Cellular stresses regulate transcriptional readthrough, whereby RNA polymerase II elongates past a gene's polyadenylation cleavage site without RNA cleavage. Readthrough has been reported in several cancer types. Here, we use long-read sequencing of nascent RNA to quantify transcriptional readthrough in chronic myeloid leukemia (CML) cells and characterize early responses to the targeted therapeutic, imatinib. We show that the amount, length, and gene specificity of readthrough increase within 1 hour, before gene expression and alternative splicing alterations emerge. Notably, imatinib-dependent messenger RNA (mRNA) isoform changes involved "readthrough chimeras," in which exons from an upstream gene are alternatively spliced to exons in a downstream gene. Altered mRNA isoforms and chimera levels were detected in imatinib-resistant K562 cells as well as cells of patients with CML. Thus, imatinib can provoke a cascade of early changes to transcription and splicing fidelity that may lead to longer-term adjustments in gene expression, cancer cell differentiation, and the development of therapy resistance.

Preoperative systemic chemotherapy plays a crucial role in enhancing the outcomes of patients with locally advanced esophageal squamous cell carcinoma (ESCC). Andrographis (major bioactive diterpenoid lactone isolated from Andrographis paniculata; PubChem ID: 5318517), a safe and cost‑effective dietary compound, has demonstrated antitumor effects against various gastrointestinal adenocarcinomas. However, its impact on squamous cell carcinoma remains unclear. The present study explored the antitumor effects of Andrographis and its potential to augment the antitumor efficacy of 5‑fluorouracil (5‑FU). A series of in vitro experiments was conducted, including cell proliferation, colony formation and apoptosis assays, using the ESCC cell lines KYSE410 and TE1. Compared with the controls, Andrographis significantly inhibited cell proliferation (P<0.05), suppressed colony formation (P<0.05), induced apoptosis (P<0.05), and upregulated the expression of ferroptosis‑related genes and proteins, such as HMOX1 (P<0.01), GCLC (P<0.05) and GCLM (P<0.001). Notably, even at a sub‑IC50 dose of 5‑FU, its combination with Andrographis resulted in additive antitumor effects (P<0.05) and further upregulation of ferroptosis‑related gene expression, particularly HMOX1 (P<0.05), compared with either mono‑treatment. The findings of the present study indicate that Andrographis exerts antitumor effects and enhances the efficacy of 5‑FU in ESCC by activating both apoptosis and ferroptosis, suggesting its potential as an adjunctive therapy for ESCC to improve efficacy and reduce 5‑FU dosage and toxicity.

Exposure to waste anesthetic gases (WAGs) is an underestimated occupational hazard in veterinary operating rooms (VORs), where insufficient ventilation and the absence of scavenging systems remain common worldwide. Veterinarians occupationally exposed to WAGs have been poorly investigated to date. Addressing this critical gap, we present the first integrative study evaluating circulating microRNAs (miRNAs), global DNA methylation, and urinary anesthetic quantification in veterinarians occupationally exposed to WAGs isoflurane and sevoflurane. In a case-control design, plasma profiling revealed 11 dysregulated miRNAs in the exposed group (n = 29) compared to the control group (n = 28) based on nominal p-values (p < 0.05), as part of an exploratory screening approach, including seven upregulated miRNAs meeting a predefined fold-change criterion (FC≥1.5). Among these, hsa-miR-1252-5p and hsa-miR-520f-3p showed robust discriminatory performance based on Receiver Operating Characteristic (ROC) curve analysis (AUC≥0.70). Functional enrichment analysis highlighted epigenetic regulators as major network hubs. For hsa-miR-1252-5p, hubs included EP300, TP53, CREBBP, HDAC1 and SIRT1, linking miRNAs modulation to histone acetylation/deacetylation, DNA damage response, apoptosis, and stress regulation. For hsa-miR-520f-3p, included MAPK1, TNRC6B, MECP2 and KMT2A, associated with cell signaling pathways, proliferation and epigenetic regulation. Global DNA methylation levels did not differ significantly between groups, suggesting that exposure under the evaluated conditions may not trigger genome-wide alterations. Occupational exposure was confirmed by urinary quantification of isoflurane and sevoflurane, indicating highly polluted workplaces. In conclusion, although global DNA methylation remained unchanged, WAG exposure was associated with modulation of circulating miRNAs, and our findings suggest that hsa-miR-1252-5p and hsa-miR-520f-3p emerge as potential biomarkers of effect associated to WAG occupational exposure in veterinarians who work in inadequately equipped VORs.

