Breast cancer is characterised by profound genetic heterogeneity, with oestrogen receptor alpha (ERα, encoded by ESR1) being a central driver in ~70% of cases. While point mutations in the ligand-binding domain of ESR1 a...Breast cancer is characterised by profound genetic heterogeneity, with oestrogen receptor alpha (ERα, encoded by ESR1) being a central driver in ~70% of cases. While point mutations in the ligand-binding domain of ESR1 are well recognised mediators of endocrine resistance, a growing body of evidence highlights ESR1 gene fusions as an emerging class of genomic alterations with important clinical implications. These rearrangements, predominantly arising from intrachromosomal events on chromosome 6, truncate the hormone-binding domain and fuse ESR1 with diverse partners, producing constitutively active chimeric proteins. Recurrent fusions such as ESR1-CCDC170, ESR1-YAP1, and ESR1-AKAP12 have been consistently associated with therapy resistance, metastatic progression, and poor clinical outcome, suggesting that they function as oncogenic drivers rather than incidental by-products of genomic instability. However, variability in oncogenic potential indicates that only a subset of fusions are biologically functional. Key determinants of functionality include reading-frame preservation, domain architecture, intrinsic disorder content, and the molecular features contributed by the partner gene, many of which encode transcription factors, signalling adaptors, or cytoskeletal regulators. Structural enrichment in intrinsically disordered regions and the potential for phase separation further implicate ESR1 fusions in the formation of aberrant transcriptional condensates that amplify ER signalling. Importantly, while many fusions are enriched in metastatic, treatment-resistant disease, a small number are present in treatment-naïve tumours, raising the possibility that some act as early drivers of oncogenesis. This review outlines the current knowledge on ESR1 fusions, evaluating their mechanisms and clinical relevance. Clarifying which ESR1 fusions act as true oncogenic drivers versus incidental events will be critical for refining diagnostics and informing future therapeutic development for oncofusion-driven cancers.
Kitisin K, Ganesan N, Tang Y
… +18 more, Jogunoori W, Volpe EA, Kim SS, Katuri V, Kallakury B, Pishvaian M, Albanese C, Mendelson J, Zasloff M, Rashid A, Fishbein T, Evans SRT, Sidawy A, Reddy EP, Mishra B, Johnson LB, Shetty K, Mishra L
Zhou W, Sun W, Yung MMH
… +29 more, Dai S, Cai Y, Chen CW, Meng Y, Lee JB, Braisted JC, Xu Y, Southall NT, Shinn P, Huang X, Song Z, Chen X, Kai Y, Cai X, Li Z, Hao Q, Cheung ANY, Ngan HYS, Liu SS, Barak S, Hao J, Dai Z, Tzatsos A, Peng W, Pei H, Han Z, Chan DW, Zheng W, Zhu W
Seijo-Vila M, Balsinde SA, Blasco-Benito S
… +30 more, Tundidor I, Rubert-Hernández M, Montero-Calle A, Barderas R, Kilpatrick LE, Platt S, Karsai N, Philps I, Antón OM, Gómez-Domínguez D, Pérez de Castro I, González-Lois C, García-Fresnadillo D, Silvestre-Egea G, Sánchez-López AJ, Ramírez-Medina E, Rivas Prieto MC, Almoguera Pérez-Cejuela B, Manso L, Zazo S, López-Ejeda N, Palomino-Duque F, Turienzo-Durán M, Martínez-Illescas NG, Salazar-Roa M, Castillo-Lluva S, Hill SJ, Guzmán M, Pérez-Gómez E, Sánchez C
Acquired or innate lack of response to standard HER2-targeted therapies remains a clinical issue in patients with HER2-positive breast cancer. Here, we investigated the role of the cannabinoid CB receptor (CBR) in trastu...Acquired or innate lack of response to standard HER2-targeted therapies remains a clinical issue in patients with HER2-positive breast cancer. Here, we investigated the role of the cannabinoid CB receptor (CBR) in trastuzumab resistance. In human breast cancer samples, a decreased expression of HER2-CBR heterodimers following neoadjuvant treatment, due to CBR downregulation, was linked to poor long-term outcomes. Using various preclinical models, we demonstrate that CBR drives trastuzumab resistance. Mechanistically, CBR loss enabled cancer cells to evade antitumor IFN-γ signaling while promoting a shift from HER2-CBR to HER2-EGFR heterodimers, thus reducing dependence on HER2 and increasing reliance on EGFR-mediated pathways. Moreover, EGFR inhibition restored trastuzumab sensitivity. In summary, we reveal an unprecedented role for CBR as a key regulator of oncogenic and immune signaling in response to anti-HER2 therapy and its potential as a predictive biomarker of therapeutic efficacy. We also propose dual HER2/EGFR targeting and non-CBR-selective cannabinoid therapies as potential strategies to overcome CBR-mediated trastuzumab resistance. Together, these findings position the endocannabinoid system as a pivotal and actionable node to elucidate, anticipate, and counteract resistance to HER2-targeted therapies.
