bims-instec 2025-04-06 papers

bims-instec

Biomed News

on Intestinal stem cells and chemoresistance in colon cancer and intestinal regeneration

Issue of 2025–04–06
nineteen papers selected by
Maria-Virginia Giolito, Université Catholique de Louvain

Research Advances in the Construction and Application of Intestinal Organoids.
C1Q+ TPP1+ macrophages promote colon cancer progression through SETD8-driven p53 methylation.
SOX9 suppresses colon cancer via inhibiting epithelial-mesenchymal transition and SOX2 induction.
Gut microbial metabolite 4-hydroxybenzeneacetic acid drives colorectal cancer progression via accumulation of immunosuppressive PMN-MDSCs.
Glycosaminoglycan modification of NRP1 exon 4-skipping variant drives colorectal cancer metastasis via endosomal-exosomal trafficking.
Revisiting lipid dysregulation in colorectal cancer: critical appraisal of pro-inflammatory bias and therapeutic implications.
SIRT5-mediated desuccinylation of PPA2 enhances HIF-1alpha-dependent adaptation to hypoxic stress and colorectal cancer metastasis.
Mast cells interact directly with colorectal cancer cells to promote epithelial-to-mesenchymal transition.
Spatial genomics reveals cholesterol metabolism as a key factor in colorectal cancer immunotherapy resistance.
High Efficiency Lentiviral Transduction of Colon Organoids Using Reversible 2D/3D Culture Techniques.
Epidemiology, pathogenesis, biology and evolving management of MSI-H/dMMR cancers.
Intestinal stearoyl-coenzyme A desaturase-inhibition improves obesity-associated metabolic disorders.
Cxcr3 promotes protection from colorectal cancer liver metastasis by driving NK cell infiltration and plasticity.
Colorectal carcinoma organoid and cancer-associated fibroblasts co-culture system for drug evaluation.
NUPR1 Promotes Radioresistance in Colorectal Cancer Cells by Inhibiting Ferroptosis.
Dipeptidase-1 knockout mice develop invasive tumors with features of microsatellite unstable colorectal cancer.
FSTL3 promotes colorectal cancer by activating the HIF1 pathway.
The colorectal cancer microenvironment governs clinical behaviors and chemotherapy response.
Understanding the cellular and molecular heterogeneity in colorectal cancer through the use of single-cell RNA sequencing.

Biomed Environ Sci. 2025 Feb 20. 38(2): 230-247

Research Advances in the Construction and Application of Intestinal Organoids.

Qing Xue Meng, Hong Yang Yi, Peng Wang, Shan Liu, Wei Quan Liang, Cui Shan Chi, Chen Yu Mao, Wei Zheng Liang, Jun Xue, Hong Zhou Lu.

  The structure of intestinal tissue is complex. In vitro simulation of intestinal structure and function is important for studying intestinal development and diseases. Recently, organoids have been successfully constructed and they have come to play an important role in biomedical research. Organoids are miniaturized three-dimensional (3D) organs, derived from stem cells, which mimic the structure, cell types, and physiological functions of an organ, making them robust models for biomedical research. Intestinal organoids are 3D micro-organs derived from intestinal stem cells or pluripotent stem cells that can successfully simulate the complex structure and function of the intestine, thereby providing a valuable platform for intestinal development and disease research. In this article, we review the latest progress in the construction and application of intestinal organoids.

Keywords:  3D culture; Colorectal cancer; Disease model; Intestinal organoids; Organ transplantation; Pluripotent stem cells; Regenerative medicine; Tumor organoids

DOI:  https://doi.org/10.3967/bes2025.010
Mol Cancer. 2025 Mar 31. 24(1): 102

C1Q+ TPP1+ macrophages promote colon cancer progression through SETD8-driven p53 methylation.

Veronica Veschi, Francesco Verona, Sebastiano Di Bella, Alice Turdo, Miriam Gaggianesi, Simone Di Franco, Laura Rosa Mangiapane, Chiara Modica, Melania Lo Iacono, Paola Bianca, Ornella Roberta Brancato, Caterina D'Accardo, Gaetana Porcelli, Vincenzo Luca Lentini, Isabella Sperduti, Elisabetta Sciacca, Peter Fitzgerald, David Lopez-Perez, Pierre Martine, Kate Brown, Giuseppe Giannini, Ettore Appella, Giorgio Stassi, Matilde Todaro.

