B-cell precursor acute lymphoblastic leukemia (preB-ALL) is characterized by pathogenic variants currently used in precision oncology. However, the mutational landscape of Mexican children with preB-ALL has not yet been thoroughly explored and defined in terms of the clinical significance. We used a custom-designed next-generation sequencing exome panel, along with high-resolution chromosome microarrays and gene fusion-targeted assays, to characterize the mutational landscape of 73 Mexican children with preB-ALL, exploring the profile of clinically significant variants. We classified the variants following the AMP-ASCO-CAP 2017 and ACMG-CG 2019 guidelines recommendations. The mutational landscape includes a broad molecular spectrum of variants affecting cell cycle regulation, B-cell development, kinase signaling, and epigenetic regulation genes. Tier 1 diagnostic variants allowed the identification of pre-B ALL genetic subtypes, diminishing the preB-ALL NOS group from 63% to 37%. Furthermore, 37% of cases presented Tier 1 variants conferring intermediate to adverse prognoses (primarily involving CRLF2 gene fusions, as well as PAX5, IKZF1, and TP53 inactivating mutations). Notably, these patients exhibited high-risk clinical features and a lower event free survival rate than patients without these variants (60% versus 41.2%; p = 0.046; 95% CI). Between 19% and 42% of patients had Tier 2 variants targetable with JAK-STAT or RAS-MAPK signaling inhibitors. These patients showed a lower overall survival rate than patients without these variants (64% versus 90%; p = 0.048; 95% CI). Tier 1 potential germline variants in cancer predisposition genes (mainly BRCA1/2 and CHEK2) were observed in 10% of patients, some of whom had a family history of cancer. This highlights the importance of genetic counseling for patients and their families. Finally, 33% of patients had Tier 3 variants that were predicted to be deleterious and potentially upgraded to pathogenic with plausible clinical relevance. In conclusion, the mutational landscape analysis revealed variants useful for oncologic management and genetic counseling of Mexican children with preB-ALL.

Alcohol drinking is a major risk factor for oropharyngeal cancer, yet the nature of risk differences across drinking patterns remains unclear.

Chondrosarcomas are malignant cartilage-forming bone tumors with heterogeneous behavior, making prognostication and clinical management challenging. Histological grading is the primary tool for predicting clinical outcomes in conventional chondrosarcoma. However, its high interobserver variability limits reliable distinction between low- and high-risk patients, potentially leading to suboptimal clinical management. Given the increasing use of DNA methylation profiling as a valuable tool in surgical pathology for tumor classification, we investigated its prognostic value in central conventional chondrosarcoma. We generated methylation data from 69 primary central conventional chondrosarcomas profiled with Illumina's Human MethylationEPIC Array (850 k sites), and identified methylation sites individually linked to patient outcome. A LASSO Cox regression model was applied to these methylation sites to identify an optimal set of eight informative sites. With the regression coefficients of these methylation sites, we constructed a risk score that predicts central conventional Chondrosarcoma Risk Outcome from Methylation (CHROME). CHROME stratifies patients into High or Low risk groups for disease recurrence, onset of metastasis and disease-specific mortality. Survival analysis in an independent validation cohort (n = 68) demonstrated strong discriminatory performance of CHROME, with complete separation of outcomes and no adverse events observed in the Low risk group. Overview of the clinico-pathological information showed that a low grade (ACT/G1) case with an event was correctly assigned to the High risk group, while nine high grade cases without events were classified as Low risk. CHROME provides accurate risk stratification in central conventional chondrosarcoma and may represent a valuable tool for improving its prognostication and clinical management.

Upper aerodigestive tract squamous cell carcinoma (UATSCC) is often easy to recognize in its conventional form. However, a subset of tumors loses histologic evidence of squamous differentiation and, instead, become sarcomatoid (spindled). These cases may be difficult or impossible to appropriately classify on biopsy material, even with immunohistochemistry. DNA methylation has emerged as a tool to help in classifying undifferentiated malignancies. We sought to explore whether conventional and sarcomatoid UATSCC harbor the same epigenetic signature.

