Marwah Al-Mathkour, Zheng Chen, Julio Poveda, Longlong Cao, Oliver G McDonald, Dunfa Peng, Mohammed Soutto, Zhibin Chen, Heng Lu, Yan Guo, Shria Kumar, Alexander Zaika, Silvia Giordano, Shoumin Zhu, Wael El-Rifai
{"title":"CDK1 drives SOX9-mediated chemotherapeutic resistance in gastric cancer.","authors":"Marwah Al-Mathkour, Zheng Chen, Julio Poveda, Longlong Cao, Oliver G McDonald, Dunfa Peng, Mohammed Soutto, Zhibin Chen, Heng Lu, Yan Guo, Shria Kumar, Alexander Zaika, Silvia Giordano, Shoumin Zhu, Wael El-Rifai","doi":"10.1186/s13046-025-03523-3","DOIUrl":"10.1186/s13046-025-03523-3","url":null,"abstract":"<p><strong>Background: </strong>Gastric carcinoma ranks as the fifth most common cause of cancer-related mortality globally. Chemoresistance remains a critical barrier to treatment efficacy, driving poor survival outcomes in gastric cancer patients. Cyclin-dependent kinase 1 (CDK1) is overexpressed in several malignancies. SOX9 transcription factor plays critical roles in gastric tumorigenesis and therapeutic resistance. This study identifies a CDK1-SOX9-BCL-xL signaling axis as an important mediator of chemoresistance in gastric cancer.</p><p><strong>Methods: </strong>Bioinformatics and computational approaches were used for analysis of human and mouse public and local data sets. Chromatin immunoprecipitation (ChIP), western blotting, quantitative PCR (qPCR), immunofluorescence, and immunohistochemistry assays were applied in the study. The study utilized a number of in vitro models including cell lines and patient-derived tumoroids. The in vivo models included patient-derived xenograft (PDX), the Tff1 knockout, and Cdk1 conditional knockout mouse models.</p><p><strong>Results: </strong>Our study identified concurrent overexpression of CDK1 and SOX9 in gastric cancer patients. Genetic knockdown and pharmacological inhibition of CDK1 suppressed SOX9 protein levels and transcriptional activity in vitro and in vivo. Mechanistically, CDK1 regulates SOX9 through a miR-145-dependent epigenetic axis: CDK1-mediated phosphorylation and activation of DNMT1 to drive methylation-dependent silencing of miR-145, thereby relieving miR-145's repression of SOX9. Strikingly, both CDK1 and SOX9 were upregulated in cisplatin-resistant gastric cancer cell lines. We further identified BCL-xL as a direct transcriptional target of SOX9, functionally mediating cisplatin resistance. CDK1 inhibition using dinaciclib re-sensitized resistant models to cisplatin by disrupting the CDK1-SOX9-BCL-xL pathway, underscoring its central role in chemoresistance. In PDX models, combining dinaciclib with cisplatin synergistically reduced tumor volume, and extended survival compared to monotherapies, highlighting the therapeutic potential.</p><p><strong>Conclusion: </strong>This study elucidates the epigenetic and transcriptional mechanisms driving the CDK1-SOX9-BCL-xL axis in gastric cancer chemoresistance. Pharmacological inhibition of CDK1 effectively disrupts this axis, restoring cisplatin sensitivity and suppressing tumor growth in gastric cancer models. The observed synergy between dinaciclib and cisplatin underscores a promising therapeutic strategy to overcome chemoresistance in gastric cancer.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"284"},"PeriodicalIF":12.8,"publicationDate":"2025-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12506374/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145253249","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Cancer-associated fibroblast-derived circKLHL24 drives perineural invasion in pancreatic cancer via dual regulation of the sec31a-CXCL12 axis.","authors":"Tingting Li, Rihua He, Qing Tian, Tianhao Huang, Honghui Jiang, Huimou Chen, Yuan Yuan, Yong Jiang, Shangyou Zheng, Chonghui Hu, Shizong Li, Guolin Li, Rufu Chen","doi":"10.1186/s13046-025-03489-2","DOIUrl":"10.1186/s13046-025-03489-2","url":null,"abstract":"<p><strong>Background: </strong>Cancer-associated fibroblasts (CAFs) are key drivers of neural invasion in pancreatic cancer, yet their regulatory mechanisms remain elusive.This study explores the role of circular RNAs (circRNAs) in CAFs and their involvement in regulating neural invasion in pancreatic cancer.