Body axial curvature is a common toxic effect of fish after exposure to environmental pollutants, but the molecular mechanism remains unclear. To investigate the underlying mechanisms, zebrafish were exposed to different persistent organic pollutants (POPs) including polychlorinated biphenyl (PCB) and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47). PCB40 was chosen as the typical POP to discover the mechanism because of its remarkable body axial developmental toxicity. The PCB40 exposure reduced the density of spinal cord cilia. We observed the upregulation of neuropeptides urotensin II-related peptide (urp2), which is downstream of cilia signaling. Further analysis revealed an upregulation of lysine (K)-specific demethylase 1a (Kdm1a, also known as LSD1), and the microscale thermophoresis and chromatin immunoprecipitation assays indicated Kdm1a directly bound PCB40 and enriched in urp2 promoter regions to enhance urp2 expression. Both interventions of Kdm1a using a specific inhibitor and CRISPR/Cas9-mediated knockout partially rescued the body axial curvature induced by PCB40. Further molecular docking suggested that the previously reported pollutants which induced zebrafish body axial curvature could interact with Kdm1a. We found that PCB95 and BDE-47 also induced body axial curvature through Kdm1a and urp2. In conclusion, the study identified a previously unrecognized molecular pathway, specific POPs induced zebrafish body axial curvature by binding to Kdm1a, with urp2 emerging as a key candidate downstream mediator via Kdm1a-dependent upregulation. The study provides a new mechanism for fish body axial curvature caused by specific POPs and provides new insights for pollutant toxicity evaluation and prevention.

The effectiveness of immunotherapy is significantly limited by the inherently low immunogenicity and immunosuppressive phenotypes of tumors. To address this challenge, we develop assemblies composed of chlorin e6 and regorafenib (designated as CeRe), which combine photodynamic therapy (PDT) with immune regulatiing functions. CeRe exhibits uniform nanoscale distribution and good stability without requiring additional carriers. Upon light activation, CeRe eradicate tumor cells via PDT-induced reactive oxygen species (ROS) while simultaneously triggering immunogenic cell death (ICD). Furthermore, CeRe downregulates PD-L1 expression in tumor cells, promotes macrophage polarization, and relieves the immunosuppressive tumor microenvironment (TME). These synergistic immunomodulatory effects substantially improve tumor responsiveness to αPD-L1 treatment, leading to the effective inhibition of both primary and metastatic tumor growth. Collectively, this work presents a carrier-free nanodrug assembly strategy with multifaceted mechanisms, offering a promising approach for precise tumor therapy and metastasis suppression.

Evidence suggests that environmental exposures induce epigenetic modifications that can have long-lasting effects on multiple health outcomes, and an in-depth review of the epidemiological evidence is urgent. We aimed to comprehensively assess the associations between long-term exposure to air pollution and epigenetic changes in adults.

Plant-specific HD2s have been characterized; HDT1 orthologs were dicot-specific. AtHDT4 interacts with AtILR3 to increase Cd tolerance. The plant-specific histone deacetylase 2 (HD2) family is crucial for growth and stress responses, yet its evolutionary origins and functional diversification remain largely unknown. Here, we systematically elucidated the evolutionary trajectories of HDT homologs. We found the emergence of HDT1 orthologs as a dicot-specific innovation, characterized by unique motif acquisition and accompanied by relaxed purifying selection. Synteny analysis indicated that the duplication events establishing the HDT1 and HDT3 lineages were associated with whole-genome duplications (WGDs) specific to the dicot lineage. In contrast, the expansion of the HDT gene family in monocots appears to rely primarily on local duplication mechanisms, such as tandem duplications. Codon usage analysis revealed distinct species-specific preferences: lycophytes, bryophytes, and algae exhibited higher frequencies of G3s, C3s, GC3, CBI, Nc, and overall GC content, suggesting potential adaptive evolution or optimization for translational efficiency. Functional validation demonstrated that AtHDT4 contributes to the plant response to cadmium (Cd) stress. Specifically, AtHDT4 expression was significantly upregulated under CdCl₂ treatment. Compared with wild-type (WT) plants, the hdt4 mutant exhibited markedly reduced activities of key antioxidant enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Inductively coupled plasma mass spectrometry (ICP-MS) analyses confirmed that AtHDT4 regulates Cd accumulation. Furthermore, AtILR3 expression was significantly downregulated in the hdt4 mutant, implicating it in the Cd stress response. As anticipated, direct protein-protein interaction between AtHDT4 and AtILR3 was verified. This study not only uncovers the critical role of AtHDT4 in mediating plant responses to Cd stress but also provides a broader evolutionary perspective on the functional diversification and specialization of HDT homologs across plant lineages.

