EGFR exon 20 insertion mutations (ex20ins) represent the third most common epidermal growth factor receptor (EGFR) mutation subtype in non-small cell lung cancer (NSCLC). Conventional EGFR tyrosine kinase inhibitors (TKIs) and chemotherapy demonstrate suboptimal efficacy in patients harboring EGFR ex20ins mutations. Furmonertinib, a highly selective EGFR-TKI with enhanced blood-brain barrier penetration, presents a potential therapeutic breakthrough. This study aims to evaluate the efficacy of double-dose furmonertinib (160 mg) in advanced EGFR ex20ins-mutant NSCLC patients and explore associations between predictive biomarkers and clinical outcomes.

Recent studies have increasingly emphasized the poorer survival outcomes and reduced treatment responses associated with right-sided colon cancer (RCC). However, the underlying molecular mechanisms remain poorly understood. This study aimed to identify and characterize key biomarkers associated with progression and treatment response in patients with RCC.

Preoperative diagnosis of the nature of PCNSL is crucial for distinguishing it from commonly confused intracranial gliomas, such as glioblastoma multiforme (GBM). The present research aims to sequence peripheral blood exosomal miRNAs from PCNSL and GBM patients, and to identify the discriminative miRNAs with high feasibility. Plasma exosomal RNAs were extracted, and miRNA sequencing was performed. The miRNAs with significant differences were validated by RT-qPCR. A total of 67 miRNAs were significantly different between the PCNSL and GBM groups. Twenty-seven PCNSL and 27 GBM patients were enrolled for the RT-qPCR validation of 10 selected differentially expressed miRNAs. The expression levels of plasma exosomal hsa-miR-148a-3p, hsa-let-7f-5p, hsa-miR-345-5p and hsa-miR-4433b-5p were upregulated in the PCNSL group compared with those in the GBM group (P = 0.019, P = 0.036, P = 0.009, and P = 0.001, respectively). The combined panel comprising hsa-miR-148a-3p, hsa-miR-345-5p, and hsa-miR-4433b-5p demonstrated significantly enhanced diagnostic performance, with an area under the ROC curve (AUC) of 0.791. Immunohistochemistry analysis revealed that the expression level of EGFR in PCNSL was significantly lower than that in GBM. Western blot and RT-qPCR analysis of EGFR expression levels in LN229 cells revealed that miR-148a-3p and miR-4433b-5p downregulated EGFR expression. The results of luciferase reporter assay showed that the relative luciferase activity of HEK293T cells transfected with EGFR-WT was notably suppressed by miR-4433b-5p (P < 0.001). In conclusion, the plasma exosomal hsa-miR-148a-3p, hsa-miR-345-5p, and hsa-miR-4433b-5p might be identified as novel biomarkers for differentiating PCNSL and GBM. The increased expression level of EGFR in GBM may be achieved by the negative regulatory effects of miR-4433b-5p. The diagnostic performance of the miRNA biomarkers still needs to be further verified in larger sample size cohorts.

Multiple myeloma (MM) is an incurable malignancy that arises from precursory conditions, specifically monoclonal gammopathy of unknown significance (MGUS) and smoldering multiple myeloma (SMM). The pathogenesis of MM remains largely elusive, particularly in the context of epigenetics.

Most patients with localized pancreatic ductal adenocarcinoma (PDAC) experience recurrence after resection. Analysis of 744 patients with resected PDAC revealed that patients with initial isolated liver-metastatic recurrence (n = 100) had significantly worse overall survival than those with initial isolated lung-metastatic recurrence (n = 31). Using single-nucleus RNA sequencing in a representative cohort, we found that transcriptional profiles of primary cancer cells with liver-metastatic recurrence and lung-metastatic recurrence were correlated with those of normal liver and lung parenchymal cells, respectively, suggesting adoption of organ-specific metastatic programs at the primary site. These signatures were confirmed in transcriptomes of PDAC lung and liver metastases, primary lung and liver tumors, and organotropic PDAC xenograft models. These signatures were independent of large genomic events, and analysis of large-scale tumor profiling data showed no genetic alterations predictive of recurrence patterns. Additional analyses suggested that metastatic recurrence may be determined early in tumorigenesis and influenced by tumor-infiltrating immune cells. Thus, pre-existing cellular states within primary tumors appear to guide organ-specific metastatic relapse.