</p><p><strong>Methods: </strong>CAF-derived circRNAs were identified through circRNA high-throughput sequencing and quantitative real-time PCR (qRT-PCR). The impact of CAF-derived circKLHL24 on perineural invasion (PNI) in tumor cells was evaluated both in vitro and in vivo. RNA sequencing, RNA pulldown, RNA immunoprecipitation, and luciferase reporter assays were conducted to identify downstream targets and elucidate the underlying mechanism of circKLHL24 in PNI.</p><p><strong>Results: </strong>CircKLHL24 (hsa_circ_0001369), a CAF-specific circRNA, is associated with PNI and poor survival in advanced PDAC. Silencing or overexpressing circKLHL24 in CAFs altered the ability of CAFs to induce tumor cell invasion and nerve infiltration via chemokine (C-X-C Motif) ligand 12 (CXCL12). Mechanistically, first, circKLHL24 binds to the membrane protein Sec31A, inhibiting its ubiquitination and degradation, thereby enhancing CXCL12 secretion. Second, circKLHL24 acts as a sponge for miR-615-5p, relieving its suppression of CXCL12 mRNA and amplifying CXCL12 expression. Moreover, high circKLHL24 levels were positively correlated with elevated serum CXCL12 levels in PDAC and poor patient survival. Targeting circKLHL24 or neutralizing CXCL12 suppresses PDAC invasion and neuronal recruitment in nude mouse and KPC models.</p><p><strong>Conclusions: </strong>The circKLHL24/Sec31A/miR-615-5p/CXCL12 axis is critical for CAF-induced PNI in PDAC. Therefore, circKLHL24 could serve as a potential therapeutic target for PDAC.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"281"},"PeriodicalIF":12.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12502155/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Co-detection of mutations and methylations in cerebrospinal fluid ctdna for minimally-invasive diagnosis of brainstem glioma.","authors":"Tian Li, Huan Wang, Yujin Wang, Yiying Mai, Pei Wang, Mingxin Zhang, Zhuang Jiang, Luyang Xie, Hang Zhou, Yi Wang, Xiaoou Li, Dan Xiao, Jingyao Geng, Wenhao Wu, Peng Zhang, Liang Wang, Zhen Wu, Junting Zhang, Dandan Cao, Changchun Pan, Liwei Zhang","doi":"10.1186/s13046-025-03455-y","DOIUrl":"10.1186/s13046-025-03455-y","url":null,"abstract":"<p><strong>Background: </strong>Genetic and epigenetic profiles are critical in managing brainstem gliomas (BSG), whose heterogeneity is far beyond the realm of the Diffuse midline glioma, H3K27 altered. Cerebrospinal fluid (CSF) circulating tumor DNA (ctDNA)-based liquid biopsy provides minimally-invasive strategies to acquire molecular information for brain tumors, whereas there is a deficiency in techniques for co-detection of genetic and epigenetic alterations due to the limited yield of ctDNA. This study aims to develop a reliable minimally-invasive approach to simultaneously detect the mutation and methylation profiles in the CSF ctDNA of BSGs, thereby enhancing diagnostic accuracy, prognostic capability, and monitoring potential.</p><p><strong>Methods: </strong>A cohort of 80 BSG cases with 138 CSF samples and 71 tissues was retrospectively established. Public tissue methylation profiles (N = 1016) were used for the development of H3K27M and IDH mutation-specific assay. The mutation and methylation co-detection classifier (BSGdiag) was trained and tested in tissue cohorts and further validated in CSF samples. CSF Methylation Risk Score (MRS) was defined and used for prognostication and monitoring.</p><p><strong>Results: </strong>The methylation assay demonstrated robust three-class (H3K27M-mut, IDH-mut and double-wildtype) classification with microAUC values of 1.00, 0.973, and 0.813 across public datasets, tissue cohorts, and CSF samples, respectively. BSGdiag achieved a sensitivity of 95.6%, specificity of 83.3%, and AUC of 0.949 for the H3K27M subtype, and a microAUC of 0.990 for the three-class classification in CSFs. MRS-stratified CSF methylation risk group was an independent prognostic factor (HR = 2.61, 95% CI: 1.09-6.25, P = 0.032). Methylation information in CSF remained even with clinical, radiological and CSF genetic indications of no disease, suggesting its utility in monitoring minimal residual disease.