bims-instec Biomed News
on Intestinal stem cells and chemoresistance in colon cancer and intestinal regeneration
Issue of 2025–11–16
27 papers selected by
Maria-Virginia Giolito, Université Catholique de Louvain



  1. iScience. 2025 Nov 21. 28(11): 113696
      The Consensus Molecular Subtype 4 (CMS4) of colorectal cancer (CRC) has the worst prognosis and the highest frequency of hepatic metastases. It is characterized by abundant cancer-associated fibroblasts (CAFs) in the tumor microenvironment and active TGFβ signaling, but the molecular drivers of metastasis remain unclear. Here, we show that TGFβ signaling in CRC patient-derived CAFs from the primary tumor induces production of IL-6 family cytokines, particularly IL-6 and IL-11. These cytokines stimulate hepatocytes to express myeloid chemoattractants, including SAA1, through gp130-dependent JAK/STAT signaling. This promotes neutrophil recruitment to the liver, potentially creating a pro-metastatic niche. This IL-6 family-JAK/STAT stromal signaling axis is active in both a murine model of CMS4 as well as in patients with human CRC in vivo. Combined, our data reveal that TGFβ-driven CAF signaling actively contributes to the formation of a neutrophil-dependent, pre-metastatic hepatic niche in the metastatic phenotype of CMS4 CRC.
    Keywords:  Microenvironment
    DOI:  https://doi.org/10.1016/j.isci.2025.113696
  2. Cancer Cell. 2025 Nov 13. pii: S1535-6108(25)00451-9. [Epub ahead of print]
      While dual KRAS and epidermal growth factor receptor (EGFR) inhibition shows promise in treating KRAS-mutant colorectal cancer (CRC), resistance remains a major challenge. Using genetically engineered mouse models, patient-derived organoids and xenografts, as well as clinical specimens, we discover that colorectal tumors surviving combined KRAS and EGFR inhibition acquire a Paneth-like cell state-a secretory lineage typically confined to the intestinal crypt. Lineage tracing reveals that CRC cells evade dual therapy by transitioning into a Paneth-like state. Through integrated transcriptomic analysis and CRISPR genetic screening, we identify SMAD1 as a key regulator of this lineage plasticity, promoting resistance by directly activating FGFR3. Genetic or pharmacological inhibition of FGFR3 prevents the Paneth-like transition, restores drug sensitivity, and synergizes with KRAS-EGFR inhibition across multiple preclinical models. These findings reveal that the SMAD1-FGFR3 axis triggers Paneth-like plasticity to drive KRAS-EGFR dual therapy resistance in CRC and highlight FGFR3 blockade as a promising strategy to overcome plasticity-driven drug tolerance.
    Keywords:  KRAS mutant colorectal cancer; Paneth-like cell state; drug resistance; dual KRAS and EGFR-targeted therapy; lineage plasticity; trans-differentiation
    DOI:  https://doi.org/10.1016/j.ccell.2025.10.010
  3. Front Cell Dev Biol. 2025 ;13 1687485
       Background: Colorectal cancer (CRC) remains a leading cause of cancer mortality, with liver metastasis being the principal determinant of poor prognosis, but the spatial mechanisms orchestrating metastatic niches remain elusive.
    Method: To dissect the molecular and spatial dynamics of CRC progression, we constructed an integrative atlas using 35 single-cell RNA-seq datasets and spatial transcriptomics from primary tumors, liver metastases, and matched normal tissues. Malignant epithelial subpopulations were stratified via inferCNV and CytoTRACE analyses. Stromal-tumor interactions were dissected using CellChat and NicheNet, with functional validation through in vitro co-culture and immunohistochemistry.
    Result: We identified a transcriptionally distinct epithelial subpopulation, termed high-malignancy CRC (High-M CRC), enriched in metastatic lesions and characterized by enhanced stemness, MYC-driven transcriptional activity, and glycolytic reprogramming. Stromal-tumor interaction analyses revealed that cancer-associated fibroblasts (CAFs), particularly matrix CAFs (mCAFs), promote malignant progression via the HGF-MET-MYC signaling axis. Spatial transcriptomic mapping confirmed the physical proximity and molecular co-localization of High-M CRC cells and mCAFs, along with enriched glycolysis and MYC expression at the cell-cell interface. In vitro functional validation demonstrated that CAF-derived HGF activates MET-MYC signaling in CRC cells, enhancing their invasion and proliferation-effects reversible by MET knockdown.
    Conclusion: We unveil a spatially organized metabolic niche driven by stromal-tumor HGF-MET-MYC signaling. These findings offer novel insights into the stromal-tumor interaction and suggest actionable targets for therapeutic intervention in CRC.
