Glioma is one of the most common types of primary brain tumors. Ivermectin (IVM), a broad-spectrum antiparasitic drug, has been identified as a novel anticancer agent due to its inhibitory effects on the proliferation of glioma cells in vitro and in vivo. However, the ability of IVM to induce autophagy and its role in glioma cell death remains unclear. The main objective of the present study was to explore autophagy induced by IVM in glioma U251 and C6 cells, and the deep underlying molecular mechanisms. In addition, we examined the effects of autophagy on apoptosis in glioma cells. In the present study, transmission electron microscopy (TEM), immunofluorescence, Western blot and immunohistochemistry were used to evaluate autophagy activated by IVM. Cell viability was measured by 3-(4,5-dimethylthiazol2-yl)-2, 5-diphenyltetrazolium bromide (MTT) and colony formation assay. The apoptosis rate was detected by flow cytometry and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL). Meanwhile, autophagy inhibition was achieved by using chloroquine (CQ). U251-derived xenografts were established for examination of IVM-induced autophagy on glioma in vivo. Taken together, the results of the present study showed that autophagy induced by IVM has a protective effect on cell apoptosis in vitro and in vivo. Mechanistically, IVM induced autophagy through AKT/mTOR signaling and induced energy impairment. Our findings show that IVM is a promising anticancer agent and may be a potential effective treatment for glioma cancers.

BACKGROUND:The PI3K/AKT pathway activation is an independent marker of poor outcome in head and neck squamous-cell carcinoma (HNSCC), and involved in resistance to cetuximab. Molecular alterations in this pathway, particularly PIK3CA mutations, represent a key event of this dysregulation. Buparlisib is a pan-Class I PI3K inhibitor that inhibits tumor growth in HNSCC xenografts. METHODS:This phase II trial evaluated buparlisib (100 mg/d) in two parallel cohorts of refractory HNSCC patients (progression after platinum and cetuximab) with or without PIK3CA mutated in exons 9/20. The primary endpoint was the 2-month Disease Control Rate (DCR2m) as per centrally reviewed. Buparlisib would be considered ineffective if DCR2m ≤ 10% and promising if ≥ 30% (α: 5% unilateral, power: 90%): 7 successes for 35 evaluable patients per cohort were required. Secondary endpoints included ORR, PFS, OS, and safety. RESULTS:From January 2013 to July 2018, 58 HNSCC patients received ≥ 1 dose of buparlisib (PIK3CA wild-type, n = 36; PIK3CA-mutated, n = 22); 78% had received ≥ 2 prior lines of systemic therapy. The PIK3CA-mutated cohort was prematurely closed because of slow accrual. The DCR2m was 38.9% (95% CI [25.5- + ∞[) in PIK3CA wild-type and 36.4% (95% CI [19.5 - + ∞[) in PIK3CA-mutated patients. A total of 53.4% of patients experienced treatment-related grade ≥ 3 adverse events. The most frequent (>5%) were hyperglycemia, lymphopenia, asthenia, sodium decrease, depression, and dermatitis. Two toxic deaths were reported. CONCLUSION:Although the trial met the primary endpoint, buparlisib monotherapy was associated with unacceptable toxicity and showed limited efficacy in heavily pretreated HNSCC patients.

BACKGROUND:Cluster of differentiation 123 (CD123) prominently overexpress in various hematological malignancies and plays a crucial role in diagnosis and prognosis of leukemia. Clinical studies have demonstrated that cell-free CD123 levels also significantly influence leukemia immunotherapy outcomes. The development of novel electrochemical immunosensors addresses the need for point-of-care detection tools, thereby facilitating advancements in clinical monitoring technologies. METHODS:This study presented a practical electrochemical immunosensor constructed using sandwich strategy for CD123 detection based on the modification of gold nanoparticles and poly (2-aminoterephthalic acid). Common proteins and tumor-related biomarkers found in human were selected as interference factors to evaluate the detection specificity of the electrochemical immunosensor. Further, the electrochemical immunosensor was utilized to directly detect CD123 in bone marrow supernatant from leukemia patients. RESULTS:The constructed electrochemical immunosensor exhibited good linearity for CD123 detection over a range of 0.02 to 2.5 µg/mL, with a detection limit of 12.8 ng/mL, alongside satisfactory specificity and repeatability. Furthermore, the immunosensor was successfully employed to detect CD123 levels in the bone marrow supernatant of leukemia patients, demonstrating results that were highly consistent with those obtained via ELISA. CONCLUSIONS:The developed approach is anticipated to provide robust technical support for the long-term monitoring of leukemia patients during their diagnosis and treatment.