SUMOylation regulates chromatin states and transcriptional programs that preserve cellular identity, yet how perturbation of the SUMOylation pathway impacts adipocyte plasticity remains unclear. Here, we show that brief pharmacologic inhibition of SUMOylation in human pre-adipocytes using TAK-981 primes stable de novo beige differentiation in the presence of the PPARG agonist rosiglitazone. Transient TAK-981 exposure produces changes in the transcriptome and metabolism of mature adipocytes, including robust induction of canonical beiging markers like UCP1 and increased mitochondrial respiration. Mechanistically, ATAC-seq and RNA-sequencing revealed immediate chromatin remodeling and early mobilization of CEBP family members, followed by stable activation of CEBPA and PPARG regulatory networks. ChIP experiments demonstrated loss of H3K27me3 and gain of H3K27ac at PPAR response elements at thermogenic enhancers, and increased PPARG occupancy across the UCP1 regulatory unit. This mechanism is enforced by enhanced cAMP-PKA-p38 signaling and stabilization of beiging transcription activators. We propose that transient relief of SUMO-mediated repression unlocks dominant regulatory units, notably the UCP1 enhancer cluster, producing a monomorphic reprogramming toward adaptive thermogenesis. These findings identify SUMOylation as a reversible epigenetic barrier to adipocyte beiging and suggest that temporally controlled SUMO pathway inhibition combined with PPARG activation could be exploited to modulate adipose tissue thermogenic capacity.

The oomycete Phytophthora capsici causes Phytophthora blight, a major constraint on global pepper production. Our previous observations indicated that pretreating plants with thiamethoxam (TMX) and imidacloprid (IMI) could reduce the incidence of pepper blight, but the underlying mechanisms remained unclear. Here, we investigated how TMX and IMI induced resistance in pepper (Capsicum frutescens) against P. capsici. Both in vitro and in vivo assays demonstrated that TMX and IMI suppressed disease, not by directly impairing pathogen virulence but by inducing systemic resistance in susceptible (Cusheng L09) and resistant (Cusheng 356) pepper cultivars. Split-plant systemic resistance assays showed that TMX/IMI-primed plants developed smaller lesions in both treated and untreated leaves following P. capsici infection. Foliar application of TMX and IMI effectively alleviated disease severity, with IMI showing superior efficacy in attenuating reactive oxygen species (ROS) accumulation, and TMX/IMI priming concomitantly altering the activities of ROS-scavenging enzymes under pathogen challenge. Reverse transcription-quantitative PCR analysis revealed time-dependent changes in defence gene expression, and whole-genome transcriptome profiling highlighted temporal reprogramming of pathogenesis-related genes. Further functional validation identified CaNEN4 as a susceptibility factor. Collectively, our findings reveal that IMI/TMX primes pepper plants with systemic resistance by modulating ROS homeostasis, defence gene expression, and susceptibility gene function, offering novel insights into chemical-induced plant immunity and genetic targets for durable blight resistance in crops.

EGFR, one of the most successful therapeutic targets, has recently been found to exert a novel function for regulating homologous recombination (HR). Activation of HR is the critical event of treatment failure of PARPi in BRCA1/2 wild-type ovarian cancer (OC). Besides, the antitumor effects of biguanides have also been a focus of attention. Here, we discovered that the new biguanide 4C inhibited HR and sensitized BRCA1/2 wild-type OC cells to PARPi by targeting EGFR. Mechanistically, EGFR promoted nuclear accumulation of both BRCA2 and Rad51, and HR activation by competitively inhibiting the binding of BRCA2 and Rad51 to E3 ubiquitin ligase c-Cbl, thereby reducing cancer cell sensitivity to PARPi following ATM-mediated DNA damage signal transmission from the nucleus to the cytoplasm. Interestingly, EGFR was downregulated by 4C, which in turn enhanced the interaction of BRCA2 and Rad51 with c-Cbl. Consequently, BRCA2 and Rad51 were then ubiquitinated and degraded to inhibit HR and increase the sensitivity of OC to PARPi. Thus, these findings reveal that the combination of 4C with PARPi leading to "synthetic lethality" is an effective strategy for treating BRCA1/2 wild-type OC.