    Keywords:  HGF-MET-MYC signaling axis; colorectal cancer liver metastasis; high-malignancy CRC subpopulation; single-cell and spatial transcriptomics; spatial stromal-tumor co-localization
    DOI:  https://doi.org/10.3389/fcell.2025.1687485
  4. Cell Mol Immunol. 2025 Nov 10.
      The microenvironment of distant organs affects the colonization and growth of disseminated tumor cells. It remains unclear how tumor-associated neutrophils are influenced by the microenvironment of distant organs. Here, we demonstrate that mature low-density neutrophils in colorectal cancer patients abnormally accumulate neutral lipids and induce the reactivation of dormant tumor cells, a process regulated by hepatic stellate cells. Mechanistically, activated hepatic stellate cells increased DGAT1/2-dependent lipid droplet synthesis in low-density neutrophils through the secretion of IL33, thereby maintaining the survival and immunosuppressive function of these neutrophils. The uptake of lipids from lipid-laden low-density neutrophils drives dormant tumor cell reactivation through the potentiation of β-oxidation and the stimulation of protumorigenic eicosanoid synthesis. In mouse models, targeting IL33 blocked neutrophil lipid synthesis, decreased the colonization of colorectal cancer cells in the liver, and enhanced the efficacy of immunotherapy. Overall, our study revealed that lipid accumulation in mature low-density neutrophils regulates the growth of dormant tumor cells and antitumor immunity to facilitate colorectal cancer liver metastasis. Targeting IL33 could be a promising therapeutic approach for colorectal cancer liver metastases.
    Keywords:  colorectal cancer; lipid droplets; liver metastasis; mature low-density neutrophils; metabolic reprogramming
    DOI:  https://doi.org/10.1038/s41423-025-01365-9
  5. Stem Cell Reports. 2025 Nov 13. pii: S2213-6711(25)00319-4. [Epub ahead of print] 102715
      The human intestinal epithelial barrier is shaped by biological and biomechanical cues, including growth factor gradients and fluid flow. While these factors are known to affect adult stem cell (ASC)-derived intestinal epithelial cells in vitro, their impact on human induced pluripotent stem cell (hiPSC)-derived cells is largely unexplored. Here, we compare the cellular composition and gene expression profiles of hiPSC-derived intestinal epithelial cells exposed to various medium compositions and cultured as organoids, in Transwell and microfluidic intestine-on-chip systems. Modulating key signaling pathways (WNT, NOTCH, bone morphogenetic protein [BMP], and mitogen-activated protein kinase [MAPK]) influenced the presence of dividing, absorptive, and secretory epithelial lineages. Upon differentiation, intestinal epithelial cells expressed genes encoding digestive enzymes, nutrient transporters, and drug-metabolizing enzymes. Notably, these pathways were most enhanced in the intestine-on-chip system, along with an expression profile that suggests a more mature state. These findings highlight the potential of hiPSC-derived intestinal cells to model important intestinal functions and guide the selection of optimal culture conditions for specific applications.
    Keywords:  Transwell; differentiation medium; gut-on-chip; human; induced pluripotent stem cell; intestinal epithelial barrier; intestine-on-chip; organ-on-chip; organoid; small intestine
    DOI:  https://doi.org/10.1016/j.stemcr.2025.102715
  6. EMBO Mol Med. 2025 Nov 10.
      Cetuximab, an EGFR-targeting monoclonal antibody, provides beneficial yet limited clinical improvement in KRAS wild-type metastatic colorectal cancer (mCRC). While circRNA dysregulation has been implicated in various cancers, the role of circ-EGFR in response to EGFR-targeted therapy in mCRC remains largely unexplored. Here, we identified circ-EGFR as a promising predictive biomarker for cetuximab response. Clinically, we first determined that tissue-based circ-EGFR biomarker effectively stratified responders from non-responders to cetuximab in mCRC, with an Area under the Curve (AUC) of 76.8%. Functional assays demonstrated that circ-EGFR enhances the sensitivity to cetuximab, whereas its depletion induces resistance in CRC. Mechanistically, we revealed that circ-EGFR functions as a sponge for miR-942-3p, resulting in the upregulation of GAS1, which activates the Hedgehog signaling pathway and promotes the efficacy of cetuximab in CRC. Importantly, we effectively translated this tissue-based biomarker into a liquid biopsy predictor for anti-EGFR response (AUC: 76.9%), highlighting its non-invasive potential. In conclusion, circ-EGFR is a significant predictor of cetuximab efficacy in mCRC, potentially aiding in patient selection and treatment management, especially for patients with low circ-EGFR expression.