</p><p><strong>Conclusions: </strong>The study de novo developed the first methylation assay for robust BSG molecular subtyping and introduced a novel methodology for co-detecting CSF ctDNA mutations and methylation in BSGs. The BSGdiag enhances the utility of ctDNA by leveraging both genetic and epigenetic information. Its comprehensiveness, minimal invasiveness, robustness, and reliability make it highly promising for future clinical applications and trial designs.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"283"},"PeriodicalIF":12.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12502398/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245560","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Long-term patient-derived ovarian cancer organoids closely recapitulate tumor of origin and clinical response.","authors":"Lucie Thorel, Enora Dolivet, Pierre-Marie Morice, Romane Florent, Jordane Divoux, Marion Perréard, Lucie Lecouflet, Guillaume Desmartin, Chloé Marde Alagama, Florence Giffard, Alexandra Leconte, Justine Lequesne, Bénédicte Clarisse, Mélanie Briand, Alimatou Traoré, Céline Villenet, Jean-Pascal Meneboo, Guillaume Babin, Léopold Gaichies, Sandrine Martin-Françoise, Jean-François Le Brun, Roman Rouzier, Emilie Brotin, Christophe Denoyelle, Nicolas Vigneron, Raphaël Leman, Dominique Vaur, Laurent Castera, Cécile Blanc-Fournier, Nicolas Elie, Benoit Plancoulaine, Florence Joly, Matthieu Meryet-Figuière, Martin Figeac, Louis-Bastien Weiswald, Laurent Poulain","doi":"10.1186/s13046-025-03537-x","DOIUrl":"10.1186/s13046-025-03537-x","url":null,"abstract":"<p><strong>Background: </strong>Ovarian cancers are the second cause of death from gynecological cancers worldwide, due to a late diagnosis combined with the development of resistance to chemotherapy. However, half of these cancers present alterations in Homologous Recombination (HR), making them sensitive to inhibitors of the PARP protein (PARPi), involved in DNA repair. Nevertheless, identifying patients who respond to chemotherapy and selecting those eligible for PARPi remains a challenge for clinicians. In this context, the use of Patient-Derived Tumor Organoids (PDTO) for predictive functional testing represents an interesting prospect for clinical decision making.</p><p><strong>Methods: </strong>Here we established a panel of 37 long-term PDTO models of various histological subtypes from 31 ovarian cancer patients. Histological and molecular profiles of PDTO were compared to tumor sample of origin using immunohistochemical analyses and global approaches (copy number variation and transcriptomic profiling). PDTO models were exposed to standard drugs for ovarian cancer patients, including PARPi, and response was assessed using viability assay. To further define the HR status of PDTO, we performed a functional assay evaluating the ability of PDTO to initiate HR (RECAP test) using automated histo-imaging quantitative analysis of RAD51 foci, as well as an NGS analysis based on the sequencing of an HR-related genes panel to obtain a Genome Instability Score (GIS).</p><p><strong>Results: </strong>We demonstrated that PDTO mimicked histological and expression of tumor markers of paired tumors. Moreover, non-negative matrix factorization approach revealed that PDTO recapitulated the transcriptomic profile of the cancer component from their sample of origin. Screening of chemotherapeutic drugs showed that PDTO exhibit heterogeneous responses, and that response of PDTO from high-grade serous ovarian carcinoma to carboplatin recapitulated patient response to first-line treatment. Additionally, the detection of HRD phenotype of PDTO using functional assay was associated with the results of the HRD test Genomic Instability Scar (GIScar).