Cancers

BackgroundHospital certification programs in Germany aim to improve the quality of cancer care. Previous research indicates that treatment in certified cancer centres leads to better overall and disease-free survival compared to non-certified hospitals for various cancers. Skin cancer, however, has not been investigated in this regard. ObjectivesTo test the hypothesis that treatment of melanoma in a certified cancer centre is related to survival benefits. MethodsData from clinical cancer registries in Germany were analysed. The analytical sample included n = 47,924 patients diagnosed with malignant melanoma between 2000 and 2022. Mixed-effects Cox regression models, adjusted for demographic and clinical confounders, were used to assess overall and disease-free survival. Hazard ratios (HR) with 95% confidence intervals (CI) are reported. ResultsThe proportion of patients treated in certified cancer centres increased over time to > 60 % from 2016 onward. Treatment in certified centres was associated with significant better overall survival (HR = 0.85, 95% CI = 0.82-0.88, p < 0.001). Results were statistically significant for stages I to III. For stage IV, the overall survival difference was not statistically significant, but subgroup analyses revealed a significant effect for cases diagnosed since 2011 (HR = 0.75, 95% CI = 0.61-0.91, p < 0.010). With regard to disease-free survival, multivariable analyses revealed better survival in certified centres (HR = 0.88, 95% CI = 0.85-0.92, p < 0.001). Similar results were observed across all subgroups stratified by stage of disease, except for stage IV. ConclusionsTreatment in certified cancer centres was associated with significant survival benefits for malignant melanoma patients, suggesting that the adherence to evidence-based quality standards improves patient outcomes. The fact that one in three patients has not been treated in certified cancer centres in recent years underscores the importance of expanding access to high-quality care. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=133 SRC="FIGDIR/small/26347792v1_ufig1.gif" ALT="Figure 1"> View larger version (27K): org.highwire.dtl.DTLVardef@7caac5org.highwire.dtl.DTLVardef@af7a1forg.highwire.dtl.DTLVardef@7aa859org.highwire.dtl.DTLVardef@c27dc4_HPS_FORMAT_FIGEXP M_FIG C_FIG Plain Language SummaryO_ST_ABSHow specialized cancer centres improve survival for melanoma patientsC_ST_ABSThis study examined whether treatment in certified cancer centres in Germany improves survival outcomes for patients with malignant melanoma, a type of skin cancer. Using data from nearly 50,000 patients diagnosed between 2000 and 2022, we compared survival rates between those treated in certified cancer centres and non-certified hospitals. Certified centres adhere to strict quality standards designed to enhance cancer care. The proportion of patients treated in certified centres increased significantly over time, from 22% in 2000 to over 60% after 2016. Patients treated in certified centres had better overall survival, with a 16% lower risk of death compared to those treated in non-certified hospitals. This survival benefit was consistent across early stages of melanoma (stages I-III) and cases with unknown stages. For patients with advanced stage IV melanoma, a positive effect of treatment in certified centres was only visible for diagnoses made since 2011. Disease-free survival, which measures the time patients remain cancer-free, was also better in certified centres, showing an 12% lower risk of recurrence or progression. The findings emphasize the critical role of certified cancer centres in improving outcomes for melanoma patients. Expanding access to these centres and ensuring adherence to high-quality treatment protocols could further enhance survival rates and reduce disease recurrence, particularly for patients diagnosed at earlier stages. Key pointsO_ST_ABSWhy was the study undertaken?C_ST_ABSExisting evidence suggests that treatment in certified centres leads to better survival compared to non-certified hospitals. Using data from clinical cancer registries in Germany, this study aims to evaluate whether these benefits extend to malignant melanoma. What does this study add?Treatment in certified cancer centres is associated with significant survival benefits for malignant melanoma patients, with effects particularly pronounced for patients diagnosed at earlier stages. What are the implications of this study for disease understanding and/or clinical care?The findings underscore the importance of expanding access to certified care and ensuring adherence to high-quality treatment protocols in order to ensure equitable care for all patients.

Invasive lobular carcinoma (ILC) is the most frequently diagnosed special histological subtype of invasive breast cancer and accounts for 10 - 15% of all cases. The pathognomonic hallmark of ILC is the genetic loss of E-cadherin (CDH1) causing the disruption of adherens junctions and resulting in discohesive, linear growth. To better understand the role of E-cadherin in ILC metastasis, we generated three ILC cell lines, MDA-MB-134-VI, SUM44PE, and BCK4, with inducible E-cadherin expression, resulting in successful restoration of functional adherens junctions. E-cadherin expression reduced growth in 2D culture, and that effect was even greater in 3D ultra-low attachment (ULA) conditions where increased cell death was consistent with the previously described role of E-cadherin in anoikis. E-cadherin expression did not rescue the lack of migration and invasion of ILC cell line models; however, it decreased haptotaxis and increased adherence to Collagen I in SUM44 cells. There was no significant effect of E-cadherin expression on primary orthotopic tumor growth, but spontaneous metastasis to the reproductive tract, brain, and GI tract was reduced. Inhibition of metastasis to the reproductive tract and brain was also seen after tail vein injection of MDA-MB-134 E-cadherin-expressing cells. In summary, overexpression of functional E-cadherin in ILC models has some, but limited, effects on 2D growth in vitro and primary tumor growth in vivo, but there are pronounced effects on 3D ULA growth and metastases in vivo, with stronger effects on metastatic sites enriched in patients with ILC, especially the reproductive and GI tracts.