Pancreatic ductal adenocarcinoma is among the most lethal malignancies, underscoring the urgent need for a deepened molecular insights and targeted strategies. While microRNAs (miRNAs) are known to regulate key oncogenic pathways in PDAC, most studies overlook the functional heterogeneity between the 3p and 5p strands derived from the same miRNA precursor. Here, we systematically investigate the strand-specific roles of miR-301a-3p and miR-301a-5p in PDAC progression. Using a multidimensional approach, including clinical tissue-based FISH, serum analysis, cell-based assays, and xenograft models, we revealed a consistent pattern of miR-301a-3p upregulation and miR-301a-5p downregulation in PDAC tissues and fluids. Importantly, the ratio of 5p to 3p is negatively correlated with TNM stage, and its combination with CA19-9 significantly improves prognostic prediction. Mechanically, the two strands exert opposing effects on tumor progression via distinct cell death pathways. miR-301a-3p promotes tumorigenesis by targeting ACSL4 to suppress ferroptosis, while miR-301a-5p induces pyroptosis by activating NF-κB pathway through CEACAM6-TNFR1 axis. These findings challenge the prevailing "dominant strand" paradigm and uncover a previously unrecognized antagonism between miRNA strands. By highlighting the risks of indiscriminate miRNA targeting, our work supports the development of strand-specific diagnostic and therapeutic strategies in PDAC. This study advances the concept of chain-level regulation as a foundation for precision oncology.

The next-generation sequencing (NGS)-based Oncomine Dx Target Test (ODxTT) is the standard tool for guiding postoperative adjuvant therapy in patients with non-small cell lung cancer (NSCLC) in Japan. To advance precision oncology, we evaluated the clinical utility of an in-house comprehensive genomic profiling (CGP) assay, Rapid-Neo, as a complementary approach to ODxTT in surgically resected NSCLC. Patients with pathological stage II-III NSCLC who underwent anatomical surgical resection between December 2019 and May 2024 were included. Resected specimens underwent genomic analysis using both ODxTT and Rapid-Neo CGP. We evaluated the mutational concordance and the frequency of additional actionable alterations identified by CGP. Among 68 eligible patients, driver mutation results were concordant in 64 (94.1%) cases. Crucially, CGP rescued one patient for targeted therapy by detecting an EGFR mutation where ODxTT failed due to insufficient DNA. CGP also identified rare EGFR variants not covered by ODxTT in two cases, although it failed to detect a RET fusion in one patient. Furthermore, CGP revealed additional actionable alterations, such as tumor suppressor gene mutations, in 57 patients (83.8%). Our in-house CGP shows high concordance with ODxTT and serves as a powerful complementary tool. These findings support a strategic testing algorithm where CGP is incorporated for patients with negative or inconclusive ODxTT results, or at the time of recurrence, to maximize opportunities for individualized therapy in resected NSCLC.

Triple-negative breast cancer (TNBC), which lacks targeted treatment options, continues to pose a major clinical hurdle. This study investigated the molecular complexities of TNBC, with a focus on m7G modifications, the immune microenvironment, and their influence on patient prognosis. An integrated analysis of m7G-related genes facilitated the creation of a risk model based on differentially expressed m7G-related genes, which effectively stratified patients with TNBC into low- and high-risk groups, with the low-risk group demonstrating a marked survival advantage. Immune cell infiltration analysis revealed substantial differences between risk groups, suggesting a potential association between m7G-related risk and immune landscape modulation. Genomic profiling identified distinct mutational patterns, with TP53 mutations predominant in the high-risk group and PIK3CA mutations in the low-risk group. Chemosensitivity analysis indicated the potential for tailored drug responses, highlighting the utility of the m7G-related risk score in guiding personalized therapeutic approaches. Additionally, a nomogram incorporating the risk score, stage, and M stage was developed, providing a valuable tool for predicting 3-, 5-, and 10-year survival probabilities.

