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.

Metastatic skin cutaneous melanomas remain a significant clinical problem. In particular, those melanomas that do not contain a gain-of-function BRAF allele remain challenging to treat because of the paucity of targets for effective therapeutic intervention. Thus, here we investigate the role of the ERBB4 receptor tyrosine kinase in skin cutaneous melanomas that contain wild-type BRAF alleles ("BRAF WT melanomas"). We have performed in silico analyses of a public repository (The Cancer Genome Atlas - TCGA) of skin cutaneous melanoma gene expression and mutation data (TCGA-SKCM data set). These analyses demonstrate that elevated ERBB4 transcription strongly correlates with RAS gene or NF1 mutations that stimulate RAS signaling. Thus, these results have led us to hypothesize that elevated ERBB4 signaling which cooperates with elevated RAS signaling to drive BRAF WT melanomas. We have tested this hypothesis using commercially available BRAF WT melanoma cell lines. Ectopic expression of wild-type ERBB4 stimulates clonogenic proliferation of the IPC-298, MEL-JUSO, MeWo, and SK-MEL-2 BRAF WT melanoma cell lines, whereas ectopic expression of a dominant-negative (K751M) ERBB4 mutant allele inhibits clonogenic proliferation of these same cell lines. Ectopic expression of a dominant-negative ERBB4 mutant allele inhibits anchorage-independent proliferation of MEL-JUSO cells and ectopic expression of a dominant-negative ERBB2 mutant alleles inhibits clonogenic proliferation of MEL-JUSO cells. These data suggest that elevated signaling by ERBB4-ERBB2 heterodimers cooperates with elevated RAS signaling to drive the proliferation of some BRAF WT tumors and that combination therapies that target these two signaling pathways may be effective against these BRAF WT tumors.

Melanoma, the most serious form of skin cancer, frequently involves the dysregulation of key signaling pathways. Treatment strategies presently target the MAPK/ERK pathway, which is overactive in melanomas due in part to BRAF and NRAS mutations, and involve inhibitors against mutated BRAF (vemurafenib or dabrafenib) or MEK kinases (cobimetinib or trametinib), or a combination of the two. Using an established biochip technology, we assessed base excision repair (BER) and nucleotide excision repair (NER) activities in a collection of BRAF mutated (A-375, Colo 829, HT-144, Malme-3M, SK-mel5, SK-mel24 and SK-mel28) and NRAS mutated (M18, MZ2 and SK-mel2) melanoma cell lines, as well as wild-type controls (A7, CHL-1). We evaluated both basal activities (i.e., without treatment) and repair capacities after treatment with vemurafenib or cobimetinib alone, or in combination. Our results indicate that globally the DNA repair capacity of the cell lines was determined by the mutation status of the BRAF and NRAS genes, indicating that the MAPK pathway participates in the regulation of both BER and NER. Treatment of BRAF mutated melanoma cells with vemurafenib alone or the vemurafenib/cobimetinib combination, but not cobimetinib alone, led to reduced DNA repair capacity in about 60% of the BRAF mutated samples, indicating that signaling pathway inhibition can alter DNA repair activity. Upregulation of some DNA repair activities was also observed in several of the treated samples, suggesting activation of compensatory signaling pathways upon treatment. The data collectively indicate that mutations in the BRAF and NRAS genes exert distinct regulatory effects on the excision/synthesis steps of the BER and NER pathways and that targeted pharmacological inactivation of the signaling mechanism can translate into specific consequences in DNA repair capacity. The heterogeneity of the responses reported herein could help define subtypes of melanoma that are associated with resistance to targeted therapies.

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.