BackgroundRecent evidence suggests that cancer can exhibit splicing alterations that give rise to tumour-specific isoforms. One example is NUMB, which produces four isoforms (p72, p71, p66, and p65) through alternative splicing of exons 3 and 9. Traditionally considered a tumour suppressor, it also has been considered an oncogene. We propose that this duality is due to isoform-specific expression. ResultsUsing public databases, we identified a tumour-associated switch in NUMB isoform expression: p72/p71 are upregulated in tumours, whereas p66/p65 are more expressed in non-tumour tissues. These isoforms correlate differently with cellular processes. NUMBL, a NUMB homolog, behaves similarly to p65. We identified two transcriptional clusters: one characterized by high expression of p72/p71, and another by p66/p65/NUMBL. Each group was associated differently with the Notch, WNT/{beta}-catenin, Hedgehog, and Hippo signalling pathways, suggesting isoform-specific regulatory roles. Analysis of breast cancer cell lines (CCLE) led to a NUMB score based on isoform expression, which classified cell lines into biologically distinct groups. The p72/p71-enriched group showed distinct signatures, pathway activity, and drug sensitivity. Applying this score to TCGA-BRCA samples revealed a significant link between high NUMB-score and poor survival, confirmed by Kaplan-Meier analysis. ConclusionsNUMB emerges as a potential oncogenic contributor and biomarker in splicing-based personalised medicine, highlighting isoform-specific expression as a clinically relevant determinant of tumour behaviour, pathway activity, and therapeutic response.

IntroductionComparison of rectal cancer characteristics in Pakistani Americans and native Pakistanis remains poorly investigated, as migrant studies have predominantly concentrated on East and Southeast Asian groups. This research aims to compare clinicopathological characteristics between the two groups. We hypothesize that significant differences will exist between these cohorts, mediated by gene-environment interactions. MethodsThis was a retrospective cohort study utilizing two multi-institutional databases to identify adult patients with rectal cancer: the National Cancer Database in the U.S (2018-2022) and the Rectal Cancer Surgery and Epidemiology Study in Pakistan (2020-2021). Non-Hispanic Whites (NHWs) were included as a reference population for comparative analysis. Clinicopathological characteristics were compared using Wilcoxon rank-sum and chi-square tests. ResultsA total of 523 Pakistani Americans and 608 native Pakistanis were included in the study. The median age at diagnosis was 57 years in Pakistani Americans (IQR 48-68), 42 years (IQR 33-54) in native Pakistanis and 63 years in NHWs (IQR 54-73) (p < 0.001). Native Pakistanis presented with early-stage disease less often than Pakistani Americans and NHWs (5.3%, 25.1%, and 20.5%, respectively; p < 0.001) and had markedly higher rates of signet cell carcinoma (20.1%, 0.6%, and 0.4%, respectively; p < 0.001) and poorly differentiated tumors (29.0%, 10.4%, and 11.4%, respectively; p < 0.001). ConclusionsThis study found that Native Pakistanis with rectal cancer presented at a younger age and with more aggressive tumor characteristics compared to both Pakistani Americans and NHWs. Notably, Pakistani Americans displayed a distinct clinical profile, intermediate between both groups.

BackgroundWilms tumour (WT) relapse occurs more frequently in patients with blastemal-type WTs. The presence of cancer stem cells (CSCs) is linked to tumour survival and relapse, and CSCs may be found in greater numbers in blastemal cell foci. CSC-associated phenotypes have been described in untreated WT, but their persistence, organisation and relevance after neoadjuvant chemotherapy is unknown. MethodsWe analysed 23 formalin-fixed paraffin-embedded blocks from 18 chemotherapy-treated patients where WTs were enriched for viable blastema, using human fetal kidney as developmental control. Immunohistochemistry and -fluorescence analysis determined progenitor (PAX2, SIX2, CITED1) and CSC-associated (NCAM, ALDH1, CD133) marker expression. We qualitatively and semi-quantitatively evaluated spatial expression patterns and co-localisation across tumour compartments. ResultsPAX2 and SIX2 were co-expressed in blastema in most cases (15/18), with PAX2 expression higher at the periphery of blastemal foci and SIX2 expression found uniformly in central aspects. CITED1 expression was also associated with SIX2 in blastema tissues (14/18). NCAM was blastema-enriched (15/18) with higher central intensity, frequently adjacent to PAX2-expressing peripheral zones. ALDH1 expression was present across blastema and epithelium while NCAM-, ALDH1-double-positive cells were rarely observed (4/18). CD133 expression was less commonly seen (2/18), localising near epithelial/nephrogenic structures. ConclusionsAfter neoadjuvant chemotherapy, WT blastema retained overlapping but non-identical progenitor/CSC-associated marker landscapes with reproducible peripheral-centre gradients. These spatial arrangements suggest a blastemal niche for CSCs that may sustain a therapy-resistant state. Our analysis provides the foundation for future functional validation and molecular profiling to define key lineage relationships and therapeutic vulnerabilities in post-chemotherapy WT. [250/250 words]