</p><p><strong>Conclusion: </strong>Although larger-scale investigations are needed to confirm the predictive potential of PDTO, these results provide further evidence of the potential interest of ovarian PDTO for functional precision medicine.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"282"},"PeriodicalIF":12.8,"publicationDate":"2025-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12502492/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245602","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Peripheral nerve injury-induced remodeling of the tumor-associated macrophages promotes immune evasion in breast cancer.","authors":"Yongxue Jiang, Wenfeng Zeng, Yaxin Feng, Xiaoting Deng, Jiayi Wang, Haiyu Liu, Boying Gao, Dexi Bi, Zifei Liu, Chaoqun Yang, Minxia Chen, Tang Li, Houying Chen, Yuxi Zhang, Luyuan Liang, Jiannan Xu, Wen Deng, Zeyu Yao, Wei Wu, Liyan Lao, Jianing Chen, Penghan Huang","doi":"10.1186/s13046-025-03545-x","DOIUrl":"10.1186/s13046-025-03545-x","url":null,"abstract":"<p><strong>Background: </strong>Peripheral nerve damage is intricately linked to the progression of various solid tumors. However, its effect on antitumor immunity and precise underlying mechanisms remain poorly understood. This study aimed to elucidate the effect of peripheral nerve damage and its subsequent immune-modulating effects influence on breast cancer progression.</p><p><strong>Methods: </strong>We analyzed nerve injury markers in the TCGA-BRCA database and clinical samples. In vivo experiments were conducted using orthotopic breast cancer models with chemical sympathetic denervation (6-OHDA) or nerve lysate/neurofilament light chain (NFL) treatment, where NFL was identified as a key effector molecule through mass spectrometry screening. The tumor microenvironment was evaluated by flow cytometry, multiplex immunohistochemistry, and single-cell RNA sequencing. In vitro co-culture systems were established to investigate the effects of NFL on macrophages and CD8<sup>+</sup> T cells, with transcriptomic profiling revealing that NFL-activated macrophage supernatants induced CD8<sup>+</sup> T cell senescence via NF-κB signal pathway activation.</p><p><strong>Results: </strong>Peripheral nerve injury was associated with poor prognosis and immune evasion in breast cancer patients. In mouse models, chemical sympathectomy (6-OHDA) and nerve lysates injection both accelerated tumor growth, suggesting that nerve damage promotes immune escape. Single-cell RNA sequencing (scRNA-seq) further revealed that nerve injury increased tumor-associated macrophages (TAMs) proportion by promoting TAMs proliferation and attracting macrophages. The key effector molecule of nerve lysates neurofilament light chain (NFL) was identified with the TAMs proliferation effect, and intratumoral NFL administration recapitulated the pro-tumor effects of nerve damage and perfomed the same immune-modulating effects as 6-OHDA and nerve lysates. Importantly, NFL-induced TAM enrichment and remodeling promoted CD8<sup>+</sup> T cell senescence, as evidenced by transcriptomic analysis showing NF-κB pathway activation and verified with NF-κB inhibitor (BAY 11-7082) in vitro, resulting in breast cancer immune escape.</p><p><strong>Conclusion: </strong>These findings underscore the critical role of peripheral nerve injury in reshaping the interplay between TAMs and antitumor immunity, via NFL-driven NF-κB activation and T cell dysfunction. Suggesting that neuroprotection could serve as a promising strategy to restore anticancer immunosurveillance.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"280"},"PeriodicalIF":12.8,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12502561/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145240137","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cristian Angeli, Demetra Philippidou, Eliane Klein, Christiane Margue, Sagarika Ghosh, Maria Lorena Cordero Maldonado, Natascia Tiso, Giovanni Risato, Fizza Irfan, Bruno Santos, Meritxell Cutrona, Joanna Patrycja Wroblewska, Stephanie Kreis
