This study characterized the genomic profile of esophageal squamous cell carcinoma (ESCC) via circulating tumor DNA (ctDNA) to identify predictive biomarkers for efficacy of definitive radiotherapy combined with systemic therapy. In this prospective cohort of 40 ESCC patients, 7 tumor tissues and 71 serial plasma samples were analyzed using a 340-gene panel. TP53 (45%) and PIK3CA (12.5%) were the most frequent SNVs, followed by NOTCH1/2 (7.5%). The 11q13.2-13.4 amplification (including CCND1/FGF19) occurred in 12.5% of cases. TP53 mutation rate was significantly higher in cT4 than cT2-3 stages (60% vs. 20%, p = 0.010). Notably, patients with persistent TP53 positivity posttreatment had significantly shorter PFS than those converting to negativity (HR = 5.090, p = 0.035); the 2-year PFS rates were 33.3% vs. 75%, respectively, with OS showing a borderline difference (p = 0.053). Posttreatment ctDNA-based minimal residual disease (MRD) positivity was 18.8%, correlating with a trend toward worse PFS (HR = 3.027, p = 0.067). Among the 31 patients with paired pre- and post-treatment plasma samples, the positive detection rate of nucleosome-positioning (left-shifted profile) decreased from 67.9% at baseline to 35.7% after treatment (χ[2] = 5.793, p = 0.016). This work based on plasma ctDNA profiling demonstrated that posttreatment clearance of TP53 mutations may represent a favorable prognostic biomarker in ESCC treated with definitive radiotherapy and systemic therapy. ctDNA-derived MRD status and nucleosome positioning constitute promising non-invasive biomarkers for treatment response monitoring and risk stratification for ESCC. Trial Registration:This study was registered with the Chinese Clinical Trial Registry (ChiCTR) on December 6, 2019 (Registration No. ChiCTR1900027936).

PABPC1, long recognized as a constitutive translational housekeeper, has emerged as a stress-responsive translational reprogramming hub in cancer. This review synthesizes recent paradigm shifts that redefine PABPC1 function: From global translation maintenance to selective oncogenic mRNA amplification via liquid-liquid phase separation; from binary protein-RNA interactions to complex regulatory networks involving lncRNAs and circRNAs that modulate PABPC1 stability, localization, and target selectivity; and from diffuse cytoplasmic distribution to functionally compartmentalized condensates. PABPC1 integrates diverse post-translational modifications, including SUMOylation, acetylation, and ubiquitination, that enable stress adaptation, and actively regulates alternative polyadenylation to influence 3'UTR length and gene expression. Positioned at the convergence point of multiple oncogenic signaling pathways (PI3K/AKT, Wnt/β-catenin, NF-κB, and etc.), PABPC1 converts upstream signals into translational outputs that drive tumor progression, metastasis, and therapy resistance across malignancies. Its functional duality, context-dependent activities, and paralog-specific roles underscore both the complexity and therapeutic potential of targeting this multifaceted RNA-binding protein.

Immune checkpoint inhibitors (ICIs) have revolutionized cancer treatment by reinvigorating antitumor immunity; however, a substantial proportion of patients show primary resistance or later relapse. To investigate T cell states associated with reduced ICI efficacy, we performed single-cell RNA sequencing (scRNA-seq) on tumor-infiltrating lymphocytes (TILs) from a patient experiencing rapid recurrence under PD-1 blockade. Our analysis identified elevated expression of the ZFP36 family of RNA-binding proteins within exhausted CD8[+] T cells (Tex cells) in this progressing tumor. The ZFP36 family is known to destabilize target mRNAs by binding to AU-rich elements in their 3' untranslated regions (3' UTRs). In EL4 T cells, Zfp36 overexpression accelerated the decay of the effector cytokine mRNAs Tnf, Il2, and Ifng, thereby impairing cytokine production. This process is exacerbated by hypoxia. Furthermore, high ZFP36 family expression correlates with poor prognosis in ICI-treated patients in a public bulk tumor transcriptomic dataset. These findings suggest that tumor microenvironment (TME)-induced hypoxia upregulates the ZFP36 family in Tex cells, which in turn suppresses cytokine production via post-transcriptional regulation, potentially contributing to impaired antitumor effector function.