Breast cancer remains the most common cancer in women worldwide. Neoadjuvant chemotherapy (NACT) is often preferred to adjuvant chemotherapy to achieve tumour shrinkage, monitor response to therapy and facilitate surgical removal in the absence of metastases. In addition, there is strong evidence that pathological complete remission (pCR) is associated with prolonged survival. In this study, we sought to identify candidate markers that signal response or resistance to therapy. We present a retrospective longitudinal serum proteomic study of 22 breast cancer patients (11 with pCR and 11 with non-pCR) matched with 21 healthy controls. Serum was analysed by LC-MS/MS after depletion of abundant proteins by immunoaffinity, trypsinisation, isobaric labelling and fractionation by reversed-phase HPLC. We observed an inverse behaviour of the serum proteins c-Met and N-cadherin after the second cycle of chemotherapy with a high predictive value (AUC 0.93). More pronounced changes were observed after the 6th cycle of NACT, with significant changes in the intensity of the proteins contactin-1, centrosomal protein, sex hormone-binding globuline and cholinesterase. Our study highlights the possibility of monitoring response to NACT using serum as a liquid biopsy.

Triple-negative breast cancer (TNBC) is the most aggressive breast cancer subgroup characterized by a high risk of resistance to chemotherapies and high relapse potential. High levels of mRNA from the Hedgehog receptor PTCH1 are associated with poor prognosis in TNBC. PTCH1 is overexpressed in many aggressive cancers. We previously reported that PTCH1 is a multidrug transporter that triggers resistance to chemotherapy of adrenocortical carcinoma and melanoma cells, and that inhibiting PTCH1 drug efflux strongly enhanced chemotherapy efficacy on these cell lines both in vitro and in vivo. In the present study, we found that PTCH1 inhibition also significantly inhibited doxorubicin efflux in three TNBC cell lines leading to a strong increase of the cytotoxic effect of doxorubicin and docetaxel, and an inhibition of cell migration. Altogether, our data highlight the therapeutic potential of targeting PTCH1 drug efflux activity using drug association strategies for the treatment of TNBC patients.

Soft tissue sarcomas (STS) histopathological classification system has several conceptual caveats, impacting prognostication and treatment. The clinical and molecular-based tools currently employed to estimate prognosis also have limitations. Clinically driven molecular profiling studies may cover these gaps. We performed DNA sequencing (DNAseq) and RNA sequencing (RNAseq), portraying the molecular profile of 102 samples of 3 of the most common STS subtypes. The RNAseq data was analyzed using unsupervised machine learning models, unravelling previously unknown molecular patterns and identifying 4 well-defined transcriptomic clusters. These transcriptomic clusters have a clear prognostic value, a finding that was externally validated. This transcriptomic cluster-based classifications prognostic value is superior to the prognostic accuracy of currently used clinical-based (SARCULATOR nomograms) and molecular-based (CINSARC) prognostication tools. The analysis of DNAseq data from the same cohort of samples revealed a plethora of unique and, in some cases, never documented molecular targets for precision treatment across different transcriptomic clusters.

PurposeWilms tumor 1 (WT1) is overexpressed in several cancers, and WT1 expression levels are associated with poor prognosis. As a host protein that functions as an oncogene, it represents an important immunotherapeutic target. This study evaluated WT1 expression in Kaposi sarcoma (KS) tumors to assess whether immunotherapy targeting WT1 is a potential therapeutic approach for KS. We also investigated the role of the causal agent of KS, Kaposi sarcoma herpesvirus (KSHV/HHV-8) in regulating WT1 expression. Experimental designImmunohistochemistry for WT1, KSHV, and B and T cells subsets, followed by image analysis, was performed in 363 KS tumor biopsies. Expression of KSHV vFLIP was evaluated by immunofluorescence. Primary endothelial cell cultures and cell lines were infected with KSHV in vitro, or transduced with an inducible vFLIP vector and induced with doxycycline, and then assessed for WT1 expression. Binding of ESK-1, a T cell receptor mimic therapeutic antibody that recognizes WT1 peptides presented on MHC HLA-A0201, was assessed using flow cytometry. ResultsWe report overexpression of WT1 in KS tumors, which was associated with increased with increasing histopathologic stage and the proportion of KSHV-infected cells. Areas with high WT1 expression showed sparse T cell infiltrates. KSHV infection in vitro resulted in WT1 upregulation, mediated by the viral protein vFLIP, which resulted in stronger binding of ESK1. ConclusionsKS lesions express high levels of WT1, a process regulated by the KSHV-encoded vFLIP. These findings suggest that immunotherapy directed against WT1 may represent a therapeutic approach for this cancer. Translational RelevanceKaposi sarcoma (KS) is a vascular neoplasm caused by the Kaposi sarcoma herpesvirus (KSHV/HHV-8). People living with HIV are not only at a significantly higher risk of developing KS, but also often have a more aggressive clinical course. Although antiretroviral therapy may cause regression of HIV-associated KS lesions, advanced cases of KS also require chemotherapy, which is rarely curative. Wilms tumor 1 (WT1) has been reported to be overexpressed in various cancers, functioning as an oncogene and associated with a poor prognosis. WT1 is also an important immunotherapeutic target, with several WT1-directed therapies showing promising results in early clinical trials for leukemias and solid tumors. Here we report high expression of WT1 in KS, especially in higher histological stages. Our findings provide pre-clinical evidence that supports conducting anti-WT1 immunotherapy trials in KS, and evaluating WT1 expression as a potential biomarker to identify individuals most likely to benefit.