    Keywords:  Cetuximab; Metastatic Colorectal Cancer; Therapeutic Response Prediction; circ-EGFR; miR-942-3p
    DOI:  https://doi.org/10.1038/s44321-025-00333-0
  7. Gut Microbes. 2025 Dec 31. 17(1): 2573045
      The colonic epithelium is a key interface between the gut microbiota and the host. How microbiota-derived signals influence epithelial cell identity and function remains incompletely understood. Here, we used single-cell transcriptomics, antibiotic-mediated microbiota depletion, germ-free mice and colonization experiments in mice to uncover cell-type-specific responses to microbiota changes, highlighting changes in the cell composition and functional diversities in enterocytes. Our analysis demonstrates that the microbiota control the absorptive profile of the colon epithelial cells and reveals non-canonical inter-crypt goblet cells as microbiota-responsive constituents that combine absorptive and secretory features and whose abundance is regulated by the gut microbiota. We found that their number is suppressed through the short-chain fatty acid butyrate and its receptor GPR109A. Analysis in mouse and humans indicates that the expansion of this hybrid population increases with age and that this expansion is driven by microbiome changes. Our work reveals a previously unrecognized level of epithelial plasticity driven by microbial triggers and highlights butyrate, acting as a signaling molecule that shapes the colon micro-anatomy.
    Keywords:  GPR109A; HCAR2; Single-cell transcriptomics; aging; butyrate; colon epithelium
    DOI:  https://doi.org/10.1080/19490976.2025.2573045
  8. Neoplasma. 2025 Oct;pii: 250610N246. [Epub ahead of print]72(5): 319-327
      Accumulating evidence highlights the critical roles of autophagy-dependent ferroptosis mediators in colorectal cancer (CRC) pathogenesis. To elucidate SIRT6's tumor-suppressive role, HT29 cells stably overexpressing SIRT6 (Oe-SIRT6) were generated via plasmid transfection. Functional assays were performed to evaluate autophagy and ferroptosis. Rescue experiments using the autophagy inhibitor 3-MA or the mTOR agonist MHY1485 were conducted. Quantitative analyses revealed marked downregulation of SIRT6 expression in CRC cell lines (SW620, SW480, and HT29) compared to normal colon epithelial cells. SIRT6 overexpression induced autophagy and activated ferroptosis. The autophagy inhibitor 3-MA blocked SIRT6-driven ferroptosis, which confirmed its dependency on autophagy. Moreover, SIRT6 was found to inactivate mTOR/STAT3 signaling, whereas the mTOR agonist MHY1485 reversed SIRT6 overexpression on autophagy-dependent ferroptosis of CRC cells. Our findings establish SIRT6 as a dual-phase regulator of CRC cell death, suppressing mTOR/STAT3 signaling to orchestrate autophagy-dependent ferroptosis.
    DOI:  https://doi.org/10.4149/neo_2025_250610N246
  9. Mol Oncol. 2025 Nov 09.
      Cancer is a highly heterogeneous disease, with many cancers containing multiple distinct subclones. While subclones are often seen as competitors (survival of the fittest), intratumor heterogeneity can also offer direct benefits to the tumor through cooperation between different clones. This has important clinical implications, as interdependent populations may present therapeutic vulnerabilities. Here, we review existing evidence for clonal cooperativity to address key questions and outline future developments based on six overarching principles: (a) secreted factors are important mediators of clonal cooperation; (b) (very) small subclones can significantly affect tumor behavior; (c) both genetic and nongenetic heterogeneity are substrates for cooperation; (d) nonmalignant cells from the tumor microenvironment can act as cooperating partners; (e) clonal cooperation occurs throughout different stages of cancer, from premalignancy to metastasis; and (f) clonal cooperation can promote therapy resistance by protecting otherwise sensitive populations. Together, these principles suggest clonal cooperation as an important mechanism in cancer. Lastly, we discuss how novel technological developments could address remaining gaps to open up new therapeutic strategies that exploit clonal cooperativity by targeting the tumor's weakest link.
    Keywords:  cancer progression; clonal cooperation; intratumoral heterogeneity; secreted factors; therapy resistance