As a T-cell receptor co-stimulatory protein, signaling lymphocyte activation molecule family 6 (SLAMF6) is a member of the immune cell activating receptor family and is expressed on T cells, B cells, natural killer cells (NK cells), and tumor cells. To date, the biological functions and clinical significance of SLAMF6 in certain solid tumors remain to be elucidated. We analyzed the transcriptional expression pattern of SLAMF6 and its relationship with clinical outcomes using clinical specimens and pancancer databases. Next, multispectral immunofluorescence was applied to assess the correlation between SLAMF6 expression and CD8[+] T-cell activation and infiltration in clinical samples, with additional validation through immune-infiltration analysis. In parallel, using a colorectal cancer xenograft model, we examined how upregulation of SLAMF6 combined with immune-checkpoint blockade (anti-PD-L1) influences immunotherapy efficacy. Ultimately, our findings indicate that SLAMF6 modulates CD8[+] T-cell activation and infiltration through the STAT1/CXCL10 axis, thereby enhancing the recruitment of these effector cells to tumor sites.

To investigate the expression pattern, biological functions and regulatory mechanisms of inhibin beta B (INHBB) in colorectal cancer (CRC), and to clarify its role in mediating CD8[+] T cell exhaustion via the INHBB-ACVR2B-SMAD signaling axis for identifying a novel therapeutic target in CRC targeted immunotherapy, we analyzed the expression profile and prognostic value of INHBB in CRC tissues using The Cancer Genome Atlas database. Stable INHBB knockdown and overexpression CRC cell lines were constructed, and the malignant phenotypes were evaluated by Transwell assays, flow cytometry and other functional experiments. The interaction between INHBB and ACVR2B was verified by co-immunoprecipitation and immunofluorescence, and the in vivo regulatory effects of this pathway were determined using mouse xenograft tumor models combined with SMAD inhibitors. INHBB was upregulated in CRC tissues and associated with poor prognosis. Functional assays demonstrated that INHBB promoted the proliferation and invasion and suppressed the apoptosis of CRC cells. Mechanistically, INHBB bound to ACVR2B on CD8[+] T cells, activated SMAD signaling, and triggered CD8[+] T cell exhaustion, whereas blockade of the SMAD pathway reversed INHBB-mediated immunosuppression and tumor progression. Collectively, INHBB dually regulates the malignant phenotypes of CRC cells and CD8[+] T cell function through the INHBB-ACVR2B-SMAD axis, and targeting this axis may represent a promising therapeutic strategy for CRC immunotherapy.

In the phase 3 AEGEAN study, perioperative durvalumab plus neoadjuvant chemotherapy significantly improved the primary endpoints of event-free survival (EFS) and pathological complete response (pCR) in patients with resectable NSCLC versus neoadjuvant chemotherapy alone. We report prespecified exploratory Japanese subgroup analyses of EFS, disease-free survival (DFS), and overall survival (OS) (interim analyses; data cut-off [DCO] May 10, 2024); pCR (final analysis; DCO November 10, 2022); and safety. In this double-blind, placebo-controlled study, 79 treatment-naïve Japanese patients (stage II-IIIB[N2]) were randomized (1:1) to neoadjuvant chemotherapy plus durvalumab/placebo (every 3 weeks [Q3W], 4 cycles) pre-surgery, then adjuvant durvalumab/placebo (Q4W, 12 cycles). Efficacy was analyzed in the modified intention-to-treat population (N = 64; durvalumab arm n = 30, placebo arm n = 34), which excluded patients with documented EGFR/ALK aberrations. At the corresponding data cutoffs, EFS benefit favored the durvalumab arm (unstratified HR, 0.63; 95% CI, 0.26-1.45; median follow-up, 42 months [censored patients]), and the pCR rate was numerically higher with durvalumab versus placebo (33.3% vs. 5.9%). A clinically meaningful DFS improvement favored the durvalumab arm (unstratified HR 0.47, 95% CI 0.16-1.21); however, no difference in OS was observed (unstratified HR 1.01, 95% CI 0.40-2.51). Grade 3/4 adverse events occurred in 62.2% and 65.9% of patients with durvalumab and placebo, respectively. Improvements in EFS and pCR with perioperative durvalumab plus neoadjuvant chemotherapy versus neoadjuvant chemotherapy alone were comparable to, or greater than, those observed in the global study population, supporting this regimen as a treatment option for Japanese patients. Trial Registration: ClinicalTrials.Gov Identifier NCT03800134.