Microbial contributions to prostate health and disease extend beyond the mere detection of organisms in urine or tissue. Rather than acting as stable colonisers, microbial influences on the prostate are better conceptual...Microbial contributions to prostate health and disease extend beyond the mere detection of organisms in urine or tissue. Rather than acting as stable colonisers, microbial influences on the prostate are better conceptualised as converging fluxes: systemically circulating gut-derived metabolites, immune education occurring in distal lymphoid compartments, and intermittent exposure to microbial products from the lower urinary tract. These inputs converge on a limited set of conserved mediator-receptor axes-including short-chain fatty acids, bile acids and indole derivatives-that calibrate epithelial barrier integrity, inflammatory thresholds, antigen-presentation capacity and myeloid cell fate. Crucially, the biological relevance of these axes is stage-dependent. In benign prostatic hyperplasia and chronic prostatitis/chronic pelvic pain syndrome, metabolite tone shapes inflammatory activation thresholds and barrier resilience. In localized prostate cancer, these same pathways intersect with antigen-processing machinery and immune exclusion. In castration-resistant disease, tumour-intrinsic metabolic plasticity and redox balance predominate, with microbial and host-derived metabolites assuming relevance when they modulate lipid remodelling and ferroptotic vulnerability. Interpretation is constrained by the intrinsically low biomass of urine and prostate tissue. Robust inference therefore requires quantitative anchoring, orthogonal validation and explicit separation of association from causality. Translational progress is most likely to emerge from calibrated measurement and stage-aware modulation rather than indiscriminate ecological manipulation. By integrating mechanistic, spatial and clinical evidence, this Review proposes a stage-aware framework for the gut-urinary-prostatic axis and delineates when microbial and metabolite signalling meaningfully conditions prostate disease biology-and when it does not.
Immune checkpoint blockade (ICB) remains ineffective in most colorectal cancers (CRC) due to intrinsic immune resistance. We identify guanylate-binding protein 2 (GBP2) as a key enhancer of ICB response through modulatio...Immune checkpoint blockade (ICB) remains ineffective in most colorectal cancers (CRC) due to intrinsic immune resistance. We identify guanylate-binding protein 2 (GBP2) as a key enhancer of ICB response through modulation of the gasdermin D (GSDMD)-Yes-associated protein (YAP) axis. Analyses of CRC cohorts, patient samples, organoids, and mouse models revealed that GBP2 directly binds GSDMD, inhibiting its cleavage-dependent activation and preventing YAP nuclear translocation. Activated GSDMD facilitates YAP nuclear accumulation, which represses CXCL9/10/11 transcription and limits CD8⁺ T-cell infiltration. Mechanistically, GBP2 disrupts this process by restraining non-pyroptotic GSDMD activity and maintaining YAP in its inactive cytoplasmic state. Genetic or pharmacologic inhibition of GSDMD restored YAP inactivation and sensitized tumors to anti-PD-L1 therapy. These findings define a GBP2-GSDMD-YAP signaling axis that inhibits immune evasion and represents a therapeutic target to overcome ICB resistance in CRC.