    DOI:  https://doi.org/10.1002/1878-0261.70160
  10. Nature. 2025 Nov 12.
      
    Keywords:  Imaging; Metabolism; Physiology
    DOI:  https://doi.org/10.1038/d41586-025-03457-y
  11. Proc Natl Acad Sci U S A. 2025 Nov 18. 122(46): e2510910122
      Metastatic outgrowth requires that cancer cells delaminate from the primary tumor, intravasate, survive in circulation, extravasate, migrate to, and proliferate at a distal site. Recurrent genetic drivers of metastasis remain elusive, suggesting that unlike the early steps of oncogenesis, metastasis drivers may be variable. We develop a framework for identifying metastasis regulators using CRISPR/Cas9-based screening in a genetically defined organoid model of colorectal adenocarcinoma. We conduct in vitro screens for invasion and migration alongside orthotopic, in vivo screens for gain of metastasis in a syngeneic mouse model. We identify CTNNA1 and BCL2L13 as bona fide metastasis-specific suppressors which do not confer any selective advantage in primary tumors. CTNNA1 loss promotes cell invasion and migration, and BCL2L13 loss promotes anchorage-independent survival and non-cell-autonomous changes to macrophage polarization. This study demonstrates proof of principle that large-scale genetic screening can be performed in tumor-organoid models in vivo and identifies novel regulators of metastasis.
    Keywords:  CRISPR screen; colorectal cancer; metastasis; organoid
    DOI:  https://doi.org/10.1073/pnas.2510910122
  12. Cell Death Discov. 2025 Nov 10. 11(1): 526
      Current therapeutic approaches for colorectal cancer (CRC) face challenges such as recurrence and drug resistance. Ferroptosis, a novel form of cell death, is a promising therapeutic approach for CRC. SPOP plays an important biological role as a substrate-binding protein of the E3 ubiquitin ligase complex CRL3, but its therapeutic effects in CRC patients and its ability to modulate ferroptosis remain largely unknown. This study demonstrated that SPOP functions as a tumor suppressor in CRC and that SPOP inhibits the proliferation and metastasis of CRC cells and increases their sensitivity to ferroptosis. Transcriptome analysis suggested that Wnt signaling may be a potential target for the function of SPOP. Further data revealed that SPOP knockdown increased β-catenin protein levels, and the clinical data indicated that SPOP expression had the opposite effect on β-catenin protein levels. Molecular biology experiments suggest that SPOP promotes polyubiquitination and degradation of the K508 site of β-catenin. Interestingly, O-GlcNAcylation of SPOP reduces its protein stability and affects SPOP binding to β-catenin, and SPOP also promotes CRC ferroptosis by inhibiting the β-catenin/SLC7A11 axis. Combined treatment with the SPOP-targeted drug maprotiline and a ferroptosis inducer has synergistic antitumor efficacy in CRC cells and xenografts. Our study reveals the multifaceted function of SPOP in CRC, and the activation of SPOP may be a feasible strategy to increase the sensitivity of CRC to ferroptosis inducers.
    DOI:  https://doi.org/10.1038/s41420-025-02832-y
  13. Int J Biol Sci. 2025 ;21(14): 6305-6325
      Background: Tumor-associated macrophages (TAMs) are pivotal mediators of the immunosuppressive tumor immune microenvironment (TIME) in colorectal cancer (CRC). However, genes of TAMs that potentiate immunotherapy remain to be explored. Methods: Single-cell RNA sequencing (scRNA-seq) data were analyzed to identify TAM molecular signatures, which were validated in patient cohorts from Huadong Hospital and TCGA to explore their clinical significance. Multidimensional characterization of CRC TIME and Dipeptidyl peptidase VII (DPP7)-positive TAMs functional state was achieved through cytometry by time-of-flight, multiplex immunofluorescence, in vitro and in vivo experiments. Mechanistic investigations integrating RNA-seq, Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)-based proteomics, and targeted lipid metabolomics have revealed the reprogramming of key metabolic pathways. Finally, the therapeutic potential of DPP7, which targets the enhancement of anti-PD-1 immunotherapy efficacy, was demonstrated. Results: DPP7 was identified as the key gene in TAMs, and DPP7+TAMs correlated with metastasis and worse overall survival in multiple clinical cohorts. Functional characterization demonstrated that DPP7+TAMs drove the immunosuppressive TIME and promoted the exhaustion of CD8+T cells, thus exhibiting M2-polarized features. Mechanistically, DPP7 reduced ubiquitination-induced degradation of Carnitine Palmitoyltransferase 1A (CPT1A) by binding to CPT1A in a mutually exclusive manner with TRIM25, thus enhancing fatty acid oxidation (FAO) in TAMs. This metabolic reprogramming consumes lipids (including triglycerides and free fatty acids), elevates adenosine triphosphate (ATP) generation, and induces an immunosuppressive phenotype. In vivo, DPP7 knockdown in bone marrow-derived macrophages (BMDMs) synergized with anti-PD-1 therapy, achieving significant suppression of subcutaneous xenograft tumor growth and liver metastatic burden by reversing the immunosuppressive TIME. Conclusions: DPP7 is mainly expressed in TAMs and DPP7+TAMs are strongly associated with adverse prognosis in CRC. Mechanistically, DPP7 enhances FAO to promote the M2-polarized phenotype in TAMs, leading to an immunosuppressive TIME. Targeting DPP7+TAMs may potentiate the efficacy of immunotherapy for CRC.
