Ewing sarcoma (ES) is an aggressive bone and soft tissue neoplasm characterized by EWSR::ETS rearrangements whose cellular origin remains unclear. EWS::FLI1 expression in human pediatric mesenchymal stem cells (MSCs) induces a transcriptional response distinct from that of human adult MSCs, but fails to form tumors. Here we show that EWS::FLI1 expression in human embryonic mesenchymal stem cells (heMSCs) results in the acquisition of an ES transcriptome, with the oncogene not preferentially binding to gene promoters, but to intronic and intergenic microsatellites. In heMSCs, EWS::FLI1 directly regulates the expression of the DNA repair protein BRCA1, although cells expressing EWS::FLI1 show DNA damage. Xenografting of EWS::FLI1-transduced heMSCs results in the formation of tumors expressing characteristic ES markers. In summary, we show that EWS::FLI1 enforces an aberrant transcriptome and solely is able to endow transforming capacity when expressed in undifferentiated, early heMSCs.

The development of traditional protein-targeted cancer therapies is a slow and arduous process, often taking years or even decades. In contrast, RNA-based therapies targeting crucial microRNA (miRNA) offer a faster alternative due to the sequence-specific nature of miRNA inhibitor binding. This, combined with the capacity of individual miRNA to influence multiple cellular pathways, makes these small RNA attractive targets for cancer therapy. While miRNA are known to be dysregulated in prostate cancer (PCa), identifying their individual contributions to disease progression and the identification of therapeutically actionable miRNA targets in PCa has been challenging due to limited profiling and lack of screening tools. To address this need, we developed miRKOv2, a miRNA-only CRISPR knockout library enabling systematic, genome-wide loss-of-function screens to identify miRNA essential for PCa cell survival. Our screens uncovered 70 potential essential miRNA candidates, with miR-483 demonstrating the most significant impact on PCa cell viability. Functional characterization revealed that miR-483 disruption potentiated apoptosis in PCa cell lines. Mechanistically, we uncovered a novel regulatory axis wherein miR-483-3p directly modulates a BCLAF1/PUMA/BAK1 apoptotic signaling network, highlighting its critical role in maintaining PCa cell survival. Our findings provide novel insights into the complex regulatory role of miRNA in PCa progression and offer a potential therapeutic strategy for targeting miRNA-mediated pathways in metastatic disease.

Castration-resistant prostate cancer (CRPC) is the advanced stage of prostate cancer (PCa) progression, characterized by limited therapeutic options and significant challenges from drug resistance development. We show that PFKFB3, an essential regulator of glycolytic metabolism, is significantly upregulated in PCa tissues and CRPC cell lines, where it plays a pivotal role in driving CRPC progression. Knockdown of PFKFB3 or inhibition by a small molecule inhibitor significantly inhibits the growth and invasion of CRPC cells, whereas overexpression promotes malignant behaviors. Mechanistically, PFKFB3 modulates the PI3K/Akt-Wnt/β-catenin pathway, resulting in enhanced tumor cell proliferation. Additionally, combining a PFKFB3 inhibitor with docetaxel produces synergistic anti-CRPC effects and reduces toxicity. Therefore, PFKFB3-mediated metabolic reprogramming underlies CRPC progression, highlighting its potential as a therapeutic target and emphasizing the need for further exploration in the development of safe and effective PFKFB3 inhibitors for precise targeted therapy in CRPC.

Metastatic uveal melanomas are highly resistant to all existing treatments. To identify actionable vulnerabilities, we conducted a CRISPR-Cas9 knockout screen using a library composed of chromatin regulators. We revealed that the lysine methyltransferase, SETDB1, plays a critical role in metastatic uveal melanoma cell proliferation and survival. Functionally, SETDB1 deficiency induces a DNA damage response, senescence-like state and growth arrest. Knockdown of SETDB1 is associated with a decreased expression of genes related to replication and cell cycle. Moreover, deficiency in CDC6, an essential regulator of DNA replication, phenocopies SETDB1 inhibition. Using a pre-clinical model, we further demonstrated that anti-SETDB1 therapy impairs tumor growth in vivo. Therefore, we not only provide evidence that SETDB1 plays a critical role in metastatic uveal melanoma cell growth, but we also identify SETDB1 as a novel relevant therapeutic target for the treatment of metastatic uveal melanoma.

