Clinical and Translational Medicine

Head and neck squamous cell carcinoma (HNSCC) is an aggressive malignancy with high mortality rates, often exhibiting resistance to conventional treatments such as radiotherapy (RT) or a combination of chemotherapy and radiotherapy (CRT). The nerve growth factor receptor (NGFR, also known as p75NTR or CD271) is a well-established cancer stem cell marker in melanoma, where it has been linked to resistance to multiple therapies. In HNSCC, NGFR has been reported as a poor prognostic marker, with its overexpression associated with disease progression. However, its contribution to therapy resistance in HNSCC remains unknown. Here, we show in a cohort of RT/CRT-treated patients that NGFR expression identifies individuals with poor prognosis and increased risk of recurrence following standard RT/CRT. Moreover, we found that NGFR is upregulated in the human Detroit 562 cisplatin (CDDP)-resistant HNSCC cell line in vitro and in vivo. Functional studies demonstrated that genetic knock out of NGFR in these cisplatin-resistant cells restored sensitivity to CDDP in vivo. These results indicate that NGFR contributes to cisplatin resistance in HNSCC. NGFR is upregulated in tumors from patients with poorer prognosis and an increased risk of recurrence after standard radiotherapy and/or RT/CRT, as well as in cisplatin-resistant models. Altogether, our findings open the way to consider NGFR as a new potential therapeutic target to overcome or mitigate cisplatin resistance in HNSCC.

BackgroundMalignant pediatric brain tumors remain the leading cause of cancer-related mortality in children. Current diagnostics rely on imaging and invasive biopsy, which may not capture tumor heterogeneity. Liquid biopsy-based biomarkers such as tumor-educated platelets have shown diagnostic value in adult cancers, but their utility in pediatric brain tumors has not been investigated. MethodsWe analyzed platelet transcriptomes of 73 blood samples from 23 pediatric brain tumor patients, classified as high-grade tumor patients or low-grade tumor patients, and 25 cancer-free controls. Platelets were isolated, CD45 depleted, and RNA was extracted for RNA sequencing. CD45 depletion efficiency was assessed using xCell-based leukocyte enrichment scores. Differential gene expression was assessed with DESeq2 and Gene Ontology over-representation analysis. Gene-level discrimination between groups was evaluated by receiver operating characteristic analysis, and a logistic regression model with patient-grouped 5-fold cross-validation was trained to classify high-grade tumor patients vs. controls. ResultsPlatelets from brain tumor patients showed transcriptional remodeling compared to controls, especially pronounced in high-grade tumor patients. We identified 398 and 649 differentially expressed genes in the brain tumor group vs. controls and high-grade tumor patients vs. control comparisons, respectively, and 85 genes in high-grade tumor patients vs. low-grade tumor patients. No genes met significance in low-grade tumor patients vs. controls. High-grade tumor patients showed consistent upregulation of cancer-associated mitochondrial genes. Similarly, gene enrichment analyses highlighted pathways related to mitotic regulation, chromosome segregation, and mitochondrial metabolism. Multiple genes demonstrated strong diagnostic performance, and logistic regression classifier based on selected platelet transcripts achieved an area under the curve of 0.89, sensitivity of 79%, and a specificity of 80% in identifying high-grade tumor patients. ConclusionsThis study provides the first evidence that platelets exhibit distinct transcriptomic signatures in pediatric brain tumor patients. Platelet RNA profiles robustly differentiate high-grade tumor patients from low-grade tumor patients and cancer-free controls, reflecting tumor presence and biological aggressiveness. These findings support the feasibility of tumor-educated platelets as a minimally invasive biomarker for pediatric malignant brain tumor detection, and longitudinal monitoring. Larger multicenter studies are warranted to validate applicability.