    Keywords:  cancer immunotherapy; dipeptidyl peptidase VII; fatty acid oxidation; tumor-associated macrophage
    DOI:  https://doi.org/10.7150/ijbs.117909
  14. Nat Rev Drug Discov. 2025 Nov 12.
      Organoids are 3D structures derived from stem cells that recapitulate key architectural and functional aspects of the corresponding tissue. Compared with conventional 2D cell lines, human organoids provide experimental models that more closely reflect human physiology. Their ability to capture the complexity and heterogeneity of human tissues enables the study of disease mechanisms, drug efficacy and toxicity. When generated from patient material, organoids also allow the assessment of individual drug responses. In this Review, we explore the utility of organoids in drug discovery. We outline current methodologies for generating and maintaining organoids, examine their applications in disease modelling, drug screening and safety evaluation, and consider regulatory aspects and the challenges for their broader adoption in drug discovery.
    DOI:  https://doi.org/10.1038/s41573-025-01317-y
  15. Cell Death Dis. 2025 Nov 10. 16(1): 823
      The tumor microenvironment in colorectal cancer (CRC) is marked by a diverse and abundant population of cancer-associated fibroblasts (CAFs), which play a crucial role in radioresistance. Nonetheless, the mechanisms through which CAFs contribute to radioresistance remain unclear. In this study, we demonstrate that CAFR, a specific subset of CAFs derived from radioresistant CRC patients, produces higher levels of transforming growth factor-β1 (TGF-β1) compared to CAFs isolated from radiosensitive CRC patients. Through long noncoding RNA (lncRNA) profiling of tumor cells treated with CAF-conditioned medium (CAF-CM), we identify WARS2-IT1 (WARS2 intronic transcript 1), whose expression is directly stimulated by TGF-β1 signaling. This lncRNA serves as a key player in promoting radioresistance and is essential for the TGFβ1-induced radioresistance pathway. Mechanistically, WARS2-IT1 interferes with the interaction between prolyl hydroxylase domain 2 (PHD2) and hypoxia-inducible factor-1α (HIF-1α), preventing the hydroxylation and subsequent degradation of HIF-1α. This process leads to the activation of glycolytic pathways, thereby enhancing radioresistance. Our findings underscore the potential of targeting CAF-driven WARS2-IT1 as a promising strategy to counteract tumor radioresistance in CRC.
    DOI:  https://doi.org/10.1038/s41419-025-08058-1
  16. Cell Death Dis. 2025 Nov 10. 16(1): 822
      Colorectal cancer (CRC) is the third most common cancer worldwide and a significant public health threat. Ferroptosis, an iron-dependent form of regulated cell death, has emerged as a promising therapeutic target in CRC treatment. Despite its significant clinical potential, the precise regulatory mechanisms underlying ferroptosis, particularly its role in ferroptosis within CRC, remain to be fully elucidated. Previous studies, including our own work, have revealed that various deubiquitinases (DUBs) are involved in regulating cellular processes; however, the specific mechanisms by which these enzymes contribute to ferroptosis in CRC remain unclear. In this study, we identify USP5 as a key regulator of ferroptosis in CRC. Traditionally recognized as a deubiquitinase, USP5 modulates cellular physiological activities through deubiquitination. However, our findings show that USP5, distinct from its conventional deubiquitination function, suppresses ferroptosis by promoting the lysosomal degradation of YBX3 (Y-box binding protein 3). Under normal conditions, YBX3 promotes the degradation of SLC7A11 (solute carrier family 7 member 11). However, USP5 facilitates the degradation of YBX3, leading to the stabilization of SLC7A11 and thereby promoting CRC cell survival and tumor progression. In patient-derived organoid and xenograft models, USP5 knockout significantly increased the sensitivity of cancer cells to ferroptosis and inhibited tumor growth. Moreover, additional knockout of YBX3 restored the stability of SLC7A11, highlighting the complex regulatory network between USP5, YBX3, and SLC7A11. Systematic functional assays and mechanistic studies further confirmed that the USP5/YBX3/SLC7A11 axis is a central pathway for ferroptosis resistance in CRC. These findings provide novel insights into therapeutic strategies for CRC, especially ferroptosis-based treatments.