Ubiquitin-like PHD and RING finger domain-containing protein 1 (UHRF1) is an important epigenetic regulatory factor that is highly expressed in various cancers and participates in tumorigenesis and progression. However, the role and molecular mechanisms of UHRF1 in ovarian cancer (OC) remain unclear. Through survival analysis, cellular functional experiments, and animal studies, we identified UHRF1 as a key gene influencing OC progression and prognosis. Hypoxia-inducible factor-1 (HIF-1α), a well-known pro-cancer molecule, undergoes classic degradation via the ubiquitin-proteasome pathway. We discovered that UHRF1 interacts with HIF-1α, affecting its hydroxylation level, thereby inhibiting HIF-1α polyubiquitination and degradation. Functional experiments revealed that knocking down HIF-1α in stable UHRF1-overexpressing cell lines significantly reversed the malignant phenotype of OC cells. Furthermore, UHRF1 can also regulate the expression of key downstream molecules such as GLUT1, HK2, LDHA, and VEGFA by modulating HIF-1α, thus influencing tumor cell metabolism and angiogenesis. In summary, our findings suggest that UHRF1 plays a crucial role in the development of OC by regulating the expression of HIF-1α.

Chordoma is a rare, slow-growing malignant tumor originating from embryonic notochord remnants and is often found in the sacrum or skull base. It is categorized into conventional, poorly differentiated, and dedifferentiated types, with the conventional type being the most common. Owing to its location near critical structures, chordoma has a high rate of local recurrence, making new therapeutic targets essential. The proteasome system, which is responsible for degrading intracellular proteins, plays a vital role in maintaining cellular function. REGγ, a proteasome activator, mediates ubiquitin-, and ATP-independent protein degradation and is overexpressed in various cancers. However, its role in chordoma remains unexplored. Ras GTPases, including RIT1, are involved in cancer progression, and understanding their involvement in chordoma could provide therapeutic insights. This study identified REGγ as a potential therapeutic target for chordoma. REGγ was found to be upregulated in chordoma, and high REGγ expression was correlated with poor clinical outcomes. It promotes cell proliferation and migration, and inhibits apoptosis, while influencing osteoclast differentiation. Mechanistically, REGγ regulates chordoma progression through the ubiquitin- and ATP-independent degradation of RIT1, which modulates the RIT1-MAPK pathway. Inhibition of RIT1 in REGγ-knockdown cells and patient-derived organoids alleviated these effects, suggesting that targeting REGγ may be a promising strategy for chordoma treatment.

Accumulation of genetic mutations in malignant myeloid precursor cells leads to an extremely poor prognosis for patients with acute myeloid leukemia (AML). Immunogenic neoantigens recognized by T cell receptor (TCR) can elicit effective immune responses against malignant cells with corresponding somatic mutations. To broaden the range of targeted treatments for AML, in this study, we explored the feasibility of immunotherapy targeting neoantigens arising from recurrent mutations, which are exclusively present on leukemic cells. We used data-driven methods to select seven neoantigens from four frequently mutated genes (NPM1, FLT3, TP53, and DNMT3A) associated with HLA-A*02:01-positive AML patients. Functional assays demonstrated that neoantigens derived from NPM1/W288fs, FLT3/D835H, and FLT3/D835Y were shown to induce specific T cell responses in AML patients. We further identified the specific TCR sequences from healthy donors capable of recognizing these neoantigens. In-depth studies of their specific T cells revealed the presence of dominant αβTCRs that could specifically recognize NPM1/W288fs and FLT3/D835H in an HLA-A*02:01-restricted manner. T cells engineered with each αβTCR selectively recognized and killed HLA-A*02:01-positive AML targets endogenously expressing corresponding mutations. Overall, our findings support the clinical translation of adoptive neoantigen-specific TCR-engineered T cells as a novel therapeutic strategy for treating AML.

