Polyunsaturated fatty acids (PUFAs) have demonstrated promising anticancer properties by inducing cancer cell death and inhibiting cancer metastasis. As dynamic organelles, lipid droplets (LDs) may protect cells from PUFA-induced lipotoxicity by sequestering excess fatty acids. However, the underlying mechanisms regulating LDs dynamics in PUFA-mediated tumor cytotoxicity remain poorly understood. In this study, we report that inhibition of LDs synthesis enhances PUFA-induced cancer cell death, suggesting that LDs formation protects against PUFA cytotoxicity. We further demonstrate that copper ions potentiate the antitumor effects of PUFAs across multiple cancer cell lines primarily by promoting apoptosis rather than cuproptosis or ferroptosis. Mechanistically, copper ions significantly reduce intracellular LDs accumulation by promoting LDs degradation via activation of ATGL-dependent lipophagy. Collectively, these findings uncover a novel mechanism whereby copper ions enhance PUFA-induced tumor cell death through promoting lipophagy, which provides valuable insights for optimizing PUFA-based cancer therapies by targeting lipid metabolism and copper homeostasis.

The mechanisms underlying the interactions between disseminated tumor cells (DTCs) and their tissue microenvironment during metastatic colonization are currently poorly understood. We integrated multimodal single-cell and spatial profiling from liver cancer mouse models and human metastases to track the spatiotemporal dynamics of DTCs and their microenvironments from single-cell seeding to overt lung metastasis. We identified a residual population of quiescent Phgdhhigh DTCs that survived initial innate immune clearance and became transiently enriched in micrometastases. These cells shaped an immune-scarce microenvironment through PHGDH-dependent, H3K27me3-mediated epigenetic silencing of chemokine transcription, thereby promoting metastatic expansion. Cx3cr1high interstitial macrophages were also transiently enriched before DTC expansion, creating an immune-privileged niche for metastatic outgrowth by recruiting immunosuppressive cells. Inactivating the PHGDH-H3K27me3 axis in DTCs or depleting interstitial macrophages restored immune surveillance and inhibited metastatic colonization. These findings provide insights into the development of micrometastasis-targeting regimens.

Bilirubin, a byproduct of heme metabolism, is traditionally recognized for its antioxidant properties. However, its accumulation under pathological conditions can result in severe clinical manifestations, including neurotoxicity. Here, we report that bilirubin acts as an activator of receptor-interacting protein kinase 3 (RIPK3), inducing necroptosis and contributing to neurotoxic effects. Unlike the canonical pathway, in which RIPK3-mediated necroptosis is dependent on RIPK1, bilirubin directly binds to the D161 and S102 residues within the kinase domain of RIPK3. This binding facilitates RIPK3 oligomerization and autophosphorylation, leading to the activation of MLKL, the primary effector of necroptosis. Notably, bilirubin-induced necroptosis does not require the RHIM domain of RIPK3 or other RHIM-containing proteins, such as ZBP1 and TRIF. Functionally, the depletion of Ripk3 reduces bilirubin-induced neurotoxicity in murine models. These findings reveal a previously unrecognized mechanism of RIPK3-mediated non-canonical necroptosis, establishing it as a key mediator of bilirubin-induced neurotoxicity.

Copper is an essential trace element that governs diverse cellular functions and influences cell fate. However, how cells adapt to copper deprivation remains poorly understood. Here, we identify a copper-ferroptosis regulatory axis mediated by the cystine transporter SLC7A11. We show that copper loss, induced either by silencing of the copper importer SLC31A1 or by pharmacological chelation, leads to a marked upregulation of SLC7A11. This adaptive response enhances glutathione synthesis, bolsters antioxidant defenses, and protects cells from ferroptosis. Mechanistically, copper deprivation activates AMPK, which stabilizes the transcription factor NRF2 to drive SLC7A11 expression. Functionally, SLC31A1 depletion diminishes ferroptosis-dependent tumor suppression in xenograft models, while dietary copper restriction alleviates ferroptosis-mediated pancreatic injury in experimental acute pancreatitis. Together, these findings reveal copper deprivation as a robust condition driving ferroptosis resistance and suggest that dietary or pharmacological copper modulation could provide new strategies to fine-tune ferroptosis in cancer and tissue injury.