Esophageal squamous cell carcinoma (ESCC) is a major global health challenge, especially in Asia, due to its high incidence, mortality and poor prognosis. As there are no reliable early - diagnosis biomarkers, ESCC is often detected at an advanced stage, when radiotherapy becomes the main treatment. However, the emergence of radioresistance significantly compromises treatment efficacy, leading to tumor recurrence and metastasis. Although some research has been done on the mechanisms of ESCC radiation resistance, a comprehensive understanding remains elusive. To address this knowledge gap and identify more molecular targets for overcoming radiation resistance, we established a radioresistant ESCC cell model and conducted systematic 4D label-free proteomic profiling. Quantitative analysis revealed 364 differentially expressed proteins, predominantly enriched in nucleotide excision repair, glutathione metabolism, and insulin resistance pathways. Functional validation identified TXNDC12 as a critical regulator of radioresistance, and its overexpression is significantly associated with enhanced glutathione synthesis and intracellular ROS scavenging. This study provides the first proteomic evidence linking redox homeostasis modulation through TXNDC12-GSH axis activation to ESCC radioresistance, offering novel therapeutic targets for overcoming radiation resistance and improving clinical outcomes in advanced ESCC management.

Early detection of juvenile clinical deterioration in acute care settings remains a significant problem in modern healthcare. This paper presents an AI-powered predictive analytics platform that combines transcriptome biomarker signals with structured vital signs, laboratory data, and unstructured clinical notes to improve early warning capabilities. The system uses ClinicalBERT to extract insights from clinical narratives, XGBoost to analyze tabular clinical information, and long short-term memory (LSTM) networks to simulate temporal dynamics. A meta-classifier combines multimodal data to produce real-time risk ratings for clinical deterioration. The performance evaluation utilizing five-fold cross-validation showed great accuracy, with an AUROC of 0.91, AUPRC of 0.83, and an average early warning lead time of 5.6 hours. Predictive markers included higher lactate levels, heart rate patterns, SpO₂ variability, and transcriptome signals indicating systemic inflammatory activation. Ablation investigations proved the importance of multimodal data fusion in increasing prediction robustness. The suggested strategy provides a scalable, interpretable, and high-performing hospital integration system that enables biomarker-informed, precision-based pediatric intervention options.

Zinc (Zn) is an essential trace element that plays a crucial role in plant growth and development, but excessive Zn can be stressful or even toxic to plants. The GLTP superfamily is critical for lipid metabolism and membrane stability maintenance, yet its function in plant Zn tolerance remains unclear. In this study, zinc stress treatment experiments were performed using transgenic apple calli, apple seedlings, Arabidopsis thaliana, and Solanum lycopersicum. Under Zn treatment, compared with the wild type (WT), the apple seedlings of the MbACD11 transgenic line exhibited significantly higher plant height and fresh weight, with increases of 5.87% and 93.21% respectively. Meanwhile, their MDA level, relative electrical conductivity, and accumulations of H2O2 and O2− were all significantly reduced, with decreases of 20.47%, 35.47%, 31.50%, and 36.78% respectively. Consistently, these data showed the same trend in calli, Arabidopsis thaliana, and tomato. These results demonstrated that the overexpression of MbACD11 significantly enhanced Zn tolerance in transgenic plants, and also verified that the function of this gene may be conserved across different species. In summary, this study establishes a molecular framework and theoretical basis for improving plant tolerance to Zn stress and paves the way for future mechanistic investigations.