Therapeutic resistance and recurrence in human epidermal growth factor receptor 2-positive breast cancer (HER2 + BC) remain critical challenges that portend poor patient outcomes. Dysregulated autophagy and lipid metabolism contribute to tumor progression, yet the crosstalk between these pathways is poorly understood. This study investigates the role of transmembrane 9 superfamily member 1 (TM9SF1) in lipophagy and lipid metabolic reprogramming in HER2 + BC under metabolic stress. Clinically, TM9SF1 was significantly upregulated in HER2 + BC tissues and correlated with poor prognosis. Functionally, its expression correlated with markers of enhanced autophagy and lysosomal lipid catabolism, and it promoted tumor cell proliferation in vitro and in vivo. Conversely, TM9SF1 knockdown suppressed lipophagy under both basal and starvation conditions, inhibiting lipid droplet (LD) hydrolysis and the conversion of triglycerides to free fatty acids. This suppression was phenotypically characterized by LD accumulation, reduced autophagosomes and lipophagosomes, and altered enzymatic and lipidomic profiles. Mechanistically, TM9SF1 sustained lipophagy by promoting the phosphorylation of AMP-activated protein kinase at Thr172 and UNC-51-like kinase 1 at Ser555. Consequently, TM9SF1 was pivotal for lipid metabolic reprogramming, maintaining energy homeostasis and enhancing adaptation to nutrient deprivation through lipophagy. Overall, our findings identify TM9SF1 as a key HER2 + BC-associated regulator that drives lipophagy via the AMP-activated protein kinase-UNC-51-like kinase 1 pathway, facilitating LD turnover and free fatty acids utilization to sustain energy homeostasis in HER2 + BC. This work establishes a critical link between malignant phenotypes and metabolic resilience. Targeting this regulatory network represents a promising strategy to dismantle the metabolic scaffolds underlying HER2 + BC aggressiveness and therapeutic resistance.

Cutaneous lesions of mastocytosis (CLM) are a hallmark for the diagnosis of mastocytosis. The nomenclature of the term "cutaneous mastocytosis" (CM) is ambiguous as it is used both for classification excluding and as a morphologic description including systemic variants of mastocytosis. Recognition of CLM may be delayed by clinically subtle and very heterogeneous presentations. Distinguishing monomorphic versus polymorphic maculopapular CM may be challenging. Less common presentations require a clearly positive Darier's sign, histopathological confirmation, and/or skin D816V kit mutational analysis. Management of patients with CLM requires multidisciplinary therapy, adequate staging, and follow-up to recognize complications.

Immune checkpoint blockade (ICB) therapy has demonstrated significant clinical potential in a variety of cancers; however, its efficacy in clear cell renal cell carcinoma (ccRCC) remains suboptimal. In ccRCC, an increased infiltration of CD8+ T cells does not necessarily correlate with improved prognosis, indicating the presence of unique immune evasion mechanisms within the tumor microenvironment (TME).

Despite progress in immunotherapy for several solid tumors, pancreatic ductal adenocarcinoma (PDAC) remains largely unresponsive, primarily due to its profoundly immunosuppressive tumor microenvironment (TME) characterized by limited CD8+ T cell infiltration. Novel strategies are needed to overcome this immune resistance and enhance the efficacy of checkpoint blockade.

Metastasis is the primary cause of poor prognosis and high mortality in lung cancer. Surgery with postoperative adjuvant chemotherapy is the standard treatment for patients with stage IIA-IIIA lung cancer with negative driver genes. However, recurrence rates remain significant. In China, traditional Chinese medicine shows potential as an adjuvant therapy to reduce treatment toxicity and improve clinical efficacy.