All individuals with colorectal cancer (CRC) should undergo genetic cancer risk assessment given its implications for personalized treatment, surveillance, risk-reduction strategies, and cascade testing. Universal screening using immunohistochemistry (IHC) for mismatch repair (MMR) proteins in tumor tissue, when combined with clinical criteria, is essential for identifying individuals at higher risk for carrying germline pathogenic variants (PVs) in resource limited countries.

Basal cell carcinoma (BCC), the most common skin cancer, is driven by UV-induced DNA damage and shaped by immune surveillance. Although GWAS has identified over 140 risk loci, their cell-type-specific effects remain obscured by tissue-level averaging.

Adrenocortical carcinoma (ACC) is a rare but aggressive malignancy with poor survival and limited treatment options. To comprehensively characterize its molecular landscape and identify clinically relevant subtypes, we performed an integrated genomic analysis - including whole-exome sequencing, RNA sequencing, and copy number variation profiling - on 61 Chinese patients with ACC. We identified recurrent mutations in TP53 (25%), CTNNB1 (15%), ZNRF3 (10%), and MEN1 (8%). Unsupervised clustering of transcriptomic data revealed four distinct molecular subtypes: cortisol-driven (CD, 14%), immune-suppressed (IS, 40%), cell cycle-altered (CCA, 22%), and immunomodulatory (IM, 24%). The CD subtype exhibited steroidogenic pathway activation; the IS subtype showed T cell receptor downregulation and the worst disease-free survival; the CCA subtype was marked by chromosomal instability and cell cycle gene overexpression; and the IM subtype displayed enriched immune signaling and favorable outcomes. Copy number analysis further uncovered focal amplifications (e.g. TERT, CDK4) and HLA-II deletions. This study establishes a novel molecular classification of ACC, providing a framework for subtype-specific therapeutic strategies, such as CDK4/6 inhibition for CCA and immunotherapy for IM tumors, while highlighting the clinical challenges of immune-cold IS tumors.

Activating mutations of Notch are drivers of the blood cell cancer, T-ALL, and some solid tumours. The negative regulatory region (NRR) of the extracellular domain (ECD) and the PEST region of the intracellular domain (ICD) are mutation hot spots which can act synergistically in T-ALL. The NRR, comprised of a heterodimerisation domain (HD) and three Lin12/Notch repeats (LNR A-C), masks the S2 cleavage site, normally only exposed following ligand binding and cleaved as the first step that ultimately leads to ICD release. Drosophila mutants have played a key role in analysing Notch structure/function, but there have been few mutational studies of the NRR. Here, we expressed, in S2 cells, over 20 cancer mutations located in the HD, LNR and LNR/HD interface, introduced into Drosophila Notch. Mutations in the HD domain core did not activate, likely due to absence in Drosophila of an S1 cleavage within the HD required for mammalian Notch activity. In contrast, mutations in the LNR/HD interface behaved similarly to T-ALL, activating constitutively with no further ligand induction and were synergistic with PEST deletion. Mutations of surface-exposed residues of LNR-C also activated constitutively but remained inducible both by ligand and by an intracellular endocytic regulator, Deltex, and were not synergistic with PEST deletions. These mutations caused elevated Notch levels and decreased turnover, suggesting a novel regulatory mechanism. Our results, therefore, uncover a variety of outcomes arising from perturbations of the NRR and will facilitate the establishment of Drosophila cancer models and the development of mutant-specific approaches to effective therapies.

Despite major advances inimmunotherapy, pancreatic ductal adenocarcinoma (PDAC) remains one of the most immunotherapy-resistant solid tumors. This review aims to summarize the key biological mechanisms underlying immune resistance in PDAC and to evaluate established and emerging biomarkers that may guide immunotherapy strategies.

Colorectal cancer (CRC) involves metabolic reprogramming alongside genetic alteration. ATP-citrate lyase (ACLY), the rate-limiting enzyme of de novo lipogenesis, is a validated target of microRNA-22 (miR-22) in tumor tissue. We asked whether both are dysregulated in the circulation of patients with CRC and whether they carry diagnostic value.