Dihuang Yinzi (DY) is a classic formula traditionally used for stroke-related disorders. Its potential therapeutic effect on post-stroke depression (PSD), however, remains to be investigated.

The mouse double minute 2 homolog (MDM2) - p53 interaction inactivates p53, a key tumor suppressor, and has been validated as a target for cancer therapy. Four triazolyl-thio-oxazines 4(a-d) were rationally designed based on a three-finger pharmacophore model, which parallels the binding conformation of p53 to suppress this oncogenic interaction. All compounds were characterized by 1H/13C NMR, IR, and mass spectrometry, whereas 4a was characterized by X-ray crystallography. All compounds bound to the MDM2 binding site at key residues (Leu54, Leu57, Ile61, Met62, Phe86, and Tyr100) with similar binding affinities (-7.88 to -8.75 kcal/mol). Further 200 ns molecular dynamics simulations and MM-GBSA analyses were performed to determine the precise differences. Complex 4a demonstrated better binding stability (RMSD <2.0 Å) and MM-GBSA Binding Score ΔGbind (-172.718 kcal/mol) than either Nutlin-3a or the p53 peptide. Its fluorobenzene dives further into the MDM2 cleft than the chlorobenzene ring in Nutlin-3a, resulting in stronger π-π stacking and hydrophobic interactions. SwissADME profiling also predicted that 4a has a good balance of pharmacokinetic properties. This was evident from the cytotoxicity studies (MCF-7 breast cancer cells), which revealed 4a to be the most potent (IC₅₀ = 4.379 μM). Further Western blot analysis showed significant upregulation of p53 and p21 in MCF-7 cells upon treatment with 4(a-d). In contrast, no significant induction of p53 or p21 was observed in MDA-MB-468 cells, confirming the upregulation of wild-type p53. These results suggest that 4a is a promising candidate for further biological evaluation as an MDM2-p53 interaction inhibitor.

Children with favorable-histology Wilms tumor (FHWT) who relapse or whose tumors show blastemal predominance post-chemotherapy often face poor outcomes. The purpose of this study is to identify mechanisms of chemotherapy resistance in FHWT. We induce a patient-derived xenograft model (KT-47) to develop blastemal predominance after chemotherapy and to become resistant to vincristine, actinomycin-D, and doxorubicin (VAD). Multi-omics analyses reveal chromatin and transcriptional changes, including increased H3K4me3 and decreased H3K27me3 at stem cell and nephrogenesis gene loci. LIN28B is the most upregulated resistance-associated gene, linked to MYCN copy gain/upregulation and chromatin remodeling. ABCB1 expression correlates with interchromosomal enhancer interactions and functions as the mediator of chemotherapy resistance in vitro. These findings are validated in additional Wilms tumor models. Overall, resistance is associated with de-differentiation to a stem-like state and is driven by ABCB1 upregulation, suggesting that therapeutic strategies targeting chromatin regulation and drug efflux may be relevant in therapy-resistant Wilms tumor.

Eradicating leukemia stem cells (LSCs) and overcoming tyrosine kinase inhibitor (TKI) resistance is urgent for chronic myeloid leukemia (CML) treatment. We find that F-box protein 3 (FBXO3) is highly upregulated in CD34+ CML stem cells from TKI-resistant patients and identify it as an innovative CML-LSC marker via single-cell RNA sequencing (scRNA-seq). FBXO3 deficiency induces apoptosis and reduces proliferation of CML cell lines and LSCs in vitro and in vivo, with minimal effects on normal CD34+ hematopoietic stem cells (HSCs). Mechanistically, FBXO3 interacts with DUSP9 to promote its ubiquitination and activate the MAPK pathway, critical for CML cell activity. DUSP9 knockdown partially reverses FBXO3-deficiency-mediated LSC elimination. Furthermore, FBXO3 inhibitor monotherapy or combination with imatinib effectively eradicates CML-LSCs, overcomes TKI resistance, and spares normal hematopoiesis. Collectively, our findings highlight FBXO3's role in CML progression and support combining FBXO3 inhibitors with TKIs for durable LSC elimination.