Ewing sarcoma is a pediatric cancer that develops in skeletal elements. The majority of Ewing sarcoma patients carry the aberrant EWSR1-FLI1 fusion gene. Despite trisomy 8 being an additional common aberration associated with a poor prognosis for patients, its induction mechanism remains unknown. When the EWSR1-FLI1 gene is formed, the cell loses one wildtype EWSR1 allele. To elucidate the induction mechanism of trisomy 8, we generated a cell line that allows for the conditional induction of EWSR1-FLI1 expression and EWSR1 knockdown (derived from a single EWSR1 allele. Specifically, the conditional cell line was generated by integrating the Tet-on EWSR1-FLI1 construct into the AAVS locus and adding a miniAID tag at the 5 end of the EWSR1 locus using auxin-degron system. A combination of the EWSR1-FLI1 expression and degradation of one allele-derived EWSR1 induced a high incidence of trisomy 8 within eight days, enhancing colony formation. Mechanistically, trisomy 8 is induced by the haploinsufficiency of EWSR1, and the remaining EWSR1 proteins are likely inhibited by interaction with EWSR1-FLI1. Our data showed that the knockout of EWSR1 alone was sufficient to increase the incidence of trisomy 8. Expression of wild-type EWSR1 in EWSR1 knockout cells rescued the high incidence of trisomy 8. In contrast, the EWSR1:R565A mutant, which lacks the ability to interact with Aurora B kinase, failed to rescue this phenotype. We propose that the combination of EWSR1-FLI1 expression and loss of EWSR1 contributes to the induction of trisomy 8 through the compromised EWSR1-Aurora B pathway. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=135 SRC="FIGDIR/small/726567v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@999891org.highwire.dtl.DTLVardef@1ef6748org.highwire.dtl.DTLVardef@65e475org.highwire.dtl.DTLVardef@179da40_HPS_FORMAT_FIGEXP M_FIG C_FIG

Importance: Non-medullary thyroid cancer (NMTC) and melanoma are associated with inherited long telomeres due to germline pathogenic/likely pathogenic variants (PV/LPV) in POT1, TINF2, and ACD resulting in long-telomere syndrome (LTS) and they commonly have somatic TERT promoter mutations. The genetic relationship between these variants and their clinical associations are defined incompletely and may inform clinical practice. Objective: To test the hypothesis that germline LTS-associated PV/LPV are exclusive from functional somatic TERT variants and assess clinical/genetic associations. Design: Retrospective observational cohort study with/without germline LTS variants, that have somatic sequencing and pathology data. Setting: Participants were enrolled through 18 cancer centers participating in the Oncology Research Information Exchange Network (ORIEN). Participants: 995 adults with NMTC and 993 with melanoma between 2013 and 2025. All adult patients at an ORIEN center were offered enrollment Exposures: All patients with NMTC or melanoma are included. There are no required exposures. Main Outcomes and Measures: The presence/absence of a germline or somatic long-telomere variant; secondary outcomes are associations with tumor stage, telomerase expression, and oncogenes. Results: Germline and somatic variants in POT1/TINF2/ACD, somatic TERT promoter variants, TERT fusions, oncogenes, and telomerase mRNA expression were evaluated in 995 NMTC and 993 melanoma patients. In NMTC, 13 (1.5%) had a germline LTS variant while 0/12 with tumor sequencing had somatic TERT promoter variants/fusions. In melanoma, 7 (0.7%) had a LTS variant; 0/2 with tumor sequencing had a TERT promoter variant/ fusion. Meta-analysis including NMTC and melanoma in the current study, a recent thyroid cancer study, and thyroid TCGA, germline LTS-associated PV/LPV and somatic TERT variants/fusions were mutually exclusive (p=0.036). High telomerase mRNA levels were associated with TERT promoter variants/fusions (p<4e-11) and larger NMTC/distant metastases (p=0.016), but not germline LTS variants. NMTCs with somatic TERT promoter variants/fusions had higher tumor mutation burden (p<0.02) versus tumors from patients with a germline LTS variant. TERT promoter mutant variant allele frequency was lower in smaller and non-metastatic vs larger/metastatic NMTC. Conclusion and Relevance: Germline LTS-associated variants appear to be exclusive from somatic TERT promoter variants/fusions but are not associated with aggressive NMTC, suggesting common roles in tumorigenesis but different biological impacts.