Aberrant alternative polyadenylation (APA) and alternative splicing (AS) contribute to numerous diseases, including cancer; however, their coordinated roles in ovarian cancer remain poorly understood. Here, we investigat...Aberrant alternative polyadenylation (APA) and alternative splicing (AS) contribute to numerous diseases, including cancer; however, their coordinated roles in ovarian cancer remain poorly understood. Here, we investigated CPSF7, an APA factor markedly upregulated in ovarian cancer and associated with poor prognosis. Silencing CPSF7 suppressed proliferation, migration, and invasion of ovarian cancer cells, while antisense oligonucleotides (ASOs) targeting CPSF7 reduced tumor growth in a patient‑derived xenograft (PDX) model. Mechanistically, knockdown of the splicing factor SNRPD2 induced exon 4 skipping in CPSF7 pre‑mRNA. Loss of exon 4 disrupted the RNA recognition motif (RRM) domain essential for CPSF7‑mediated pre‑mRNA cleavage and polyadenylation, and introduced premature termination codons (PTCs) that generated noncoding transcripts subject to nonsense‑mediated decay (NMD), thereby reducing CPSF7 expression. Thus, efficient splicing mediated by SNRPD2 is crucial for sustaining high CPSF7 levels in ovarian cancer cells. Functional assays showed that CPSF7 knockdown reduced proliferation and metastatic potential in cells with elevated SNRPD2, suggesting that CPSF7 is a key mediator of SNRPD2-driven oncogenesis. Moreover, CPSF7 governed specific APA events to maintain transcript stability, with UBE2K identified as a critical downstream target. CPSF7 preferentially bound distal polyadenylation signals (PASs) within the predominant UBE2K transcript (UBE2K-201), thereby increasing its mRNA stability and maintaining high functional UBE2K expression. Collectively, these findings reveal that AS and APA are interconnected in ovarian cancer via the SNRPD2-CPSF7-UBE2K axis, which drives disease progression and represents a promising target for therapeutic intervention.
Development of castration resistance and distant metastasis remain two major clinical challenges in prostate cancer (PCa) treatment. By analyzing multiple public cancer datasets, we found that ribosomal protein S15 (RPS1...Development of castration resistance and distant metastasis remain two major clinical challenges in prostate cancer (PCa) treatment. By analyzing multiple public cancer datasets, we found that ribosomal protein S15 (RPS15) is overexpressed in PCa and related to its metastasis. Beyond its canonical role as a structural component of the ribosome, emerging evidence has highlighted the extraribosomal functions of RPS15 in disease progression. Our study demonstrates that RPS15 significantly promotes proliferation and migration in PCa through the establishment of RPS15 knockdown cells and xenograft models. Mechanistically, RPS15 interacts with the functional domain of DExD-box helicase 21 (DDX21) and facilitates the binding of DDX21 to the transcription start region of stearoyl-CoA desaturase-1 (SCD1), thereby enhancing its transcriptional activity and protein expression to drive the growth, ferroptosis-resistance, and metastasis of PCa cells. Moreover, analysis of clinical samples revealed that RPS15, DDX21, and SCD1 are concomitantly upregulated and exhibit strong positive correlations in PCa tissues. Collectively, our findings uncover the significance of the RPS15-DDX21-SCD1 axis in PCa development, expanding the understanding of noncanonical functions of ribosomal proteins and providing new insights for PCa management.