Atrazine, a widely used chlorotriazine herbicide, persists in aquatic environments and poses potential carcinogenic risks. While epidemiological studies link atrazine exposure to malignancies, its intrinsic molecular mechanisms across organ systems remain incompletely understood. This study employed integrated network toxicology and transcriptomic analyses to clarify atrazine-associated oncogenic pathways in liver hepatocellular carcinoma (LIHC), kidney renal clear cell carcinoma (KIRC), lung adenocarcinoma (LUAD), and sarcoma (SARC). Transcriptomic data from The Cancer Genome Atlas (TCGA) for these cancers were analyzed to identify atrazine-related genes. Protein-protein interaction networks were constructed and analyzed to identify hub genes. Functional enrichment, immune microenvironment analyses, and survival analysis were performed. Molecular docking validated atrazine-target binding, and independent datasets were used for hub gene expression and pan-cancer relevance validation. We identified 92 (LUAD), 136 (LIHC), 137 (KIRC), and 161 (SARC) atrazine-associated targets. Hub genes including CDC6, MCM5/7, UBE2C, FEN1, CDCA8, and VIM were differentially expressed across these cancers. Enrichment analyses revealed atrazine disruption of core pathways, including cell cycle progression and chromosomal instability, epithelial-mesenchymal transition, metabolic reprogramming, and senescence-associated secretory pathways. Molecular docking confirmed high-affinity binding between atrazine and key targets. Pan-cancer validation implicated these hub genes in multiple additional malignancies. Transcription factor analysis nominated HSD17B8 as a key regulatory node. This study demonstrates that atrazine promotes carcinogenesis by dysregulating conserved networks governing genomic stability, cell proliferation, metabolic adaptation, and immune microenvironment remodeling, providing a mechanistic framework linking aquatic atrazine exposure to multi-organ carcinogenesis and nominating HSD17B8-associated pathways for therapeutic intervention. These findings underscore the imperative for enhanced environmental monitoring of atrazine contamination.

BACKGROUND:Tissue-resident macrophages (TRMs) exhibit dual roles in tumor progression, yet their functional reprogramming within the tumor microenvironment (TME) remains a critical unresolved question. METHODS:We integrated single-cell and spatial transcriptomics from a pan-cancer atlas of 1.39 million cells across five malignancies with 2,318 bulk RNA-seq samples to investigate macrophage states. A TRM inflammatory remodeling signature (TIR-Sig) was developed for clinical biomarker validation. RESULTS:We identified a conserved inflammatory TRM subtype (iTRM) characterized by CXCL8/IL1B/IL6 co-expression that correlates with poor clinical outcomes. Crucially, both TRMs and monocyte-derived tumor-associated macrophages (Mono-TAMs) underwent convergent differentiation into functionally similar inflammatory phenotypes, establishing iTRM as a universal tumor-educated state. Further integration analysis revealed an iTRM-enriched TME subtype which featured coordinated infiltration of neutrophils and cancer-associated fibroblasts (CAFs), forming a 'cold tumor' ecosystem associated with immune checkpoint blockade (ICB) resistance and poor prognosis. The derived TRM inflammatory remodeling signature (TIR-Sig) demonstrated dual clinical utility: it predicted patient survival (HR = 19.86, p < .001) and stratified ICB responders (AUC = .706). CONCLUSION:This study establishes phenotypic links between tissue-resident and recruited macrophages through inflammatory reprogramming within TME, provides a unifying framework for pan-cancer macrophage plasticity in TME, delivers a clinically actionable biomarker suite (TIR-Sig), and provides potential therapeutic targets for TME remodeling. KEY POINTS:Cross-tissue single-cell atlas of tissue-resident macrophages (TRMs). Identification of conserved inflammatory TRM phenotype (iTRM) in pan-cancer. Dynamic convergence of TRM and monocyte-derived macrophage lineages. TRM inflammatory remodelling signature (TIR-Sig) with clinical potential.