The contrast in therapy sensitivity and response across triple negative breast cancer (TNBC) patients suggest underlying genotypic heterogeneity. Using publicly available data, we found significant associations between DNA-level copy number alterations of 1q21.3 locus and therapy sensitivity. We show that in spite of their aggressive nature, 1q21.3 amplified tumors are more responsive to commonly used cytotoxic therapies, highlighting the relevance of 1q21.3 copy number status as a genetic marker for risk stratification, therapy selection and response.

The quality of life for many cancer survivors is compromised due to severe, long-lasting side effects of chemotherapy. As part of a preliminary prospective, non-interventional study to examine the side effects of chemotherapy in patients with breast cancer, we examined the change in protein expression levels in blood collected from patients after treatment with taxanes for 12 weeks (n=7). Protein expression levels were measured with reverse-phase proteomic arrays. Here, we examine changes in proteins related to apoptosis, senescence, and calcium signaling in adjuvant vs. neoadjuvant-treated patients The largest change identified was BCL2 (B-cell lymphoma 2), a founding member of the BCL2 family of proteins that regulate apoptosis. Other proteins regulated by BCL2, including RB1 (retinoblastoma protein 1) and NLRP3 (NLR family pyrin domain containing 3) changed significantly over the course of treatment. These differences are relevant to calcium signaling dysregulation and an increased senescent response, both contributors to cancer recurrence. To validate the observations in this small sample, comparisons were made using Kaplan-Meier plots generated from The Cancer Proteome Atlas (TCPA) breast cancer data. The analysis of the TCPA data also shows a large population with upregulation of BCL2 and that elevated BCL2 is associated with a lower survival probability. Once further validated, these findings indicate that the long-term regulation of BCL2 and related proteins should be considered to optimize patient health and prevent recurrence after taxane-based treatment for breast cancer patients.

Given the fundamental importance of T lymphocytes in the effectiveness of cancer immunotherapy, and potentially in the response to chemotherapy and radiotherapy, immune cell markers have been evaluated as predictive and prognostic biomarkers, with conflicting results in the different subtypes of breast cancer and in different studies. In this study, we report the association of T lymphocyte markers with overall survival in a cross-sectional cohort of Asian breast cancer patients. Formalin-fixed, paraffin-embedded tumour samples from 576 breast cancer patients from a Malaysian hospital were stained with CD3, CD4, CD8 and PD-L1 antibodies, and area-based positive staining was quantified from digitized whole slide images. At a median follow up of 68 months, we report that CD3, CD4 and CD8 scores, but not PDL1 scores, were associated with overall survival, with the strongest associations seen in triple-negative breast cancer (TNBC). CD3, CD4, and CD8 scores were also compared to genomic and transcriptomic data from previous studies, and found to have a negative association with copy number aberrations, but no association with tumour mutational burden or neoantigen load. Taken together, T lymphocyte markers may be associated with good prognosis in Asian breast cancer patients.