Chemotherapy resistance is a major factor contributing to the failure of nasopharyngeal carcinoma (NPC) treatment. Migrasomes can export damaged mitochondria out of the cell, and the timely removal of damaged mitochondri...Chemotherapy resistance is a major factor contributing to the failure of nasopharyngeal carcinoma (NPC) treatment. Migrasomes can export damaged mitochondria out of the cell, and the timely removal of damaged mitochondria is key to cancer cell resistance. However, whether migrasomes regulate tumor resistance remains unknown. Here, we elucidated the role and mechanism of migrasomes in chemoresistance of NPC. We found that the formation of migrasomes was increased in cisplatin-resistant NPC cells, and inhibiting migrasome formation reduced cisplatin resistance. PinX1 was lowly expressed in tumor tissues of patients with high migrasome scores. Upstream mechanism analyses showed that TP53 was effectively bound to the promoter of PinX1, thereby enhancing its transcriptional activity. Knockdown of PinX1 facilitated migrasome formation via its telomerase inhibitory domain 252-328aa region binding to Rab11a, which relied on serine residues at the N-terminal 25aa site for promoting migrasome formation. Mechanistically, PinX1 recruited RanBP2 to induce the SUMOylation of Rab11a, leading to the degradation of Rab11a at the K207 site. Furthermore, PinX1 reduced cancer cell energy metabolism by inhibiting the export of damaged mitochondria via migrasomes. Collectively, TP53-activated PinX1 recruits RanBP2 to Rab11a, triggering Rab11a K207 SUMOylation and degradation, leading to impaired migrasome formation and mitochondrial transfer, and ultimately suppresses cisplatin resistance in NPC. Our study provides a new target for clinical reversal of chemotherapy resistance in patients with NPC.
Prostate cancer (PC) is one of the most common malignancies in men, and the emergence of androgen receptor-low/negative castration-resistant PC (ARL/- CRPC) following androgen receptor signaling inhibitor (ARSI) therapy...Prostate cancer (PC) is one of the most common malignancies in men, and the emergence of androgen receptor-low/negative castration-resistant PC (ARL/- CRPC) following androgen receptor signaling inhibitor (ARSI) therapy remains a critical clinical challenge. The RNA-binding protein DEAD-box helicase 3 X-linked (DDX3X) has been implicated in the translational regulation of androgen receptor (AR) mRNA; however, the underlying binding mechanisms are not well defined. Here, we show that DDX3X colocalizes with AR mRNA in ARL/- CRPC cells and selectively recognizes non-canonical RNA G-quadruplex (rG4) motifs within the sequence of AR mRNA. RNA immunoprecipitation sequencing (RIP-seq) revealed enrichment of DDX3X-AR mRNA interactions in ARL/- CRPC cells. Fluorescence imaging confirmed the colocalization of DDX3X and AR mRNA within cytoplasmic granules, and biochemical assays confirmed the ability of selected AR mRNA fragments to form rG4 structures bound by DDX3X. Proteomic profiling of DDX3X-Ras GTPase-activating protein-binding protein 1 (G3BP1) complexes identified several RNA-binding proteins, including IGF2BP1, PUM2, and UBAP2, which may act as candidate cofactors. Together, these findings shed light on the interaction between AR mRNA and DDX3X and identify putative protein partners, offering insights into future therapeutic strategies.