BackgroundMetabolic dysfunction-associated steatotic liver disease (MASLD) is the fastest-growing etiology of hepatocellular carcinoma (HCC). A mechanistic understanding of the metabolic heterogeneity of MASLD-driven tumors is crucial to inform strategies for future treatment options. MethodsPaired tumor (n=8) and adjacent non-tumor tissue (n=8) were collected from patients with steatohepatitic HCC at the University of Kentucky Markey Cancer Center. Hematoxylin and eosin (H&E) staining was used for pathological determination of tumor and adjacent nontumor tissue by a board-certified pathologist. Lipidomic, metabolomic, and transcriptomic analyses were performed, and data were integrated across platforms to identify novel relationships across tumor and adjacent nontumor tissue. ResultsHistological analysis by H&E showed significant lipid vacuole accumulation and inflammatory foci in HCC tumors relative to nontumor tissue. Across omics platforms, we identified 1,679 genes, 1,696 metabolites, and 292 lipids that were significantly (padj<0.01) increased or decreased in tumors relative to nontumor tissue. We identified significant reductions in total ceramides and increases in fatty acyl chain saturation in tumor tissue. Furthermore, metabolites involved in amino acid and fatty acid metabolism were largely decreased in tumors relative to nontumor tissue. We also identified a total of 303 highly significant and novel transcript-metabolite associations (117 gene-metabolite; 186 gene-lipid) across tumor and nontumor tissue. ConclusionsTaken together, this integrative analysis reveals novel relationships between steady-state gene transcripts and specific metabolites in steatohepatitic tumors, thereby identifying new pharmacological targets that may be exploited for therapeutic benefit.

ObjectiveProstate cancer (PCa) constitutes a considerable public health concern worldwide, primarily attributable to its elevated mortality rates. Changes in exosome are shown to significantly influence tumor development. This study aimed to investigate the prognostic value of exosome-related genes (ERGs) in PCa. MethodsPCa single-cell RNA sequencing (scRNA-seq) and transcriptome datasets were obtained from public databases, with ERGs extracted from existing literature. Candidate genes were identified by overlapping 6,004 PCa-related differentially expressed genes (DEGs) and 121 ERGs. Multiple algorithms screened prognostic genes to construct and validate a risk model. Function enrichment, immune infiltration, and drug sensitivity analyses were performed for high/low-risk groups, while scRNA-seq determined cell types via prognostic genes. ResultsA sum of 36 candidate genes was discovered at the intersection of 6,004 DEGs and 121 ERGs. NOC2L, RPS10, POSTN, and BIRC5 were selected as the prognostic genes. The survival status of PCa patients was effectively predicted by a risk model. The majority of pathways identified as significantly enriched between the 2 groups were related to cellular functions. Additionally, 7 differential immune cell types were identified between the 2 groups. RPS10 demonstrated the most significant negative correlation with immature dendritic cells. Chemotherapy drugs were more effective for PCa patients classified as low-risk group. Finally, epithelial cells, endothelial cells, and T cells were considered as key cells and played a critical role in PCa. ConclusionNOC2L, RPS10, POSTN, and BIRC5 were identified associated with exosome in PCa, providing a strong reference for exosome mechanisms in PCa.

Chronic myelomonocytic leukemia (CMML) is a clonal myelodysplastic/myeloproliferative neoplasm characterized by persistent monocytosis and heterogeneous risk of progression to acute leukemia. Mutations within the RAS/MAPK signaling pathway, particularly involving KRAS, are linked to a proliferative disease phenotype and adverse prognosis. We report the first Colombian CMML case harboring two concurrent activating KRAS mutations (p.G12S and p.G13D). Both variants were detected at variant allele frequencies of 17% and 21% in a female patient in her late 50s presenting with marked leukocytosis, anemia, and thrombocytopenia. The coexistence of these mutations suggests synergistic hyperactivation of the RAS/MAPK pathway, likely driving clinical aggressiveness and therapeutic resistance. This case delineates a rare molecular subgroup within CMML and highlights the critical role of comprehensive genomic profiling to improve prognostic accuracy and inform precision medicine approaches.