Breast cancer shows plasticity in terms of classification. Upon drug treatment and metastasis some tumors switch to another subtype leading to loss of response to therapy. In this study, we ask the question which molecular subclasses of breast cancer are more switchable upon drug therapy and metastasis. We used in silico data to classify breast cancer tumors in PAM50 molecular classes before treatment and after treatment using gene expression data. Similar analysis was performed for primary tumors and their metastatic growth. Our analysis showed that in both scenarios some breast tumors shift from one class to another. This suggests that patients who underwent chemotherapy but resulted in relapse or/and metastasis should be retyped for molecular subclass so that treatment protocol should be adopted according to those subtypes. Additionally, 20 genes were identified as biomarkers for metastasis in breast cancer.

The Notch signaling pathway is frequently altered in oral squamous cell carcinoma (OSCC), the most common malignant neoplasm of the oral mucosa. This study aimed to elucidate the functional role of this pathway in both the initiation and progression of OSCC. Using transgenic animal models, advanced imaging, and next-generation-sequencing techniques, we analyzed Notch-dependent changes driving OSCC. We found specific expression patterns of Notch1 and Delta-like-4 confined to the malignant tissue, while Jagged1 was downregulated in OSCC. This mutually exclusive expression of Delta-like-4 and Jagged1 occurs at the early hyperplastic stage and persists until more advanced stages of the developing tumor. Transcriptomic analyses confirmed the dysregulation of the Notch pathway circuitry and of the genes associated with the undifferentiated state of OSCC cells. Furthermore, pharmacological Notch inhibition significantly impaired cancer cell motility. Taken together, these results reveal the pivotal importance of the Notch1/Delta-like-4 signaling axis as a central oncogenic driver in OSCC. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/576524v1_ufig1.gif" ALT="Figure 1"> View larger version (15K): org.highwire.dtl.DTLVardef@dcaf6corg.highwire.dtl.DTLVardef@811036org.highwire.dtl.DTLVardef@14f5aecorg.highwire.dtl.DTLVardef@da6087_HPS_FORMAT_FIGEXP M_FIG C_FIG

It is now recognized that mitochondria play a crucial role in tumorigenesis, however, it has become clear that tumor metabolism varies significantly between cancer types. The failure of recent clinical trials aimed at directly targeting tumor respiration through oxidative phosphorylation inhibitors underscores the critical need for further studies providing an in-depth evaluation of mitochondrial bioenergetics. Accordingly, we comprehensively assessed the bulk tumor and mitochondrial metabolic phenotype in murine HER2-driven mammary cancer tumors and benign mammary tissue. Transcriptomic and proteomic profiling revealed a broad downregulation of mitochondrial genes/proteins in tumors, including OXPHOS subunits comprising Complexes I-IV. Despite reductions in tumor mitochondrial proteins, mitochondrial respiration was several-fold higher compared to benign mammary tissue, which persisted regardless of normalization method (wet weight, total protein content and when corrected for mitochondrial content). This upregulated respiratory capacity could not be explained by OXPHOS uncoupling, suggesting HER2 signaling regulates intrinsic mitochondrial bioenergetics. In further support, lapatinib, an EGFR/HER2 tyrosine kinase inhibitor, attenuated mitochondrial respiration in NF639 murine mammary tumor epithelial cells. Together, this data highlights that the typical correlation between mitochondrial content and respiratory capacity may not apply to all tumor types and implicates HER2-linked activation of mitochondrial respiration supporting tumorigenesis in this model.

