Cancers

BackgroundColorectal carcinoma (CRC) remains a significant cause of cancer morbidity and mortality worldwide. Right- and left-sided tumours differ in clinical, morphological, and molecular features. Microsatellite instability-high (MSI-H) tumours, often right-sided, are associated with distinct histopathological characteristics and prognostic implications. In Sri Lanka, molecular MSI testing is currently unavailable, highlighting the need for alternative predictive approaches. ObjectivesGeneral Objective: To describe the clinical and histopathological characteristics of right- and left-sided colorectal cancers in a Sri Lankan cohort and evaluate their usefulness in predicting MSI-H tumours. Specific ObjectivesTo compare clinicopathological features between right- and left-sided colorectal cancers. To predict MSI-H tumours based on clinicopathological features, including assessment of the MsPath score and histological parameters. To determine interobserver agreement for MsPath score application in selecting cases for MSI assessment. MethodsA retrospective analytical study was conducted on 156 colorectal carcinoma resections diagnosed between 2019 and 2021 at the National Hospital of Sri Lanka. Histopathological evaluation included tumour differentiation, mucinous and medullary features, tumour-infiltrating lymphocytes (TILs), and Crohn-like reaction. MsPath scores were calculated based on age, tumour site, and histological parameters. Immunohistochemistry (IHC) for PMS2 and MSH6 was performed on 46 selected cases to assess mismatch repair (MMR) status. ResultsOf 156 cases, 41 (26%) were right-sided and 115 (74%) left-sided. The majority were moderately differentiated adenocarcinomas (89%). Histological features suggestive of MSI-H including TILs (29%) and Crohn-like lymphoid reaction (23%) were more frequent in right-sided tumours. MsPath scores ranged from 0.0 to 5.9, with 50% of cases scoring below 1. Among the 46 cases evaluated by IHC, MMR deficiency was observed predominantly in higher MsPath score categories, and a significant association was found between MsPath score category and MMR status ({chi}{superscript 2} = 13.76, df = 2, p = 0.001). Interobserver agreement for MsPath scoring was substantial (Kappa = xx, indicating reproducibility). ConclusionRight-sided colorectal carcinomas in this Sri Lankan cohort more frequently exhibited histological features suggestive of MSI-H, including mucinous differentiation, TILs, and Crohn-like lymphoid reaction. MsPath scoring correlated with MMR deficiency in the IHC-tested subset, but its predictive value is limited without immunohistochemical confirmation. IHC using a two-antibody panel (PMS2 and MSH6) proved to be a feasible, cost-effective, and reliable method for MSI screening in resource-limited settings. This study is the first comprehensive evaluation of right-versus left-sided colorectal carcinomas and MsPath utility in Sri Lanka, underscoring the need for expanded IHC capacity, larger cohorts, and integration of molecular testing for MSI validation.

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.

BackgroundAcute lymphoblastic leukemia is the most common childhood malignancy; however, survival remains highly heterogeneous across regions, particularly in low- and middle-income settings. In geographically vulnerable areas such as the Brazilian Amazon, structural barriers related to health system organization, centralization of specialized services, and continuity of care may substantially influence treatment trajectories and mortality. This study aimed to examine health system and epidemiological determinants of mortality among patients with acute lymphoblastic leukemia in Amazonas, Brazil. MethodsThis retrospective cohort study included 393 patients diagnosed with acute lymphoblastic leukemia between 2016 and 2021 at the state referral center for hematologic diseases in Amazonas, Brazil. Sociodemographic, clinical, laboratory, and health system-related variables were analyzed. Multivariate logistic regression models were used to identify factors associated with mortality. Spatial analyses described the geographic distribution of cases, and overall survival was evaluated using Kaplan-Meier curves. ResultsThe cohort was predominantly pediatric, with a substantial proportion of adolescents and young adults. Most patients presented with high-risk clinical and laboratory features at diagnosis. Mortality occurred in 48.5% of patients and was strongly associated with age at diagnosis, with higher odds of death among adolescents, young adults, and individuals aged 51-60 years. Geographic concentration of specialized services and treatment-related trajectories were closely linked to survival patterns. ConclusionMortality from acute lymphoblastic leukemia in the Brazilian Amazon remains high and is primarily influenced by age-related vulnerability and health system factors rather than baseline sociodemographic characteristics. These findings underscore persistent regional inequalities in access to specialized care and highlight the need for health system strengthening to improve cancer survival in geographically vulnerable settings.