   BACKGROUND: In many tumors, the tumor suppressor TP53 is not mutated, but functionally inactivated. However, mechanisms underlying p53 functional inactivation remain poorly understood. SETD8 is the sole enzyme known to mono-methylate p53 on lysine 382 (p53K382me1), resulting in the inhibition of its pro-apoptotic and growth-arresting functions.
METHODS: We analyzed SETD8 and p53K382me1 expression in clinical colorectal cancer (CRC) and inflammatory bowel disease (IBD) samples. Histopathological examinations, RNA sequencing, ChIP assay and preclinical in vivo CRC models, were used to assess the functional role of p53 inactivation in tumor cells and immune cell infiltration.
RESULTS: By integrating bulk RNAseq and scRNAseq approaches in CRC patients, SETD8-mediated p53 regulation resulted the most significantly enriched pathway. p53K382me1 expression was confined to colorectal cancer stem cells (CR-CSCs) and C1Q+ TPP1+ tumor-associated macrophages (TAMs) in CRC patient tissues, with high levels predicting decreased survival probability. TAMs promote p53 functional inactivation in CR-CSCs through IL-6 and MCP-1 secretion and increased levels of CEBPD, which directly binds SETD8 promoter thus enhancing its transcription. The direct binding of C1Q present on macrophages and C1Q receptor (C1QR) present on cancer stem cells mediates the cross-talk between the two cell compartments. As monotherapy, SETD8 genetic and pharmacological (UNC0379) inhibition affects the tumor growth and metastasis formation in CRC mouse avatars, with enhanced effects observed when combined with IL-6 receptor targeting.
CONCLUSIONS: These findings suggest that p53K382me1 may be an early step in tumor initiation, especially in inflammation-induced CRC, and could serve as a functional biomarker and therapeutic target in adjuvant setting for advanced CRCs.

Keywords:  C1Q+ TPP1+ macrophages; CRC; Cancer stem cells; IBD; P53K382me1 ; SETD8

DOI:  https://doi.org/10.1186/s12943-025-02293-y
J Clin Invest. 2025 Apr 03. pii: e184115. [Epub ahead of print]

SOX9 suppresses colon cancer via inhibiting epithelial-mesenchymal transition and SOX2 induction.

Ying Feng, Ningxin Zhu, Karan Bedi, Jinju Li, Chamila Perera, Maranne Green, Naziheh Assarzadegan, Yali Zhai, Qingzhi Liu, Veerabhadran Baladandayuthapani, Jason R Spence, Kathleen R Cho, Eric R Fearon.

  The Wnt/β-catenin pathway regulates expression of the SOX9 gene, which encodes SRY-box transcription factor 9, a differentiation factor and potential β-catenin regulator. Because APC tumor suppressor defects in ~80% of colorectal cancers (CRCs) activate the Wnt/β-catenin pathway, we studied SOX9 inactivation in CRC biology. Compared to effects of Apc inactivation in mouse colon tumors, combined Apc and Sox9 inactivation instigated more invasive tumors with epithelial-mesenchymal transition (EMT) and SOX2 stem cell factor upregulation. In an independent mouse CRC model with combined Apc, Kras, and Trp53 defects, Sox9 inactivation promoted SOX2 induction and distant metastases. About 20% of 171 human CRCs showed loss of SOX9 protein expression, which correlated with higher tumor grade. In an independent group of 376 CRC patients, low SOX9 gene expression was linked to poor survival, earlier age at diagnosis, and increased lymph node involvement. SOX9 expression reductions in human CRC were linked to promoter methylation. EMT pathway gene expression changes were prominent in human CRCs with low SOX9 expression and in a mouse cancer model with high SOX2 expression. Our results indicate SOX9 has tumor suppressor function in CRC; its loss may promote progression, invasion, and poor prognosis by enhancing EMT and stem cell phenotypes.

Keywords:  Colorectal cancer; Gastroenterology; Genetics; Mouse models; Oncology; Tumor suppressors

DOI:  https://doi.org/10.1172/JCI184115
J Clin Invest. 2025 Apr 03. pii: e181243. [Epub ahead of print]

Gut microbial metabolite 4-hydroxybenzeneacetic acid drives colorectal cancer progression via accumulation of immunosuppressive PMN-MDSCs.

Qing Liao, Ximing Zhou, Ling Wu, Yuyi Yang, Xiaohui Zhu, Hangyu Liao, Yujie Zhang, Weidong Lian, Feifei Zhang, Hui Wang, Yanqing Ding, Liang Zhao.