{"title":"High-throughput drug screening in advanced pre-clinical 3D melanoma models identifies potential first-line therapies for NRAS-mutated melanoma.","authors":"Cristian Angeli, Demetra Philippidou, Eliane Klein, Christiane Margue, Sagarika Ghosh, Maria Lorena Cordero Maldonado, Natascia Tiso, Giovanni Risato, Fizza Irfan, Bruno Santos, Meritxell Cutrona, Joanna Patrycja Wroblewska, Stephanie Kreis","doi":"10.1186/s13046-025-03539-9","DOIUrl":"10.1186/s13046-025-03539-9","url":null,"abstract":"<p><strong>Background: </strong>Despite significant advances in targeted (BRAFi + MEKi) and immune (anti-PD1/PD-L1, anti-CTLA4, and anti-LAG3) therapies, treatment options for NRAS<sup>mut</sup> melanoma remain limited. Currently, NRAS<sup>mut</sup> patients rely on immune checkpoint inhibitors, classical chemotherapy, and off-label MEK inhibitors, with over 50% experiencing rapid disease progression. One of the key challenges in developing effective targeted therapies is the lack of preclinical models that accurately recapitulate the tumor microenvironment (TME) and the intrinsic resistance of melanoma cells bearing NRAS mutations.</p><p><strong>Methods: </strong>To address this, we performed high-throughput screening (HTS) of over 1,300 compounds in 3D NRAS<sup>mut</sup> melanoma spheroids. A multi-step analysis was performed to identify hits, which were further tested by performing drug-response curve (DRC) analysis. Most promising compounds were further validated using mono- and co-culture 3D in vitro models that mimic three main metastatic sites in melanoma, such as skin/dermal, lung, and liver, utilizing spheroid and hydrogel systems. Ultimately, validation was conducted using zebrafish xenograft models to enable a more refined and accurate assessment of drug response.</p><p><strong>Results: </strong>High-throughput drug screening of NRAS<sup>mut</sup> melanoma spheroids identified 17 candidate compounds, which were subsequently validated through DRC analyses. Among the most promising drugs, Daunorubicin HCl (DH) and Pyrvinium Pamoate (PP) were selected for further investigation, demonstrating potent anti-melanoma activity in advanced 3D co-culture systems and zebrafish xenograft models. Notably, PP demonstrated higher cytotoxicity compared to Trametinib, the off-label MEK inhibitor, with an inhibitory effect on AKT and invasive behavior in the patient-derived metastatic melanoma cell lines. Additionally, combinatorial treatment with Trametinib resulted in additive effects on cell proliferation and viability. Importantly, both compounds showed similar efficacy in NRAS<sup>mut</sup> and BRAF<sup>wt</sup>/NRAS<sup>wt</sup> melanoma cell lines that were resistant to Trametinib (MEK inhibitor).</p><p><strong>Conclusions: </strong>Using advanced 3D melanoma models that incorporate key TME elements and zebrafish xenograft models, this study highlights the potential of Daunorubicin HCl and Pyrvinium Pamoate as novel first-line therapies for NRAS<sup>mut</sup> melanoma, with a noteworthy effect also on MEKi-resistant cells. These findings support drug repurposing strategies and underscore the importance of physiologically relevant preclinical models in identifying effective therapies.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"278"},"PeriodicalIF":12.8,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12487241/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145208037","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Yongjin Fang, Xiaofan Xu, Rihui Lu, Ye Huang, Xinshen Dai, Pucheng Huang, Xuefeng Fu, Pan Zhuge