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]

BackgroundWnt/{beta}-catenin signalling is dysregulated in acute myeloid leukaemia (AML), where it lacks effective targeting strategies. Previously, we discovered that {beta}-catenin interacts with several RNA-binding proteins (RBP), indicating post-transcriptional influence which is yet to be therapeutically interrogated in AML. MethodsCo-immunoprecipitation confirmed protein interactions, and TCF/LEF reporters were used to assess Wnt signalling output in leukaemia cells. Regulatory crosstalk was assessed using immunoblotting and RT-qPCR approaches following lentiviral transduction of myeloid cell lines. Targeting of {beta}-catenin and LIN28B was tested through combinations of genetic and pharmacological inhibition in AML cells. ResultsThe most frequent RBP-binding motif amongst {beta}-catenin-bound mRNAs was the GGAG motif targeted by oncofetal miRNA-regulating RBP; LIN28B. {beta}-Catenin:LIN28B interactions were detected in lymphoid and myeloid cell lines, plus primary human CD34 fetal-liver HSCs. LIN28B positively regulated Wnt signalling output through LEF1 regulation involving a post-transcriptional let7 miRNA mechanism. Further miRNA sequencing of {beta}-catenin- and LIN28B-depleted myeloid cells revealed potential cooperative and antagonistic function in miRNA regulation. Finally, dual-targeting both {beta}-catenin and LIN28B through either genetic and/or pharmacological means preferentially reduced AML cell viability. ConclusionThe {beta}-catenin:LIN28B axis could represent a novel synthetically lethal relationship in AML which could be exploited in rare subtypes where LIN28B expression becomes reactivated.

Tunneling nanotubes (TNTs) are actin-based cytoplasmic connections that can mediate intercellular transfer of various cellular cargo and have been implicated in cancer progression and chemoresistance. However, the signalling mechanisms driving their formation remain poorly understood. Given the frequent dysregulation of EGFR and c-Met signalling in non-small cell lung cancer (NSCLC), and prior evidence of TNTs in lung adenocarcinoma patient samples, we investigated the role of EGFR and c-Met receptor signalling crosstalk in TNT induction in A549 lung adenocarcinoma cells. Stimulation with EGF, HGF, or in combination induced a concentration dependent increase in the formation of TNTs. TNTs exhibited typical characteristics, including F-actin expression, non-adherence to the substratum and facilitated intercellular trafficking of lysosomes, mitochondria, and lipid vesicles. EGFR was identified as a novel component of TNTs, but had little co-localisation with the c-Met receptor. Co-stimulation with HGF and EGF did not produce consistent additive or synergistic effects on TNT formation, suggesting shared downstream signalling. Furthermore, although EGFR and c-Met inhibition blocked EGF- and HGF-induced TNTs respectively, inhibition of both receptors was required to suppress TNTs following dual HGF/EGF treatment. Interestingly, blocking the EGF receptor alongside c-Met resulted in a more potent inhibition of HGF-induced TNTs, indicating crosstalk. Furthermore, inhibition of downstream MEK and PI3K pathways reduced HGF- or EGF- induced TNT formation, but dual inhibition was required to completely block TNT formation in HGF+EGF co-stimulated cells. These findings reveal a novel convergence of EGFR and c-Met and their downstream MAPK/PI3K pathways in TNT regulation, which can have important clinical implications in combinatorial receptor and cell signalling pathway targeting in NSCLC.