BackgroundMeningioma-induced hyperostosis (MIH) is a frequent radiographic finding, yet its underlying mechanisms remain poorly understoodWhile hyperostosis has traditionally been treated as a binary phenomenon, the aim of this study was to determine whether MIH represents a heterogenous process with distinct radiological subtypes associated with genetic associations. MethodsWe retrospectively reviewed the records and imaging of patients with meningiomas resected between 2021-2024 at a single institution. Somatic mutations identified through next-generation sequencing were analyzed. CT images were analyzed for bone involvement and hyperostosis subtype. Type I hyperostosis was defined by destruction of cortical architecture while Type II hyperostosis was defined by the preservation of cortical structure. Associations with TRAF7 mutations were assessed using univariate testing, multivariable logistic regression, and supervised machine-learning models. Quantitative bone density analysis was performed using region-of-interest grayscale histogram analysis. ResultsAmong 384 tumors, 54 (14.1%) exhibited hyperostosis--23 Type I and 31 Type II. TRAF7 mutations were significantly enriched in Type I hyperostosis compared with Type II and non-hyperostotic tumors (78.3% vs 25.8% vs 17.0%, p<0.001). Type I hyperostosis independently predicted TRAF7 mutations (OR:18.73, p=0.001), along with skull base location, smaller tumor size, homogeneous contrast enhancement, and extensive T2 hyperintensity. Gradient boosting achieved the highest predictive accuracy (AUC=0.854). Quantitative bone density analysis demonstrated preserved cortical-cancellous architecture in Type II hyperostosis, whereas Type I showed architectural disruption. ConclusionsMIH is a radiographically heterogenous phenomenon. Hyperostosis with disrupted cortical architecture is strongly associated with TRAF7 mutations and may represent a key feature of this mutations radiographic phenotype.

BackgroundInflammatory breast cancer (IBC) is a rare and highly aggressive form of breast cancer with an increased propensity to metastasize to distant organs including the brain. Higher serum levels of the calcium-binding proteins S100A8/A9, particularly of S100A9, have emerged as a clinically and biologically significant factor in aggressive breast cancers that are associated with poorer prognosis, tumor progression, and resistance to therapy. However, its contribution in IBC specifically remains undefined. Here, we investigated whether serum levels of S100A8/A9 predict outcomes in patients with IBC. MethodsSerum S100A8/A9 levels were measured in a cohort of 304 IBC patients using ELISA assay. S100A8/A9 levels were categorized by their third quartile value (S100A8/A9-low [&le;] 3rd quartile; S100A8/A9-high > 3rd quartile). Overall survival (OS) and breast cancer-specific survival (BCSS) were analyzed with Kaplan-Meier curves, log-rank tests, and Cox proportional hazard regression models. The cumulative incidence of any metastases and the cumulative incidence of brain metastases were analyzed using Aalen-Johansen method, Gray test, and Fine-Gray models. ResultsThe median follow-up time was 64 months. Forty-six percent of patients had estrogen receptor (ER)-negative tumors, 61.3% were stage III-IV, 77% high grade, 16.8% received adjuvant chemotherapy and 53.6% received adjuvant radiation. On univariate analysis, S100A8/A9 levels, disease stage, ER status, PR status, HER2 status, adjuvant chemotherapy, and adjuvant radiation therapy were significantly associated with OS and BCSS. Patients with high S100A8/A9 serum levels had poor OS (P=0.01) and BCSS (P=0.007) and had a higher risk of developing brain metastasis (P=0.01) but not other metastasis. On multivariate analysis, high S100A8/A9 serum levels were independently associated with reduced OS (hazard ratio [HR]=1.7, 95% CI 1.1 to 2.6, P=0.01), reduced BCSS (HR=1.8, 95% CI 1.2 to 2.8, P=0.006), and increased cumulative incidence of developing brain metastasis (subdistribution hazard ratio (sHR)=1.8, 95% CI 1.1 to 3.0, P=0.03). ConclusionsIn patients with IBC, high serum levels of S100A8/A9 are an independent prognostic factor for brain metastasis and poor clinical outcomes. These findings support the potential of S100A8/A9 as predictive biomarker for identifying increased risk of brain metastasis and unfavorable prognosis in patients with IBC.