Lynch syndrome (LS) is a common hereditary cancer syndrome caused by heterozygous germline pathogenic variants in DNA mismatch repair (MMR) genes. Splicing defect constitutes one of the major mechanisms for MMR gene inactivation. Using RT-PCR based RNA analysis, we investigated 24 potential spliceogenic variants in MMR genes and determined their pathogenicity based on refined splicing-related American College of Medical Genetics and Genomics/Association for Molecular Pathology (ACMG/AMP) criteria. Aberrant transcripts were confirmed in 19 variants and 17 of which were classified as pathogenic including 11 located outside of canonical splice sites. Most of these variants were previously reported in LS patients without mRNA splicing assessment. Thus, our study provides crucial evidence for pathogenicity determination, allowing for appropriate clinical follow-up. We also found that computational predictions were globally well correlated with RNA analysis results and the use of both SPiP and SpliceAI software appeared more efficient for splicing defect prediction.

BACKGROUND AND PURPOSE:In this manuscript we describe the academic French multicentric molecular analysis platforms including PROFILER, promoted by Centre Léon Berard, and the multicentric personalized medicine trials MOST, MOST Plus and MEGAMOST. PATIENTS/MATERIAL AND METHODS:MOST, MOST Plus and MEGAMOST comprise 14 cohorts with different targeted agents and immunotherapies. RESULTS AND INTERPRETATION:PROFILER has recruited 5,991 patients in 10 years, MOST and MOST Plus 875 patients since 2014 and MEGAMOST 172 patients since 2020, and are still ongoing. We provide a description of the local, national and international implications of these initiatives, and we review the results of the sorafenib and olaparib cohorts.

Background:V-raf murine sarcoma viral oncogene homolog B1 (BRAF) inhibitor (BRAFi) therapy resistance affects approximately 15% of cancer patients, leading to disease recurrence and poor prognosis. The aim of the study was to develop a machine-learning based method to identify patients who are at high-risk of BRAFi resistance and potential biomarker. Methods:From Cancer Cell Line Encyclopedia (CCLE) and Genomics of Drug Sensitivity in Cancer (GDSC) databases, we collected RNA sequencing and half maximal inhibitory concentration (IC50) data from 235 pan-cancer cell lines and then identified 37 significant differential expression genes associated with BRAFi resistance. Employing machine learning (ML) models, we successfully classified cell lines into resistant and sensitive groups, achieving robust performance in external validation datasets. Results:AOX1 may play a vital part in BRAFi metabolism and resistance. Further, we found that higher mRNA expression of OXTR, H2AC13, and TBX2, and lower mRNA of SLC2A4, as detected by PCR in WM983B and SKMEL-5 cell lines, were independent risk factors for BRAFi resistance and were associated with poor prognosis. Conclusions:We established a gene-expression model using ML methods, consisting of 37 variables based on RNA-seq database, which was externally validated and could be used to predict BRAFi resistance. Meanwhile, our findings provide valuable insights into the molecular mechanisms of BRAFi resistance, enabling the identification of high-risk patients.

A substantial number of hereditary colorectal cancer (CRC) and colonic polyposis cannot be explained by alteration in confirmed predisposition genes, such as mismatch repair (MMR) genes, APC and MUTYH. Recently, a certain number of potential predisposition genes have been suggested, involving each a small number of cases reported so far. Here, we describe the detection of rare variants in the NTLH1, AXIN2, RNF43, BUB1, and TP53 genes in nine unrelated patients who were suspected for inherited CRC and/or colonic polyposis. Seven of them were classified as pathogenic or likely pathogenic variants (PV/LPV). Clinical manifestations of carriers were largely consistent with reported cases with, nevertheless, distinct characteristics. PV/LPV in these uncommon gene can be responsible for up to 2.7% of inherited CRC or colonic polyposis syndromes. Our findings provide supporting evidence for the role of these genes in cancer predisposition, and contribute to the determination of related cancer spectrum and cancer risk for carriers, allowing for the establishment of appropriate screening strategy and genetic counseling in affected families.