BackgroundLeptomeningeal disease (LMD) is a serious complication of metastatic breast cancer (MBC) with poor survival. This single-institution retrospective study compares overall survival (OS) among MBC patients with LMD based on CSF parameters (glucose, protein, and WBC count) MethodologyMBC patients who were diagnosed with LMD between 2010-2023 at Wilmot Cancer Institute were screened for eligibility. Only those with available data on CSF glucose, protein, and WBC count were included. OS was assessed via the Kaplan-Meier method and compared using the log-rank test. Cox models were used for multivariate analysis. ResultsOut of 69 patients with MBC LMD, 28 had CSF data and were included in the final analysis. The CSF cytology-positive cohort had significantly lower glucose levels vs the CSF cytology-negative cohort [median (IQR) 40 (18-58) vs 64 (53-92) mg/dl, p=0.006]. Median CSF WBC count was significantly higher in the CSF cytology positive cohort vs the CSF cytology negative cohort [median (IQR) 13 (6-44) vs. 2(2-4)cells/mm3, p=0.001]. When stratified by CSF cytology results and CSF glucose levels, the CSF cytology negative, glucose-low group was associated with the worst OS, while the CSF cytology negative, normal/high glucose group was associated with the best OS(p=0.03) in an unadjusted analysis. Multivariate analysis confirmed that low CSF glucose was independently associated with poorer survival [HR 4.64 (1.71, 13.2)]. Neither CSF protein levels nor CSF WBC counts were significantly associated with OS in unadjusted and multivariate analyses. ConclusionLow CSF glucose was associated with worse OS than normal/high CSF glucose. There was insufficient evidence to suggest that CSF protein or CSF WBC counts were associated with OS.

Breast cancer is the most common cancer diagnosis in women. Clinical studies with triple-negative breast cancer (TNBC) are encouraging for immunotherapy combined with chemotherapy (anti-PD-1 with paclitaxel and/or carboplatin). However, additional clinical advances may be pursued more rapidly with assistance from preclinical TNBC models including syngeneic mammary tumor cell lines. Here, we report two mammary tumor cell lines that exhibit differential responsiveness to immunotherapy in vivo. Spontaneous mammary tumors from C57BL/6J MMTV-Cre Trp53fl/+ animals were passaged serially in cell culture and in vivo in the mammary fat pad of fully wildtype animals. The resulting lines, MM001i and MM008i, lost Trp53 and formed 1000 mm3 tumors in the mammary fat pad within 21-28 days. Despite originating from the same genetic background, these lines exhibit differential responses to immunotherapy. For anti-PD-1 therapy, MM001i is poorly responsive and MM008i is strongly responsive with near-complete tumor regression. In comparison, both MM001i and MM008i respond rapidly to anti-CTLA-4 therapy. Both models express unique tumor antigens as evidenced by immunity to subsequent engraftments. Primary MM008i tumors exhibit greater T cell infiltration, and CD8-positive T lymphocytes are required for anti-PD-1 responses. These TNBC models are promising for further mechanistic studies and testing future single and combinatorial therapies.

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.

Somatic mutations and the tumor immune microenvironment in breast tumors are important predictors of treatment response and survival, yet data for Hispanic/Latina (H/L) women are limited. Here we analyzed whole exome sequencing data from tumor/normal pairs and RNAseq data from 748 H/L women and 388 non-Hispanic White (NHW) women. Overall, the somatic profiles in tumors from H/L women were similar to NHW women. However, somatic mutations in genome organizer CTCF were significantly more common in H/L women. We also found that tumor microenvironment immune ecotypes CE9 and CE10, characterized by increased lymphocyte infiltration and more favorable prognosis, were more common among women with higher Indigenous American ancestry. Finally, we found that a germline APOBEC3A/B copy-number deletion was more prevalent in H/L than in NHW and was associated with the COSMIC APOBEC mutational signatures and with CE10 ecotype. Overall, these results suggest that ancestry differences may provide insights into specific mutation and immune profiles.