BackgroundLung cancer remains the deadliest malignancy. Although PD-1/PD-L1 immune checkpoint inhibitors have improved survival, their benefit is limited by persistent immune resistance. Identifying systemic regulators of tumor immune microenvironment may enhance therapeutic efficacy. We previously showed that RBC-derived miR-93-5p promotes PTEN loss and PI3K-AKT activation. This study examined whether RBC-derived miR-93-5p is associated with immune checkpoint activity and features of T-cell exhaustion in lung cancer. MethodsRBCs, RBC-derived exosomes, and plasma were isolated from 80 lung cancer patients and 30 controls, and tumor and matched noncancerous lung tissues were collected. miR-93-5p expression was quantified by droplet digital PCR. PD-1, PD-L1, CD8+ T-cell infiltration, CD69, IFN-{gamma}, and TNF- were evaluated by immunohistochemistry. Associations with clinicopathologic features and survival were statistically analyzed. ResultsmiR-93-5p levels in RBCs, their exosomes, and tumor tissues were significantly elevated in lung cancer. Higher miR-93-5p was associated with increased PD-1 and PD-L1 expression and reduced CD69, IFN-{gamma}, and TNF-, consistent with T-cell exhaustion. Elevated miR-93-5p correlated with advanced disease and reduced survival. ConclusionsRBC-derived miR-93-5p is associated with immune checkpoint activation, T-cell exhaustion, and poor clinical outcomes, suggesting impaired antitumor immunity. Targeting this axis may improve the efficacy of immunotherapy in lung cancer.

Lung cancer remains the leading cause of cancer-related deaths worldwide, with tumor recurrence a major contributor to its high mortality. The genetic and molecular mechanisms of recurrence remain poorly understood. Using whole-exome sequencing of 155 primary non-small cell lung cancers, we studied the mutational landscape and driver alterations associated with recurrence. Primary tumors that developed recurrence had higher mutational burden, including hypermutated tumors explained by mutations in DNA polymerase or mismatch repair pathways. Mutational signatures of reactive oxygen species were associated with recurrence. Combined mutations in TP53 and CDKN2A were enriched in non-recurrent tumors, while ATRNL1 mutations were enriched in recurrent tumors. Pathway analyses implicated DNA repair and cilium organisation processes with tumor recurrence and highlighted 50 additional candidate genes including BRCA2. Recurrence-associated genes showed essentiality in lung cancer cell lines and included known therapeutic markers, indicating their functional and translational relevance. This analysis provides insights into the molecular basis of lung cancer recurrence and informs experiments to develop diagnostic and therapeutic strategies.

In this report, experiments were performed to assess how Arylsulfatase B (ARSB; N-acetylgalactosamine-4-sulfatase) treatment might interact with Pembrolizumab and improve therapeutic responses in melanoma. ARSB acts to remove 4-sulfate groups from chondroitin 4-sulfate (C4S; chondroitin sulfate A; CSA). ARSB is required for the degradation of C4S, identified as an oncofetal, tumor-agnostic antigen. Previous reports showed that in syngeneic B16F10 mouse melanomas, ARSB inhibited progression of subcutaneous and metastatic pulmonary melanomas and improved survival by direct effects on melanoma cells. ARSB enhanced apoptosis, which was mediated by increased expression of Constitutive Photomorphogenic (COP)1, an E3 ubiquitin ligase. Combined treatment by recombinant human ARSB, directed at melanoma cells, and Pembrolizumab, directed at infiltrating cytotoxic lymphocytes, can lead to increased apoptosis by different mechanisms, to declines in metalloproteinases and invasiveness, and to altered expression of cytokines. Synergism between effects of ARSB and Pembrolizumab may further inhibit the progression of melanoma and improve treatment outcomes.