Background: Breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death in women worldwide, and the great majority of these deaths are caused by metastatic disease. Whether the immunohistochemical (IHC) phenotype of breast cancer is associated with the anatomical site of metastasis has been characterized mainly in high-income, registry-based populations, while data from ecologically stressed and medically under-served regions such as the Lower Aral Sea basin are lacking. Methods: We retrospectively reviewed 652 women diagnosed with breast cancer at the Khorezm Branch of the Republican Specialized Scientific-Practical Medical Center of Oncology and Radiology (Uzbekistan) between 2020 and 2024, of whom 213 had metastatic disease (306 metastatic foci). Histological type was assessed on hematoxylin-eosin and van Gieson-stained sections; quantitative morphometry was performed in Fiji/ImageJ; and HER2, estrogen receptor (ER), progesterone receptor (PR) and Ki-67 were assessed by IHC. The association between marker expression and metastatic site (liver, lung, lymph node) was tested in 187 foci with adequate tissue using the chi-square test, with significance at p < 0.05. Results: Invasive ductal carcinoma predominated. Metastatic site was significantly associated with the IHC phenotype. Liver metastases showed the highest frequency of HER2 3+ (45.7%), ER-negativity (65.2%), PR-negativity (69.6%) and high proliferation (Ki-67 [&ge;] 60%; 47.8%), whereas lymph-node metastases were more often hormone-receptor-positive (ER+ 58.7%; PR+ 52.4%) with lower HER2 3+ (22.2%); lung metastases were intermediate (all p < 0.05). The combination of HER2 3+ and Ki-67 [&ge;] 60% was associated with multi-organ spread. Morphometry corroborated these patterns: liver lesions had larger atypical cells (up to 132.8 m), a higher nuclear-to-cytoplasmic ratio (0.76 vs 0.51) and more extensive necrosis and microvascularity than lymph-node lesions. A pragmatic 5-criterion morphological score (histological type, Ki-67, HER2, ER/PR status, atypical-cell size) stratified metastatic risk into three tiers. Conclusions: In this regional cohort, the IHC phenotype of breast cancer tracked the anatomical site of metastasis, with an aggressive HER2-driven, hormone-receptor-negative profile concentrated in liver metastases and a hormone-receptor-positive profile in lymph-node metastases. These findings reproduce established organotropism patterns in a previously uncharacterized population and support phenotype-aware, site-specific surveillance together with a low-cost morphological risk score for resource-limited settings.

Chemotherapeutic treatment of breast cancer with Doxorubicin (DOX) can induce tumor and stromal cell senescence leading to therapy-resistance. Senescence-associated secretory phenotype (SASP) promotes secretion of pro-inflammatory and tumorigenic factors causing systemic inflammation. Combined, this can result in immune suppression, tumor growth and secondary spread of cancer. Targeting and removing senescent and cancerous cells using a combination of chemotherapeutic and senolytic drugs may reduce systemic inflammation, improve therapeutic efficacy, and prevent metastasis. Exposure of triple-negative breast cancer (MDA-MB-231), hormone-responsive (MCF-7) and HER2+ (MDA-MB-453) cells, and primary spine osteoblasts to DOX showed significant induction of p21-positive senescent cells. DOX and senolytics (RG-7112, o-Vanillin) treatment of co-culture spheroids showed a significant additive effect in reducing tumor sphere viability and growth, indicating reduced metastatic potential. This was correlated with reduced SASP in triple-negative and hormone responsive lines and decreased levels of senescent cells in all subtypes and primary stromal cells, while proliferation was decreased, and apoptosis increased across all breast cancer subtypes. Future chemotherapeutic treatment in breast cancer models may be optimized by adding senolytic drugs to more effectively clear senescent tumor and stromal cells, reducing risk for relapse and metastatic potential, while allowing for tissue regeneration in the bone metastatic environment. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/724653v1_ufig1.gif" ALT="Figure 1"> View larger version (24K): org.highwire.dtl.DTLVardef@c4cb8forg.highwire.dtl.DTLVardef@105219org.highwire.dtl.DTLVardef@17e0517org.highwire.dtl.DTLVardef@802bd2_HPS_FORMAT_FIGEXP M_FIG C_FIG Senolytics selectively eliminate senescent cancer and stromal cells and enhance Doxorubicin efficacy in a 3D bone-like tumor microenvironment model.