This is the brief protocol for seeding and culturing human neuroblastoma SH-SY5Y cells.

BACKGROUND Computed tomography (CT) imaging features are associated with risk stratification of gastric gastrointestinal stromal tumors (GISTs). AIM To determine the multi-slice CT imaging features for predicting risk stratification in patients with primary gastric GISTs. METHODS The clinicopathological and CT imaging data for 147 patients with histologically confirmed primary gastric GISTs were retrospectively analyzed. All patients had received dynamic contrast-enhanced CT (CECT) followed by surgical resection. According to the modified National Institutes of Health criteria, 147 lesions were classified into the low malignant potential group (very low and low risk; 101 lesions) and high malignant potential group (medium and high-risk; 46 lesions). The association between malignant potential and CT characteristic features (including tumor location, size, growth pattern, contour, ulceration, cystic degeneration or necrosis, calcification within the tumor, lymphadenopathy, enhancement patterns, unenhanced CT and CECT attenuation value, and enhancement degree) was analyzed using univariate analysis. Multivariate logistic regression analysis was performed to identify significant predictors of high malignant potential. The receiver operating curve (ROC) was used to evaluate the predictive value of tumor size and the multinomial logistic regression model for risk classification. RESULTS There were 46 patients with high malignant potential and 101 with low-malignant potential gastric GISTs. Univariate analysis showed no significant differences in age, gender, tumor location, calcification, unenhanced CT and CECT attenuation values, and enhancement degree between the two groups (P > 0.05). However, a significant difference was observed in tumor size (3.14 +/- 0.94 vs 6.63 +/- 3.26 cm, P < 0.001) between the low-grade and high- grade groups. The univariate analysis further revealed that CT imaging features, including tumor contours, lesion growth patterns, ulceration, cystic degeneration or necrosis, lymphadenopathy, and contrast enhancement patterns, were associated with risk stratification (P < 0.05). According to binary logistic regression analysis, tumor size [P < 0.001; odds ratio (OR) = 26.448; 95% confidence interval (CI): 4.854-144.099)], contours ( P = 0.028; OR = 7.750; 95% CI: 1.253- 47.955), and mixed growth pattern (P = 0.046; OR = 4.740; 95%CI: 1.029-21.828) were independent predictors for risk stratification of gastric GISTs. ROC curve analysis for the multinomial logistic regression model and tumor size to differentiate high-malignant potential from low-malignant potential GISTs achieved a maximum area under the curve of 0.919 (95%CI: 0.863-0.975) and 0.940 (95%CI: 0.893-0.986), respectively. The tumor size cutoff value between the low and high malignant potential groups was 4.05 cm, and the sensitivity and specificity were 93.5% and 84.2%, respectively. CONCLUSION CT features, including tumor size, growth patterns, and lesion contours, were predictors of malignant potential for primary gastric GISTs.

PMS2 germline pathogenic variants are one of the major causes for Lynch syndrome and constitutional mismatch repair deficiencies. Variant identification in the 3' region of this gene is complicated by the presence of the pseudogene PMS2CL which shares a high sequence homology with PMS2. Consequently, short-fragment screening strategies (NGS, Sanger) may fail to discriminate variant's gene localization. Using a comprehensive analysis strategy, we assessed 42 NGS-detected variants in 76 patients and found 32 localized on PMS2 while 6 on PMS2CL. Interestingly, four variants were detected in either of them in different patients. Clinical phenotype was well correlated to genotype, making it very helpful in variant assessment. Our findings emphasize the necessity of more specific complementary analyses to confirm the gene origin of each variant detected in different individuals in order to avoid variant misinterpretation. In addition, we characterized two PMS2 genomic alterations involving Alu-mediated tandem duplication and gene conversion. Those mechanisms seemed to be particularly favored in PMS2 which contribute to frequent genomic rearrangements in the 3' region of the gene.