Genetic testing in ovarian carcinoma (OC) patients is very important for patients and their relatives. The Tumor-First workflow uses a tumor DNA test to stratify germline testing for hereditary cancer predisposition as well as treatment options with PARP inhibitors. This workflow is adopted and successfully implemented nationwide. Here, we evaluated recent tumor DNA testing rates in the Netherlands to identify untested OC patient groups and optimize tumor DNA testing rates. OC patients diagnosed in 2023 or 2024 were selected from the Netherlands Cancer Registry. We analyzed patient characteristics associated with the likelihood of tumor DNA testing using multivariable logistic regression. Tumor-First testing was performed for 1765 out of the 2221 (79%) OC patients. Patients diagnosed with advanced stage OC were more likely to receive tumor DNA testing (OR = 2.5, p < 0.001) than those with low-stage disease. Compared with patients who underwent surgery as primary treatment, those who received chemotherapy or no primary treatment were less likely to be tested (OR = 0.21 and OR = 0.07, respectively, p < 0.001). Patients who died within 100 days were less likely to receive testing (OR = 0.57, p = 0.003). Patients without resection after diagnosis on biopsy/cytology had the lowest Tumor-First testing rates (58% vs. 87-95%, χ2 p < 0.001). Tumor-First testing rates are high. However, OC patients that do not undergo a resection are less likely to undergo testing. With these data strategies to identify hereditary cancer predisposition in patients and their relatives can be further improved.

Circular RNAs (circRNAs) are a class of endogenous non-coding RNAs that regulate gene expression and participate in various biological processes. circRNAs play a critical role in bladder cancer (BCa) development. circ_0001495, generated by back-splicing of exons 6 and 7 of the CCNB1 pre-mRNA, remains uncharacterized in BCa pathogenesis. This study explored the functional role and molecular mechanism of circ_0001495 in BCa. Using qRT-PCR, we detected circ_0001495 expression in BCa tissues and analyzed its correlation with clinical parameters. Sanger sequencing validated its circular structure, while fluorescence in situ hybridization (FISH) revealed predominant cytoplasmic localization. Bioinformatics analyses predicted its potential as a microRNA sponge, confirmed by dual-luciferase reporter assays showing direct binding to miR-1184. CCK-8, EdU incorporation, and colony formation assays evaluated cell proliferation and clonogenic capacity. Results showed that circ_0001495 correlated with tumor grade (p = 0.016) and was significantly upregulated in BCa tissues, exhibiting preliminary diagnostic potential for early-stage BCa (AUC = 0.739, sensitivity = 0.906, specificity = 0.562). Functional experiments demonstrated that circ_0001495 silencing inhibited BCa cell proliferation and colony formation. Mechanistically, circ_0001495 acted as a sponge for miR-1184, thereby activating key components of the MAPK signaling pathway (p-ERK, p-JUN, and p-JNK). Collectively, these findings indicate that circ_0001495 may serve as a potential diagnostic/prognostic biomarker for BCa and promotes tumor progression by activating the miR-1184/MAPK signaling axis.

To clarify the role of glutathione peroxidase 4 (GPX4)-mediated ferroptosis in curcumin-treated papillary thyroid carcinoma (PTC) cells and explore the underlying mechanisms. GPX4 expression and clinical relevance were analysed in 54 PTC tissues by immunohistochemistry. B-CPAP cells were used to assess proliferation, migration, apoptosis and cell cycle after curcumin treatment. Ferroptosis was evaluated by detecting malondialdehyde (MDA), reactive oxygen species (ROS), Fe2⁺ and glutathione (GSH) levels, with ferrostatin 1 (Fer-1) and agonists (RSL3) used for validation. Molecular docking and Western blot were performed to analyse GPX4 interaction and expression. A nude mouse xenograft model was established to evaluate in vivo effects. Elevated GPX4 expression correlated with poor prognosis in PTC, including increased lymph node and distant metastasis and reduced progression-free survival. In vitro studies showed that curcumin downregulated GPX4 and induced ferroptotic features, including increased MDA and ROS, elevated Fe2⁺ and decreased GSH. These changes were accompanied by inhibited proliferation and migration, as well as G2/M arrest. Importantly, the ferroptosis inhibitor Fer-1 reversed curcumin-induced cell death and partially restored migration, confirming ferroptosis as the primary anticancer mechanism. Molecular docking analysis suggested a possible interaction between curcumin and the GPX4 protein at the Gln-55 and Pro-155 residues; however, this prediction was not experimentally validated in the present study. Importantly, GPX4 overexpression partially rescued curcumin-induced loss of cell viability and lipid peroxidation, supporting a functional role for GPX4 in curcumin-induced ferroptosis. In vivo experiments showed that curcumin significantly suppressed tumour growth in the xenograft model without observable organ toxicity and reduced GPX4 expression in tumour tissues. This study shows that curcumin inhibits PTC progression via GPX4-mediated ferroptosis; GPX4 overexpression partially reversed these effects, supporting its functional role. These findings identify curcumin as a promising therapeutic agent for PTC treatment and suggest GPX4 as a potential therapeutic target.