  Colorectal cancer (CRC) is characterized by an immune-suppressive microenvironment that contributes to tumor progression and immunotherapy resistance. The gut microbiome produces diverse metabolites that feature unique mechanisms of interaction with host targets, yet the role of many metabolites in CRC remains poorly understood. In this study, the microbial metabolite 4-hydroxybenzeneacetic acid (4-HPA) promoted the infiltration of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) in the tumor microenvironment, consequently inhibiting the anti-tumor response of CD8+ T cells and promoting CRC progression in vivo. Mechanistically, 4-HPA activates the JAK2/STAT3 pathway, which upregulates CXCL3 transcription, thereby recruiting PMN-MDSCs to the CRC microenvironment. Selective knockdown of CXCL3 re-sensitized tumors to anti-PD1 immunotherapy in vivo. Chlorogenic acid (CGA) reduces the production of 4-HPA by microbiota, likewise abolishing 4-HPA-mediated immunosuppression. The 4-HPA content in CRC tissues was notably increased in patients with advanced CRC. Overall, the gut microbiome uses 4-HPA as a messenger to control chemokine-dependent accumulation of PMN-MDSC cells and regulate anti-tumor immunity in CRC. Our findings provide a scientific basis for establishing clinical intervention strategies to reverse the tumor immune microenvironment and improve the efficacy of immunotherapy by reducing the interaction between intestinal microbiota, tumor cells and tumor immune cells.

Keywords:  Cancer immunotherapy; Colorectal cancer; Gastroenterology; Immunology; Oncology

DOI:  https://doi.org/10.1172/JCI181243
Cancer Lett. 2025 Mar 27. pii: S0304-3835(25)00249-6. [Epub ahead of print] 217683

Glycosaminoglycan modification of NRP1 exon 4-skipping variant drives colorectal cancer metastasis via endosomal-exosomal trafficking.

Yiwei Gu, Qing Ye, Xiuping Huang, Yanan Cao, Luksana Chaiswing, Qing-Bai She.

  Neuropilin-1 (NRP1) is a transmembrane glycoprotein that functions as a co-receptor with various cellular functions. Our previous studies identified the NRP1 exon 4-skipping (NRP1-ΔE4) splice variant as an aggressive metastasis driver by activating endosomal signals. Here, we demonstrate the critical role of glycosaminoglycan (GAG) modification in regulating NRP1-ΔE4's cellular trafficking and oncogenic activity. NRP1-ΔE4 undergoes constitutive internalization into endosomes and subsequent exosomal release from colorectal cancer (CRC) cells. Exosomal NRP1-ΔE4 enhances the migration and invasion of both donor and recipient CRC cells. Genetic or pharmacological inhibition of exosome secretion, or immunodepletion of exosomal NRP1-ΔE4, markedly reduces its metastatic potential. Notably, GAG modification at the O-glycosylation site Ser612 is essential for NRP1-ΔE4's endosomal trafficking and exosomal release. This modification also promotes the formation of a trimeric complex with Met and β1-integrin, leading to their co-internalization and accumulation in endosomes, which activates FAK signaling and drives CRC metastasis. These findings reveal GAG modification as a key regulatory process that governs the endosomal-exosomal trafficking of NRP1-ΔE4 to facilitate CRC cell dissemination.

Keywords:  NRP1; NRP1-ΔE4; colorectal cancer; endosomes; exosomes; glycosaminoglycan modification; metastasis

DOI:  https://doi.org/10.1016/j.canlet.2025.217683
Gut. 2025 Apr 02. pii: gutjnl-2025-335153. [Epub ahead of print]

Revisiting lipid dysregulation in colorectal cancer: critical appraisal of pro-inflammatory bias and therapeutic implications.

Xudong Zhu.



Keywords:  COLORECTAL CANCER; INFLAMMATION; LIPID METABOLISM

DOI:  https://doi.org/10.1136/gutjnl-2025-335153
EMBO J. 2025 Mar 31.

SIRT5-mediated desuccinylation of PPA2 enhances HIF-1alpha-dependent adaptation to hypoxic stress and colorectal cancer metastasis.

Xiang Zhang, Yuqin Di, Youpeng Wang, Jiale Qin, Lvlan Ye, Xiangqiong Wen, Zunfu Ke, Ziyang Wang, Weiling He.