Anaplastic thyroid carcinoma (ATC) is one of the most lethal malignancies. Immune dysregulation is believed to play an important role in ATC. Here, we aimed to characterize the systemic inflammation and the function of circulating immune cells of patients with ATC. First, we retrospectively assessed biochemical parameters of patients with ATC and observed that high systemic inflammation correlated with worse survival. Next, we prospectively investigated the inflammatory proteome, single-cell peripheral blood mononuclear cell transcriptome and epigenetic changes. Circulating concentrations of proinflammatory cytokines were increased in ATC patients. This proinflammatory profile was apparent at the level of gene transcription and chromatin accessibility, especially in monocytes. These findings were substantiated by an increased capacity of peripheral blood mononuclear cells of ATC patients to produce IL-6, IL-8 and lactate. As IL-6 is known to promote tumor cell survival, we assessed its capacity to influence ATC cell proliferation. Blocking IL-6/gp130/Jak/STAT3 pathway inhibited proliferation of ATC cell lines in vitro. In conclusion, these findings show that ATC is characterized by inappropriate systemic inflammation and epigenetic and transcriptional reprogramming of circulating monocytes. Proinflammatory cytokines released by monocytes support survival and proliferation of ATC tumor cells, suggesting a therapeutic potential of targeting this pathway in ATC patients.

Objective To determine if it is possible to assess individual patient risk of the development of colorectal cancer (CRC) in people in high-risk groups due to their family history. Design/Method Retrospective observational study of prospectively collected data from consecutive patients referred for a colonoscopy. 2,478 consecutive patients were referred to a single colorectal surgical practice in Sydney, Australia between 1977 and 2018 for a colonoscopy because of a family history of CRC. Of these, 1,963 have been followed for more than 10 years and are the subject of this paper. Histopathological findings categorised as normal (N), non-advanced adenoma (NAA) or advanced neoplasia (AN) with AN proven to be the precursor to CRC. Intervention Colonoscopic screening on the basis of contemporary practice to 2006 and subsequently according to Australian National Health and Medical Research Council guidelines. Results Participants with normal or low-risk findings in the first decade remain at lower risk of CRC for 30 years from the commencement of screening. Conclusion It is possible to stratify individual patients in a high relative risk cohort into those with high or low personal risk of CRC based on colonoscopic findings in the first 10 years of surveillance. Those with no AN in the first ten years have a lower 30-year risk of developing AN than the general community. This offers the possibility of structuring surveillance programs around individual risk rather than group risk, lessening the need for multiple surveillance colonoscopies in the majority of such patients and improving the cost effectiveness of CRC screening at the population level.

INTRODUCIONTumor cell infiltration in regional lymph nodes is a strong prognostic marker, guiding treatment decisions in breast cancer. While the immune cell composition in primary tumors has been more widely explored in later years, the immune cell composition of the sentinel node (SN) and axillary lymph nodes (ALN) remains understudied. A better understanding of how primary tumor and metastatic tumor cells alter the nodal immune microenvironment can shed light on metastasis and cancer progression to unveil new treatment strategies. MATERIALS AND METHODSFrom a prospective clinical cohort of 458 treatment-naive patients with primary operable breast cancer, we performed comprehensive immunophenotypic analysis using mass cytometry analysis of non-metastatic (SN-) and metastatic (SN+) and ALN (ALN+) lymph nodes. RESULTSAs expected, patients with ALN+ cases had a shorter time to distant metastases than SN+ and SN- cases. We identified an exhausted T-cell phenotype and an increase in Germinal Center B (GC B) cells and plasma cells in ALN+ samples compared to SN- samples, both in the whole cohort as well as when investigating estrogen-receptor positive (ER+) patients only. There were no differences in immune cell composition across breast cancer (BC) subtypes within SN-samples. SN+ samples from triple negative BC (TNBC) showed a trend towards increased abundance of GC B and plasma cells, similar to more advanced ALN+, suggesting that smaller TN metastases may trigger an immune activation at an early stage of dissemination. Further analysis of SN- samples from ER+ patients revealed a subset of patients where the immune response had a more exhausted T-cell phenotype. This group was enriched for lymph nodes that were deemed negative by ordinary pathology examination (microscopy) but had detectable tumor cells by CyTOF analysis. CONCLUSIONThe immune profiles of SN and ALN samples from breast cancer patients are highly diverse, showing limited associations to BC subtype, clinical parameters or patient outcome. Metastatic tumor cells play a significant role in driving T-cell exhaustion and immunosuppression. Notably, in approximately 50% of the ER+ samples, T-cell exhaustion was detectable. This coincides with the presence of tumor cells identified by CyTOF, which were likely missed by conventional pathological examination. These findings suggest that small tumor deposits alter the immune composition, and the immune profile reveals the presence of tumor cells.