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 & AimsPerihilar cholangiocarcinoma is an aggressive malignancy with clinical heterogeneity and poor long-term outcomes after resection. Current prognostic assessment relies mainly on anatomical staging and pathological features, which incompletely capture the entire postoperative risk. We aimed to determine whether integrative analysis of clinical, surgical, pathological and tumor genomic data could improve time-resolved, individualized recurrence-risk prediction after curative-intent resection. MethodsWe performed a multicenter retrospective study including patients undergoing curative-intent resection for perihilar cholangiocarcinoma in ten Spanish hospitals (2003-2023). Overall and disease-free survival were analyzed using Cox models. Outcome-agnostic clinical phenotypes were derived by unsupervised clustering of clinical and surgical features. Targeted tumor sequencing of cancer-associated hotspot regions and selected genes was performed. Prognostic models integrating clinical and genomic data were trained and evaluated in independent training/test sets using penalized and latent-component Cox frameworks, with time dependent discrimination. ResultsThe final cohort comprised 142 patients, with a median follow-up of 26.4 months. Recurrence occurred in 61.3% of patients, and 53.5% died during follow-up. Classical pathological factors were strongly associated with survival and recurrence. Unsupervised outcome-agnostic clustering identified three reproducible clinical phenotypes with markedly different recurrence patterns and survival, only partially explained by anatomical staging. Integrative clinical-genomic modelling further improved recurrence-risk prediction, achieving high discrimination in independent validation (time-dependent AUC [~]0.8). Moreover, the integrative model assigned higher risk over time to patients who relapsed. Patients combining unfavorable clinical phenotype with high genomic-derived risk exhibited a high probability of early recurrence. ConclusionsIntegrated clinical phenotyping and targeted genomic profiling substantially refine recurrence-risk stratification after resection of perihilar cholangiocarcinoma beyond anatomical staging alone. This provides a pragmatic framework for risk-adapted postoperative surveillance and therapeutic decision-making. Impact and ImplicationsThis study provides a data-driven framework integrating clinical, surgical and targeted genomic information to refine prognostic stratification after resection of perihilar cholangiocarcinoma, addressing the limitations of anatomy-based staging in capturing biological heterogeneity. The results are particularly relevant for clinicians managing postoperative surveillance and adjuvant strategies, as they identify patient subgroups with markedly different risks of early recurrence despite similar conventional staging. In practical terms, the combination of unsupervised clinical phenotyping and a targeted, biologically informed genomic panel could support risk-adapted follow-up intensity, selection for adjuvant or experimental therapies, and enrolment into clinical trials. While prospective validation is required before routine implementation, this approach offers a feasible and interpretable pathway toward precision postoperative management in a highly aggressive malignancy.

PurposePrognosis in metastatic non-seminomatous germ cell tumors (mNSGCT) is currently guided by the IGCCCG classification, which incorporates tumor markers, organs involved with metastatic disease, and primary site but not histologic subtype. We aimed to evaluate whether specific histological components provide additional prognostic information in a large international mNSGCT cohort. Patient and MethodsWe analyzed clinical, pathologic, and outcome data from 662 patients with mNSGCT across multiple international centers. Cox regression and multivariable stepwise models were used to evaluate the impact of age, tumor histology, serum markers, primary site of disease, chemotherapy, IGCCCG, and post-chemotherapy surgery on overall survival. Analyses were performed using both complete-case and imputed datasets to account for missing values. ResultsThe presence of any percentage of embryonal carcinoma (EC) was independently associated with improved overall survival HR 0.603 (95% CI: 0.37-0.98, p=0.040), whereas yolk sac tumor (YST) predicted worse prognosis in complete-case analysis HR 2.27 (95% CI: 1.43 - 3.61 p = 0.001). Choriocarcinoma was also associated with a HR 1.58 (95% CI: 1.08 - 2.32 p= 0.019) adverse outcomes. IGCCCG risk classification remained a strong predictor of mortality HR up to 8.9 for Poor vs Good risk, (95% CI: 4.63 - 17.09 p < 0.001), but histologic components added significant independent prognostic value. Post-chemotherapy retroperitoneal lymph node dissection (RPLND) conferred a substantial survival benefit HR 0.44 (95% CI: 0.258 - 0.754 p=0.003). Interestingly, teratoma was not associated with mortality but was linked to younger age, testicular primaries, and higher likelihood of residual disease requiring surgery. ConclusionsHistological composition, particularly the presence of EC or YST, has a significant and independent impact on survival in mNSGCT, beyond established risk classifications. Integration of histological subtypes may enhance prognostic accuracy and guide individualized treatment strategies in advanced germ cell tumors.