Adrenocortical carcinoma (ACC) is a rare malignancy with dismal prognosis. Deregulated microRNA (miRNA) expression has been implicated in ACC aggressiveness. Nevertheless, the mechanisms underlying such deregulations remain unknown. Aberrant Wnt/{beta}-Catenin signaling has been reported in about 40% of ACC and is associated with poor outcome. Here, we investigated the link between constitutive activation of Wnt/{beta}-Catenin pathway and miRNA expression alterations in ACC. Inducible shRNA-mediated gene silencing of {beta}-Catenin ({beta}-Cat) was performed in ACC cells expressing constitutively active {beta}-Catenin. The miRnome of ACC cells was analyzed using RNA-Sequencing. Selected miRNAs and mRNAs were validated using quantitative PCR and functional experiments with an emphasis on miR-139-5p, its host gene phosphodiesterase 2A (PDE2A) and its target gene N-Myc Downstream-Regulated Gene 4 (NDRG4). Prognostic values of Wnt/{beta}-Catenin pathway components or mutational status and their correlations with miRNA/mRNA expressions were determined in COMETE-ENSAT and TCGA cohorts. We carried out the first miRnome analysis in {beta}-Catenin-deficient ({beta}-Cat-) ACC cells. Twelve upregulated miRNAs and 42 downregulated miRNAs among which miR-139-5p and miRNAs of the 14q32 locus were identified in {beta}-Cat- cells. Downregulation of selected poor prognosis-associated miRNAs was confirmed using RT-qPCR. Remarkably, the expression of the intronic miR-139-5p was decreased by 90% in {beta}-Cat- cells with a concomitant repression of its host gene PDE2A and upregulation of its target gene NDRG4. In ACC patients, miR-139-5p levels were highly correlated with the levels of PDE2A and anti-correlated with those of NDRG4. MiR-139-5p and PDE2A expressions were higher in patients with mutations in components of Wnt/{beta}-Catenin signaling pathway or high expression of LEF1, with LEF1 proving a better predictor of prognosis than Wnt/{beta}-Catenin signaling pathway mutational status. Our findings indicate that in addition to inducing protein-coding genes in ACC, constitutively active Wnt/{beta}-Catenin signaling upregulates the expression of a subset of miRNAs involved in tumour aggressiveness and poor clinical outcome.

Triple-negative breast cancer (TNBC) is an aggressive form of breast cancer defined by the lack of expression of estrogen receptor, progesterone receptors, and of the human epithelial growth factor receptor 2. Neoadjuvant chemotherapy has proven efficacy in the treatment of TNBC, and a pathological complete response (pCR) is predictive of improved long-term survival. The immune response exerts a vital role in response to neoadjuvant chemotherapy, as indicated by the relation between the percentage of stromal tumor-infiltrating lymphocytes (TILs) in pre-treated tumor tissue samples and the likelihood of achieving pCR. Despite this, the relationship between the systemic immune response and the tumor microenvironment is unclear. In this prospective study, we determined the systemic plasma immune profile of TNBC patients before treatment using a panel of 27 immune mediators and measured the percentage of TILs from the same patients. Patients who demonstrated pCR had significantly higher systemic immune mediators; GM-CSF, FGF-basic, VEGF, IL-2, and IL-5, than the non-responders. Moreover, responders displayed a strong positive correlation between the cytokines IFN-{gamma} and IL-7 with the percentage of TILs, while non-responders had a negative or no correlation. Finally, systemic immune mediator levels before treatment predict pCR (AUC range 0.64 - 0.71), and the combination of immune mediators and TILs improved pCR prediction (AUC 0.71 - 0.82). In conclusion, increased systemic immune mediators reflect increased TILs percentage and act as potential predictive biomarkers of pCR for TNBC patients submitted to neoadjuvant chemotherapy.