Background: Immune checkpoint inhibitors have transformed cancer treatment, yet a large number of patients fail to respond. Identifying molecular characteristics that predict response before treatment initiation remains an unmet need. Towards that end, this study presents a large-scale integrative analysis of existing single-cell and bulk tissue datasets, aimed at identifying predictive features while providing insights into their cellular origin and potential function within the tumor microenvironment. Methods: A stepwise analysis was performed using single-cell RNA-sequencing data from 60 melanoma patients at baseline, separated into discovery (n=41) and validation (n=19) sets. An integrated bulk transcriptomics dataset (n=128) from melanoma patients and a bladder cancer dataset (n=298) were used for further validation. Results: Integrative analysis of melanoma single-cell datasets revealed that responders exhibit distinct molecular profiles across multiple cell types compared to non-responders. Notably, these included downregulation of the TNFR superfamily and other immunosuppressive genes (TNFRSF18, TNFRSF9, TNFRSF4, LGALS1, BATF, IL12RB2, LINGO1, DUSP4, SDC4, VCAM1) in T-cells. By investigating the findings from the immune cell populations in the bulk tumor context, 13 transcripts were found to be consistently associated with response across all cohorts. These were differentially expressed in T-cells (SELL, EPB41, CD96, UHFR2, LINGO1, LGALS1), B-cells (ALDH5A1), NK cells (PLEC, PDGFRB) and Monocytes (TLR10, ST6GAL1, IKZF1, MPRIP). A predictive model based on these features effectively discriminated responders from non-responders in melanoma (AUC=0.73). The model maintained significant predictive power in an independent bladder cancer dataset (IMvigor210; AUC=0.64). Of high clinical relevance, it demonstrated enhanced performance in identifying responders among patients with low tumor mutational burden (AUC=0.75). Conclusion: Our study reveals pre-treatment molecular features related to immune-cancer crosstalk that are associated with response to immunotherapy. A 13-gene model demonstrates potential added clinical value in stratifying responders, particularly in patients with low tumor mutational burden, meriting further validation.

Abstract Background Tumour hypoxia is a major determinant of treatment resistance and poor prognosis in cervical cancer but remains difficult to assess in clinical practice. Gene expression signatures offer a potential means to characterise hypoxia-related biology. This study aimed to develop and validate a hypoxia-associated gene expression signature for cervical cancer. Methods RNA sequencing was performed on five cervical cancer cell lines exposed to normoxia (21% O?) and hypoxia (1% O?). Differentially expressed genes were mapped to The Cancer Genome Atlas cervical cancer cohort (TCGA-CESC) to train a 55-gene hypoxia classifier using k-means clustering and Prediction Analysis for Microarrays. The model was validated in an institutional Manchester cohort (n=153) and two public datasets from Seoul (n=300) and Oslo (n=283). Results The Manchester 55-gene signature was enriched for canonical hypoxia pathways. In the Manchester cohort, hypoxia classification correlated with advanced FIGO stage, nodal involvement, tumour size ? 4 cm, and hydronephrosis (adjusted p<0.05). Hypoxic tumours showed reduced overall survival (OS) and progression-free survival (PFS) in all cohorts. In multivariable models, the signature remained independently prognostic for OS in both TCGA (HR 1.70, 95% CI 1.10-2.60, p=0.012) and Manchester (HR 1.95, 95% CI 1.08-3.51, p=0.026). A direct comparison with a published 6-gene hypoxia signature in the Oslo cohort demonstrated 71% concordance in classification. Conclusions Our 55-gene signature should be tested prospectively in trials to assess its ability to stratify patients for hypoxia-targeted therapies.

PurposeBladder cancer (BLCA) is a heterogeneous tumor type. Only one third of muscle-invasive (MIBC) patients respond to immune checkpoint inhibitors (ICIs). Reliable resistance markers are needed to guide clinical decisions. We investigated the nerve growth factor receptor (NGFR) in BLCA and analyzed its correlation with disease progression and response to immunotherapy. Experimental DesignWe analyzed NGFR expression in BLCA cell lines, organoids, mouse models and patient samples. The cohorts used were The Cancer Genome Atlas (TCGA), enriched in muscle-invasive bladder cancer (MIBC) (n=407); IMvigor210, representing MIBC patients treated with ICIs (n=348); and UROMOL2, as a non-muscle-invasive bladder cancer (NMIBC)-specific cohort (n=535). IMvigor010 was also included (n=728). Patients were stratified by NGFR expression quartiles. We analyzed survival and tumor subtypes and performed stromal deconvolution and functional profiling. We assessed stemness- and invasion-related features in SCaBER cells. ResultsNGFR marks a basal tumor cell subcluster and is independently associated with poor prognosis in TCGA and IMvigor210. NGFR-high tumors show stromal content enriched in cancer-associated fibroblasts, lower neoantigen burden, higher CD8+ T effector signature together with an immune-excluded phenotype, and a CAF-specific TGF{beta} signature. In the immunotherapy-treated cohort, high NGFR expression was also associated with poorer outcome. Functionally, NGFR appears to promote a stem-like/pro-invasive program in BLCA cells. ConclusionsNGFR identifies a basal-like BLCA subpopulation linked to poor survival, while its association with immunotherapy response requires further validation. In addition, our in vitro analyses support a role of NGFR in stem-like and invasive traits, highlighting its relevance as a biomarker in BLCA.