    DOI:  https://doi.org/10.1038/s41419-025-08146-2
  17. Cell Commun Signal. 2025 Nov 13. 23(1): 496
      Cancer stem cells were prominent responsible for cancer initiation, metastasis, and invasion as well as therapeutic resistance in colorectal cancer (CRC). The extracellular axon guidance factor netrin-1 has been found to be overexpressed in several malignant cancers such as glioma, lung cancers, and colorectal cancer. However, the role of netrin-1 on cancer stemness in CRC remains unveiled. Our study revealed high expression of netrin-1 in colorectal cancer tissues and its ability to promote cancer stemness by interacting with receptors UNC5B and neogenin on murine colorectal cancer cell. Mechanistically, the netrin-1-UNC5B/neogenin axis activates the downstream NF-κB and ERK1/2 signaling pathways, reinforcing the stemness properties of tumor cells, and further exacerbating tumor progression. Clinically, netrin-1 expression associated with poor survival and high CD133 expression in patients with CRC. Taken together, these results suggest that netrin-1 blockade could be a compelling therapeutic strategy to improve the poor outcomes and trigger cancer stemness inhibition in CRC treatment.
    Keywords:  Cancer stemness; Colorectal cancer; ERK1/2; NF-κB; Neogenin; Netrin-1; UNC5B
    DOI:  https://doi.org/10.1186/s12964-025-02489-1
  18. Biomed Pharmacother. 2025 Nov 11. pii: S0753-3322(25)00935-7. [Epub ahead of print]193 118741
      Colorectal cancer (CRC) remains a leading cause of cancer-related deaths globally, though targeted therapies have transformed treatment and brought renewed hope. However, resistance to these therapies is a persistent challenge, driven by genetic mutations, epigenetic changes, tumor microenvironment (TME) adaptations, and disruptions in key signaling pathways. These mechanisms allow tumor cells to evade treatment, causing disease progression and limiting long-term success. This review highlights recent breakthroughs and innovative strategies aimed at overcoming resistance in CRC. A deeper understanding of tumor heterogeneity, exosome-mediated communication, and the evolving TME is central to this effort. Promising approaches include biomarker-guided combination therapies and the use of epigenetic modulators to disrupt resistance mechanisms and restore treatment efficacy. Emerging studies also reveal the vulnerabilities of cancer persister cells. Targeting these cells, along with addressing cell cycle dysregulation and DNA damage repair (DDR) pathways, offers new opportunities for durable responses. This review serves as a roadmap for tackling resistance in CRC, with the ultimate goal of improving patient outcomes.
    Keywords:  Antibody-drug conjugates (ADC); Colorectal cancer; Combination therapy; Drug-tolerant persister cells (DTP); Precision oncology; Targeted therapy
    DOI:  https://doi.org/10.1016/j.biopha.2025.118741
  19. BMC Cancer. 2025 Nov 14. 25(1): 1764
      Normal epithelial cells detach from extracellular matrix during normal homeostasis by a selective process called epithelial extrusion. Thereafter, they undergo apoptosis, in a phenomenon called anoikis. Metastatic tumor cells, however, detach from normal epithelial tissues or primary tumors and survive in circulation, sometimes long enough to disseminate and colonize metastatic sites. The thirty years since the discovery of anoikis, exciting progress has been made in understanding anoikis mechanisms and the diverse mechanisms by which tumor cells evade or subvert them. Previous reviews have summarized earlier aspects of anoikis, including integrin-apoptosis signaling, Epithelial-Mesenchymal Transition and intracellular metabolism. Based on recently emerging work, we focus here on four issues that advance our conceptual understanding significantly: How do cytoskeleton and signaling interface so as to control anoikis? How is epithelial extrusion programmed in normal vs. cancer cells, upstream of anoikis? What is the role of translational regulation in anoikis? How do tumor cell-blood cell interactions affect anoikis? This review summarizes recent observations that inform these issues, and provides perspective on future problems to be investigated.
    Keywords:  Anoikis; Apoptosis; Epithelial extrusion; Mechanotransduction; Translational control
    DOI:  https://doi.org/10.1186/s12885-025-15178-6
  20. Gene. 2025 Nov 10. pii: S0378-1119(25)00680-8. [Epub ahead of print] 149890
      The miR-302 cluster, a key pluripotency-associated non-coding RNA, has been implicated in stem cell homeostasis and tumourigenesis. However, its regulatory mechanisms in cancers, including colorectal cancer (CRC) remain poorly understood. Here, we demonstrate that the β-catenin/TCF4 complex significantly enhances miR-302 expression through direct promoter activation in CRC cells. We hypothesized that the β-catenin/TCF4 complex directly activates the miR-302 promoter and cooperates with NANOG in a transcriptional feedback loop sustaining stem-like traits in CRC cells. Using a combination of promoter-driven luciferase reporter assays, chromatin immunoprecipitation (ChIP), and molecular dynamics simulations, we identify a regulatory axis involving Wnt signalling and the transcription factor NANOG. Our data show that individual members of the miR-302 cluster activate the NANOG promoter, while both NANOG and β-catenin/TCF4 synergistically enhance miR-302 promoter activity, suggesting the presence of a positive feedback loop. Structural simulations further elucidate the binding interactions between TCF4, NANOG, and the miR-302 promoter, corroborating our experimental observations. Together, these findings position miR-302 as a downstream effector of Wnt/β-catenin signalling and an integral component of NANOG-mediated transcriptional networks in CRC stem-like cells. This work advances our understanding of non-coding RNA regulation in cancer and highlights potential therapeutic opportunities for targeting stemness-associated pathways.