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.

PurposePDPK1 functions downstream of PI3K and is essential for activating AKT and other AGC kinases. Although PDPK1 has a central role in the PI3K/AKT/mTOR signaling pathway, there has been limited evaluation of it as a target for cancer therapy. Anaplastic thyroid cancer (ATC) has one of the highest mortality rates of all human malignancies. Although combined BRAF and MEK inhibition in BRAF V600E-mutant ATC (45% of cases) results in response, resistance is common, and there is no curative treatment. The majority (up to 95.8%) of ATC cases have activation in the PI3K/AKT/mTOR and RAS/RAF/MEK/MAPK pathways due to genetic alterations (including driver mutations and genomic gains/losses), involved in these pathways. In this study, we investigated PDPK1 as a therapeutic target for ATC. Experimental designWe used in vitro, ex vivo, and in vivo ATC models to evaluate the effect of targeting PDPK1 (BX795) alone and in combination with mutated BRAF V600E inhibition (dabrafenib), and the mechanism of action that resulted in ATC cell death. ResultsBX795 monotherapy significantly reduced ATC cell proliferation, invasion, colony formation, and spheroid size. The combination of BX795 with dabrafenib produced strong synergistic antitumor activity in BRAF V600E-mutant ATC models. Dual inhibition led to simultaneous and sustained suppression of PDPK1/AKT and MAPK signaling, preventing the compensatory pathway reactivation observed with single-agent treatment. This integrated blockade induced pronounced oxidative stress, DNA damage, and G2-phase cell-cycle arrest, accompanied by mitochondrial dysfunction and robust activation of apoptotic cascades. These effects translated into marked tumor regression in in vitro, ex vivo, and in vivo experimental systems. ConclusionsOur findings identify PDPK1 as a critical and therapeutically tractable vulnerability in anaplastic thyroid cancer. Co-targeting PDPK1 and BRAF V600E produces potent synergistic antitumor activity by shutting down convergent oncogenic signaling nodes and amplifying apoptotic stress responses. These data support PDPK1 inhibition--alone and in combination with BRAF blockade acts as a promising strategy to improve outcomes for patients with BRAF V600E-mutant ATC.

Immune checkpoint inhibitors such as anti-PD1 antibodies are essential in cancer therapy. Emerging data suggest that lower doses may be effective and more economical, though further evidence is needed. We conducted a retrospective study at Centre Leon Berard to assess the efficacy and safety of low-dose nivolumab (20 mg every three weeks) in patients with advanced cancer, mainly squamous cell carcinomas (SCC). Between 2023 and 2024, 53 patients were treated, with a median age of 74 years; 39.6% were over 80. Most were male (64%) and had ECOG >1 (69.9%). Primary tumor sites included cutaneous SCC (34%), head and neck SCC (32%), and soft tissue sarcoma (15%). After a median follow-up of 8.3 months, median overall survival was 7.5 months. The objective response rate (ORR) was 20.8% overall, rising to 35.3% in cutaneous SCC and 23.5% in head and neck SCC-comparable to standard-dose nivolumab. Toxicity was manageable: 18.7% experienced immune-related adverse events, with only 3.7% grade 3. Low-dose nivolumab demonstrates encouraging efficacy and tolerability in a frail population, supporting its potential role in resource-limited settings. Prospective trials are warranted to confirm these findings in broader populations.