Intracellular redox regulation is increasingly recognized as compartment-specific, with each organelle maintaining a distinct redox environment. Mitochondria are a major source of reactive oxygen species (ROS), and many cancer cells exhibit elevated basal ROS levels, making mitochondrial redox modulation an attractive strategy for redox-based cancer therapy. However, efficient mitochondrial delivery of redox-active agents remains challenging. Small-molecule redox agents often require hydrophobicity that compromises their water solubility and increases their non-specific toxicity. Although synthetic polymers improve solubility and biocompatibility, they are predominantly taken up via endocytosis, which often requires additional elements for endosomal escape to achieve efficient cytosolic access and mitochondrial delivery. In this study, we developed mitochondria-directed phospholipid polymers based on 2-methacryloyloxyethyl phosphorylcholine (MPC) and redox-active ferrocene using a simple copolymerization strategy. We leveraged the cell-penetrating characteristics of amphiphilic MPC copolymers bearing hydrophobic ferrocene units to access the cytosol and introduced a mitochondria-directed motif by copolymerizing a triphenylphosphonium (TPP)-containing monomer. The TPP-bearing polymer exhibited higher in vitro anticancer activity than the non-directed polymer in CT26 mouse colon cancer cells. Confocal co-localization analysis revealed that the non-directed polymer was widely distributed throughout the cells, whereas the TPP-bearing polymer showed significantly higher mitochondrial co-localization. The TPP-bearing polymer also induced higher intracellular ROS levels than the non-directed polymer. Moreover, intratumoral administration of the TPP-bearing polymer significantly suppressed CT26 tumor growth in mice. Overall, this study provides a simple design strategy for organelle-directed redox-active polymers toward redox-based cancer therapy.

In advanced non-squamous non-small cell lung cancer (non-sq NSCLC), companion diagnostic testing (CDx) identifies actionable driver alterations and guides initial targeted therapy. However, disease progression inevitably occurs through acquired resistance, including activation of bypass signaling pathways. Although comprehensive genomic profiling (CGP) can identify emerging resistance alterations, its clinical utility in patients with known actionable driver alterations remains unclear. We retrospectively analyzed 227 patients who underwent CGP between June 2019 and August 2025. Patients with actionable driver alterations identified by CDx before CGP formed the known driver group. We also analyzed the nationwide C-CAT registry to assess registry-recorded therapeutic implementation. Among 227 patients, 74 (32.6%) had known actionable driver alterations. CGP identified additional clinically relevant genomic findings, including ESCAT Tier I-III or V alterations, in 34 patients (45.9%), comparable to that in patients without known actionable driver alterations (77/153, 50.3%). In the EGFR-mutated (EGFRm) subgroup, CGP frequently identified alterations consistent with potential bypass resistance mechanisms, including MET amplification, RET fusions, and BRAF V600E mutations. Overall, 10 patients (13.5%) received CGP-guided therapy based on additional findings, all within the EGFRm subgroup, whereas no patients with non-EGFR driver alterations did. In the C-CAT registry, CGP-associated therapy was documented in 21 of 381 (5.5%) in the EGFRm subgroup, 6 of 155 (3.9%) in the non-EGFR driver subgroup, and 195 of 2132 (9.1%) without registry-defined driver alterations. These findings indicate that CGP provides molecular re-evaluation during disease progression, particularly by identifying alterations consistent with potentially actionable resistance or bypass mechanisms in EGFRm NSCLC.

Cancer-associated fibroblasts (CAFs) have been recognized as key contributors to tumor progression. Chimeric antigen receptor (CAR) T-cell therapy targeting fibroblast activation protein (FAP), a marker of CAFs, has gained attention and is being evaluated in both preclinical and clinical studies. Cord blood (CB)-derived CAR natural killer (NK) cell therapy has shown efficacy in hematologic malignancies. In this study, we generated FAP-targeting CAR NK cells (FAP-CAR NK cells). CD3-depleted CB mononuclear cells were stimulated with irradiated K562 cells expressing 4-1BB ligand (tumor necrosis factor ligand superfamily member 9) and membrane-bound interleukin-15 and interleukin-21 to expand NK cells, into which FAP-CAR was introduced. FAP-CAR NK cells produced cytokines and exerted cytotoxic activity when co-cultured with FAP-transduced HT1080 cells and CAFs isolated from human lung cancer tissue. In a xenograft model established by intrathoracic co-injection of A549 lung cancer cells and luciferase-expressing CAFs, injection of FAP-CAR NK cells, but not control CD19-CAR NK cells, resulted in their robust expansion and persistence in vivo for 5 weeks. However, CAFs were not eliminated by FAP-CAR NK cells, although CAR NK cells were clearly detected in the tumors. The levels of cytotoxicity-associated molecules (such as granzymes) were higher in FAP-CAR NK cells persisting in the spleens of mice compared with those in pre-infusion FAP-CAR NK cells. These results indicate that FAP-CAR NK cells have the potential to respond specifically to the target antigen and expand robustly in vivo, although further optimization is required to maintain cytotoxic function for a prolonged duration in vivo.