Resistance to immune checkpoint blockade (ICB) remains a major clinical challenge in lung cancer. Osteopontin (OPN) is frequently overexpressed in lung tumors and correlates with poor prognosis, yet its mechanistic role in shaping the immunosuppressive tumor microenvironment remains incompletely defined.

Colorectal cancer (CRC) development in Lynch syndrome (LS) has long been regarded to follow an adenoma-carcinoma sequence accelerated in comparison to microsatellite-stable (MSS) CRC development. Yet, several clinical observations challenged this hypothesis, most notably the persistently high CRC incidence under colonoscopy surveillance, a non-measurable benefit from shorter screening intervals, and substantial differences in cancer risk between carriers of the different mismatch repair (MMR) genes. Advances in the last decade have produced new hypotheses, highlighting the distinct biology of LS CRC and unravelling several co-existing pathways to cancer. Alongside the genomic heterogeneity, the immunogenic load created by the accumulation of frameshift mutations imposing selective pressure and leading to immune evasion, appears to be a critical step for cancer manifestation. We discuss potential clinical implications of these advances in understanding CRC pathogenesis for cancer prevention in LS and outline open questions that remain.

Pleural mesothelioma (PM) is an aggressive cancer with limited therapeutic options and poor prognosis, necessitating comprehensive model systems for mechanistic studies and drug discovery.

IntroductionFirst-generation EGFR-TKIs in NSCLC frequently lose efficacy as a result of the secondary EGFR-T790M mutation and a "persistent-STAT3" prosurvival pathway that sustains STAT3-survivin signaling in the face of EGFR inhibition. Preclinical evidence demonstrates that phytochemicals in the JI017 herbal formulation (2:1:1, Angelica gigas: processed Aconitum carmichaeli: Zingiber officinale) can reinstate erlotinib sensitivity in T790M-positive NSCLC, and relevant molecular mechanisms have been examined.MethodsAnti-proliferative activities were evaluated in A549 (EGFR-WT), HCC827 (EGFR Δ19), and H1975 (EGFR L858R/T790M) cells utilizing MTT assays, colony formation, and Annexin V/7-AAD flow cytometry. Mechanistic analyses included immunoblotting for p-EGFR (Tyr1068/1173), p-JAK2, p-STAT3 (Tyr705), PARP, Bcl-2, survivin, and AXL, complemented by RT-qPCR for BIRC5 and AXL transcripts. Drug interaction effects were determined using the Chou-Talalay combination index (CompuSyn). Anti-tumor efficacy was assessed in H1975 xenografts treated for 14 days with vehicle, JI017, erlotinib, or combination therapy; tumors underwent H&E staining and IHC for p-STAT3, survivin, and Ki-67.ResultsJI017 inhibited proliferation in all NSCLC cell lines tested, showing greatest effectiveness in H1975, where it triggered PARP cleavage and suppression of Bcl-2 and survivin expression. In H1975, co-treatment with JI017 and erlotinib led to synergistic growth inhibition, eradicated colony growth, and significantly elevated apoptotic cell populations compared to single treatments. While erlotinib alone reduced p-EGFR and p-JAK2, it left p-STAT3 largely unaltered, reflecting persistent-STAT3 activity. The combination regimen abrogated p-STAT3, further lowered p-EGFR and p-JAK2 levels, diminished BIRC5 mRNA, and decreased both AXL protein and transcript levels. In vivo, the drug combination achieved sustained tumor stasis relative to controls or monotherapy; combination group tumors displayed widespread necrosis and substantial decreases in p-STAT3, survivin, and Ki-67.DiscussionThese data support the suppression of STAT3-survivin as the primary mechanism by which JI017 sensitizes EGFR-T790M models to erlotinib. The consistent down-regulation of AXL indicates the inhibition of an AXL-mediated bypass that may maintain STAT3 signaling during EGFR blockade, although causality has yet to be confirmed. The marked in-vivo tumor inhibition without observable toxicity underscores the translational promise as a low-toxicity therapeutic adjunct.ConclusionsJI017 restores erlotinib sensitivity in EGFR-T790M NSCLC by inhibiting STAT3-survivin signaling and possibly reducing AXL-mediated resistance, resulting in durable antitumor effects both in vitro and in vivo. Additional preclinical studies and early-phase clinical assessment of JI017 in combination with erlotinib are justified.