Spellecy O, Qadir J, Han R
… +20 more, Zhu K, Xu B, Zhang Y, Aryal I, Lin RL, Lin AH, Faisal ASM, Napier D, Piecoro D, Scott T, Lin P, Chen L, Brewer LD, Wang C, Kyprianou N, Guo Z, Guo R, Thibault O, Yang BB, Yang XH
Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. Here, we investigated how integrin-dependent signaling pathways regulate tumor metabolism and therapeutic vuln...Triple-negative breast cancer (TNBC) is an aggressive breast cancer subtype with limited therapeutic options. Here, we investigated how integrin-dependent signaling pathways regulate tumor metabolism and therapeutic vulnerability in TNBC. Pharmacological inhibition of the integrin/FAK axis and/or BRD4 induced cell cycle arrest, autophagy, and senescence in highly proliferative cells, consistent with a metabolic stress phenotype. Metabolomic analyses using [U-¹³C]-glucose revealed a marked suppression of glycolytic carbon flux, accompanied by an approximately 30-47% reduction in intracellular NAD⁺ levels and coordinated alterations in NADH and tricarboxylic acid (TCA) cycle intermediate α-ketoglutarate. Mechanistically, we identified nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme in NAD⁺ biosynthesis, as a central metabolic node integrating signaling/function of the two axes. NAMPT expression/activity was sustained transcriptionally or post-translationally, including sirtuin-associated deacetylation and neddylation-dependent proteasomal turnover. In BRCA1/2-deficient TNBC, integrin-FAK and NAMPT/NAD pathways converged on Wnt/β-catenin signaling to regulate DNA repair, and response to PARP1/2 inhibitors. Co-inhibiting FAK and NAMPT synergistically suppressed tumor growth by approximately 80%. Elevated stromal NAMPT expression was associated with a trend toward favorable clinical outcomes. Collectively, these findings uncover a previously unrecognized crosstalk between integrin/FAK and NAMPT/NAD⁺ pathways in TNBC and identify a synthetic lethal-like therapeutic vulnerability that warrants further evaluation in clinically relevant models.
The immunosuppressive tumor microenvironment (TME) is a major issue in the malignant progression of glioma patients. The membrane spanning four domains A4A (MS4A4A) has a relationship with M2 polarization of macrophages,...The immunosuppressive tumor microenvironment (TME) is a major issue in the malignant progression of glioma patients. The membrane spanning four domains A4A (MS4A4A) has a relationship with M2 polarization of macrophages, and participates in the malignant progression of various cancers. Therefore, exploration of the key role of MS4A4A contributing to glioma biological processes is urgently needed. We performed the bioinformatics analysis of M2 gene expression and built a model predicting the prognosis of glioma patients. Knocking down or overexpressing MS4A4A was achieved in macrophages, and we identified the polarization of macrophages with different MS4A4A expression levels. In vitro and in vivo experiments were used to investigate the role of MS4A4A in regulating M2 polarization and contributing to malignant behaviour in glioma. We found that MS4A4A was associated with the macrophages' M2 scores and the prognosis of GBM patients. MS4A4A had a higher expression level in M2 polarization macrophages. MS4A4A regulates macrophage M2 polarisation through NF-κB and JAK-STAT6 signalling pathways. Macrophages with MS4A4A overexpression promoted the proliferation, invasion, and TMZ-resistance of glioma cells in vitro and in vivo experiments. The treatment targeting the MS4A4A/ NF-κB/STAT6 axis could improve the prognosis and TMZ-resistance in the glioma mouse model. The present study revealed the novel mechanism of the MS4A4A regulating macrophages M2 polarization, contributing to the formation of immunosuppressive tumor microenvironment in glioma through NF-κB/STAT6 signaling pathways, which promotes the malignant biological process of glioma cells. Our results provided new evidence that NF-κB and STAT6 inhibitors might be a potential adjuvant agent in overcoming MS4A4A-mediated chemotherapy resistance in glioma.
Small cell lung cancer (SCLC) is prone to developing chemoresistance, which is associated with epigenetic reprogramming. While LSD1-mediated histone demethylation has been reported, a systematic investigation into the ro...Small cell lung cancer (SCLC) is prone to developing chemoresistance, which is associated with epigenetic reprogramming. While LSD1-mediated histone demethylation has been reported, a systematic investigation into the role of histone demethylases in SCLC chemoresistance is lacking. In this study, we established nine chemoresistant cell lines from parental cells via gradual dose escalation and conducted RNA sequencing, followed by linear regression and random effects meta-analysis to identify genes associated with chemoresistance among 23 histone demethylases. We subsequently validated our findings using two neuroendocrine SCLC cell lines and their corresponding chemoresistant counterparts. We identified KDM4B as a key driver of chemoresistance, with immunohistochemical analysis revealing its elevated expression in chemoresistant tissues from SCLC patients, although its association with neuroendocrine subtypes warrants further investigation. KDM4B promoted chemoresistance through the Hedgehog pathway by enhancing cell proliferation and stemness. Mechanistically, KDM4B interacts with DHX9 and corecruits to the MYCN promoter to promote its transcription and activate the Hedgehog signaling pathway. An inhibitor of DHX9 had synergistic antitumor effects with cisplatin and etoposide, and effectively rescued the chemosensitivity of SCLC both in vitro and in vivo. These findings provide valuable insights for future studies aimed at developing therapeutic strategies to overcome chemoresistance in SCLC.