Tumor metastasis is an important risk factor for death in patients with colorectal cancer (CRC). This study aims to explore the effect of CXCL9 and SPP1 (CS) polarity alteration of tumor-associated macrophages (TAMs) on CRC progression and its associated molecular mechanisms. The heterogeneity of cellular subsets between the CRC and Normal groups was analyzed using single-cell RNA sequencing (scRNA-seq) technology. Developmental trajectories of the cellular subsets were plotted using pseudotime analysis, and the differences in enrichment scores among the cellular subsets were analyzed by combining gene set variation analysis (GSVA). Mouse CRC models were constructed, and SPP1- TAMs or SPP1+ TAMs were isolated from mouse tumor tissues by flow cytometry sorting. MC38 cells were treated with the JAK/STAT3 inhibitor WP1066 and co-cultured with SPP1+ TAMs. MC38 cell viability was detected by the cell counting kit-8 (CCK-8) assay. The apoptosis rate of MC38 cells was detected by TdT-mediated dUTP nick end labeling (TUNEL) staining. The expression of JAK/STAT3 pathway proteins was detected using western blot (WB), and the expression of epithelial-mesenchymal transition (EMT)-related proteins was detected using immunofluorescence. There were significant differences in SPP1+ TAMs between the CRC and Normal groups by scRNA-seq, and JAK/STAT3 signaling pathway had significant scores in each cell subset. In vitro assays showed that compared with the Negative group, MC38 cells in the Positive group exhibited higher viability and lower apoptosis rate, protein levels of p-JAK2 and p-STAT3 were significantly up-regulated, and the EMT levels were increased. In contrast, after the cells were co-treated with WP1066 on the basis of co-culture with SPP1+ TAMs, MC38 cell viability was decreased, and apoptosis was increased; the levels of JAK2, STAT3, and their phosphorylation were decreased, and the EMT process was inhibited. Therefore, SPP1+ TAMs promote CRC cell proliferation and EMT by activating JAK2/STAT3 signaling pathway.

One of the less invasive methods in cancer treatment is the use of Extremely Low-Frequency Pulsed-Electromagnetic Fields (ELF P-EMF). In this study, after culturing and proliferating DU-145 prostate cancer cells, they were exposed to ELF P-EMF with different intensities and times. Then, the cells' viability was examined by applying the MTT test, and their level of apoptosis/ necrosis was analyzed using a flow cytometry test. Then, for the studied groups, RNA extraction steps were performed, and cDNA was subsequently synthesized. Finally, using Real-time PCR, the expression levels of key genes such as PTEN, BAX, BCL-2, and MIR-21 in the targeted cancer cells were examined, and statistically, the significance of their expression differences with the control group was measured using SPSS software. The results indicated that by increasing the ELF P-EMF intensity from 22.6 to 35 mT and the duration of exposure, the mortality rate of cancer cells increased significantly. In addition, exposing cells of this line to ELF P-EMF could induce apoptosis in these cells. Also, applying fields with intensities of 22.6 and 35 mT led to a significant increase in the expression of the tumor suppressor gene PTEN and an increase in the expression of the apoptosis-inducing gene BAX. In addition, applying the field significantly reduced the expression of the oncogene MIR-21 and, to some extent, reduced the expression of the BCL-2 gene, which blocks apoptosis.