A cell's fate is shaped by its inherited state, or lineage, and the ever-shifting context of its environment. CRISPR-based recording technologies are a promising solution for mapping the lineage of a developing system; however, challenges remain regarding single-cell recovery, engineering complexity, and scale. Here, we introduce BASELINE, which uses base editing to generate high-resolution lineage trees in conjunction with single-cell profiling. BASELINE uses the Cas12a adenine base editor to irreversibly edit nucleotides across target arrays built from 50 synthetic target sites, which are integrated multiple times into a cell's genome. We demonstrate that BASELINE accumulates lineage-specific marks over a wide range of biologically relevant intervals, recording more than 4300 bits of information in a model of pancreatic cancer, a 50-fold increase over existing technologies. Single-cell sequencing reveals high-fidelity capture of these recorders, averaging 29 cell divisions captured per lineage, within the estimated range of mammalian development. We expect BASELINE to apply to a wide range of lineage-tracing projects in development and disease, especially those in which cellular engineering makes small, more distributed systems challenging.

Ferroptosis is an iron-dependent form of programmed cell death driven by lipid peroxidation, distinct from apoptosis and necrosis. It has been confirmed as a key regulator in hepatocellular carcinoma (HCC). This study systematically elucidates three interconnected regulatory networks of ferroptosis-metabolic, epigenetic, and microenvironmental-and proposes a closed-loop regulatory model integrating these dimensions. By integrating metabolic biomarkers, epigenetic indicators, and microenvironmental features, we also summarize emerging strategies to enhance ferroptosis sensitivity. By summarizing the regulatory nodes as well as clinical translation progress, this work provides a comprehensive roadmap for overcoming therapeutic bottlenecks in HCC and realizing ferroptosis-based precision medicine.

The fat mass and obesity-associated protein (FTO), an RNA demethylase acting on both internal m6;A and cap-proximal m6;Am, functions in cancer as a context-dependent epitranscriptomic regulator whose net effect cannot be reduced to an oncogene-tumor-suppressor dichotomy. Its biological output is shaped by tumor lineage, subcellular localization, upstream signaling, and competing m6;A reader activities, predominantly YTHDF2-mediated decay and IGF2BP-mediated stabilization, although both reader families display additional non-canonical functions and are themselves modulated by post-translational modifications. Building on the now well-established context-dependence of FTO biology, which we do not claim as a novel observation, this review synthesizes current evidence on FTO's roles at the intersection of tumor immune contexture, immune checkpoint regulation, metabolic reprogramming, and therapeutic resistance. We examine how FTO may contribute to immune exclusion through metabolic competition, exosomal signaling, and stromal reprogramming; modulate PD-L1 expression through direct and indirect mechanisms; and influence response to chemotherapy, targeted therapy, radiotherapy, and CNS-directed treatment. Emerging FTO inhibitors, FTO-degraders, and combination strategies with immune checkpoint blockade, ferroptosis inducers, or glycolytic inhibitors are evaluated against their underlying preclinical evidence base. The contribution of this review lies less in proposing a new framework than in three forms of integration typically addressed in isolation: explicit calibration of mechanistic claims to evidence tier, systematic separation of tumor-intrinsic from immune-cell-intrinsic FTO functions across lymphoid and myeloid compartments, and translation of reader-network biology into biomarker-stratified trial design. Technical limitations of epitranscriptomic methods are addressed as constraints on inference. To our knowledge, no FTO-targeted strategy has yet entered Phase I oncology evaluation; current combination rationales therefore remain preclinically supported rather than clinically established.