Effective therapy for relapsed or refractory central nervous system lymphoma (r/r CNSL) remains an unmet medical need. Meanwhile, developing antitumor drugs for CNS malignancies faces the dual challenge of achieving effective blood‒brain barrier (BBB) penetration and potent tumor cell killing. To address these challenges, a comprehensive predictive system was established to support decision-making during the discovery of HZ-A-018, a potent and BBB-permeable Bruton tyrosine kinase (BTK) inhibitor. This study further presents key preclinical results for HZ-A-018, as well as efficacy/safety data from a multicenter Phase 1 trial in r/r CNSL patients. HZ-A-018 demonstrated manageable safety, with only 19.2% of patients experienced grade 3 or higher adverse events according to the Common Terminology Criteria for Adverse Events version 5.0. Treatment with HZ-A-018 at the recommended phase II dose (RP2D) of 600 mg achieved an overall response rate (ORR) of 72.7% (95% CI, 39.0–94.0) and a 12-month survival rate of 90.5%. The Center for Drug Evaluation in China has authorized the initiation of this single-arm Phase II study as a pivotal registrational clinical trial for accelerated approval of HZ-A-018 for monotherapy in patients with r/r PCNSL. This trial has been registered under the identifiers ChiCTR2400091821 at www.chictr.org.cn and CTR20210181 at www.chinadrugtrials.org.cn.

The conserved non-coding RNA 7SK is a well-established global transcriptional repressor, yet its context-specific functions in cancer and therapy resistance remain paradoxical. Here, we resolve this paradox by uncovering a dual-axis mechanism through which 7SK drives colorectal cancer (CRC) resistance. By integrating single-cell multi-omics with functional assays, we demonstrate that 7SK not only selectively activates the JUN transcriptional network to fuel tumor proliferation but also reduces global transcriptional entropy to stabilize an immunosuppressive microenvironment and promote immune escape. This “local activation-global suppression” paradigm is conserved across multiple cancer types, positioning 7SK as a potential pan-cancer therapeutic target. Our findings reveal 7SK as a dynamic modulator that balances oncogene-specific transcription with global transcriptional suppression across cancers, providing a new framework for understanding and targeting ncRNA-mediated resistance.

Relapsed/refractory (r/r) angioimmunoblastic T-cell lymphoma (AITL) is associated with a dismal prognosis, with historical overall survival (OS) < 6 months, underscoring the urgent need for novel therapies. We conducted the RCLARITY trial to evaluate the efficacy and safety of a chemotherapy-free regimen in this patient population. This single-arm, multicenter, prospective Phase II trial enrolled adult patients with r/r AITL. Patients received rituximab (375 mg/m2 intravenously on Day 1), lenalidomide (15 mg orally on Days 1-21), and chidamide (30 mg orally twice weekly) in 28-day cycles for up to 6 cycles. The primary endpoint was progression-free survival (PFS). Secondary endpoints included the overall response rate (ORR), OS, duration of response (DoR), and safety. In total, 28 patients were enrolled between August 2019 and June 2024. The median age was 64 years (range, 44-70), and 71.4% had refractory disease. The ORR was 71.4% (complete remission: 32.1%), median PFS was 5.5 months (95% confidence interval [CI], 3.5-7.6), median OS was 17.6 months (95% CI, 10.3-24.8), and median DoR among responders was 10.7 months (95% CI, 2.7-16.0). Notably, 50% of patients with clonal Ig heavy- or light-chain rearrangements achieved PFS > 16 months. All nine patients with baseline serum EBV-DNA positivity converted to undetectable levels after two cycles. No Grade 5 adverse events or venous thromboembolic events occurred. The most common hematological adverse events per cycle were leukopenia (14.3%), thrombocytopenia (12.4%), and neutropenia (11.4%). Overall, the RLC regimen induced clinically meaningful anti-tumor activity with manageable toxicity in patients with r/r AITL. Trial Registration: ClinicalTrials.gov identifier: NCT04319601.