The adenoma-adenocarcinoma pathway represents a crucial mechanism underlying the development of colorectal precancerous lesions, encompassing approximately 85%-90% of colorectal cancer (CRC). Elucidating the molecular mechanisms underlying colorectal cancer progression is of paramount importance for achieving early and accurate diagnosis as well as effective treatment. We collected peripheral blood mononuclear cells (PBMC) from healthy controls, adenoma patients, and adenocarcinoma patients, and performed transcriptomic profiling to characterize dynamic gene expression during carcinogenesis. Diagnostic potential was assessed using receiver operating characteristic (ROC) analysis, and a random-forest model was trained to classify disease status. The screening identified genes with consistent expression changes as potential early diagnostic markers, and further exploration of their functions and significance in CRC is conducted through analysis of the TCGA database. The findings revealed that the progression of precancerous lesions in the "Normal-Adenoma-Cancer" (N-A-C) sequence was accompanied by a sustained enhancement of the immune response. Notably, HECW2, WARS1, SLC16A3, SECTM1, IFITM3, ADAMTSL4, FCGR1A, F2RL1, OPLAH, SERPINA1, FCGR1CP showed consistent upregulation with promising diagnostic performance. In our PBMC cohort, the random-forest classifier achieved an accuracy of 93.62%, indicating potential for distinguishing cancer from precancerous lesions. The bioinformatics analysis revealed a significant association of these genes with DNA methyltransferase, DNA mismatch repair, m6A regulator, tumor mutational burden (TMB) and microsatellite instability (MSI). Furthermore, a detailed analysis was further performed on WARS1. In the "N-A-C" sequence, WARS1 exhibited a significant upregulation in both blood and tissues, demonstrating a positive correlation with augmented infiltration of immune cells, activation of stromal and immune responses, as well as heightened activity during the cancer immune cycle. However, it demonstrates a declining trend in the progression of CRC from stage I to IV, which may be intricately associated with the metastasis of CRC. The WARS1 can serve as a reliable indicator of the immune response in CRC, thereby demonstrating its potential to impede tumorigenesis or metastasis.

Neuroendocrine neoplasms (NEN) are thought to originate from diffuse neuroendocrine networks and therefore most frequently arise in the gastrointestinal tract and lungs. The liver is a frequent site of metastasis of NEN but also the existence of primary hepatic NEN has been proposed. Due to the impact on disease management, it is urgently required to discriminate the origin of hepatic NEN metastases and to identify clinically relevant subgroups. Using a comprehensive set of NEN (N = 212) from two independent cohorts, we show that the DNA methylation profiles of NEN of distinct anatomical localizations differ significantly and primary tumor-metastasis pairs cluster together, enabling the identification of the tumor origin. Furthermore, the subgroup of hepatic NEN without clinically detectable primary tumor, thus classified as primary hepatic NEN, does not form a distinct cluster by DNA methylation analysis but colocalizes with various subgroups of extrahepatic NEN. Organ-specific subtyping of NEN delineates a foregut-like epigenetic profile for hepatic NEN with unknown primary. We propose a classifier with high prediction accuracy for each of the different organ sites. In conclusion, our results demonstrate that DNA methylation profiling enables precise prediction of NEN origin and suggests that a substantial proportion of presumed primary hepatic NEN may in fact represent misclassified secondary hepatic NEN of unknown primary.