IntroductionObstructive sleep apnea (OSA) is characterized by recurrent upper airway obstruction during sleep, leading to intermittent hypoxia, sleep fragmentation, and autonomic dysregulation. These disturbances contribute to nocturnal blood pressure (BP) surges and increased cardiovascular risk. While sleep-stage-dependent BP modulation is well established, high-resolution data on sleep-stage- specific systolic BP variability (BPV) in OSA are limited. This study examined beat-to-beat systolic BPV during N2 and rapid eye movement (REM) sleep and its relationship with sleep fragmentation indices in patients with moderate-to-severe OSA. MethodsClinically suspected OSA patients aged 18-65 years underwent overnight level I video polysomnography. Patients with apnea-hypopnea index (AHI) <15 events/h, central sleep apnea >35%, beta-blocker use, or excessive artifacts were excluded. Continuous systolic BP was estimated using a validated pulse transit time (PTT) method calibrated against cuff BP. Artifact-free 20-minute continuous segments were extracted separately from N2 and REM sleep. Sleep fragmentation metrics included number and duration of respiratory events, arousals, and area under the curve of oxygen saturation (AUC SpO2). BPV indices included mean systolic BP, standard deviation (SD), and coefficient of variation (COV). ResultsSixteen patients contributed 16 N2 and 16 REM segments. N2 sleep showed a higher number of respiratory events (p = 0.005) and arousals (p = 0.01) than REM sleep, while event duration and AUC SpO2 were comparable. Mean systolic BP was 126 {+/-} 12.5 mmHg during N2 and 130 {+/-} 14.9 mmHg during REM, with REM significantly higher than N2 (mean difference -3.62 mmHg; p = 0.01). BP variability was highest during REM (SD 7.12 [4.91-9.25] mmHg; COV 5.95 [3.89-6.84%), intermediate during N2 (SD 5.25 [4.02-6.75] mmHg; COV 4.42 [3.17-4.83%), and lowest during wake (p < 0.001). In N2 sleep, arousal duration predicted mean systolic BP (R{superscript 2} = 0.48, p = 0.0025), while AUC SpO2 strongly predicted SD and COV (R{superscript 2} = 0.74-0.79, p < 0.0001). REM-stage correlations were weaker and not predictive. ConclusionSystolic BP variability in OSA is strongly sleep-stage dependent, with REM sleep exhibiting exaggerated BP instability despite fewer respiratory events. Stage specific mechanisms linking arousals and hypoxia to BP regulation may underlie cardiovascular vulnerability in OSA.

BackgroundInterstitial lung diseases (ILDs), including idiopathic pulmonary fibrosis (IPF) and connective tissue disease-associated ILD (CTD-ILD), share similar features that complicate diagnosis. Tumor markers are often elevated in ILD, yet their diagnostic utility remains unclear. MethodsThis retrospective study included ILD patients hospitalized between 2018 and 2025. Serum levels of alpha-fetoprotein, carcinoembryonic antigen (CEA), carbohydrate antigen (CA) 199, CA125, CA153, neuron-specific enolase (NSE), and cytokeratin 19 fragment (CYFRA 21-1) were analyzed. Arterial blood gases and erythrocyte sedimentation rates (ESRs) were also collected. Statistical analyses involved the Kruskal-Wallis test, Dunns post hoc test, Spearmans correlation, logistic regression, and receiver operating characteristic (ROC) curve analysis. ResultsCEA, CA199, and CA125 levels varied significantly among ILD subtypes (all p < 0.05). NSE differed among CTD-ILD subgroups (p = 0.0409). In IPF, CEA and NSE correlated inversely with PaO2 (r = -0.1556, p = 0.0380; r = -0.2205, p = 0.0031). In CTD-ILD, NSE correlated negatively with PaCO2 (r = -0.1811, p = 0.016), and CYFRA 21-1 with PaO2 (r = -0.1999, p = 0.0078). A diagnostic model incorporating CEA, CA199, sex, age, smoking, PaO2, and ESR differentiated IPF from CTD-ILD with an AUC of 0.833 (95% CI: 0.790-0.876), showing 73.6% sensitivity and 82.4% specificity at a cutoff of 0.569, outperforming single markers. ConclusionCEA, CA199, and CA125 aid in distinguishing ILD subtypes, while CEA, NSE, and CYFRA 21-1 correlate with impaired gas exchange. The combined clinical and biomarker model demonstrated superior performance in discriminating IPF from CTD-ILD, highlighting its clinical potential.

Pulmonary arterial hypertension (PAH) is a rare, life-limiting disease where deficiency of the TGF/BMP pathways have causal roles in hereditary and idiopathic forms. It is an attractive candidate for therapeutic intervention but there is an unmet need for clinically-relevant and practical biomarkers that can measure target engagement. A major challenge has been the inaccessibility of lung tissue in disease for molecular profiling. Here we explore the surrogate capacity of peripheral blood BMP pathway-specific markers. We demonstrate that BMPR-II in flow cytometrically characterised white blood cell subsets is reduced in a proportion of patients, however proteomic analysis demonstrates pleiotropic alterations of TGF{beta}/BMP modulators. Downstream BMPR-II canonical and non-canonical signalling is impacted and measurable in whole blood. We present discovery and international replication cohorts for a transcriptomic BMPR-II signalling signature. The composite biomarker panel is repeatable, reproducible, longitudinally stable and expressed in correlated gene modules in PAH which associate with clinical outcomes. The assay performance characteristics of the biomarker panel make it feasible for early phase, target engagement clinical trials and an adaptive three arm study of two pre- clinically validated modulators of BMPR-II is underway. One Sentence SummaryThe first demonstration of clinically-relevant BMP biomarker panels validated in international populations of patients with PAH.