Gallbladder cancer (GBC) is a highly lethal disease which is usually diagnosed at advanced stage owing to unavailable screening tools and effective therapies. Persistent inflammation induced by various risk factors is th...Gallbladder cancer (GBC) is a highly lethal disease which is usually diagnosed at advanced stage owing to unavailable screening tools and effective therapies. Persistent inflammation induced by various risk factors is the main cause of GBC, however, the molecular program remains elusive. Unveiling the molecular trajectory and events during gallbladder epithelium malignant transformation contribute to prevention and drug discovery for GBC. Single cell RNA sequencing was performed by using 4 gallbladder adenoma and cancer samples. Pseudotime trajectory analyses were employed to reconstruct epithelium transformation track in order to identify crucial genes which promoted GBC development. Functional and mechanism studies were performed to validate the regulatory network and cell behavior in vitro and in vivo. Three clusters of gallbladder epithelium were identified among GBC microenvironment which were characterized by distinct epithelial mesenchymal transition (EMT) and inflammation states. Cell adhesion molecular binding was the most significant GO term between EMT high and low states, in which OLFM4 was differentially expressed gene participated. Further studies implicated that OLFM4 could promote GBC metastasis and activate EMT through CEACAM6/AKT signaling cascade, and the CEACAM6 expression was regulated by TGF-β/Smad3 pathway. Interestingly, we disclosed that TGFβR1 was the functional receptor of OLFM4, through which the tumorigenic signaling of OLFM4 was transduced. These findings suggest the oncogenic role of OLFM4 during GBC carcinogenesis which can be a candidate biomarker of GBC, and OLFM4, TGFβR1 and downstream signaling elements are promising therapeutic targets for GBC.
Wang J, Wee Y, Jacob T
… +15 more, Rogers A, Sorensen LK, Brooks DM, Gupta P, Tay JKH, Wilson EC, Liu T, Smith EA, Gopal YNV, Davies MA, McMahon M, Holmen SL, Judson-Torres RL, Wolff RK, Grossmann AH
The intrinsic ability of cancer cells to evade death underpins tumorigenesis, progression, metastasis, and the survival of drug-tolerant persister (DTP) cells. Herein, we discovered that the small GTPase ARF6 plays a cen...The intrinsic ability of cancer cells to evade death underpins tumorigenesis, progression, metastasis, and the survival of drug-tolerant persister (DTP) cells. Herein, we discovered that the small GTPase ARF6 plays a central role in tumor survival by fortifying RAF oncoprotein levels. ARF6 activation was sufficient to increase BRAF, ARAF, and CRAF proteins through a post-transcriptional mechanism, while sustained inhibition of ARF6 eventually led to decay. In a genetically engineered model of aggressive melanoma, tumor-specific Arf6 deletion attenuated BRAF protein expression and MAPK signaling and prevented rapid tumor progression. In human melanoma cells, pharmacologic inhibitors of BRAF uniformly induced swift activation of ARF6, driving a positive feedback loop that restored MAPK-driven anti-apoptotic signaling and supported drug-tolerant survival and growth. Furthermore, in patient-derived melanoma xenografts with innate or clinically acquired resistance to MAPK inhibitors, ARF6 silencing alone significantly suppressed tumor growth in vivo. When combined with BRAF and MEK targeted therapy in vitro, inhibition of ARF6 markedly reduced survival and drug-tolerant growth. Collectively, these findings reveal a previously unknown mechanism of maintaining BRAF protein expression that preserves the MAPK pathway during targeted therapy. This ARF6-dependent mechanism may be exploited in BRAF driven cancers as a therapeutic vulnerability.
