 fusions. Notably, we observed a predilection for intrachromosomal distribution in common FGFR1/2/3 fusions. In contrast, most novel fusions (12/15) exhibited an interchromosomal distribution pattern, indicating variations in the fusion formation mechanism. Importantly, our study demonstrates the substantial incremental value of RNA-NGS or other orthogonal methods in confirming the functionality of FGFR rearrangements initially identified by DNA sequencing. In our cohort, 46% (6/13) of rare FGFR1/2/3 fusions lacked detectable RNA transcripts; however, this does not definitively indicate non-functionality as factors such as low RNA quality, expression below detection limits, or nonsense-mediated decay may contribute. Therefore, RNA-based validation is critical for accurately identifying potentially targetable FGFR fusions and guiding therapy. CONCLUSION:Our findings offer critical novel insights into functional FGFR fusions and bear considerable clinical implications for identifying individuals whose tumors are most likely to respond favorably to FGFR-targeted therapies.

BACKGROUND & AIMS: Aquaporin-7 (Aqp7) is an aquaglyceroporin that provides transmembrane gateway of water, glycerol, and hydrogen peroxide (H2O2). Analysis of the Gene Expression Omnibus (GEO) database revealed upregulation of hepatic AQP7 expression in liver fibrosis patients. This study aimed to elucidate the role of Aqp7 in the pathogenesis of liver fibrosis. METHODS: The GEO database analysis and TGFb-induced human hepatic stellate cell (HSC) line LX-2 cells were used to study the relevance of AQP7 to human liver fibrosis. Bile duct ligation-induced and carbon tetrachloride-induced liver fibrosis models were employed to investigate the role of Aqp7 in liver fibrosis formation in conventional and HSC-specific Aqp7 knockout mice. Primary mouse HSCs were isolated to explore the role of Aqp7-mediated glycerol and H2O2 transport in HSC activation and proliferation. RESULTS: AQP7 mRNA and protein levels are remarkably upregulated in TGFb-induced LX-2, as well as in primary mouse HSCs isolated from liver fibrosis models induced by bile duct ligation and peritoneal injection of carbon tetrachloride. Liver fibrosis formation was significantly alleviated in both conventional and HSC-specific Aqp7 knockout mice compared with their respective wild-type littermates, as evidenced by significantly decreased deposition of fibrous extracellular matrix. Aqp7 deletion resulted in the accumulation of intracellular glycerol, an increase in triglyceride content, the retention of intracellular lipid droplets, and dilatory activation of HSCs. Moreover, Aqp7 deficiency led to elevated intracellular H2O2 levels during activation, which impaired autophagy, proliferation, and survival of HSCs by disrupting relevant cell signaling pathways. Virus-mediated replacement with glycerol- or H2O2-transporting aquaporins Aqp3 or Aqp8, but not the strictly water-selective channel Aqp4, effectively rescued the impaired activation and proliferation in primary cultured Aqp7-/-HSCs. CONCLUSIONS: Our findings suggest that Aqp7 plays a crucial role in the activation of HSCs and the formation of liver fibrosis by regulating triglyceride catabolism and maintaining reactive oxygen species homeostasis in HSCs.

Aims Ovarian cancer (OV) is one of the most prevalent and life-threatening malignancies among women worldwide. Resistance to conventional therapies poses a major challenge in OV treatment. Ferroptosis, a type of programmed cell death driven by iron accumulation and marked by lipid peroxidation, has garnered significant attention in cancer research. The regulation of ferroptosis involves intricate epigenetic, transcriptional, and post-translational modification (PTM) processes. O-GlcNAcylation, a reversible PTM occurring on serine/threonine hydroxyl groups of proteins, has been connected with the regulation of apoptosis, autophagy, and necroptosis. However, its role in ferroptosis is still poorly understood. Materials and methods O-GlcNAcylation levels and ferroptosis-associated markers were compared between normal and OV tissues. OV cells were subjected to ferroptosis induction using Erastin or RSL3, while O-GlcNAcylation was modulated via the OGT inhibitor OSMI-1 or the OGA inhibitor Thiamet-G. Subsequent analyses were performed to assess ferroptotic phenotypes and the c-MYC/xCT/GSH/GPX4 signaling pathway. In vitro findings were validated using a nude mouse xenograft model. Key findings In this study, we observed elevated O-GlcNAcylation, higher protein levels of xCT, GPX4, and FTH1, as well as increased antioxidant capacity in ovarian cancer tissues compared with normal ovarian tissues. Modulating O-GlcNAcylation levels in OV cells revealed that its downregulation enhanced ferroptosis, whereas upregulation inhibited it. Further investigation revealed that c-MYC protein levels were regulated by O-GlcNAcylation. The O-GlcNAcylation-mediated stabilization of c-MYC led to increased xCT expression, thereby enhancing the xCT/GSH/GPX4 antioxidant axis and suppressing ferroptosis in OV. Significance Our research may provide intervention strategies for the treatment of OV.