BackgroundThe MLH1 gene is one of the DNA mismatch repair genes (MMR), implicated in Lynch syndrome (LS), an autosomal dominant hereditary tumor susceptibility disease. The advent of next-generation sequencing (NGS) technologies has accelerated the diagnosis of inherited diseases and increased the percentage of diagnosis of inherited cancers. However, some complex genomic alterations require the combination of several analytical strategies to allow correct biological interpretations. Here, we describe a novel MLH1 deletion and its pathogenicity determination in a patient suspected of LS. MethodsThe index case was a French 73-year-old man diagnosed with colorectal cancer displaying microsatellite instability and the loss of MLH1 and PMS2 expression. NGS analysis was used as the primary method for MMR genes screening. Long-range PCR and reverse transcriptase polymerase chain reaction (RT-PCR) were used for breakpoints and pathogenicity determinations. ResultsA large genomic deletion was detected which removed the last six nucleotides of MLH1 exon 11 together with a large part of intron 11. It was initially considered as a variant of unknown significance (VUS). Genomic breakpoints were subsequently characterized defining the deletion as c.1033_1039-248del. Further RNA analysis demonstrated that this variant activated a cryptic donor splice site at the 5 & PRIME; of the breakpoint, leading to a premature truncated protein: p.Thr345Alafs*13. ConclusionOur finding suggested that although NGS technologies have increased variant detection yield, combined approaches were still needed for complex variant characterization and pathogenicity assessment.

The PMS2 gene is one of the DNA mismatch repair genes (MMR) implicated in Lynch syndrome (LS). A subset of PMS2 pathogenic variants (PVs) are splice variants mostly affecting canonical GT/AG splicing sequences. However, the majority of the intronic variants outside canonical splice sites remained as variants of unknown significance, even though some of them would alter the splicing process. In this report, we describe the analysis of such an intronic variant (c.251-5T > C) detected in an 82-year-old patient diagnosed with endometrial cancer displaying microsatellite instability and the loss of PMS2 expression displayed. RNA analysis demonstrated that this variant lead to the complete exon 4 skipping, resulting in the synthesis of a truncated protein. This finding shows the relevance of functional RNA analysis in the non-canonical intronic variant assessment and the importance of systematic evaluation of MSI/loss of expression of MMR genes for LS screening in patients with endometrial cancers.

The POLD1 gene is involved in DNA proofreading to ensure accurate DNA replication. Some germline alterations in its exonuclease domain are associated with predisposition to cancers and colonic polyps. Only a few pathogenic variants have been clearly identified so far. Here we report a novel variant: c.1458G>T p.(Lys486Asn) that we classified as pathogenic, detected in two putatively unrelated families. The cancer spectrum was very similar to Lynch syndrome, implying an overlapped tissue susceptibility. The common presence of colonic polyps in carriers and the MMR proficient phenotype in tumors were distinctive features suggesting POLD1 implication. Some clinical characteristics observed in the carriers of this variant differed from those reported previously, suggesting a potential genotype/phenotype correlation, and very likely in relation to the functional importance of affected residues. Our findings provide further insight into understanding the role of POLD1 in cancer-related risk.

INTRODUCTION:Epithelial-to-mesenchymal transition (EMT) is associated with tumor aggressiveness, drug resistance, and poor survival in non-small cell lung cancer (NSCLC) and other cancers. The identification of immune-checkpoint ligands (ICPLs) associated with NSCLCs that display a mesenchymal phenotype (mNSCLC) could help to define subgroups of patients who may benefit from treatment strategies using immunotherapy. METHODS:We evaluated ICPL expression in silico in 130 NSCLC cell lines. In vitro, CRISPR/Cas9-mediated knockdown and lentiviral expression were used to assess the impact of ZEB1 expression on CD70. Gene expression profiles of lung cancer samples from the TCGA (n = 1018) and a dataset from MD Anderson Cancer Center (n = 275) were analyzed. Independent validation was performed by immunohistochemistry and targeted-RNA sequencing in 154 NSCLC whole sections, including a large cohort of pulmonary sarcomatoid carcinomas (SC, n = 55). RESULTS:We uncover that the expression of CD70, a regulatory ligand from the tumor necrosis factor ligand family, is enriched in mNSCLC in vitro models. Mechanistically, the EMT-inducer ZEB1 impacted CD70 expression and fostered increased activity of the CD70 promoter. CD70 overexpression was also evidenced in mNSCLC patient tumor samples and was particularly enriched in SC, a lung cancer subtype associated with poor prognosis. In these tumors, CD70 expression was associated with decreased CD3+ and CD8+ T-cell infiltration and increased T-cell exhaustion markers. CONCLUSION:Our results provide evidence on the pivotal roles of CD70 and ZEB1 in immune escape in mNSCLC, suggesting that EMT might promote cancer progression and metastasis by not only increasing cancer cell plasticity but also reprogramming the immune response in the local tumor microenvironment.