  Metastasis is the primary cause of death in patients with colorectal cancer (CRC). Hypoxia is a hallmark of solid tumors that promotes cellular metabolic adaptation and dissemination. However, the mechanisms linking hypoxia-regulated metabolic adaptation to CRC metastasis remain unclear. Here, we found that inorganic pyrophosphatase 2 (PPA2) suppresses metastatic progression of CRC via its phosphatase function. PPA2 expression levels are reduced in CRC specimen and correlate with enhanced response to hypoxia by promoting hypoxia-inducible factor-1 (HIF-1) signaling to promote CRC cell glycolysis and dissemination. Mechanistically, PPA2 decreases HIF-1alpha stability through non-canonical ubiquitin-mediated proteasomal degradation via recruitment of E3 ligase NEDD4. Furthermore, PPA2 directly dephosphorylates NEDD4 at threonine 758 residue, resulting in its activation. Under hypoxic stress, NAD-dependent protein deacetylase sirtuin-5 promotes the dissociation of PPA2 and NEDD4 by inducing PPA2 desuccinylation at lysine 176, contributing to the improved stability of HIF-1alpha under hypoxic conditions. Our findings reveal a tumor-suppressive role of PPA2 in HIF-1alpha-dependent colorectal cancer, providing a potential therapeutic target and prognostic strategy.

Keywords:  HIF-1α; Hypoxia; Metastatic Colorectal Cancer; NEDD4; PPA2 Succinylation

DOI:  https://doi.org/10.1038/s44318-025-00416-1
bioRxiv. 2025 Mar 19. pii: 2025.03.19.644113. [Epub ahead of print]

Mast cells interact directly with colorectal cancer cells to promote epithelial-to-mesenchymal transition.

Rosie Lanzloth, Nicole L Harris, Anthony M Cannon, Mark H Kaplan, Heather M O'Hagan.

Mast cells (MCs), a type of granulocytic immune cell, can be both pro- and anti-tumorigenic in colorectal cancer (CRC). We hypothesized that these contrasting findings may be in part due to di_erential interactions of MCs with CRC subtypes. BRAF mutant CRC uniquely contains intestinal secretory cell types. In this study, we demonstrated that MCs are enriched in BRAF mutant CRC, likely because they are recruited by factors released from cancer secretory cells. To investigate the functional consequences of MC-CRC cell interactions, we performed direct coculture experiments. We demonstrated that MCs promote epithelial-to-mesenchymal transition (EMT) in CRC cells in a calcium- and contact-dependent fashion. Furthermore, inhibiting LFA-1 and ICAM1 integrin binding reduced the coculture-induced EMT-related marker expression in CRC cells. The MC-CRC cell interaction facilitates the transfer of biological materials, including mRNA molecules, from MCs to CRC cells. This study is the first to report a contact-dependent, pro-tumorigenic role of MCs in CRC, as well as the transfer of molecules encoded by MCs to CRC cells. These findings enhance our comprehension of cell-cell communication between immune and cancer cells. Furthermore, this work suggests that targeting MC-CRC interactions, particularly through modulating integrin pathways, could o_er new therapeutic strategies for aggressive CRC subtypes.

DOI: https://doi.org/10.1101/2025.03.19.644113
Front Oncol. 2025 ;15 1549237

Spatial genomics reveals cholesterol metabolism as a key factor in colorectal cancer immunotherapy resistance.

Andrew J Kavran, Yulong Bai, Brian Rabe, Anna Kreshock, Andrew Fisher, Yelena Cheng, Anne Lewin, Chao Dai, Matthew J Meyer, Konstantinos J Mavrakis, Anna Lyubetskaya, Eugene Drokhlyansky.

  Immune checkpoint inhibitors (ICIs) have transformed the treatment landscape across multiple cancer types achieving durable responses for a significant number of patients. Despite their success, many patients still fail to respond to ICIs or develop resistance soon after treatment. We sought to identify early treatment features associated with ICI outcome. We leveraged the MC38 syngeneic tumor model because it has variable response to ICI therapy driven by tumor intrinsic heterogeneity. ICI response was assessed based on the level of immune cell infiltration into the tumor - a well-established clinical hallmark of ICI response. We generated a spatial atlas of 48,636 transcriptome-wide spots across 16 tumors using spatial transcriptomics; given the tumors were difficult to profile, we developed an enhanced transcriptome capture protocol yielding high quality spatial data. In total, we identified 8 tumor cell subsets (e.g., proliferative, inflamed, and vascularized) and 4 stroma subsets (e.g., immune and fibroblast). Each tumor had orthogonal histology and bulk-RNA sequencing data, which served to validate and benchmark observations from the spatial data. Our spatial atlas revealed that increased tumor cell cholesterol regulation, synthesis, and transport were associated with a lack of ICI response. Conversely, inflammation and T cell infiltration were associated with response. We further leveraged spatially aware gene expression analysis, to demonstrate that high cholesterol synthesis by tumor cells was associated with cytotoxic CD8 T cell exclusion. Finally, we demonstrate that bulk RNA-sequencing was able to detect immune correlates of response but lacked the sensitivity to detect cholesterol synthesis as a feature of resistance.