IntroductionNiraparib and bevacizumab are two principal maintenance therapies for newly diagnosed advanced ovarian cancer (AOC) patients with BRCA wild-type (BRCAwt) status, regardless of homologous recombination deficiency (HRD). In China, however, a considerable proportion of BRCAwt patients have unknown or untested HRD status, complicating treatment selection.MethodsTo evaluate and compare the efficacy of niraparib and bevacizumab as maintenance therapy for BRCAwt AOC, we conducted a retrospective cohort study using real-world clinical data. Descriptive statistics were used to summarize clinical and demographic characteristics. Progression-free survival (PFS) was estimated using Kaplan-Meier analysis and compared using a stratified Cox proportional hazards model. A multivariable Cox regression was performed to adjust for potential confounding variables. Exploratory subgroup analyses were conducted, and propensity score matching (PSM) was applied as a sensitivity analysis.ResultsA total of 94 patients were included, with 51 receiving niraparib and 43 receiving bevacizumab. The median PFS was not reached in the niraparib group versus 13.77 months (95% CI, 4.12-23.41) in the bevacizumab group (HR = 0.240, 95% CI, 0.128-0.451; P < .001). After covariate adjustment, the median PFS was 19.55 months (95% CI, 9.40-NA) with niraparib and 8.64 months (95% CI, 4.53-NA) with bevacizumab, with an adjusted HR of 0.282 (95% CI, 0.136-0.587; P = .001). In the PSM sensitivity analysis, the median PFS was not reached (95% CI, 19.55-NR) in the niraparib group and was 18.33 months (95% CI, 8.90-25.26) in the bevacizumab group (HR = 0.360, 95% CI, 0.176-0.736; P = .005).ConclusionThis analysis suggests that niraparib may provide a progression-free survival advantage compared with bevacizumab in BRCAwt AOC patients, with both regimens appearing to be generally well tolerated in the real-world setting. These findings offer preliminary reference value for maintenance treatment selection in patients with newly diagnosed BRCAwt AOC.

The epigenetic landscape and tumor microenvironment (TME) interactions of non-functioning pituitary adenomas (NFPAs), benign tumors with high morbidity and recurrence rates, are not well characterized. We completed single-nucleus (sn) multiomics assays on 4 gonadotrope NFPAs (34,819 cells) and 11 non-diseased postmortem control pituitaries (51,535 cells), finding decreased proportions of tumor-associated endothelial cells and pericytes and increased proportions of macrophages. We identified bidirectional tumor-macrophage crosstalk comprising nine ligand-receptor interactions and experimentally validated the macrophage-initiated SFRP1-FZD6 interaction, whose predicted target genes CCND1, CDK6, SGK1, and TGFBR2 were linked to tumorigenesis. We uncovered coordinated gene expression and chromatin accessibility programs, which distinguished adenoma cells from gonadotropes. Integrated transcriptome-chromatin modeling revealed gene regulatory circuits (GRCs) that showed altered activity in adenoma cells and were regulated by transcription factors (TFs), including PBX3 and MEF2C. Our study provides insight into the altered epigenetic gene control landscape and TME processes of the NFPA tumor phenotype. Our data are freely available at https://rstudio-connect.hpc.mssm.edu/nfpa_browser/.

Abstract Introduction ILC2s (group 2 Innate lymphoid cells) are involved in type 2 immune responses during both phases of innate and adaptive response. The transcription factor Bhlhe40 has been reported to regulate the balance between inflammatory and anti-inflammatory reactions in CD4 T cells. However, although Bhlhe40 is also expressed by ILC2s, its functions in these cells are unknown. Methods By challenging Bhlhe40-germline knockout mice and ILC2-specific Bhlhe40 knockout mice with papain acutely, we found that eosinophils were reduced in these mice compared to their wild type controls, indicating that Bhlhe40 is required for ILC2 function in recruiting eosinophils in response to papain-induced lung damage. Results Interestingly, the expression of IL-5 known for recruiting eosinophils was not altered in Bhlhe40 deficient ILC2s. Instead, we found a reduction in GM-CSF expression but an increase in IL-10 production in the absence of Bhlhe40. To get a clearer picture of how Bhlhe40 regulates ILC2 function, we performed scRNA-seq and scATAC-seq of WT and Bhlhe40 knockout ILC2s, and found Bhlhe40 affected ILC2 subsets and regulated gene expression in distinct clusters of ILC2s. Further analysis of the Il10 locus identified several interesting elements at the Il10 promoter/enhancer regions that had differential gene accessibility between WT and knockout. Furthermore, we confirmed that Bhlhe40 can directly bind to Il10 locus in ILC2s by ChIP-Seq. On the other hand, no obvious changes in DNA accessibility were noted at the Csf2 (encoding GM-CSF) locus, suggesting Bhlhe40 may indirectly regulate GM-CSF expression. Conclusion Overall, our results demonstrate that just as its function in T cells, Bhlhe40 also regulates the balanced expression of inflammatory and anti-inflammatory cytokines in innate lymphoid cells. Funding Source n/a Topic Categories Innate Immune Responses and Host Defense: Molecular Mechanisms (INM)