Cancer dormancy refers to a reversible state where cancer cells enter a quiescent phase, allowing them to evade therapeutic interventions and remain undetected. This state can lead to potential reactivation years later, resulting in relapse and metastasis. This phenomenon presents a significant challenge in cancer treatment, as dormant cells often exhibit resistance to conventional therapies. Recent studies emphasize the crucial role of metabolic reprogramming in regulating cancer dormancy, closely interacting with the tumor microenvironment. Dormant cancer cells undergo metabolic adaptations that enable their survival in a hostile tumor microenvironment. These adaptations include a decreased reliance on glycolysis and an increased dependence on oxidative phosphorylation and fatty acid oxidation. Exosomes, extracellular matrix, and cancer-associated fibroblasts dynamically regulate these metabolic states by mediating intercellular communication and modulating the biochemical and mechanical properties of the tumor microenvironment. In parallel, epigenetic regulation fine-tunes metabolic gene expression, reinforcing the dormant phenotype and enabling plastic transitions between dormancy and proliferation. Additionally, these cells utilize autophagy to recover nutrients and manage microenvironmental stress. These metabolic changes help dormant cells maintain a low metabolic state while preserving their ability to reactivate when conditions become favorable. Understanding the relationship between dormancy and metabolism offers new therapeutic opportunities aimed at targeting metabolic pathways to prevent relapse and metastasis. This review explores the mechanisms of metabolic reprogramming in dormancy induction, maintenance, and escape, providing insights into potential therapeutic strategies.

Tumor protein p53 (TP53) plays a crucial role in regulating apoptosis and maintaining genomic stability, making TP53-mediated cell death an important diagnostic marker for breast cancer therapy. This review compiles laboratory methodologies for the application of TP53 apoptosis assays to standard clinical specimens. Modern diagnostic techniques enhance the management of formalin-fixed paraffin-embedded (FFPE) and fresh-frozen (FF) samples, as well as liquid biopsy systems, to preserve nucleic acid integrity for subsequent analysis. Analytical platforms, including targeted next-generation sequencing (NGS), droplet digital polymerase chain reaction (ddPCR), and multiplex reverse transcription (RT), exhibit high sensitivity, achieving detection limits below 0.5 % variant allele fraction (VAF), and are supported by orthogonal confirmation methods. Immunohistochemical (IHC) assays employing proteins such as p53, BCL2-associated X protein (BAX), and cleaved caspase-3 provide quantifiable indicators of apoptosis in standard pathological sample. Integrating genomic variants with pathway-level signatures enhances the clinical interpretation of prognosis, therapeutic decision-making, and monitoring of minimal residual disease (MRD). Achieving laboratory accreditation necessitates the implementation of standardized preanalytical controls, validation of methods with clearly defined analytical performance criteria, and establishment of consistent reporting systems. In metastatic disease cases, there is an 87 % concordance between tissue and circulating tumor DNA (ctDNA) across various platforms, supporting its clinical application. This approach translates TP53-apoptosis data from research tools into validated tests for clinical application in breast cancer treatment and future research initiatives.

Neuroblastoma is one of the highest incidence solid tumors of childhood. Therapy for neuroblastoma with MYCN proto-oncogene (MYCN) amplification is of great challenge, due to its aggressive invasion and chemoresistance. In our study, differentially expressed genes (DEGs) between MYCN-amplified and non-amplified neuroblastoma tissues were analyzed using "limma" R package. Then, DEGs were screened by silencing or enhancing expression of MYCN in neuroblastoma cells, and distal-less homeobox 5 (DLX5) was identified to be the downstream target gene. The positive correlation between DLX5 and MYCN expression was confirmed in neuroblastoma tissues and cell lines. Overall survival analysis suggested that high expression of DLX5 predicted poor clinical outcome.Then, DLX5 was demonstrated to promote proliferation and migration of neuroblastoma. Gene set enrichment analysis revealed that base excision repair and nucleotide excision repair processes were enriched in samples with DLX5 high expression. Thus, the half-maximal inhibitory concentration was measured and cisplatin treatment assay was carried out. The results revealed DLX5 promoted cisplatin chemoresistance of neuroblastoma. To explore the mechanism, mRNA sequencing was conducted and AKT signaling was suggested to be the mainly regulatory pathway, which was further verified in cells and tumor tissues. Then, we proved that heart and neural crest derivatives expressed 2, paired like homeobox 2B, GATA binding protein 3, and MYCN coregulated the expression of DLX5 at the transcriptional level, and identified the direct binding site of MYCN on DLX5 promoter. Finally, we demonstrated that DLX5 which was transcriptionally activated by MYCN, promoted growth, metastasis and chemoresistance of neuroblastoma through enhanced AKT phosphorylation. The findings in our study provided new insight for progression and chemoresistance of neuroblastoma.