Environmental stressors such as radiation, pH shifts, temperature variations, and electromagnetic fields can trigger intracellular oxidative stress, upregulating voltage-gated ion channel (VGIC) gene expression. This paper presents a hybrid modeling framework integrating Hodgkin-Huxley-based electrophysiological simulations with redox-sensitive transcriptional feedback to investigate how reactive oxygen species (ROS) modulate calcium signaling and drive electrophysiological reprogramming. In healthy epithelial cells (MCF-10A), sustained oxidative perturbations induce non-voltage-gated calcium influx, mitochondrial ROS generation, and VGIC transcription, shifting membrane potential from non-excitable to excitable states. Repeated ROS or thermal pulses promote progressive VGIC expression, depolarization, mRNA accumulation, and genomic instability. A Transformer-Long Short-Term Memory (LSTM) model, trained on simulated ROS-VGIC-Vm-mutation trajectories and human datasets (GSE45827), achieved >90% accuracy in predicting tumorigenic transformation. This framework enables simulation-guided drug target identification, ion channel parameter optimization, and AI-assisted screening of VGIC-modulating compounds, bridging systems biology with predictive oncology and informing electrophysiology-based therapeutic design.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most aggressive and lethal cancers, with limited therapeutic options and a dismal prognosis. A critical driver of its progression is mutant p53 (mutp53), which alters the tumor microenvironment (TME) by influencing crucial pro-tumoral signaling factors. Given the potential of secretome profiling to reveal novel biomarkers and druggable targets, we investigated the role of the mutp53-driven secretome in PDAC cells and its implications for disease progression. Through mass-spectrometry (MS) analysis, we identified a set of secreted proteins modulated by mutp53, with the nuclear high mobility group A1 (HMGA1) serving as a central regulator. HMGA1 is a transcription factor involved in several cellular processes and found to be upregulated in different tumors, but its extracellular role in cancer remains largely unexplored. We demonstrate that mutp53-driven HMGA1 secretion promotes PDAC cell hyperproliferation, where HMGA1 deficiency significantly impairs tumor growth highlighting a critical role of this protein in tumor aggressiveness. Notably, we discovered that chemotherapy enhances HMGA1 secretion specifically in TP53-mutant PDAC cells through a mechanism dependent on Casein Kinase 2 (CK2) activity. To unravel the downstream oncogenic signaling triggered by secreted HMGA1, we conducted phosphoproteomic analysis, identifying hyperphosphorylation of Nucleophosmin 1 (NPM1), as a pivotal event that further amplifies tumor cell proliferation. Collectively, our findings reveal that a panel of chemotherapeutic agents stimulate a novel mutp53-dependent CK2-HMGA1-NPM1 axis that fuels PDAC proliferation in an autocrine/paracrine manner. Targeting this pathway at multiple levels emerges as a promising therapeutic strategy to counteract mutp53-driven tumor progression and improve patient outcomes.

Osteosarcoma is an extremely aggressive malignant tumor, which is quite common among children and has a high rate of disability and mortality. Tripartite Motif Containing 17 (TRIM17) is a member of the TRIM protein family and exhibits E3 ubiquitin ligase activity. In recent years, TRIM17 has been implicated in the development of various tumors, particularly in cancer cell clonability and survival potential and drug resistance; however, its regulatory mechanism in osteosarcoma progression remains poorly understood. We found that TRIM17 was significantly upregulated in osteosarcoma tissues and cells. Survival analysis revealed that TRIM17 was associated with poor prognosis in osteosarcoma patients. The higher the expression level of TRIM17, the worse the prognosis. Its expression was an independent prognostic factor for osteosarcoma patients. The effects of TRIM17 on osteosarcoma cell clonability and survival potential, migration, and invasion were assessed through phenotypic assays. The results showed that the downregulation of TRIM17 significantly inhibited osteosarcoma cell clonability and survival potential, migration, and invasion, whereas its overexpression promoted these processes. FTO is an m6A methyltransferase and has been identified as a new target for TRIM17. Mechanistically, TRIM17 promotes the ubiquitination and degradation of FTO protein, enhances PDK1 mRNA stability via N6-methyladenosine (m6A) modification, and subsequently promotes phosphorylation-dependent activation of the AKT/mTOR signaling pathway, thereby driving osteosarcoma malignancy. In summary, our findings suggest that TRIM17 may serve as a potential prognostic marker and therapeutic target for osteosarcoma.

Pluripotent cancer stem cells play a pivotal role in inducing phenotypic plasticity across various cancer types, including bladder cancer. This plasticity, crucial for cancer progression, is largely regulated by epigenetic modifications including N6-methyladenosine (m6A) in RNAs. However, the role of the m6A reader protein YTHDC2 in this process remains poorly understood. In this study, we uncovered that the depletion of YTHDC2 significantly increased the pool of bladder cancer stem cells (BCSCs), resulting in a phenotypic shift towards a more invasive subtype of bladder cancer. This shift was characterized by enhanced proliferation, migration, invasion, and self-renewal capabilities of cancer cells, highlighting YTHDC2's function as a tumor suppressor. Mechanistically, YTHDC2 recognized and bound to m6A-modified SOX2 mRNA, resulting in translational inhibition of SOX2. In conclusion, our study identifies YTHDC2 as a tumor suppressor in bladder cancer through inhibiting SOX2-mediated cell pluripotency and underscores the therapeutic potential of targeting the YTHDC2-SOX2 axis in bladder cancer.