Colorectal cancer (CRC) is one of the most commonly diagnosed and globally spread malignant diseases. Cancer-associated fibroblasts (CAFs) are key architects of the tumor microenvironment, yet their origin, stability, an...Colorectal cancer (CRC) is one of the most commonly diagnosed and globally spread malignant diseases. Cancer-associated fibroblasts (CAFs) are key architects of the tumor microenvironment, yet their origin, stability, and interconvertibility remain poorly understood. Using transcriptomic profiling of fibroblasts from colorectal cancer (CRC) patients, we identify highly expressed (HEX) markers that define fibroblast subpopulations and uncover mechanisms governing their plasticity. We find that ADH1B marks normal colon-associated fibroblasts (NAFs), which consist of PI16-NAFs and ADAMDEC1-NAFs. ITGA3 delineates the total CAF population, which comprises myofibroblastic CAFs (myCAFs), whose characterizing markers were associated with poor prognosis and proteolytic inflammatory CAFs (piCAFs), characterized by markers not associated with prognosis. An AGT/TGM2-expressing fibroblast subset is present in both healthy and tumor tissues, suggesting alternative trajectories to the classical NAF-to-CAF transition model. While PI16-NAFs, AGT/TGM2-fibroblasts, and myCAFs maintain stable identities in long-term culture, the ADAMDEC1-NAF and piCAF phenotypes are lost in vitro. ITGA3-CAFs demonstrate dynamic plasticity, with TGF-β stably inducing myCAF formation and TNF-α or inhibition of DNA methylation promoting transient piCAF emergence. These findings redefine fibroblast heterogeneity in CRC and reveal a coexisting stable and plastic fibroblast network that may be amenable to modulation and provides a framework for future functional and translational studies. We identified highly expressed markers (HEX markers) to distinguish CAFs, NAFs and corresponding subpopulations in CRC. ADH1B characterized NAFs, which consisted of stable (solid outline) PI16-NAFs and unstable (dashed outline) ADAMDEC1-NAFs. ITGA3 identified CAFs consisting of stable myCAFs associated with poor prognosis and unstable piCAFs not associated with prognosis. AGT/TGM2 fibroblasts did not express ADH1B or ITGA3, were stable in culture and could be detected in both healthy colon and CRC. Treatment of PI16-NAFs with LPS or IFN-γ induced ADAMDEC1-NAFs, TGF-β the formation of myCAFs, while treatment with TNF-α led to the formation of piCAFs. Reduced DNA methylation converted myCAFs and PI16-NAFs into piCAFs.
Hepatocellular carcinoma (HCC) is a prevalent and aggressive malignancy, notorious for its high recurrence, substantial drug resistance, and poor prognosis. Although METTL1, a key regulator of RNA mG modification, is imp...Hepatocellular carcinoma (HCC) is a prevalent and aggressive malignancy, notorious for its high recurrence, substantial drug resistance, and poor prognosis. Although METTL1, a key regulator of RNA mG modification, is implicated in the progression of various cancers, its role and underlying mechanisms in HCC remain poorly understood. This study identified that METTL1 was significantly upregulated in HCC tissues, correlating with advanced stages and poor survival. Knockdown of METTL1 inhibited cell proliferation, migration, and invasion, which was reversed by restoring METTL1 expression. Multi-omics analysis indicated that METTL1 regulated gene expression through mG modification, particularly in the Wnt, mTOR signaling pathways, and amino acid metabolism (especially asparagine metabolism). Further analysis revealed that METTL1 increased the stability and upregulated the expression levels of asparagine synthetase (ASNS) mRNA through mG modification, thereby reprogramming asparagine metabolism and activating the mTOR pathway, ultimately promoting HCC progression. In conclusion, METTL1 regulated ASNS mRNA stability and expression via mG modification, driving HCC malignancy through reprogramming of asparagine metabolism and activation of the mTOR signaling pathway.