Double-hit or triple-hit lymphoma (DH/THL) is an aggressive subtype with poor prognosis. This study evaluated the efficacy and safety of selinexor, a first-in-class oral inhibitor of exportin 1 (XPO1), combined with R-CHOP (S-RCHOP) as first-line therapy for newly diagnosed DH/THL. This single-arm, prospective phase Ⅱ trial (NCT 05974085) enrolled 13 patients between May 2022 and August 2024. Patients received up to six 21-day cycles of S-RCHOP (selinexor 60 mg on days 1, 8, 15). The primary endpoint was overall response rate (ORR). Secondary endpoints included progression free survival (PFS), overall survival (OS), central nervous system (CNS) relapse rate within 2 years, and adverse events (AEs). Exploratory analyses included next-generation sequencing (NGS) and circulating cell-free DNA (cfDNA) monitoring. 13 patients (9 DHL, 4 THL) were enrolled. The ORR was 100% (CR: 76.9%, PR: 23.1%). At a median follow-up of 25.4 months, the 2-year PFS and OS were 67.7% and 67.1%, respectively. One patient developed CNS relapse. The most common all-grade AEs included leukopenia/neutropenia (55.1% each), febrile neutropenia and fatigue (43.5% each), and thrombocytopenia (33.7%). NGS and cfDNA revealed frequent BCL6 and IGLL5 mutations. Post-treatment cfDNA negativity was achieved in 72.7% of patients, all in CR by PET-CT. In this exploratory phase Ⅱ study, S-RCHOP achieved high response rates in newly diagnosed DH/THL but was associated with frequent hematologic AEs. cfDNA monitoring provided valuable insights into treatment response. These preliminary findings support further investigation of XPO-1 inhibition with R-CHOP, with priority given to dose optimization. NCT05974085

Obesity and its associated metabolic complications represent a global health crisis, yet effective microbiota-targeted pharmacotherapies remain limited. Here, we report that GV-971 (sodium oligomannate), a marine-derived oligosaccharide originally developed for Alzheimer's disease, exerts potent anti-obesity and metabolic benefits by reprogramming gut microbial and host signaling networks. In high-fat diet-induced obese mice, GV-971 reduced adiposity, improved glucose homeostasis, and alleviated hepatic steatosis without affecting food intake. Multi-omics and causal intervention experiments revealed that GV-971 selectively decreased the abundance of Clostridium scindens, a keystone bacterium responsible for secondary bile acid synthesis. This decrease downregulated the expression of the baiF gene encoding 7α-hydroxysteroid dehydrogenase, leading to reduced intestinal deoxycholic acid (DCA) levels and inhibition of intestinal farnesoid X receptor (FXR) signaling. Restoration of C. scindens abundance, baiF expression, or DCA supplementation abrogated the metabolic benefits of GV-971, confirming the causal role of the C. scindens-DCA-FXR axis. Mechanistically, inhibition of intestinal FXR promoted thermogenic gene expression and white adipose tissue browning, thus enhancing systemic energy expenditure. These findings uncover a bacterium-metabolite-host signaling pathway underlying the effects of GV-971 and establish microbiota-directed FXR modulation as a promising therapeutic approach for obesity and metabolic disease.

The transcription factor Foxp3 in mice dictates the development and function of regulatory T cells (Tregs) to maintain self-tolerance. The mutation of amino acid 370 (M to I) of FOXP3 protein was identified in IPEX patients with type-2 autoimmunity. We generated an M370I-knock in (KI) mouse by CRISPR-Cas9. The homozygous female KI (Foxp3M370I/M370I) mice and the hemizygous male KI (Foxp3M370I/Y) mice showed T cell activation with multi-organ inflammation. The KI Tregs were capable to produce effector T cell cytokines (IL-2, IFN-γ, IL-4, and IL-17A). We also generated heterozygous KI female (Foxp3M370I/GFP) mice which were healthy in general without type-2 inflammation. However, the KI Tregs still possessed the ability of producing IL-2/IFN-γ. IP-MS of Foxp3 showed an altered pattern of Foxp3 interacting proteins between WT and KI Tregs, including NFAT and RUNX1. Structural modelling indicated that M370I may disrupt head-to-head dimerization of FOXP3. While another amino acid substitution F374C that sits in the center of H4 helix, may form an intra-domain disulfide to favor swap-domain dimer. We further generated mice carrying FOXP3-F374C mutation and these mice indeed had similar phenotypes. Thus, our results indicate that M370I and F374C KI Tregs displayed effector-phenotypes, possibly resulting from enhanced formation of domain-swap FOXP3 dimerization. This work was supported by the Division of Intramural Research of NIAID, NIH. NIH/NIAID 1ZIAAI001169 Immune Response Regulation: Molecular Mechanisms (IRM)