Carbapenem- and colistin-resistant Gram-negative bacteria have become one of the most severe public health issues worldwide. The development of advanced antibacterial agents that can outpace microbial adaptation is imperative. The thioredoxin (Trx) and glutaredoxin (Grx) systems play important roles in maintaining redox homeostasis within Gram-negative bacterial cell membranes, with thioredoxin reductase (TrxR) and glutathione reductase (GR) being classical antibacterial targets. In this study, we found that Au(III) Schiff base complexes Au10 and Au17 exhibited potent activities against carbapenem- and colistin-resistant Gram-negative bacteria, as demonstrated in evaluations both in vitro and in vivo. Mechanistic studies revealed that Au10 and Au17 exert their antibacterial effects through multiple pathways: irreversibly inhibiting TrxR and GR activities via targeting redox-active motifs, impairing bacterial energy metabolism even at low concentrations, degrading deoxyribonucleic acid (DNA), causing reactive oxygen species (ROS) generation and intracellular redox imbalance. This study provides the first evidence that Au(III) Schiff base complexes possess strong activity against carbapenem- and colistin-resistant Gram-negative bacteria and can simultaneously inhibit the oxidoreductase in carbapenem- and colistin-resistant Gram-negative bacteria, establishing a new paradigm for antibacterial strategies and guiding future innovations in antibacterial therapy.

GL-V9, a derivative of wogonin, has shown potent antitumor effects in various cancers, yet its impact on chronic myeloid leukemia (CML) remains unexplored. In this study, we found that GL-V9 significantly decreased the viability of CML cells. Annexin V/PI staining demonstrated that GL-V9 induced apoptosis in a concentration-dependent manner. The JC-1 assay indicated a significant reduction in mitochondrial membrane potential (ΔΨm) in cells treated with GL-V9. Additionally, GL-V9 altered reactive oxygen species (ROS) levels in CML cells. Through transcriptomic sequencing and Western blot analysis, we further revealed that GL-V9 activated the MAPK pathway. These results suggest that GL-V9 is a promising therapeutic candidate for CML.

Background: The regenerative capacity of the liver is pivotal for mitigating various forms of liver injury and requires the rapid proliferation of hepatocytes. Aquaporin-9 (AQP9) provides vital support for hepatocyte proliferation by preserving hydrogen peroxide (H2O2) oxidative balance and glucose/lipid metabolism equilibrium within hepatocytes. Our previous study demonstrated that Radix Astragali (RA) decoction promotes liver regeneration by upregulating hepatic expression of AQP9, possibly via two major active constituents: astragaloside IV (AS-IV) and cycloastragenol (CAG). Purpose: To verify that upregulated AQP9 expression in hepatocytes maintains liver oxidative balance and glucose/lipid metabolism homeostasis, and is the main pharmacological mechanism by which AS-IV and CAG promote liver regeneration. Study Design/Methods: Effects of AS-IV and CAG on liver regeneration were scrutinized using a mouse model of 70 % partial hepatectomy (PHx). AQP9-targeted liver regeneration mediated by AS-IV and CAG was verified using AQP9 gene knockout mice (AQP9- /- ). The AQP9 protein expression pattern in hepatocytes was determined using tdTomato-tagged AQP9 transgenic mice (AQP9-RFP). Potential mechanisms of AS-IV and CAG on liver regeneration were studied using real-time quantitative PCR, immunoblotting, staining with hematoxylin and eosin, oil red O, and periodic acid-Schiff, and immunofluorescence, immunohistochemistry, HyPerRed fluorescence, and biochemical analyses. Results: AS-IV and CAG promoted substantial liver regeneration and increased hepatic AQP9 expression in wildtype mice (AQP9+/+) following 70 % PHx, but had no discernible benefits in AQP9- /- mice. Both saponin compounds also helped maintain oxidative homeostasis by reducing levels of oxidative stress markers (reactive oxygen species [ROS], H2O2, and malondialdehyde) and elevating levels of ROS scavengers (glutathione and superoxide dismutase) in AQP9+/+ mice post-70 % PHx. This further activated the PI3K-AKT and insulin signaling pathways, thereby fostering liver regeneration. Furthermore, AS-IV and CAG both promoted hepatocyte glycerol uptake, increased gluconeogenesis, facilitated lipolysis, reduced glycolysis, and inhibited glycogen deposition, thus ensuring the energy supply required for liver regeneration. Conclusion: This research is the first to demonstrate AS-IV and CAG as major active ingredients of RA that promote liver regeneration by upregulating hepatocyte AQP9 expression, improving hepatocyte glucose/lipid metabolism, and reducing oxidative stress damage, constituting a crucial pharmacological mechanism underlying the liver-protective effects of RA. The augmentation of hepatocyte AQP9 expression underscores an important aspect of the Qi-tonifying effect of RA. This study establishes AQP9 as an effective target for regulation of liver regeneration and provides a universal strategy for clinical drug intervention aimed at enhancing liver regeneration.