The TGIF1 transcription factor gene is present on chromosome 18, which is subject to whole chromosome copy number reduction in colon cancer. Despite this, TGIF1 expression is significantly higher in cancer than in normal. In mice complete deletion of Tgif1 reduced tumor burden in an Apc mutant model of intestinal cancer. Here we show that reducing TGIF1 expression in a human colon cancer cell line slows proliferation and reduces growth of orthotopic xenografts. To ask if additional genes with copy number loss are more highly expressed in tumors we identified chromosomal regions subject to copy number reductions from ten TCGA cancer datasets. Within these regions a small proportion of genes, generally less than 10%, are expressed at higher levels in the tumor than in corresponding normal samples. Enrichment analysis using a set of 435 genes that have copy number reduction and increased expression identified mitosis as the most enriched gene set and FOXM1 and E2F family transcription factors as potential regulators. For mitotic genes, the average expression increase in tumor compared to normal is independent of copy number. In contrast, while DepMap common essential genes are generally more highly expressed in cancer than normal tissue, the relative increase in expression tracks well with copy number. Similarly, expression differences for gene sets such as S-phase, rRNA processing and DNA repair show increased expression in cancer versus normal, but changes also track with copy number. Thus, genes with increased expression despite copy number reduction may represent the output of key pro-tumorigenic transcriptional programs and could be potential therapeutic targets.

Background Cancer treatment response is highly variable, even among patients with the same tumor type and treatment. Exceptional responders (ERs), who are individuals who experience unusually favorable outcomes, provide critical insights into the biological factors driving treatment success. While prior studies have highlighted the role of somatic changes, the contribution of germline rare variants remains underexplored. This study aimed to uncover the genetic underpinnings of exceptional responses by identifying rare, non-silent and predicted deleterious germline mutations enriched among ERs compared to typical cancer patients. Methods The Network of Enigmatic Exceptional Responders (NEER) project collected clinical and germline whole-genome sequencing (WGS) data from 53 ERs. After quality control procedures and ancestry background checks, 51 ERs were left for final analysis. While non-silent mutations were identified based on allele frequencies and mutation types, multiple pathogenicity predictors were applied for predicted deleterious variants. These were compared to a harmonized and comparable subset from the Pan-Cancer Analysis of Whole Genomes (PCAWG) cohort (n=414) using Fisher's exact tests. Kaplan-Meier survival analysis applied to evaluate prognostic associations in PCAWG patients. Additionally, Fisher's exact tests were conducted stratified by cancer type and treatment regimen to identify potential associations between rare germline variants and therapeutic responses. Results Variants in immune-related genes such as CCL26 and GPRC5D were prevalent, suggesting enhanced immune regulation among ERs. Fourteen genes with non-silent and eight with predicted deleterious mutations showed significantly different frequencies between NEER and PCAWG cohorts (FDR < 0.05). IRX3 emerged as a protective gene enriched in ERs, whereas OR6B2 was associated with poor survival in PCAWG lung cancer patients. Moreover, rare non-silent germline variants in drug target genes were enriched among ERs treated with cisplatin and doxorubicin, implicating altered DNA repair and drug-binding mechanisms in their remarkable outcomes. Conclusions This study reveals a distinctive germline mutation landscape in exceptional cancer responders, marked by immune-related and drug-target-associated variants that may enhance therapy response and prolong survival. The findings highlight potential novel prognostic biomarkers, such as IRX3 and OR6B2, providing a foundation for developing personalized cancer treatments informed by rare genetic variation.

High-Grade Serous Ovarian Cancer (HGSOC) is the most lethal gynecological malignancy due to aggressive growth, widespread metastases, and high intra-tumoral heterogeneity. Poor prognosis is largely due to late diagnosis, hence there is an urgent need to identify novel biomarkers for screening, diagnosis, and monitoring. Here, we propose the voltage-dependent calcium channel hCaV1.2 encoded by CACNA1C as a potential biomarker and therapeutic target in HGSOC. Using IHC analysis for ten ovarian cancer patients, cytotoxicity assay, TCGA gene expression and survival analyses, homology modeling, molecular docking, Calcium channel membrane assembly and molecular dynamics simulations, we tested CACNA1Cs role in HGSOC progression and the effect of blocking on cancer cell survival. We show that nifedipine (NIFE), a calcium channel blocker (CCB), had a tumor suppressive effect based on binding models predicted by three-dimensional computer assisted molecular modeling and in vitro validation using human HGSOC cell line. Using The Cancer Genome Atlas ovarian public cohort, we found CACNA1C mRNA expression strongly correlated with poor patient survival for late-stage and metastasis than primary. We also show strong correlation of CACNA1C protein expression using immunohistochemistry correlating with COH ovarian carcinomas patients disease progression. This research demonstrates that targeting HGSOC via CCBs may be therapeutically beneficial. By establishing further in vitro, in vivo, and clinical trials using FDA approved NIFE may be repurposed to target CACNA1C for HGSOC. Novelty and ImpactHigh-grade serous ovarian cancer (HGSOC) remains lethal due to late diagnosis and drug resistance. This study identifies CACNA1C (Cav1.2) as a novel prognostic biomarker and therapeutic target in HGSOC, showing that elevated expression correlates with metastatic/recurrent disease and poor survival. Using molecular dynamics and in vitro models, we demonstrate that the FDA-approved calcium channel blocker nifedipine binds stably to Cav1.2 and suppresses tumor cell growth more effectively than cisplatin. These findings support repurposing nifedipine for biomarker-driven HGSOC therapy. Translational RelevanceLate diagnosis and progressive relapses significantly contribute to the poor prognosis of ovarian cancer. Identification of a tumor biomarker that can be used for screening, diagnosis, and monitoring is critical for improving clinical outcome. Our findings demonstrate that CACNA1C is a viable diagnostic marker for HGSOC and that its blockade with CCBs reduces tumor progression, highlighting their therapeutic potential.