Triple-negative breast cancer (TNBC) is an aggressive subtype of invasive breast cancer characterized by limited treatment options and a poor prognosis. While ferroptosis, an iron-dependent form of regulated cell death, plays a role in tumor suppression, its specific molecular mechanisms in TNBC remain largely unexplored. In this study, we identify deubiquitinase USP24 as the most significantly altered enzyme among key deubiquitinating enzymes during ferroptosis in human TNBC cells. Silencing USP24 enhances ferroptosis-mediated tumor suppression in TNBC cells. Mechanistically, USP24 interacts directly with dihydroorotate dehydrogenase (DHODH) and deubiquitinates it, a process critical for maintaining coenzyme Q reduction and protecting cells from lipid peroxidation. Consistently, pharmacological inhibition of USP24 synergizes strongly with ferroptosis inducers in both in vitro and in vivo models via a DHODH-dependent pathway. These findings highlight USP24 as a potential therapeutic target to enhance ferroptosis sensitivity in TNBC.

Background The current treatment management for primary central nervous system lymphoma (PCNSL) relies on high-dose methotrexate (HD-MTX)-based regimens, followed by consolidation therapy (1-3), including whole-brain radiotherapy (WBRT) (4). However, concerns persist regarding WBRT-related neurotoxicity and cognitive impairment, particularly in older patients. Whether dose-reduced WBRT in patients achieving complete remission (CR) can mitigate these risks, or whether WBRT alone is sufficient for patients ineligible for systemic therapy, remains uncertain. Patients and methods Consecutive patients with biopsy-proven PCNSL who received cranial irradiation between January 2000 and December 2022 were identified. Dates of symptom onset, diagnosis, first hospital visit, first chemotherapy cycle, radiotherapy initiation, relapse or progression, death, and last follow-up were recorded to compute overall response rate (ORR), progression-free survival (PFS) and OS. Patients completed Minimum Mental State Examination (MMSE) evaluation. Survival analyses were performed using Kaplan-Meier method. Results Of 244 biopsy-proven PCNSL patients, 62 who received brain irradiation were screened. The median age was 64 years (range 28-81), with 32.26% (20/62) were ≥60 years old. Males comprised 56.45% (35/62). Disease was unifocal in 41.94% (26/62) and multifocal in 58.06% (36/62). Deep brain involvement was observed in 54.84% (34/62) of patients, with the most common tumor location of frontal lobe (28/62, 25.69%). Resection was performed in 61.29% (38/62), and stereotactic biopsy in 38.71% (24/62). First-line chemotherapy was administered to 82.26% (51/62) patients. 29 patients (46.77%) received lumbar puncture and MTX or Ara-c plus dexamethasone were used as intrathecal chemotherapy agents. WBRT was used for consolidative intention in 53.23% (33/62) of patients, as palliative treatment after surgery/biopsy in 32.26% (20/62), and palliative treatment after progression of chemotherapy in 14.52% (9/62). Median overall treatment time was 17 days (range 13-22). Pre-WBRT disease status was CR in 32 patients (51.61%), PR in 6 patients (9.68%), and sable or progressive disease (SD/PD) in 24 patients (38.71%). After WBRT, 45 patients (72.58%) achieved CR and 17 patients (27.42%) still had residual tumor. WBRT doses were 23.4Gy (n=21, 33.87%), 30Gy (n=20, 32.26%), 36Gy (n=2, 3.22%), 40Gy (n=7, 11.29%), 50Gy (n=7, 11.29%), <20Gy (n=5, 8.06%), respectively. A boost to the tumour bed was delived in 18 patients (29.03%). 7 patients (11.29%) received oral lenalidomide (10mg) as maintenance after WBRT. A total of 42 patients (≤30Gy) completed MMSE test at 6-month intervals after completion of WBRT. The mean MMSE scores stratified by educational attainment were 23 (illiterate), 24 (primary school), and 26.5 (secondary education or higher), indicating preserved cognition across literacy levels. The median follow-up was 77.8 months, with a mean time between initial symptoms occur and the date of first visit to hospital of 27.4 days (range 0-183). The relapse or disease progression occurred in 41.94% (26/62) of patients, while 48.39% (30/62) patients died. The median OS was 77.03 months (95% CI, 52.01-102.05), the 1-year OS rate was 85.2%, the 2-year OS rate was 71.6%, and the 5-year OS was 58.6%. In univariate analysis, the variables associated with increased OS were receiving systemic chemotherapy (p=0.048), consolidiative WBRT after chemotherapy (p=0.037), no residual disease before WBRT (p=0.001), CR after WBRT (p=0.001) and intrathecal chemotherapy (p=0.008). Gender (p=0.375), age>50 years (p = 0.086), the ECOG (p=0.794), elevated LDH level (p=0.653), elevated protein level of CSF (p=0.508), deep brain involvement (p=0.101), multifocal disease (p=0.364), non-GCB phenotype (p=0.186), the radiotherapy dose of the whole brain less than 30Gy (p=0.149), a boost to the tumour bed (p=0.589), IELSG score (p=0.537), MSKCC score (p=0.193), and receiving salvage chemotherapy (p=0.777) did not show statistically significant association with OS. Conclusion OS was superior when WBRT was given as consolidation after systemic therapy and when pre-WBRT CR was achieved. Low-dose WBRT (≤30Gy) could not negatively affect OS, with no evidence of significant cognitive decline, especially in older patients. Prospective trials evaluating reduced-dose WBRT with neurocognitive endpoints are warranted.