TRAF1 is a pro-survival signaling adaptor that contributes to NF-{kappa}B activation downstream of a subset of TNFR superfamily members. TRAF1 is overexpressed in many cancers of mature B cells, including chronic lymphocytic leukemia (CLL). Previous studies have established that TRAF1 S146 is a target of phosphorylation by the kinase PKN1 and that PKN1 is required to prevent cellular inhibitor of apoptosis protein (cIAP)-dependent degradation of TRAF1 in the CD40 signaling complex. The kinase inhibitor OSST167 inhibits PKN1 in the nm range and its addition to primary CLL cells was shown to induce dose-dependent loss of TRAF1 and concomitant increases in activated caspase 3 and cell death. These studies identified PKN1 as a target for therapy of CLL. To identify more potent and specific PKN1 inhibitors for therapy of B cell cancers it is important to measure a direct target of PKN1, such as phospho-TRAF1. To this end, here we use overexpression of an S146A mutant of human TRAF1 in 293 cells to validate a recently generated phospho-TRAF1 S146-specific antibody and to confirm that this phosphorylation is lost upon treatment with OTSSP167. Using Cas/Crispr knockout in RAJI cells we also show that both PKN1 and the closely related family member PKN2 can phosphorylate TRAF1 S146. We further show that TRAF1 S146 is constitutively phosphorylated in primary human CLL cells, including those with p53 mutations and that this phosphorylation is sensitive to inhibition with OTSSP167. These findings provide support the development of more potent PKN1/2 inhibitors for CLL.

Mucociliary clearance is a key component of the pathophysiology of bronchiectasis but cilia function is poorly defined. This study aims to characterize nasal ciliary function in bronchiectasis and examine associations with disease severity, infection, inflammation and outcome. Adults with bronchiectasis and healthy volunteers were recruited to the international observational study EMBARC-BRIDGE. Individuals with a known diagnosis of Primary Ciliary Dyskinesia (PCD) were excluded. Nasal respiratory epithelium was sampled by brush biopsy. Ciliary function was assessed by high-speed video microscopy in primary samples and following re-differentiation in air-liquid interface (ALI) culture. Ciliary parameters (cilia length, angle, amplitude, clearance, frequency and ciliation) were quantified and compared with disease severity, microbiology, inflammation and future risk of exacerbations. 171 participants with bronchiectasis were recruited (54% female, age 68years (59-74)). Bronchiectasis nasal brushings showed greater epithelial disruption compared to healthy volunteers (p=0.0006). Six individuals with previously undiagnosed PCD were identified and excluded. In the remaining bronchiectasis cohort, ciliary beat frequency and length were similar to healthy controls. In contrast ciliary beat amplitude, angle, amplitude per second and clearance capacity, were significantly reduced (all p<0.001). These parameters were restored following ALI culture. Regenerated epithelia from bronchiectasis donors exhibited reduced ciliated area. Ciliary dysfunction was strongly associated with future risk of severe exacerbations. The upper airway epithelium is disrupted in bronchiectasis; ciliary movement is impaired and is associated with future risk of exacerbation. Ciliary dysmotility is reversible following ALI culture. This indicates that impaired ciliary function is secondary to the airway environment and therapeutically targetable.

BackgroundGlioblastoma multiforme (GBM) is an aggressive brain tumor that is notoriously resistant to treatment, with an average survival time of 17 months. While the overall outcome is poor for both males and females, sex differences in GBM incidence and outcome suggest sex-specific biological mechanisms underlie tumorigenesis. In contrast, low-grade glioma (LGG) is a less aggressive brain tumor that tends to have a better prognosis and a longer survival time. MethodsTo understand mechanisms contributing to treatment resistance in GBM in both males and females, we inferred gene regulatory networks (GRNs) for males and females with LGG and GBM using RNA-seq data from The Cancer Genome Atlas (TCGA). We analyzed these to identify both sex-specific and sex-stratified gene regulation in GBM. ResultsWe found few sex-specific differences in gene regulation in individuals with LGG, consistent with the lack of evidence for significant clinical endpoints dependent on sex. However, in GBM-we found sex-specific differential targeting of several pathways, including hypoxia and related pathways (carbohydrate metabolism, innate immune processes, and extracellular matrix pathways) known to be dysregulated in hypoxic conditions. In comparing between individuals with GBM, we found that females exhibited a greater degree of co-regulation between hypoxia with the aforementioned downstream pathways than did males. ConclusionsOur results suggest that dysregulation of hypoxia-related pathways in GBM plays a female-specific role in resistance to treatment and overall outcomes.