Keywords:  MC38; PD-1; Visium; cholesterol; colorectal cancer; immunotherapy resistance; spatial genomics; spatial transcriptomics

DOI:  https://doi.org/10.3389/fonc.2025.1549237
Methods Mol Biol. 2025 ;2905 245-254

High Efficiency Lentiviral Transduction of Colon Organoids Using Reversible 2D/3D Culture Techniques.

Florian Rathje, Martina M Sykora, Fritz Aberger, Peter W Krenn.

  Organoids are a promising research tool for studying tissue development and disease in vitro. While organoids are frequently considered a replacement or complementary model for in vivo mouse experiments, exploiting their full potential often requires genetically engineered mice as a source of transgenic stem cells, also because genetic manipulation of organoids is rather inefficient and cumbersome. Here, we describe an alternative and optimized murine colon organoid manipulation protocol that reversibly and temporarily interrupts the 3D organoid structure for short-term 2D monolayer culture. This approach allows highly efficient viral transduction and genetic manipulation of stem cells in a 2D setting, followed by 3D stem cell embedding and restoration of the original organoid architecture. This method greatly improves the efficiency of lentiviral-mediated genetic manipulation of organoids and increases their potential applications in CRISPR/Cas9 and compound screens, immune-competent co-cultures, and disease modeling.

Keywords:  2D/3D organoid culture; Colon organoids; Genetic manipulation; Stem cells; Viral transduction

DOI:  https://doi.org/10.1007/978-1-0716-4418-8_16
Nat Rev Clin Oncol. 2025 Apr 03.

Epidemiology, pathogenesis, biology and evolving management of MSI-H/dMMR cancers.

Margherita Ambrosini, Paolo Manca, Vincenzo Nasca, Carolina Sciortino, Filippo Ghelardi, Jenny F Seligmann, Julien Taieb, Filippo Pietrantonio.

Deficiency in DNA mismatch repair (dMMR) is a common pathway of carcinogenesis across different tumour types and confers a characteristic microsatellite instability-high (MSI-H) molecular phenotype. The prevalence of the MSI-H/dMMR phenotype is highest in endometrial and colorectal cancers, and this phenotype is associated with a distinct tumour biology, prognosis and responsiveness to various anticancer treatments. In a minority of patients, MSI-H/dMMR cancers result from an inherited pathogenic variant in the context of Lynch syndrome, which has important implications for familial genetic screening. Whether these hereditary cancers have a different biology and clinical behaviour to their sporadic counterparts remains uncertain. Interest in this tumour molecular subtype has increased following the discovery of the high sensitivity of metastatic MSI-H/dMMR cancers to immune-checkpoint inhibitors (ICIs) in a histology-agnostic manner, which reflects the genomic hypermutation resulting from dMMR that renders these tumours highly immunogenic and immune infiltrated. This vulnerability is now also being exploited in early stage disease settings. Despite this common biological foundation, different MSI-H/dMMR cancers have histotype-specific features that correspond to their particular cell or tissue of origin, which might be associated with differences in prognosis and sensitivity to ICIs. In this Review, we provide an overview of the epidemiology, biology, pathogenesis, clinical diagnosis and treatment of MSI-H/dMMR tumours as a histology-agnostic cancer phenomenon. We also highlight peculiarities associated with specific pathogenetic alterations and histologies of MSI-H/dMMR tumours.

DOI: https://doi.org/10.1038/s41571-025-01015-z
Acta Pharm Sin B. 2025 Feb;15(2): 892-908

Intestinal stearoyl-coenzyme A desaturase-inhibition improves obesity-associated metabolic disorders.

Yangliu Xia, Yang Zhang, Zhipeng Zhang, Nana Yan, Vorthon Sawaswong, Lulu Sun, Wanwan Guo, Ping Wang, Kristopher W Krausz, Oksana Gavrilova, James M Ntambi, Haiping Hao, Tingting Yan, Frank J Gonzalez.