Endurance exercise induces multisystem adaptations that improve performance and benefit health. Gene regulatory circuit responses within individual skeletal muscle cell types, which are key mediators of exercise effects, have not been studied. Here, we map transcriptome, chromatin, and regulatory circuit responses to acute endurance exercise in muscle using same-cell RNA-seq/ATAC-seq multiome assays. High-quality data were obtained from 37,154 nuclei comprising 14 cell types in vastus lateralis samples collected before and 3.5 h after either 40 min cycling exercise at 70% VO2max or 40 min supine rest. Both shared and cell-type-specific regulatory programs were identified. Differential gene expression and accessibility sites are largely distinct within nuclei for each cell type and muscle fiber, with the largest numbers of regulatory events observed in the three muscle fiber types (slow, fast, and intermediate) and lumican (LUM)-expressing fibro-adipogenic progenitor cells. Single-cell regulatory circuit triad reconstruction (transcription factor, chromatin interaction site, regulated gene) also identifies largely distinct gene regulatory circuits modulated by exercise in the three muscle fiber types and LUM-expressing fibro-adipogenic progenitor cells, involving a total of 328 transcription factors acting at chromatin sites regulating 2025 genes. This web-accessible single-cell data set and regulatory circuitry map serve as a resource for understanding the molecular underpinnings of the metabolic and physiological effects of exercise and for guiding interpretation of the exercise response literature in bulk tissue.

Alternative splicing (AS), a crucial driver of proteomic diversity, is a fundamental source of cellular heterogeneity alongside gene expression levels. AS is closely linked to various physiological and pathological processes, including tumor progression and embryonic development. Single-cell RNA sequencing (scRNA-seq) technologies capture AS events through junction reads at cellular resolution, enabling the identification of core AS events that regulate specific cell types or states. However, single-cell sequencing technologies and their data are plagued by inherent limitations, such as shallow sequencing depth, high dropout rates, and batch effects. Furthermore, previous clustering approaches have overlooked the crucial interplay between AS and gene expression in defining distinct "cell types," posing ongoing challenges in this field. In this study, we present a novel method called Alternative Splicing-Gene Expression Network (AEnet), which combines gene expression levels with AS patterns to profile cellular heterogeneity and define what we term "cell subpopulations." AEnet also identifies key AS events and infers the regulatory mechanisms underlying these events. By applying AEnet to tumor cells, pan-cancer immune cells, and embryonic cells, we demonstrate enhanced cell clustering, the identification of novel AS events with potential functional importance, and the discovery of the key splicing factors involved in cell state transitions. The application of AEnet provides new insights into cellular heterogeneity and its role in both physiological and pathological processes.