    Keywords:  Cancer stem-like cells; Colorectal cancer; NANOG; Non-coding RNA; TCF4; Transcriptional regulation; Wnt/β-catenin; miR-302 cluster
    DOI:  https://doi.org/10.1016/j.gene.2025.149890
  21. Methods Mol Biol. 2025 Nov 12.
      High-throughput drug testing combined with synergy evaluation in patient-derived tumor organoids (PDOs) represents a robust methodological approach to identify effective therapeutic combinations. PDOs retain tumor heterogeneity and the three-dimensional microenvironment, offering a physiologically relevant platform for rational drug testing. In this chapter, we provide a methodological guide to synergy testing in PDOs, including experimental design, statistical frameworks, and troubleshooting strategies. Emphasis is placed on practical aspects of drug combination screening, interpretation of synergy scores, and considerations for reproducibility. This practical guide aims to support researchers in applying organoid-based synergy assays as a translational tool in oncology.
    Keywords:  CalcuSyn; CompuSyn; Drug synergy; Drug testing; Patient-derived organoids; Synergy Finder R; Viability assays
    DOI:  https://doi.org/10.1007/7651_2025_676
  22. Cancer Lett. 2025 Nov 08. pii: S0304-3835(25)00695-0. [Epub ahead of print] 218123
      Although metastasis-initiating cells drive metastasis, only a certain subpopulation of these cells can successfully disseminate from the primary tumor to colonize metastatic sites. The identification and characterization of this subpopulation remain poorly studied. We designated this specific subpopulation as pre-metastatic state (PMS) cells. Identifying biomarkers and understanding the mechanisms underlying PMS formation are vital for preventing metastasis. In this research, we employed machine learning and bioinformatics techniques, integrating data from human cell lines, bulk RNA sequencing, single-cell sequencing, and spatial transcriptomics, to pinpoint PMS and classify its subtypes (PMS1 and PMS2). We revealed that PMS1 exhibits high epithelial-mesenchymal transition (EMT) characteristics and is associated with poor prognosis, while PMS2 is associated with tumor stemness. Notably, KLF7 was identified as a key biomarker of PMS, highly expressed in PMS1 and colorectal liver metastasis (CRLM) lesions. Through dual-luciferase reporter assays and both in vitro and in vivo experiments, we demonstrated that KLF7 promotes EMT characteristics in PMS1 via TGFβ autocrine signaling, thereby facilitating CRLM. Moreover, it was determined that the small molecule drug epigallocatechin gallate (EGCG) can inhibit the transcriptional activity of KLF7, thereby suppressing the TGFβ autocrine and EMT-related proteins. This result highlights EGCG as a promising compound for the treatment of colorectal cancer (CRC) with high KLF7 expression. Through this research, we established a systematic framework defining two PMS subtypes and identified KLF7 as a pivotal driver of CRLM in PMS1 through TGFβ autocrine activation.
    Keywords:  Drug sensitivity; Metastasis-initiating cells (MICs); Transcriptional regulation multi-omics; Tumor evolutionary trajectory
    DOI:  https://doi.org/10.1016/j.canlet.2025.218123
  23. Eur J Cancer. 2025 Nov 05. pii: S0959-8049(25)00983-9. [Epub ahead of print]231 116097
       INTRODUCTION: Patient-derived tumor organoids (PDTOs) have histological, molecular and clinical (drug sensitivity) characteristics comparable to those of their originating tumors. However, little is known about their ability to replicate the release of tumor-derived DNA.
    METHODS: Supernatants from 21 colorectal cancer PDTO cultures, established from 13 patients, were prospectively collected. The presence, structure, and mutational landscape of nuclear (cf-nDNA) and mitochondrial (cf-mtDNA) cell-free DNA as well as extracellular mitochondria (ex-Mito) were analyzed using qPCR, fragmentomics and shallow whole-genome sequencing. Mutation profiling was performed via IntPlex qPCR and whole-exome sequencing (WES).