BackgroundPATZ1 fusion-positive central nervous system (CNS) tumors frequently harbor MN1::PATZ1 fusions as driver mutations, provisionally classified as a rare DNA methylation class of low-grade neuroepithelial tumors. Radiographically, they resemble pilocytic astrocytomas with tumor and cystic components, but their supratentorial cortex location and higher recurrence rates are distinguishing features. An intermediate clinical course, despite focal high-grade histopathology, underscores the need for longitudinal molecular and immune analyses to refine classification and standard therapy. Case SummaryA female pediatric patient presented with neurological symptoms, including headache and right upper extremity weakness. MRI revealed a large cystic lesion in the left frontal lobe, leading to a differential diagnosis of low-grade glioma and ependymoma. Genomic analysis identified an MN1::PATZ1 fusion. The tumor recurred after gross total resection prompting a second resection. Transcriptomic and histopathologic assessments identified multiglial lineage, and high-grade features closely related to adult glioblastoma alongside pro-inflammatory activity in the primary tumor. The recurrent tumor showed reduced malignancy, and oligodendroglioma-like features. Increased MHC gene expression, immune checkpoint receptors (PDCD1, CTLA4, TIGIT,TIM3), T cell regulators (CXCR6), and elevated macrophage frequency, coupled with reduced PD-L1 in the recurrent tumor, suggest a complex anti-tumor immune response constrained by T cell dysregulation. This case, along with two other MN1::PATZ1 fusion-positive tumors, identifies a distinct transcriptomic subtype separate from circumscribed astrocytic glioma, highlighting upregulation of growth factor receptor pathways, like PI3K/AKT, and immune dysfunction linked to recurrence. ConclusionLongitudinal multi-omics analyses of recurrent MN1::PATZ1 fusion-positive CNS tumors revealed tumor maturation, immune dysfunction, and potential therapeutic targets. Introductory ParagraphPATZ1 fusion-positive central nervous system (CNS) tumors are rare, predominantly pediatric and frequently recurrent neoplasms provisionally classified as neuroepithelial tumors. Their pronounced histopathological and clinical heterogeneity, along with limited immunological characterization complicates their treatment standardization. We report a new case of an MN1::PATZ1 fusion-positive CNS tumor with recurrence, highlighting its radiographic similarities to low-to-intermediate grade pediatric glioma. Longitudinal multi-omics analyses of this case, along with additional MN1::PATZ1 fusion-positive CNS tumors, however, delineates a transcriptome subtype resembling adult high-grade glioma, with activated oncogenic and pro-inflammatory programs. The recurrent tumor exhibits features of decreased malignancy and enhanced glial differentiation, phenotypically shifting towards oligodendroglioma, suggesting tumor maturation. This was accompanied by increased antigen presentation programs, indicating immune engagement, while increased immune checkpoint expression and microglia/macrophage frequency indicate T cell exhaustion and immunomodulation, respectively. This longitudinal study highlights potential therapeutic strategies targeting both the tumor and its immune environment in MN1::PATZ1 fusion-positive CNS tumors.

IntroductionSalvage surgery is known for an increased surgical complication rate. However, currently available patient, tumor and therapy factors are not reliable enough to predict surgical site complications (SSC). This exploratory study tests skin acquired by neck dissection for several histochemical and immunohistochemical stains to explore tissue changes related to wound healing. Patients and methodsFrom an existing database of 232 isolated neck dissections (ND), seventeen patients with skin resection during ND were included - five primary and twelve salvage procedures. Skin specimens were tested for ten histochemical and immunohistochemical stains, with emphasis on the stages of wound healing; inflammation, proliferation and remodeling. ResultsThree stains (CD-68, -SMA, and Vimentin), showed significant different expressions between the primary surgical and salvage groups (respectively p = 0.013, p = 0.044 and p = 0.015), without a clear pattern of decreased or increased expression. None of the ten stains showed a significant difference related to SSC. ConclusionThe three significantly different staining patterns found in this exploratory study all are related to macrophage and (myo-)fibroblast transition, possibly representing an imbalance of macrophage/stroma interplay. Future studies should focus on the pivotal role of macrophages in wound healing after salvage surgery.