  Stearoyl-coenzyme A desaturase 1 (SCD1) catalyzes the rate-limiting step of de novo lipogenesis and modulates lipid homeostasis. Although numerous SCD1 inhibitors were tested for treating metabolic disorders both in preclinical and clinic studies, the tissue-specific roles of SCD1 in modulating obesity-associated metabolic disorders and determining the pharmacological effect of chemical SCD1 inhibition remain unclear. Here a novel role for intestinal SCD1 in obesity-associated metabolic disorders was uncovered. Intestinal SCD1 was found to be induced during obesity progression both in humans and mice. Intestine-specific, but not liver-specific, SCD1 deficiency reduced obesity and hepatic steatosis. A939572, an SCD1-specific inhibitor, ameliorated obesity and hepatic steatosis dependent on intestinal, but not hepatic, SCD1. Mechanistically, intestinal SCD1 deficiency impeded obesity-induced oxidative stress through its novel function of inducing metallothionein 1 in intestinal epithelial cells. These results suggest that intestinal SCD1 could be a viable target that underlies the pharmacological effect of chemical SCD1 inhibition in the treatment of obesity-associated metabolic disorders.

Keywords:  High-fat diet; Intestinal epithelium; MT1; Metabolic disorders; Obesity; Oxidative stress; SCD1; Steatosis

DOI:  https://doi.org/10.1016/j.apsb.2024.11.022
J Clin Invest. 2025 Apr 01. pii: e184036. [Epub ahead of print]

Cxcr3 promotes protection from colorectal cancer liver metastasis by driving NK cell infiltration and plasticity.

Eleonora Russo, Chiara D'Aquino, Chiara Di Censo, Mattia Laffranchi, Luana Tomaipitinca, Valerio Licursi, Stefano Garofalo, Johann Promeuschel, Giovanna Peruzzi, Francesca Sozio, Anna Kaffke, Cecilia Garlanda, Ulf Panzer, Cristina Limatola, Christian A J Vosshenrich, Silvano Sozzani, Giuseppe Sciumè, Angela Santoni, Giovanni Bernardini.

  The anti-metastatic activity of NK cells is well established in several cancer types, but the mechanisms underlying NK cell metastasis infiltration and acquisition of anti-tumor characteristics remain unclear. Herein, we investigated the cellular and molecular factors required to facilitate the generation of an ILC1-like CD49a+NK cell population within the liver metastasis (LM) environment of colorectal cancer (CRC). We show that CD49a+NK cells had the highest cytotoxic capacity among metastasis-infiltrating NK cells in the MC38 mouse model. Furthermore, the chemokine receptor CXCR3 promoted CD49a+NK cell accumulation and persistence in metastasis where NK cells co-localize with macrophages in CXCL9 and CXCL10 rich areas. By mining a published scRNA-seq dataset of a cohort of treatment-naïve CRC patients, we confirmed the accumulation of CXCR3+NK cells in metastatic samples. Conditional deletion of Cxcr3 in NKp46+ cells and antibody-mediated depletion of metastasis-associated macrophages impaired CD49a+NK cell development, indicating that CXCR3 and macrophages contribute to efficient NK cell localization and polarization in LM. Conversely, CXCR3neg NK cells maintained a CD49a- phenotype in metastasis with reduced parenchymal infiltration and tumor killing capacity. Furthermore, CD49a+NK cell accumulation was impaired in an independent SL4-induced CRC metastasis model, which fails to accumulate CXCL9+ macrophages. Together, our results highlight a role for CXCR3/ligand axis in promoting macrophage-dependent NK cell accumulation and functional sustenance in CRC LM.

Keywords:  Cell migration/adhesion; Immunology; Innate immunity; NK cells; Oncology

DOI:  https://doi.org/10.1172/JCI184036
In Vitro Model. 2025 Feb;4(1): 31-44

Colorectal carcinoma organoid and cancer-associated fibroblasts co-culture system for drug evaluation.

Yan Wang, Zilin Zhang, Xiaoran Li, Xiaobing Lu, Xuemei Zhuansun, Qiwei Li, Jing Zhang, Xi Xu, Xueqiang Liu, Yuan Wei, Feng Hua, Runda Wu, Zaozao Chen.

  Patient-derived organoids (PDO) have the potential to be used as preclinical cancer models for testing anti-cancer drug efficiency. Cancer-associated fibroblasts (CAFs), which have been closely linked with colorectal carcinoma (CRC) progression and drug resistance, however, are generally not included (or gradually lost during culture) in the PDO models, leading to a major limitation in this cancer model. In this study, we established a new in vitro model with CRC organoids and co-cultured with CAFs and compared it with the organoid-only model. Through testing with anti-cancer drug, we demonstrated a significant difference in drug sensitivity between the two models, and the co-culture model showed higher drug resistance. RNA and whole exome sequencing were performed to reveal gene expression profiles in organoids and organoids co-culture with CAFs to assess interactions between drug sensitivity and gene copy number variation. We found that the expression levels of several pathway protein genes, which are highly expressed in original surgical specimens of colorectal carcinomas, were downregulated in organoids but restored in organoids by co-culturing with CAFs. In summary, the PDO-CAF joint model for CRC can recapitulate a more biomimetic tumor microenvironment and the drug resistance lead by changes in multiple signaling pathways that we discovered; thus, it could be a suitable model for future usage in drug discovery and precision medicine research.