Abstract Background Immune-related adverse events (irAEs) involving the breast remain rarely reported. Purpose To characterize clinical and imaging features of camrelizumab-associated breast lesions (CABLs). Materials and Methods This retrospective dual cohort study (October 2019 to February 2026) included 196 female patients. Cohort A comprised 180 non-breast cancer patients; Cohort B comprised 16 breast cancer patients receiving neoadjuvant camrelizumab. Baseline characteristics, treatment response, and CT/MRI features were compared between CABL-positive and CABL-negative groups using Mann-Whitney U and chi-square tests. Results CABLs developed in 34.4% (62/180) of Cohort A and 93.8% (15/16) of Cohort B. CABL-positive patients were younger (median 50.5 vs 54.5 years; P = 0.006) and more often premenopausal (46.8% vs 26.3%; P = 0.009). The objective response rate was relatively high among patients with positive lesions; in Group A, the disease progression rate was lower in the CABL-positive group than in the CABL-negative group (3.2% vs 17.8%), whilst in Group B, the pathological complete response rate was as high as 53.3% (8/15). On CT/MRI, CABLs were predominantly multiple (62.5%), with well-defined margins and unrestricted diffusion. The predominant time-intensity curve (TIC) pattern was washout (46.7%). Median time to onset was 2-3 cycles (the second MRI scan); most lesions disappeared (40.3%) and shrank (46.8%) during follow-up. ADC values of lesions were significantly higher than those of primary tumors (1.847+/-0.284 vs 0.976+/-0.055 x10[-3] mm[2]/s; P < 0.001). Histopathology of four lesions revealed lymphocytic infiltration and fibrosis without malignancy. Conclusion CABLs are benign reactive changes driven by multiple factors. Their recognition prevents misinterpretation as disease progression, thereby avoiding unnecessary treatment discontinuation or biopsy.

BackgroundThe tumor immune microenvironment likely plays a central role in progression of thyroid cancer. As for most other solid tumors, it is unknown if immune dysregulation contributes to earlier, subclinical stages of thyroid tumor development, or whether thyroid tumor heterogeneity might involve differential expression of pro-inflammatory mediators. MethodsThe time course of tumor-associated inflammation was studied in Tg-CreERT2;Braf CA/+ mice representing a model of BRAFV600E-driven papillary thyroid carcinoma (PTC). Tumor growth was estimated by histological examination and magnetic resonance imaging. Cytokine expression was monitored by quantitative RT-PCR, RNAScope and Western blot analyses. ResultsBased on spontaneous BrafCA activation due to leaky Cre activity in a minority of targeted cells tumors developed within a preserved thyroid tissue architecture to multifocal papillary thyroid carcinoma (PTC) over a period of 12 months. Tumorigenesis was accompanied by a gradually increased mRNA and protein expression of interleukin-1beta (IL-1{beta}), interleukin-6 and tumor necrosis factor-alpha (TNF-) starting already before Braf mutant cells commenced neoplastic growth. RNAScope revealed that both follicular cells and stromal cells expressed Il1b whereas Il6 and Tnfa transcripts were mostly confined to neoplastic epithelia. Early cytokine expression was associated with oncogene-induced senescence, whereas during tumor development (3-6 months) and in advanced tumor stages (at 12 months) the cytokine expression pattern differed among glands and tumor foci of the same gland accompanied by a highly variable locoregional lymphocytic infiltration. Oral treatment of mutant mice for 1 month with PLX4720, a vemurafenib prodrug, partially reduced cytokine expression along with inhibited tumor growth and redifferentiation of thyroid function. The magnitude of reduced cytokine expression differed much between glands and among mice of both sexes. ConclusionsThese findings indicate that oncogenic BRAFV600E targeted to the thyroid both stimulates endogenous production of IL-1{beta}, IL-6 and TNF- and recruits inflammatory cells to foci of early tumor development. PTCs of different clonal origin are distinguished by differential expression of pro-inflammatory cytokines. The anti-inflammatory effect of mutant Braf kinase inhibition varies presumably related to heterogeneous tumor development, which evolves from stochastic BrafCA activation suggesting there are clonally different probabilities of acquiring drug resistance among Braf mutant thyroid follicular cells.