BackgroundIsocitrate dehydrogenase wildtype (IDHwt) histologic grade 2 adult diffuse gliomas represent a highly heterogeneous entity, and the effects of postoperative adjuvant temozolomide (TMZ) therapy, as well as the predictive value of chemotherapy-related biomarker O6-methylguanine-DNA methyltransferase promoter (MGMT-p) methylation, remain to be further investigated. MethodsA Discovery dataset, comprising 108 IDHwt histologic grade 2 gliomas obtained from three public resources, was constructed to investigate the impact of TMZ on patient survival. Furthermore, an independent Validation dataset, consisting of 123 IDHwt grade 2 gliomas, was created to validate the effect of TMZ on patient survival. Kaplan-Meier overall survival (OS) analyses and Cox proportional hazard models were used. ResultsMultivariable analysis in the validation dataset demonstrated that temozolomide (TMZ) chemotherapy was an adverse independent prognostic factor for survival in patients with histologic grade 2 IDH-wildtype (IDHwt) gliomas (HR = 2.19, P = 0.033). Subgroup analyses further revealed that the detrimental effect of TMZ was mainly confined to MGMT-p unmethylated tumors (Discovery cohort: TMZ vs noTMZ, median overall survival [OS]: 37.0 vs 130.0 months, log-rank P = 0.005, HR = 1.89; Validation cohort: TMZ vs noTMZ, median OS: 34.4 vs >120 months, log-rank P = 0.004, HR = 3.39). Moreover, in the validation cohort, TMZ therapy remained associated with significantly poorer survival in the NEC (Not Elsewhere Classified) subgroup (TMZ vs noTMZ, median OS: 73 vs >120 months, P = 0.017), while in molecular GBMs it did not reach statistical significance but still showed a trend toward worse survival (TMZ vs noTMZ, median OS: 20 vs 30 months, P = 0.37). By comparing multi-omics differences between patient groups, we observed that MMR-related genes were specifically downregulated in IDHwt grade 2 gliomas at both the single-cell and bulk transcriptomic levels. DiscussionThe therapeutic benefit of TMZ in IDHwt histologic grade 2 gliomas appears to be limited, with even a potential adverse impact on survival. Therefore, postoperative use of TMZ as a recommended chemotherapy should be approached with caution in these patients, particularly in cases with unmethylated MGMT-p, where alternative treatment strategies are warranted. High MGMT expression and specific downregulation of MMR-related genes may represent key factors underlying the limited efficacy of TMZ in IDHwt, MGMT-p unmethylation grade 2 gliomas. Level of evidenceIII.

BackgroundMetabolomic changes related to colorectal cancer (CRC) may serve as diagnostic markers to identify patients may develop or have developed CRC. MethodsUntargeted lipidomics were performed on serum from CRC cases and clean-colon controls from the Chicago Colorectal Cancer Consortium (CCCC) and the University of Arizona Cancer Center (UACC). ResultsUntargeted lipidomics in the CCCC CRC series revealed significant alterations in sphingolipids. Targeted lipidomics revealed a signature of five sphingomyelins (SMs) were significantly decreased in CRC patients in CCCC and UACC CRC series. Circulating SMs are degraded primarily by S-SMase and serum S-SMase activity was significantly higher in UACC cases as compared to controls. Serum S-SMase activity was also measured in two series of adenoma patients to determine if S-SMase may serve as a biomarker for development of colorectal neoplasia. While S-SMase activity was significantly higher in adenoma patients compared to controls in the mostly white UACC series, S-SMase activity in samples from the Chicago Black series (CCCC) were indistinguishable from each other and significantly higher than UACC controls. ConclusionsTogether, these studies suggest the potential for S-SMase activity to serve as a biomarker for colorectal neoplasia, with potential implications in some but perhaps not all populations.