Keywords:  Cancer-associated fibroblasts; Co-culture; Colorectal carcinoma organoid; Drug resistance

DOI:  https://doi.org/10.1007/s44164-025-00084-9
J Cell Mol Med. 2025 Apr;29(7): e70519

NUPR1 Promotes Radioresistance in Colorectal Cancer Cells by Inhibiting Ferroptosis.

Yimin Fang, Haiyan Chen, Yunhua Liu, Kai Jiang, Yucheng Qian, Jingsun Wei, Dongliang Fu, Hang Yang, Siqi Dai, Tian Jin, Tongtong Bu, Kefeng Ding.

  Radioresistance is a major clinical challenge and the underlying mechanism has not been thoroughly elucidated. In this study, a radioresistant (RR) cell line is established to explore the transcriptomic signatures of radioresistance in colorectal cancer (CRC). KEGG enriched pathway analysis demonstrated that ferroptosis is inactivated in RR cells. Further detection confirmed that radiotherapy can promote ferroptosis, and ferroptosis inactivation is one of the hallmarks of radioresistance in CRC. What's more, induction of ferroptosis can restore the radiosensitivity of CRC cells. Then, we performed RNA sequencing to compare gene expression between parental and RR cells, and cells pretreated with or without RSL3. Via high-throughput screening, NUPR1 was identified as a potential candidate for ferroptosis-mediated radioresistance in CRC. CRC cells can acquire radiation resistance by NUPR1-mediated ferroptosis suppression in the NUPR1-overexpressing cell line. More importantly, ZZW-115, an NUPR1 inhibitor, can sensitise RR cells to radiotherapy. Overall, our findings identify ferroptosis inactivation linked with resistance to radiotherapy. Besides, NUPR1 can promote radiation resistance by inhibiting ferroptosis, and targeting NUPR1 may be a potential strategy to relieve radioresistance associated with ferroptosis in CRC.

Keywords:  NUPR1; colorectal cancer; ferroptosis; radiotherapy resistance

DOI:  https://doi.org/10.1111/jcmm.70519
JCI Insight. 2025 Apr 03. pii: e186938. [Epub ahead of print]

Dipeptidase-1 knockout mice develop invasive tumors with features of microsatellite unstable colorectal cancer.

Sarah E Glass, Matthew E Bechard, Zheng Cao, Radhika Aramandla, Ping Zhao, Samuel T Ellis, Emily H Green, Elizabeth G Fisher, Ryan T Smith, Chelsie K Sievers, Maria Johnson Irudayam, Frank Revetta, M Kay Washington, Gregory D Ayers, Cody N Heiser, Alan J Simmons, Yanwen Xu, Yu Wang, Annika Windon, Martha J Shrubsole, Nicholas O Markham, Qi Liu, Ken S Lau, Robert J Coffey.

  Dipeptidase-1 (DPEP1) is highly upregulated in colorectal cancer (CRC), with its enzymatic function linked to invasion and metastasis. More recently, DPEP1 was found to serve as a receptor for neutrophils when expressed by activated endothelial cells. It is unknown whether neutrophils bind to DPEP1-expressing CRC cells and whether this impacts features of CRC. Neutrophils have been shown to be tumor-promoting in cancers including CRC, where they act to exclude CD8+ T cells. Herein, we show that neutrophils bind DPEP1-expressing CRC cells. In addition, DPEP1 is preferentially expressed in microsatellite stable (MSS) CRC, in which there are a paucity of CD8+ T cells, whereas DPEP1 is negatively correlated with microsatellite unstable (MSI-H) CRC, which are T cell-rich and are more responsive to immunotherapy. Remarkably, carcinogen-treated Dpep1 null mice develop multiple, large, plaque-like, locally invasive adenocarcinomas and squamous cell cancers in the distal colon. These adenocarcinomas exhibit a marked reduction of neutrophils and an influx CD8+ T cells, along with reduced expression of mismatch repair proteins, consistent with features of MSI-H CRC. These results establish DPEP1's importance in maintaining MSS CRC and its ability to shape the tumor microenvironment.