Summary: The NLRP3 inflammasome plays a pivotal role in various chronic inflammation-driven human diseases. However, no drugs specifically targeting NLRP3 inflammasome have been approved by the Food and Drug Administration (FDA) of the United States. In our current study, we showed that dimethyl fumarate (DMF) efficiently suppressed the activation of the NLRP3 inflammasome induced by multiple agonists and covalently modified Cys673 of NLRP3, thereby impeding the interaction between NLRP3 and NEK7. The inhibitory effect of DMF was nullified by anaplerosis of the Cys673 mutant (but not the wild-type) NLRP3 in Nlrp3−/− THP-1 cells. In vivo experiments, DMF demonstrated protective effects in the dextran sodium sulfate (DSS)-induced ulcerative colitis of WT mice, but not in Nlrp3−/− mice. In summary, our study identified DMF as a direct covalent inhibitor of NLRP3 and a potential candidate for the treatment of NLRP3 inflammasome-mediated diseases.

Bacterial antimicrobial resistance (AMR), particularly multidrug resistance (MDR) in gram-negative bacterial strains, has emerged as a formidable challenge of substantial consequence, necessitating an urgent pursuit of a sustainable and efficacious strategic response. Repurposing nonantibiotic drugs as potential antibiotics or antibiotic adjuvants is a valuable approach to targeting MDR bacteria. A total of 1,750 FDA-approved drugs (APExBIO, USA) were screened to test their antimicrobial activities against MDR bacteria using the broth microdilution method according to the standard of the Clinical and Laboratory Standards Institute (CLSI). Microscale thermophoresis (MST) analysis was performed to detect the Fty720-LPS interactions. Fty720-indcued lipid changes were measured by untargeted lipidomic analysis. Isothermal titration calorimetry (ITC) analysis was used to determine the Fty720-lipid binding affinities. DNA degradation was assessed via agarose gel electrophoresis with ethidium bromide (EB) staining and visualized using a gel imaging system. Galleria mellonella larvae infection model and Mouse peritonitis infection models were used to evaluated the antibacterial ability of Fty720 in vivo. In this study, we identified Fty720, a pharmaceutical agent for treating multiple sclerosis, as a potent inhibitor of carbapenem-resistant Acinetobacter baumannii (CRAB). We demonstrated that Fty720 exerts antibacterial effects through multiple strategies, including disruption of the structural integrity of the membranes by interacting with LPS and glycerophospholipids, as well as degradation of bacterial DNA. Furthermore, through judicious structural modification, the pivotal role of the positively charged moiety (NH2) in Fty720's antibacterial activity was substantiated. Intriguingly, the translation of Fty720's antibacterial efficacy was demonstrated in vivo, substantiating its pronounced influence on elevating survival rates among models afflicted with MDR gram-negative bacterial infections. Fty720 targets CRAB via multiple pathways, including disruption of outer and inner membrane integrity and DNA degradation. This investigation unveils the multifaceted antibacterial mechanisms of Fty720 while concurrently delineating a prospective therapeutic avenue to counteract MDR gram-negative bacterial strains.