Breast cancer is a leading cause of cancer-related mortality in women worldwide. Aberrant alternative splicing contributes to BRCA progression, but the role of splicing factors such as QKI and their regulation of specific splicing events, such as those in the NIN gene are poorly understood. Through multi-omics analysis of TCGA-BRCA data, we identified splicing factor QKI as a key regulator of the exon-skipping event NIN_ES_27495, which was significantly associated with poor patient prognosis. Mechanistically, QKI directly bound to NIN pre-mRNA and promoted exon 18 inclusion, generating the oncogenic isoform exon 18-included NIN isoform. Functional experiments demonstrated that depletion of exon 18-included NIN isoform suppressed proliferation, migration, and invasion in breast cancer cells, induced S-phase arrest using dysregulation of the CDK2/Cyclin A axis, and significantly inhibited tumor growth in vivo. These findings revealed that the QKI/exon 18-included NIN isoform axis is a critical driver of breast cancer progression, highlighting its potential as a therapeutic target for BRCA treatment.

Radiation therapy for cancer takes advantage of the higher sensitivity of tumor cells to radiation compared to normal tissue, but some cancers, such as glioblastoma (GBM) and malignant melanoma, acquire radiation resistance (radioresistance), rendering treatment ineffective. Radioresistance is characterized by strong activation of DNA repair mechanisms in response DNA damage induced by radiation, together with possession of malignant property such as enhanced invasiveness and metastasis, though the molecular mechanisms involved remain to be fully established. Here, we show that P2X7 receptor-specific inhibitors suppress the γ-irradiation-induced DNA damage response (DDR) and enhance cell death of A172 GBM cells. In contrast, ATP, a P2X7 receptor ligand, promotes the DDR and suppresses cell death. Irradiation immediately induced ATP release from cells, and P2X7 receptor inhibitor suppressed the release of ATP. Furthermore, P2X7 receptor inhibitors suppress the release of high mobility group box 1 (HMGB1), which is known to promote cancer cell migration. Inhibitors of the receptor for advanced glycation end products (RAGE) also suppress ATP-induced cell motility, indicating that the P2X7-HMGB1-RAGE pathway contributes to radiation-induced malignant transformation. These data indicate firstly that the P2X7 receptor promotes the γ-irradiation-induced DDR, leading to increased resistance of GBM cells to γ-radiation-induced death, and secondly that the P2X7 receptor and extracellular ATP may be involved in the γ-irradiation-induced acquisition of malignant property such as cytoskeletal changes and enhanced motility in GBM cells.

Eleven Flavored Shenqi Tablets (EFST) is a classical multi-herbal prescription in traditional Chinese medicine, traditionally prescribed for kidney tonification and systemic regulation. Despite its reported anti-inflammatory and antiproliferative properties, the therapeutic value of EFST in clear cell renal cell carcinoma (ccRCC) remains unclear.

First-line osimertinib is one of the standards of care for EGFR-mutant advanced non-small cell lung cancer (NSCLC), but most patients eventually progress. After progression, carboplatin plus pemetrexed remains the most widely used second-line therapy, yet robust real-world data in this setting are scarce.

N-4 cytidine acetylation (ac4C), facilitated by N-acetyltransferase 10 (NAT10), is essential in regulating mRNA stability, rRNA biogenesis, cell proliferation, and epithelial-mesenchymal transition. However, the prognostic implications of NAT10-mediated ac4C modifications in colon adenocarcinoma (COAD) remains unclear. Our study demonstrated that NAT10 facilitates tumor growth and modulates an immune response in COAD. Moreover, we identified 2156 ac4C-related genes from the literature and obtained COAD patient information from TCGA. We constructed a prognostic risk model based on NAT10-mediated ac4C-related genes, thoroughly examining biological pathways, immune infiltration, tumor mutation burden, and drug sensitivity. The NAT10-related genes risk model developed in this study accurately predicts clinical outcomes in COAD, offering potential prognostic biomarkers and therapeutic targets. It helps identify candidates for immune checkpoint inhibitors (ICIs) and targeted therapies, offering insights for personalized precision treatment of COAD.