Keywords:  Cancer; Cell biology; Oncology

DOI:  https://doi.org/10.1172/jci.insight.186938
Gene. 2025 Mar 26. pii: S0378-1119(25)00223-9. [Epub ahead of print]954 149435

FSTL3 promotes colorectal cancer by activating the HIF1 pathway.

Xiang-Rong Luo, Li-Zhe Huang, Jie Yin, Zu-Ming Xiong, Wen-Xin Li, Cun Liao, Ming-Lin Lin, Wei Huang, Sen Zhang.

  Follistatin-like 3 (FSTL3) is a glycoprotein known to promote tumor growth, invasion, and angiogenesis in various cancers. However, its role in Colorectal Cancer (CRC), particularly concerning the hypoxia-inducible factor 1α (HIF1α) signaling pathway, remains unclear. The HIF1α pathway is critical in CRC progression, enabling tumor cells to adapt to hypoxia through angiogenesis, Epithelial-Mesenchymal Transition (EMT), and metabolic reprogramming. Analysis of The Cancer Genome Atlas (TCGA) and GSE39582 datasets revealed that FSTL3 is significantly upregulated in CRC tissues and correlates with poor Overall Survival (OS), Progression-Free Survival (PFS), and aggressive features such as venous, lymphatic, and perineural invasion. In vitro experiments demonstrated that FSTL3 overexpression in HCT15 and HCT116 cells promoted proliferation, migration, and cell cycle progression, whereas knockdown in LOVO and Caco2 cells suppressed these processes and induced apoptosis. Transcriptome sequencing and western blot analysis indicated that FSTL3 activated the HIF1α pathway by upregulating HIF1α, ANGPT2, and HK3, which are key regulators of angiogenesis and glycolysis. Importantly, treatment with the HIF1α inhibitor KC7F2 reversed the oncogenic effects of FSTL3 overexpression both in vitro and in vivo. In xenograft and tail vein metastasis models, KC7F2 suppressed tumor growth, reduced pulmonary metastasis, and restored lung tissue integrity, further downregulating FSTL3 and HIF1α expression. These findings suggest that FSTL3 promotes CRC progression via the HIF1α pathway and highlight its potential as a prognostic biomarker and therapeutic target for CRC treatment.

Keywords:  Biomarker; Colorectal cancer; FSTL3; HIF1 pathway; Oncogenesis; Prognosis

DOI:  https://doi.org/10.1016/j.gene.2025.149435
Cancer Lett. 2025 Apr 01. pii: S0304-3835(25)00253-8. [Epub ahead of print] 217687

The colorectal cancer microenvironment governs clinical behaviors and chemotherapy response.

André Holdfeldt, Simona Salerno, Anders Ståhlberg, Elinor Bexe Lindskog, Göran Landberg.

DOI: https://doi.org/10.1016/j.canlet.2025.217687
Transl Oncol. 2025 Mar 30. pii: S1936-5233(25)00105-6. [Epub ahead of print]55 102374

Understanding the cellular and molecular heterogeneity in colorectal cancer through the use of single-cell RNA sequencing.

Yuemiao Chen, Jian Huang, Yufang Fan, Lifeng Huang, Xiaoping Cai.

  The very prevalent nature, genetic variability, and intricate tumor microenvironment (TUME) of colorectal cancer (COREC) are its defining features. In order to better understand the molecular and cellular make-up of COREC, this work used single-cell RNA sequencing (SRNAS) to isolate and characterize important cell types as well as their interactions within the TUME. Our analysis of 51,204 cells yielded six distinct types: epithelial, fibroblast, endothelial, T&NK, B, and myeloid. C3 B cells were shown to be the most active in immunological regulation, according to chemokine signaling study, which was one of seven clusters of B cells that were thoroughly subtyped. The examination of copy number variation (CONUV) revealed a great deal of genetic variability, especially in epithelial cells. We traced the activity of three key transcription factor clusters (M1, M2, and M3) across all B cell subtypes using transcription factor analysis. We created a predictive model that correctly sorts patients according to survival results by using marker genes from C3 B cells. In addition, the relationship between genetic changes and the immune system was better understood by tumor mutational burden (TUMUB) and immune infiltration studies. Our research sheds light on the genetic complexity and cellular variety of COREC, which in turn opens up new possibilities for targeted treatments and individualized approaches to patient care.

Keywords:  B cell subtypes; Colorectal cancer; Genetic heterogeneity; Single-cell RNA sequencing; Tumor microenvironment

DOI:  https://doi.org/10.1016/j.tranon.2025.102374