    RESULTS: Cf-nDNA was detected in 95 % of PDTO supernatants with concentrations ranging from 0.009 to 209 ng/mL. Cf-nDNA fragment size analysis revealed patterns consistent with circulating DNA, including mononucleosome-associated profile. Cf-mtDNA was present in all samples (0.27-89.2 pg/mL) and extracellular mitochondria was also detected (0.009-17.4 pg/mL). A strong concordance (>85 %) was observed between oncogenic mutations in cfDNA and the molecular alterations detected in PDTOs and patient tumors.
    CONCLUSION: PDTOs release both nuclear and mitochondrial cfDNA into their culture medium displaying high similarity with patient-derived circulating DNA (cirDNA), including fragmentation patterns and oncogenic mutations. This study strengthens the relevance of the PDTOs as patient tumors models and highlights the potential of analyzing PDTO-derived cfDNA as a non-invasive approach to investigate tumor evolution and as a valuable tool to support functional precision oncology.
    Keywords:  Cell-free DNA (cfDNA); Circulating DNA; Fragmentomics; Patient-derived tumor organoids (PDTOs)
    DOI:  https://doi.org/10.1016/j.ejca.2025.116097
  24. Cell Genom. 2025 Nov 13. pii: S2666-979X(25)00318-0. [Epub ahead of print] 101062
      Tumor-specific antigens (TSAs) are crucial for activating T cells against cancer, but traditional discovery methods focusing on exonic mutations overlook non-canonical TSAs from non-coding regions. We employed an integrative proteogenomic strategy combining whole-genome and RNA sequencing with immunoprecipitation mass spectrometry to comprehensively explore TSA generation in colorectal cancer patients. Analysis of 10 paired tumor samples identified 96 mutated major histocompatibility complex class I-presented neo-epitopes, with 80.21% originating from non-coding regions. In hypermutated tumors with high mutational burden, neo-epitopes predominantly arose from intergenic and intronic areas, while in non-hypermutated tumors with low mutational burden, they mainly stemmed from coding variations and alternative splicing events. Functional validation in mouse models demonstrated that mutated non-canonical neo-epitopes effectively activated CD8+ T cells and significantly suppressed tumor growth. These findings underscore the importance of considering the entire genomic landscape in TSA discovery, suggesting new avenues for personalized immunotherapy.
    Keywords:  colorectal cancer; immunopeptidome; immunotherapy; neo-epitope; proteogenomics; tumor-specific antigens
    DOI:  https://doi.org/10.1016/j.xgen.2025.101062
  25. Drug Resist Updat. 2025 Nov 04. pii: S1368-7646(25)00124-4. [Epub ahead of print]84 101321
       AIMS: To investigate the regulatory role of NANOG in genes associated with stemness, symmetric division, and therapeutic resistance in colorectal cancer stem-like cells (CRC-SCs), with a focus on ERK/GSK-3β/β-catenin signalling and epithelial-mesenchymal transition (EMT), in order to evaluate the translational potential of targeting NANOG-associated signalling pathways.
    METHODS: Stemness, signalling activity, and cell division modes were analysed using 3D colonospheres enriched for CRC-SCs. Drug responses to the MEK inhibitor U0126 and the GSK-3β inhibitor TDZD-8 were assessed in CRC patient-derived organoids (PDOs), alongside molecular assays, immunohistochemistry with H-score quantification in xenograft models, and molecular dynamics simulations.
    RESULTS: NANOG overexpression enhanced the expression of stemness-associated genes, promoted symmetric cell division, and activated ERK/GSK-3β signalling, contributing to increased sphere formation. Inhibition of MEK and GSK-3β reduced EMT, cell proliferation, and symmetric division in CRC-SCs. NANOG-mediated dysregulation of ERK/GSK-3β altered β-catenin signalling and disrupted E-cadherin-dependent cell-cell adhesion. Molecular simulations and drug assays demonstrated that TDZD-8 and U0126 interfere with NANOG-DNA binding and β-catenin/E-cadherin interactions.
    CONCLUSIONS: NANOG drives CRC-SC maintenance via ERK/GSK-3β/β-catenin signalling and EMT modulation. This study offers significant insights into the translational impact of targeting NANOG and its downstream pathways with small-molecule inhibitors U0126 and TDZD-8 and presents a promising strategy to reduce CRC-SCs stemness, functionality, and tumourigenicity.
    Keywords:  Colorectal cancer stem cells (CRC-SCs); DNA-binding inhibitors; ERK/GSK-3β/β-catenin signalling; Epithelial-mesenchymal transition (EMT); NANOG; Symmetrical and asymmetrical cell division
    DOI:  https://doi.org/10.1016/j.drup.2025.101321