BackgroundDopamine, a frequently used therapeutic agent for critically ill patients, has been shown to be implicated in clinical infections recently, however, the precise mechanisms underlying this association remain elusive. Klebsiella quasivariicola, a novel strain belonging to the Klebsiella species, exhibits potential pathogenic attributes. The impact of dopamine on K. quasivariicola infection has aroused our interest.ObjectiveConsidering the contribution of host immune factors during infection, this study aimed to investigate the intricate interactions between K. quasivariicola, dopamine, and macrophages were explored.MethodsRAW264.7 cells and C57/BL6 mice were infected with K. quasivariicola, and the bacterial growth within macrophage, the production of inflammatory cytokines and the pathological changes in mice lungs were detected, in the absence or presence of dopamine. ResultsDopamine inhibited the growth of K. quasivariicola in the medium, but promoted bacterial growth when co-cultured with macrophages. The expression of proinflammatory cytokines increased in RAW 264.7 cells infected with K. quasivariicola, and a significant rise was observed upon the addition of dopamine. The infection of K. quasivariicola in mice induced an inflammatory response and lung injury, which were exacerbated by the administration of dopamine. ConclusionsOur findings suggest that dopamine may be one of the potential risk factors associated with K. quasivariicola infection. This empirical insight provides solid references for clinical precision medicine. Furthermore, an in vitro model of microbes-drugs-host immune cells for inhibitor screening was proposed to more accurately replicate the complex in vivo environment. This fundamental work had contributed to the present understanding of the crosstalk between pathogen, dopamine and host immune cells.

The BCR:ABL T315I mutation presents a significant challenge in the current management of Chronic Myeloid Leukemia (CML), highlighting the need to identify novel targets and drugs. In our study, we observed the elevated expression of UBE2Q1 in KBM5-T315I cells compared to KBM5 cells, where it interacted with DDX3, regulating its ubiquitination. Furthermore, we found that Bortezomib (BTZ) targeted UBE2Q1, reducing its protein level expression. Consequently, BTZ dose-dependently inhibited the growth vitality of KBM5-T315I cells, inducing increased ROS production, mitochondrial membrane potential collapse, cytochrome C release, and expression of apoptosis-related proteins. These events collectively induced apoptosis in KBM5-T315I cells. Moreover, BTZ enhanced the therapeutic effects of anti-PD-1 treatment. In NOD/SCID mice bearing KBM5-T315I cell line xenografts, BTZ administration (2 mg/kg, ip, every other day for 4 weeks) significantly inhibited the growth of KBM5-T315Iderived xenografts and extended survival. In conclusion, our study sheds new light on the BTZ-induced apoptosis mechanism, suggesting the potential of BTZ as a promising chemo-immunotherapy agent against BCR:ABL T315I mutation CML.

BackgroundBuilding on our prior work that RNA alternative splicing modulates the druggability of kinase fusions, this study probes the clinical significance of sole reciprocal fusions. These rare genomic arrangements, despite lacking kinase domains at the DNA level, demonstrated potential RNA-level druggability in sporadic cases from our prior research.MethodsUtilizing the large-scale multicenter approach, we performed RNA sequencing and clinical follow-up to evaluate a broad spectrum of kinase fusions, including ALK, ROS1, RET, BRAF, NTRK, MET, NRG1, and EGFR, in 1943 patients.ResultsOur findings revealed 51 instances (2.57%) of sole reciprocal fusions, predominantly in lung (57%), colorectal (14%), and glioma (10%) cancers. Comparative analysis with an MSKCC cohort confirmed the prevalence in diverse cancer types and identified unique fusion partners and chromosomal locales. Cross-validation through RNA-NGS and FISH authenticated the existence of functional kinase domains in subsets including ALK, ROS1, RET, and BRAF, which correlated with positive clinical responses to targeted kinase inhibitors (KIs). Conversely, fusions involving EGFR, NRG1, and NTRK1/2/3 generated nonfunctional transcripts, suggesting the need for alternative therapeutic interventions.ConclusionThis inaugural multicenter study introduces a novel algorithm for detecting and treating sole reciprocal fusions in advanced cancers, expanding the patient population potentially amenable to KIs.