Background: Although thin, T1 melanomas have an excellent cure rate with surgery alone, >25% of melanoma deaths originate from thin melanomas (TMs). There is, therefore, an urgent need to improve the identification and management of patients with TMs at high risk of recurrence. Methods: Patients with T1 melanoma and recurrence [&le;] 2 years of diagnosis (T1 rapid group) were compared to patients with T1 melanoma and recurrence [&ge;]10 years after diagnosis (T1 late group). Results: 442 patients from 14 sites were included: 310 and 132 patients in the T1 rapid and late groups, respectively. Median age at primary melanoma diagnosis was 51 years [15-85], 272 (62%) male, 254 (58%) superficial spreading and 101 (23%) head/neck primary. The majority (73%) of recurrences in the T1 rapid group were locoregional. Using univariable logistic regression analysis, age >65 years (p<0.0001), lentigo maligna (LM) melanoma subtype (p=0.025), head/neck primary site (p=0.0065), mitoses [&ge;]1/mm2 (p=0.0181) and ulceration (p=0.0087) were significantly associated with T1 rapid recurrence compared to T1 late recurrence. Using multivariable analysis, age >65 years (p=0.0010), mitoses [&ge;]1/mm2 (p=0.049) and ulceration (p=0.037) remained significant. Conclusions: Rapid recurrence of TM is associated with age >65 years, LM subtype, head/neck primary site, mitoses [&ge;]1/mm2 and ulceration.

The equilibrium between cell death and cell division is crucial for maintaining tissue homeostasis in a multicellular organism. Apoptosis plays an essential role in preserving homeostasis and hence occurs in a coordinated manner. However, inhibition of apoptosis is one of the hallmarks of cancer. Apoptosis Inhibitor 5 (Api5), an anti-apoptotic protein, is upregulated in various cancers, including ovarian, bladder, cervical, and lung cancers. Studies have demonstrated that altered expression of Api5 leads to the transformation of non-tumorigenic breast epithelial cells. However, the mechanism regulating this process is not well-elucidated. Our study demonstrates that overexpression of Api5 increased FGF2 (Fibroblast Growth Factor 2) levels both at protein and transcript levels. We studied the mechanistic details of changes in morphology, proliferation, and polarity observed upon FGF2/FGFR1 deregulation in Api5-overexpressing cells. Deciphering the signalling mechanism underlying Api5-FGF2-mediated breast tumorigenesis revealed that the PDK1/Akt and Ras/MAPK/ERK pathways regulated multiple transformation phenotypes. PDK1/Akt enhanced proliferation and altered morphology during initial stages, whereas Ras/MAPK/ERK regulated polarity disruption, proliferation, and reduced apoptosis during later stages of morphogenesis. In conclusion, this study provides insights into the signalling mechanism regulating the transformation phenotypes associated with Api5 overexpression in a non-tumorigenic breast epithelial cell line.

I.Cutaneous skin melanomas with wild-type BRAF alleles ("BRAF-WT melanomas") remain relatively difficult to treat, even though they typically possess driver mutations in a RAS gene or NF1. For example, these tumors respond relatively poorly to combinations of MEK and BRAF inhibitors, and their response to ICIs is muted compared to the response of BRAF-mutant melanomas. ERBB2 and ERBB4, which encode receptor tyrosine kinase genes, are necessary and sufficient for the proliferation of multiple BRAF-WT melanoma cell lines. Consequently, we have postulated that ERBB4-ERBB2 heterodimerization drives BRAF-WT melanomas. This mechanism is consistent with the observation that elevated ERBB4 transcription or ERBB4 mutations are found in a significant fraction of BRAF-WT melanoma tumor samples. Moreover, a subset of ERBB4 mutations found in BRAF-WT melanoma samples increases proliferation in a BRAF-WT melanoma cell line. Because the elevated ERBB4 transcription observed in BRAF- WT melanomas is typically insufficient to cause ligand-independent ERBB4 signaling, we have postulated that ligands for ERBB family receptors drive the elevated ERBB4-ERBB2 heterodimerization responsible for the proliferation of BRAF-WT melanoma cell lines. We have explored this hypothesis by analyzing data found in the Broad Institutes Cancer Cell Line Encyclopedia. These data suggest that some EGF family hormones are required for the proliferation of BRAF-WT melanoma cell lines. Likewise, the G11/Gq pathway, which can stimulate cleavage and maturation of EGF family hormones, is also required for the proliferation of BRAF-WT melanoma cell lines. Thus, these data suggest additional therapeutic targets in BRAF-WT melanomas. Moreover, because many uveal (ocular) melanomas possess elevated G11/Gq signaling, these data suggest that ligand stimulation of ERBB receptor signaling may contribute to uveal melanomagenesis or progression.