Osimertinib resistance poses a major challenge in treating advanced EGFR-mutant lung adenocarcinoma (LUAD). Exosomes, key mediators of intercellular communication, may contribute to drug resistance, but their specific role in osimertinib resistance remains unclear. This study aimed to elucidate the function and mechanisms of hypoxia-induced exosomes (HExo) in promoting osimertinib resistance in LUAD, with the goal of identifying potential diagnostic biomarkers and therapeutic targets. Cell survival under osimertinib treatment was analyzed using CCK8 and colony formation assays, while exosomal LUCAT1 was identified via RNA-seq and validated by qRT-PCR. Biological roles of LUCAT1 were assessed through in vitro and in vivo experiments, including immunoblotting, RNA immunoprecipitation (RIP)-qPCR, xenograft tumor models, and organoid models. Results demonstrated that hypoxia-induced exosomal LUCAT1 reduced the sensitivity of recipient LUAD cells to osimertinib. Mechanistically, LUCAT1 promoted osimertinib resistance by preventing c-MET degradation and activating downstream AKT/mTOR and ERK pathways. Targeting c-MET effectively restored osimertinib sensitivity in resistant cells. Moreover, higher levels of exosomal LUCAT1 were significantly associated with poor therapeutic responses to osimertinib in patients. In conclusion, hypoxia-induced exosomal LUCAT1 drives osimertinib resistance by stabilizing c-MET and activating its downstream pathways. Plasma exosomal LUCAT1 levels are closely linked to osimertinib resistance and may serve as an ideal liquid biopsy target for monitoring patient response.

Colorectal cancer (CRC) is one of the most common malignant diseases which imposing a substantial burden on global health and economies. We previously identified a novel molecule, FAM227A, through functional screening using an shRNA retroviral library.

Breast cancer (BC) is the most commonly diagnosed cancer in women globally, highlighting the need for new therapeutic targets. This study aims to clarify the role of pleckstrin-2 (PLEK2) in BC progression. We particularly focus on how the transcription factor Homeobox D9 (HOXD9) regulates PLEK2 and its influence on the AKT signaling pathway. We employed quantitative PCR, Western blotting, gene knockdown and overexpression techniques, scratch assays, Transwell assays, and biochemical analyses to investigate the effects of PLEK2 on cell migration, invasion, and metabolic activity in BC cells. We found that PLEK2 was significantly overexpressed in BC tissues compared to normal tissues in the database, and this overexpression was associated with poor prognosis. Functional assays showed the modulatory effect of PLEK2 on glycolytic activity, migration, and invasion of breast cancer cells. Mechanistically, PLEK2 regulates the PI3K/AKT/mTOR pathway, HIF-1α protein levels, and the expression of its target glycolytic genes, thereby promoting glycolysis critical for cancer progression. Notably, HOXD9 was identified as a key regulator of PLEK2 expression, confirmed by luciferase assays and ChIP-qPCR. Overexpression of HOXD9 restored the migratory and invasive abilities of PLEK2-silenced cells, indicating a significant interplay between HOXD9 and PLEK2 in breast cancer. In conclusion, our study highlights the pivotal role of PLEK2 in BC cell dynamics and introduces HOXD9 as a novel regulator of PLEK2. Targeting the HOXD9/PLEK2/AKT signaling axis may offer a promising therapeutic strategy for treating breast cancer.

Positive pleural lavage cytology (PLC) indicates microscopic pleural dissemination and is associated with poor prognosis; however, it is not incorporated into the current TNM staging system and rarely guides adjuvant therapy. The prognostic importance of PLC positivity in patients with EGFR-mutated non-small cell lung cancer (NSCLC) remains unclear.