{"title":"TEAD3 + high-risk melanoma cells crosstalk with GAS6 + macrophages via the GAS6-TYRO3 ligand-receptor axis to modulate propionate metabolism and drive melanoma progression.","authors":"Yongjin Fang, Xiaofan Xu, Rihui Lu, Ye Huang, Xinshen Dai, Pucheng Huang, Xuefeng Fu, Pan Zhuge","doi":"10.1186/s13046-025-03542-0","DOIUrl":"10.1186/s13046-025-03542-0","url":null,"abstract":"<p><strong>Background: </strong>Melanoma, a highly heterogeneous malignancy, remains refractory to conventional therapies due to poorly defined molecular and metabolic drivers. Short-chain fatty acid (SCFA) metabolism influences tumor progression, yet its role in melanoma subtypes and clinical outcomes is unclear. This study aims to delineate melanoma subgroups driven by SCFA metabolic dysregulation and identify mechanisms underlying their aggressiveness.</p><p><strong>Methods: </strong>Using non-negative matrix factorization (NMF), we clustered 468 TCGA melanoma samples into six subgroups based on SCFA-related gene sets (GO:0019745, GO:0019746, GO:0006085). Survival, differential expression, and pathway analyses were performed to characterize high-risk subgroups. Key drivers were validated via CRISPR/Cas9, siRNA knockdown, and immunohistochemistry. Single-cell RNA-seq (GSE215120) and spatial transcriptomics elucidated tumor-microenvironment crosstalk. Metabolic profiling, Seahorse assays, and myeloid-specific GAS6 knockout models were employed to dissect mechanisms.</p><p><strong>Results: </strong>NMF clustering revealing a high-risk subtype (Group 6) with dysregulated short-chain fatty acid (SCFA) metabolism and poor survival. Group 6 exhibited upregulation of GLTP and RAPGEFL1, enrichment in melanogenesis, Hippo signaling, and skin/lipid metabolism pathways. Through integrative analysis, TEAD3 emerged as a key risk driver, with high expression correlating with poor prognosis. Functional validation demonstrated that TEAD3 knockout suppressed melanoma proliferation, migration, and epithelial-mesenchymal transition (EMT) in vitro and in vivo. Single-cell RNA sequencing of acral melanoma revealed TEAD3-enriched tumor cells interacting with M2 macrophages via the GAS6-TYRO3 axis. Mechanistically, GAS6 + macrophages exhibited hypermetabolic phenotypes (elevated glycolysis/OXPHOS) that fueled GAS6 secretion. GAS6-TYRO3 signaling in TEAD3 + cells drove tumor aggressiveness by rewiring propionate metabolism, inducing methylmalonic acid accumulation via Mmut upregulation. Targeting this axis in myeloid-specific GAS6 knockout mice enhanced anti-PD-1 efficacy, boosting CD8 + T cell infiltration and survival.</p><p><strong>Conclusion: </strong>We define a TEAD3-driven melanoma subtype reliant on SCFA metabolic reprogramming and M2 macrophage crosstalk. The GAS6-TYRO3 axis and Mmut-mediated methylmalonic acid accumulation represent actionable targets. Combining myeloid-GAS6 ablation with immune checkpoint blockade overcomes therapy resistance, offering a precision strategy for high-risk melanoma.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"279"},"PeriodicalIF":12.8,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12486507/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145208125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gemma Di Pompo, Thimios A Mitsiadis, Pierfrancesco Pagella, Alessandro Pasquarelli, Giuliano Bettini, Silvia Sabattini, Alberto Righi, Sofia Avnet, Nicola Baldini
{"title":"Mesenchymal stroma drives axonogenesis and nerve-induced aggressiveness in osteosarcoma.","authors":"Gemma Di Pompo, Thimios A Mitsiadis, Pierfrancesco Pagella, Alessandro Pasquarelli, Giuliano Bettini, Silvia Sabattini, Alberto Righi, Sofia Avnet, Nicola Baldini","doi":"10.1186/s13046-025-03532-2","DOIUrl":"10.1186/s13046-025-03532-2","url":null,"abstract":"<p><strong>Background: </strong>Osteosarcoma (OS), the most common primary bone malignancy, is a leading cause of cancer-related mortality in children and adolescents. Besides genomic abnormalities, several features of tumour microenvironment (TME), including cancer-associated mesenchymal stromal cells (MSC), have been recognized to play a key role in OS progression. The pathogenetic function of de novo innervation in TME has been extensively studied in carcinomas but is still an unexplored area of investigation in sarcomas, including OS.