Background: a specific subset of metastatic triple-negative breast cancers (mTNBC) is characterized by homologous recombination deficiency (HRD), leading to enhanced sensitivity to platinum-based chemotherapy. Apart from mutations in BRCA1/2 genes, the evaluation of other HRD-related alterations has been limited to date. As such, we analyzed data from mTNBC patients enrolled in the ProfiLER-01 study to determine the prevalence of alterations in homologous recombination-related (HRR) genes and their association with platinum sensitivity. Methods: next-generation sequencing and promoter methylation of BRCA1 and RAD51C were performed on tumors from patients with mTNBC, using a panel of 19 HRR genes. Tumors were separated into three groups based on their molecular status: mutations in BRCA1/2, mutations in other HRR genes (BRCA1/2 excluded) or BRCA1/RAD51C promoter methylation and the absence of molecular alterations in HRR genes (groups A, B and C, respectively). Sensitivity to platinum-based chemotherapy was evaluated through the radiological response. Results: mutations in BRCA1/2 were detected in seven (13.5%) patients, while alterations in other HRR genes or hypermethylation in BRCA1 or RAD51C were reported in 16 (30.7%) patients; furthermore, no alteration was found in the majority of patients (n = 29; 55.8%). Among 27 patients who received platinum-based chemotherapy, the disease control rate was 80%, 55% and 18% (groups A, B and C, respectively; p = 0.049). Regarding group B, patients with disease control exhibited mutations in FANCL, FANCA and the RAD51D genes or RAD51C methylation; Conclusion: mutations in HRR genes and epimutations in RAD51C were associated with disease control through platinum-based chemotherapy. As such, apart from well-characterized alterations in BRCA1/2, a more comprehensive evaluation of HRD should be considered in order to enlarge the selection of patients with mTNBC that could benefit from platinum-based chemotherapy.

Glioblastoma (GBM) is one of the most widespread and lethal types of cancer. However, there are currently no drugs or therapeutic strategies that can completely cure GBM. Doramectin (DRM) has a broad range of activities against endoparasites and ectoparasites, and is extensively used in livestock. In the present study, the effect of DRM on the induction of autophagy in U87 and C6 GBM and glioma cell lines, as well as the mechanism of autophagy, were examined. First, transmission electron microscopy, plasmid transfection and western blot analysis demonstrated that DRM could induce autophagy in U87 and C6 cells in vitro. Next, MTT and colony formation assays revealed that DRM-induced autophagy prevented U87 and C6 cell viability and colony formation ratio. In addition, DRM-induced autophagy promoted U87 and C6 cell apoptosis, as indicated by DAPI analysis and flow cytometry. Furthermore, transcriptome analysis demonstrated that DRM modulated a number of genes and pathways involved in autophagy. In a nude mouse xenograft model, immunohistochemical staining and the TUNEL assay demonstrated that the effect of DRM on the tumor was consistent with that in vivo. These data indicated that DRM induced autophagy mainly by blocking the PI3K/AKT/mTOR signaling pathway in GBM cells. DRM-induced autophagy promoted the inhibition of GBM cell proliferation and apoptosis in vitro and in vivo. The present study suggested that DRM may be an effective drug for the treatment of GBM.