Aging is a long-term and complex process characterized by cellular changes, including cell cycle arrest, increased production of the senescence-associated secretory phenotype (SASP), and nuclear membrane disintegration. To accelerate the aging process, various animal models are made using chemical inducers, such as D-galactose (D-Gal). D-Gal has been extensively applied to induce aging in experimental animals; however, its effects on the gastrointestinal system, particularly the colon, remain underexplored. Observation of molecular aging markers will provide a valuable approach to assess these cellular changes. This study aimed to observe the expression of genes related to cell cycle arrest (p53 and Cdkn1a), SASP production (Il-6 and Il-1β), nuclear membrane disintegration (Lmnb1), as well as histological inflammation process in the colon of D-Gal-induced aging rat models. Twelve-week-old male Sprague-Dawley rats (n = 12) were divided into the control and D-Gal (100 mg/kg/day for 6 weeks) groups. The expression levels of aging-related genes (p53, Cdkn1a, Il-6, Il-1β, and Lmnb1) were assessed by reverse-transcription quantitative polymerase chain reaction. The D-Gal group exhibited inflammatory cell infiltration. Il-1β immunohistochemical score along with Il-1β expression was significantly elevated in D-Gal group (p < 0.01), suggesting SASP activation and pro-inflammatory signaling. No statistically significant differences were observed in the expression levels of p53, Il-6, Cdkn1a, or Lmnb1 expression. Molecular and histological results demonstrated that D-Gal administration induces colonic inflammation, characterized by increased Il-1β expression, which subsequently promotes potential cellular senescence.

Cancer persists as a major health burden, fueled not only by genetic mutations but also by profound epigenetic instability that rewires transcriptional programs and DNA repair networks. Among the most intensively studied epigenetic regulators are histone deacetylases (HDACs) and poly (ADP-ribose) polymerases (PARPs), whose dysregulation fosters genomic instability, unchecked proliferation, and therapeutic resistance. Pharmacological inhibition of HDACs or PARPs alone has achieved meaningful advances, yet intrinsic and acquired resistance, limited tumor selectivity, and relapse remain formidable barriers to durable efficacy. To address these challenges, attention has shifted toward rational combination strategies and, more recently, to the development of dual inhibitors. By integrating key pharmacophoric features from both HDAC and PARP inhibitors, these hybrids are designed to achieve balanced target engagement within a single scaffold, thereby maximizing synergy while reducing pharmacokinetic complexity. Mechanistically, dual blockade disrupts DNA repair fidelity, induces chromatin relaxation, and amplifies apoptotic signaling, thereby producing antitumor effects that exceed those of monotherapy. While this paradigm offers substantial promise, it is not without limitations, including potential off-target toxicity, challenges in optimizing linker chemistry, and the need for precise structure-activity relationship (SAR) refinement. This review consolidates structural, mechanistic, and SAR insights, emphasizing how dual HDAC/PARP inhibition represents a next-generation therapeutic strategy poised to overcome resistance and broaden the spectrum of effective cancer interventions.

Aging is a primary risk factor for disease progression in multiple sclerosis (MS). Because of this, treatments that can reduce the consequences of molecular aging, like senescence, have been proposed as a strategy to address disease progression. However, the effects of senolytics, a class of drugs which selectively ablate senescent cells, on the central nervous system are largely unknown. Here, we examined the effects of senolytic treatment on myelination and oligodendrocyte function in vivo using C57BL6/J mice and in vitro using primary rat oligodendrocyte cultures. Initial data showed that naïve young (3 to 4 mo) and aged (22 mo) C57BL6/J mice treated with dasatinib and quercetin (D+Q) developed significant demyelination compared to vehicle-treated controls, though no cell death was observed in the brain. In vitro, oligodendrocyte progenitor cells treated with D+Q in differentiation media exhibited significantly reduced myelin basic protein protein and morphological complexity, also without inducing cell death. Bulk RNA sequencing and ingenuity pathway analysis of D+Q treated oligodendrocytes identified differentially expressed genes associated with endoplasmic reticulum stress. These data suggest that D+Q evokes the unfolded protein response in oligodendrocytes, causing oligodendrocyte dysfunction and myelination failure. Due to the resemblance between oligodendrocytes treated with D+Q and those found in MS lesions, D+Q treatment offers a potential method to model an aspect of oligodendrocyte dysfunction relevant to MS. Therefore, understanding the mechanism by which D+Q perturb oligodendrocyte function may provide insight into some of the pathological features contributing to disease progression in MS.