</p><p><strong>Methods: </strong>We evaluated nerve infiltration in tissue samples from a small cohort of human OS (n = 5) and from canine OS (n = 11), a translational model for the human disease, by βIII-tubulin immunostaining. We then analysed nerve-stroma-tumour crosstalk using direct and indirect co-cultures of dorsal root ganglion (DRG) neurons with OS/tumour-associated mesenchymal stromal cells (MSC and cancer-associated fibroblasts, CAF), both under standard and microfluidic conditions. In particular, we investigated the effects of tumour and stromal cells on axonal tropism and outgrowth by measuring neurite recruitment, length, and branches and, vice versa, the impact of neuron-derived secretome on OS cell proliferation and migration. Finally, we assessed the secretion of pro-neurotrophic mediators, including brain-derived neurotrophic factor (BDNF), interleukin-6 (IL-6), and nerve growth factor (NGF), by MSC, CAF, and OS cells. The functional roles of IL-6 and BDNF were also verified by the blocking antibody Tocilizumab (TCZ) and the neutralizing Anti-BDNF antibody.</p><p><strong>Results: </strong>We provided evidence of OS innervation within and surrounding the tumour in association with mesenchymal stroma that also corresponded to the most proliferative area of the tumour (Ki-67+). In vitro, both MSC and, to a lesser extent, OS cells promoted axonal growth through cytokine (IL-6) and neuromodulator (BDNF) secretion. Extracellular acidosis - a hallmark of OS aggressiveness - amplified IL-6 release by stromal cells, and its pro-neurogenic effect was prevented by IL-6 blockade. In turn, tumour-associated innervation stimulated OS cell proliferation and migration, eventually driving tumour aggressiveness.</p><p><strong>Conclusions: </strong>We showed, for the first time, that bone-associated nerves, fostered by the OS microenvironment, promote tumour aggressiveness. Interfering with the nerve-tumour axis, particularly with the signalling associated with mesenchymal stroma, offers novel opportunities for OS treatment.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"276"},"PeriodicalIF":12.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12487423/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145201995","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Marco Bocchetti, Alessia Maria Cossu, Manuela Porru, Maria Grazia Ferraro, Carlo Irace, Rossella Tufano, Giovanni Vitale, Gabriella Misso, Nicola Amodio, Marianna Scrima, Ines Simeone, Michele Ceccarelli, Ugo Chianese, Lucia Altucci, Vincenzo Desiderio, Tarik Regad, Michele Caraglia, Silvia Zappavigna
{"title":"MiR-423-5p is a metabolic and growth tuner in hepatocellular carcinoma via MALAT-1 and mitochondrial interaction.","authors":"Marco Bocchetti, Alessia Maria Cossu, Manuela Porru, Maria Grazia Ferraro, Carlo Irace, Rossella Tufano, Giovanni Vitale, Gabriella Misso, Nicola Amodio, Marianna Scrima, Ines Simeone, Michele Ceccarelli, Ugo Chianese, Lucia Altucci, Vincenzo Desiderio, Tarik Regad, Michele Caraglia, Silvia Zappavigna","doi":"10.1186/s13046-025-03524-2","DOIUrl":"10.1186/s13046-025-03524-2","url":null,"abstract":"<p><strong>Background: </strong>MicroRNAs (miRNAs) and long non-coding RNAs (lncRNAs) are key regulators of gene expression and play a crucial role in cancer progression. Recent studies have highlighted miR-423-5p as a potential modulator in hepatocellular carcinoma (HCC), especially in patients responding to sorafenib treatment. A functional interaction with the oncogenic lncRNA MALAT-1 has been hypothesized, suggesting a regulatory mechanism that may influence tumor aggressiveness.</p><p><strong>Methods: </strong>To investigate this interaction, we analyzed in silico patient datasets to correlate miR-423-5p and MALAT-1 expression with overall survival (OS) and disease free survival (DFS). Stable overexpression of miR-423-5p and MALAT-1 was achieved in HCC cell lines (HepG2, Hep3B, and SNU387) using a lentiviral transduction system. Functional assays were performed to assess proliferation, migration, invasion, and clonogenic potential. The interaction between miR-423-5p and MALAT-1 was confirmed by RNA immunoprecipitation (RIP), followed by transcriptomic analysis using next-generation sequencing (NGS). Mitochondrial activity was evaluated using the Seahorse Mito Stress Test to measure oxygen consumption rate (OCR) and ATP production. In vivo experiments in orthotopic mouse models were performed to assess tumor growth.