PurposeTriple-negative breast cancer (TNBC) has a higher incidence, a younger age of onset, and a more aggressive behavior in African-ancestry women. Biological disparities have been suggested as an important factor influencing the ancestry-associated TNBC discrepancy. In this study, we sought to identify ancestry-associated differential gene and protein expression between African-ancestry and White TNBC patients, controlling for patients menopause status and pathological staging at diagnosis. MethodsDifferential gene expression analyses (DGEA) were performed using RNA-sequencing data from The Cancer Genome Atlas (TCGA). Gene set enrichment analysis (GSEA) and Ingenuity Pathway Analysis (IPA), with focus on network design, were performed to highlight candidate genes for further validation through immunohistochemistry of TNBC samples from patients followed in Portugal. ResultsWith 52 African-American and 90 White TNBC patients included, TCGAs data corroborate that African-American patients have a higher TNBC incidence (28.42% vs 11.89%, p<0.0001). Particularly, premenopausal and stage II disease African-American patients also have significantly lower survival probability, comparing with White patients (log-rank p=0.019 and 0.0038, respectively). DGEA results suggest that expression profile differences are more associated with TNBC staging than with patients menopause status. Hippo pathway and cellular community gene sets are downregulated, while breast cancer gene set is upregulated in African-Americans, comparing with White TNBC patients. Furthermore, MAPK pathway gene set is upregulated when controlling for stage II disease. Due to their central role in highly scored networks resulted from IPAs network design, EGFR, Myc and Bcl2 genes were selected for further validation through immunohistochemistry. We also included {beta}-Catenin in the validation study as it is consensually reported to be required in TNBC tumorigenesis. Although patients used in the DGEA and in the immunohistochemistry experiments are geographically and culturally distinct, both groups of African-ancestry patients are mostly of western-African ancestry and, interesting, differential gene and protein expression matched. ConclusionsWe found ancestry-associated gene expression patterns between African-ancestry and White TNBCs, particularly when controlling for menopause status or staging. EGFR, Myc, Bcl2 and {beta}-catenin gene and protein differential expression matching results in distinct populations suggest these markers as being important indicators of TNBCs ancestry-associated development.

Chromosomal alterations have recurrently been identified in Wilms tumors (WTs) and some are associated with poor prognosis. Gain of 1q (1q+) is of special interest given its high prevalence and is currently actively studied for its prognostic value. However, the underlying mutational mechanisms and functional effects remain unknown. For 30 primary WTs, we integrated somatic SNVs, CNs and SVs with expression data and distinguished four clusters characterized by affected biological processes: muscle differentiation, immune system, kidney development and proliferation. We identified 1q+ in eight tumors that differ in mutational mechanisms, subsequent rearrangements and genomic contexts. 1q+ tumors were present in all four expression clusters and individual tumors overexpress different genes on 1q. Through integrating CNs, SVs and gene expression, we identified subgroups of 1q+ tumors reflecting differences in the functional effect of 1q gain, indicating that expression data is likely needed for further risk stratification of 1q+ WTs.

The incidence of urothelial carcinoma of the bladder is four times higher in males than females; however, females tend to present with a more aggressive disease, a poorer response to immunotherapy and suffer worse clinical outcomes. Recent findings have demonstrated sexual dimorphism in the tumor immune microenvironment of non-muscle invasive bladder cancer and associated clinical outcomes. However, a significant gap in knowledge remains with respect to the current pre-clinical modeling approaches and more precisely recapitulating these differences towards improved therapeutic design. Based on the similarities in mucosal immune physiology between humans and mice, we evaluated the sex and age-related immune alterations in healthy murine bladders. Bulk-RNA sequencing and multiplex immunofluorescence based spatial immune profiling of healthy murine bladders from male and female mice of age groups spanning young to old showed a highly altered immune landscape that exhibited sex and age associated differences, particularly in the context of B cell associated responses. Spatial profiling using markers specific to macrophages, T lymphocytes, B lymphocytes, activated dendritic cells, high endothelial venules, myeloid cells and the PD-L1 immune checkpoint showed sex and age associated differences. Bladders from healthy older female mice showed a higher number of tertiary lymphoid structures (TLSs) compared to both young female and male equivalents. Spatial immune profiling of N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) carcinogen exposed male and female bladders from young and old mice revealed a similar frequency of TLS formation, sex differences in the bladder immune microenvironment and, age differences in latency of tumor induction. These findings support the incorporation of sex and age as factors in pre-clinical modeling of NMIBC and will potentially advance the field of immunotherapeutic drug development to improve clinical outcomes in NMIBC.