Immunotherapy targeting the programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) (PD-1/PD-L1) axis has revolutionized cancer treatment, yet its efficacy in hepatocellular carcinoma (HCC) remains limited....Immunotherapy targeting the programmed cell death-1 (PD-1)/programmed cell death-ligand 1 (PD-L1) (PD-1/PD-L1) axis has revolutionized cancer treatment, yet its efficacy in hepatocellular carcinoma (HCC) remains limited. Emerging evidence suggests that gut microbiota plays a pivotal role in modulating tumor immune microenvironments (TIME), offering a novel avenue to enhance immunotherapy outcomes. This study investigates the regulatory effects of Lactobacillus kefiranofaciens (LK) on the TIME in HCC, focusing on its modulation of Suppressor of cytokine signaling 3 (SOCS3) expression and the Janus-activated kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway, with the goal of improving responses to anti-PD-1/PD-L1 therapy. LK was isolated from kefir grains and identified through genomic sequencing. In vitro assays, including Cell Counting Kit-8 (CCK-8), 5-Ethynyl -2'- deoxyuridine (EdU) staining, colony formation, Transwell, and apoptosis detection, were conducted using Hepa1-6 HCC cells. In vivo, subcutaneous and orthotopic HCC mouse models were treated with LK to assess tumor progression. Single-cell RNA sequencing (scRNA-seq) and Bulk RNA-seq analyses were performed to identify key signaling pathways and therapeutic targets. SOCS3 expression was manipulated via lentiviral transfection to validate its role in immunotherapy enhancement. LK significantly inhibited HCC cell growth, migration, and invasion, while promoting apoptosis in vitro. In vivo, LK treatment reduced tumor size and improved immune cell infiltration, particularly T cells and NK cells. Transcriptomic analysis revealed that LK upregulates SOCS3, suppresses the JAK-STAT signaling pathway, and reduces PD-L1 expression, enhancing T cell-mediated immune responses. This study highlights the potential of gut microbiota modulation, specifically through LK, to enhance the efficacy of anti-PD-1/PD-L1 immunotherapy in HCC by targeting SOCS3 and the JAK-STAT pathway. These findings provide a new therapeutic approach for improving immunotherapy outcomes in HCC. Gut probiotics modulate the immune microenvironment to enhance the response of liver cancer patients to anti-PD-1/PD-L1 immunotherapy: molecular mechanisms.
Ecotropic viral integration site 1 (EVI1) is essential for hematopoietic stem cell maintenance, and its aberrant expression is a significant adverse prognostic indicator in myeloid leukemia. EVI1 overexpression typically...Ecotropic viral integration site 1 (EVI1) is essential for hematopoietic stem cell maintenance, and its aberrant expression is a significant adverse prognostic indicator in myeloid leukemia. EVI1 overexpression typically occurs due to chromosomal rearrangement involving 3q26. However, aberrant EVI1 expression is still observed in numerous cases without 3q26 abnormalities, leading to similarly poor outcomes, while the mechanism behind EVI1 overexpression in these cases remains largely unknown. Here, we performed genome-wide CRISPR screening using cells with GFP knock-in at the EVI1 locus and identified zinc finger protein 91 (ZFP91) was the leading activator of EVI1. ZFP91 knockout significantly reduced EVI1 expression and cell proliferation. We also showed that ZFP91 binds to the EVI1 promoter, enhancing H3K4me3/H3K27ac and chromatin accessibility. Our data showed that the ZFP91-EVI1 axis plays a critical role for activation of EVI1 in myeloid leukemia. Our screening approach represents a powerful and unbiased method for identifying expression regulators that can be broadly applied across a range of contexts.