</p><p><strong>Results: </strong>Patient data analysis revealed that high miR-423-5p expression correlated with a less aggressive tumor phenotype and improved survival, while MALAT-1 was associated with poorer prognosis. In vitro, miR-423-5p overexpression reduced MALAT-1 levels and significantly impaired proliferation, migration, and invasion. NGS analysis identified transcriptomic changes linked to tumor progression and metabolic shift. The Seahorse Mito Stress Test demonstrated decreased cellular respiration and ATP production upon miR-423-5p overexpression. In vivo, both tumors derived from miR-423-5p-overexpressing cells and MALAT-1 downregulation by ASO GapmeR evidenced a significantly reduced growth compared to controls.</p><p><strong>Conclusion: </strong>These findings suggest, for the first time, that miR-423-5p acts as a tumor suppressor affecting mitochondrial metabolism through MALAT-1 downregulation in HCC. This regulatory axis represents a potential therapeutic target for precision medicine approaches in liver cancer.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"270"},"PeriodicalIF":12.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12487375/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145201967","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"O-GlcNAcylation of UBAP2L regulates stress granule formation and sunitinib resistance in clear cell renal cell carcinoma.","authors":"Jiajun Xing, Baochao Li, Songbo Wang, Zengjun Wang, Chenkui Miao","doi":"10.1186/s13046-025-03534-0","DOIUrl":"10.1186/s13046-025-03534-0","url":null,"abstract":"<p><strong>Background: </strong>Sunitinib resistance is one of the main reasons for the poor prognosis of clear renal cell carcinoma (ccRCC). Moreover, Stress granules (SGs) was found to enhance the stress adaptation capability of tumor cells, becoming an important mechanism for drug resistance in various cancers.</p><p><strong>Methods: </strong>We developed sunitinib-resistant patient-derived xenograft (PDX) and organoid (PDO) models to investigate sunitinib resistance in ccRCC. Proteomic analysis identified UBAP2L as a key mediator of this resistance. To explore its role in stress granule formation and sunitinib resistance, we conducted both in vitro and in vivo studies. We further elucidated the regulatory mechanisms of UBAP2L O-GlcNAcylation using immunoprecipitation, mass spectrometry, modification-based proteomics, RNA sequencing (RNA-seq), and RNA immunoprecipitation sequencing (RIP-seq).</p><p><strong>Results: </strong>In this study, enrichment of UBAP2L was elucidated to be significantly associated with sunitinib-resistant ccRCC patient-derived xenograft (PDX) model. Functional experiments showed that UBAP2L protected ccRCC from apoptosis and promoted ccRCC prolifecation and angiogenesis upon sunitinib treatment, thus enhancing drug resistance of ccRCC cells. Furthermore, mechanistic investigation demonstrated that O-GlcNAcylation of UBAP2L promoted its protein stability via inhibiting TRIM37-mediated ubiquitination and it regulated stress granule formation, thereby enhancing the mRNA stability of Melk and activating the PI3K signaling pathways.</p><p><strong>Conclusions: </strong>These results validated the significant roles of O-GlcNAcylation of UBAP2L in ccRCC sunitinib resistance, which provided an innovative theoretical basis for the clinical diagnosis and therapy of ccRCC.</p>","PeriodicalId":50199,"journal":{"name":"Journal of Experimental & Clinical Cancer Research","volume":"44 1","pages":"273"},"PeriodicalIF":12.8,"publicationDate":"2025-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12487280/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145201973","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}