Huntington's disease (HD) is a progressive neurodegenerative disorder caused by a CAG-repeat expansion in the Huntington gene (HTT). Herein, we describe the discovery of a series of HTT pre-mRNA-splicing modulators that promote the inclusion of a cryptic stop codon that in turn lowers levels of mutant Huntington protein (mHTT). Optimization of the starting thienopyridine amide core resulted in the discovery of the potent, CNS-penetrant, selective, and orally bioavailable HTT-splicing modulator BIO-6553. This lead compound is structurally distinct from existing splicing modulators, demonstrated significant HTT-lowering in both human cells and mouse YAC128 models, and has an attractive off-target profile from RASL- and RNA-seq analysis.

Glioblastomas (GBMs) are lethal brain tumors characterized by rapid growth and resistance to standard treatment. Resveratrol (RES) increases the levels of reactive oxygen species (ROS) and induces cell death in sensitive GBM cells. However, the death patterns induced by RES and their relevance to NFE2-related factor 2 (NRF2) remain unclear. The current study aimed to address these issues using RES-sensitive U251 and less-sensitive LN428 GBM cell lines, as well as an orthotopic GBM xenograft rat model. In silico analysis revealed high NRF2 expression in GBM tissues and a strong correlation with tumor progression in the TCGA dataset. After 48 h of 100 µM RES treatment, NRF2 levels remained stable in LN428 cells but significantly decreased by 2.3-fold in U251 cells, accompanied by suppressed growth and NRF2-regulated and ferroptosis-related xCT and GPX4 downregulation. Elevated Fe2+, ROS levels, lipid peroxidation, and ferroptotic frequency were evidenced in RES-treated U251 cells; meanwhile, apoptosis and reduced NRF2-HO-1 expression were also evident in U251 cells. Combined treatment with Ferrostatin-1 and Z-VAD FMK rescued 60% of U251 cells compared to RES-treated counterparts. In vivo, lumbar puncture (LP) administration of RES induced both ferroptosis and apoptosis in rat orthotopic GBM xenografts. These findings highlight the dual cell death induced by RES in sensitive GBM cells and identify NRF2 signaling status as a novel determinant of cellular response to RES treatment.

Intrahepatic cholangiocarcinoma (iCCA), a highly lethal subtype of liver cancer with increasing incidence and poor prognosis, displays profound molecular heterogeneity that limits therapeutic efficacy. Combining multi-omics with pathological subtyping in iCCA, we identified a strong association between the most aggressive molecular subtype and the large-duct pathological type, characterized by aberrant mucin overexpression and prominent myeloid cell infiltration. Through integrative analysis, MUC16 was identified as a key molecular marker specifically enriched in large-duct type intrahepatic cholangiocarcinoma (L-iCCA). Functional modulation of MUC16 markedly inhibited L-iCCA cell proliferation and migration. Furthermore, high-throughput drug screening identified the small-molecule compound OSMI-1 as a potent suppressor of MUC16 expression. Using primary iCCA cell lines and patient-derived organoids (PDOs), we confirmed that OSMI-1 inhibits L-iCCA cell proliferation and migration by downregulating MUC16. Mechanistically, OSMI-1 suppresses MUC16 expression by disrupting the transcriptional complex formed between OGT and IRF1. Employing a genetically engineered L-iCCA mouse model, we further validated the crucial role of MUC16 in tumor progression and neutrophil infiltration, demonstrated that OSMI-1 synergized with gemcitabine, effectively inhibiting L-iCCA growth. This study presents a pathological subtype-based precision therapeutic strategy for L-iCCA, thereby providing a foundation for novel translational approaches to the personalized management of this disease.