We examined the association between serum uric acid (UA) levels and cancer incidence through a conventional observational and two-sample Mendelian randomization (MR) study in the same population (triangulation). We followed 13,695 individuals aged 35-69 years for cancer incidence (total, gastric, colorectal, lung, prostate, and breast cancers) over a median period of 10.6 years. The conventional observational dataset consisted of two subsets sampled from the full cohort using different sampling procedures. Odds ratios were computed using logistic regression for one type of single-nucleotide polymorphism (SNP) array, and hazard ratios were computed using the Cox proportional hazards model for another type of SNP array, combining them to calculate conventional relative risks (RRs). For two-sample MR, 42 UA-related SNPs were identified from Biobank Japan genome-wide association study data (109,029 participants). Associations between these SNPs and cancer were estimated using J-MICC Study data. MR RRs of cancer incidence per one standard deviation (SD) increase in genetically predicted UA were predicted using the inverse variance weighted (IVW) method and other MR approaches. In the conventional analysis, the highest UA concentration group (≥ 6.4 mg/dL) showed a decreased risk of lung cancer (RR: 0.64, 95% confidence interval [CI]: 0.41-0.98) compared with the lowest UA concentration group (< 4.0 mg/dL). The two-sample MR (IVW method) also suggested that UA was inversely (although marginally) associated with lung cancer risk (RR per one SD: 0.65, 95% CI: 0.40-1.05). In conclusion, lower, rather than higher, UA concentrations may be associated with an increased risk of lung cancer.

This review aims to systematically summarize the mechanisms of action, clinical evolution, and application prospects of radiotherapy in neoadjuvant therapy for stage III non-small cell lung cancer. Focusing on three core techniques: spatially fractionated radiotherapy, stereotactic body radiotherapy, and low-dose radiotherapy, key study data were screened across different treatment eras. Data extraction covers the unique mechanisms of the three radiotherapy modalities, as well as the efficacy and safety of treatment regimens before and after the immunotherapy era, including pathological response rate, survival benefit, adverse reactions, and other core endpoints. Comprehensive data analysis indicates that the three radiotherapy techniques have distinct characteristics; radioimmunotherapy synergy and triple regimens in the immunotherapy era significantly improve pathological response rates and long-term disease control. Spatially fractionated radiotherapy demonstrates unique advantages due to its low toxicity and dual mechanisms of action. In conclusion, radiotherapy shows prominent value in neoadjuvant therapy for stage III non-small cell lung cancer, and spatially fractionated radiotherapy is expected to become an important therapeutic option. However, its optimal dose-fractionation regimen, standardized procedures, and other issues still require further validation by multicenter randomized controlled trials to promote the development of more precise, safe, and effective treatments.

Real-world achievement of guideline-concordant treatment in osteosarcoma remains unclear. This study aimed to determine stage-specific rates of guideline-concordant treatment, evaluate their association with survival outcomes, and identify factors associated with non-concordance. Patients with osteosarcoma between 2011 and 2020 were identified from the Japanese Bone and Soft Tissue Tumor Registry. Rates of guideline-concordant treatment and disease-specific survival (DSS) were investigated. Among 1149 patients, 969 (84%) received guideline-concordant treatment. Achievement rates were 85% for localized low-grade, 83% for localized high-grade, and 90% for metastatic osteosarcoma. In localized low-grade disease, guideline-concordant treatment was associated with superior 5-year DSS (97% vs. 68%, p = 0.007); trunk location was associated with non-concordance. In localized high-grade disease, guideline-concordant treatment was also associated with superior 5-year DSS (78% vs. 50%, p < 0.001); age > 40 years and trunk location were associated with non-concordance. The association between guideline-concordant treatment and superior DSS was observed in both patients ≤ 40 years (82% vs. 50%; p < 0.001) and > 40 years (61% vs. 50%; p = 0.030). However, in a sensitivity analysis restricted to localized high-grade extremity osteosarcoma, this association remained significant among patients aged ≤ 40 years. In metastatic disease, guideline-concordant treatment was associated with superior 2-year DSS (54% vs. 16%; p < 0.001). Overall, guideline-concordant treatment was associated with superior survival outcomes in osteosarcoma. Treatment feasibility appeared to substantially influence treatment achievement and survival, particularly among older patients. No significant geographic disparities in achievement rates were observed across Japan, indicating a high and uniform standard of care nationwide. Wide/radical surgical resection and perioperative chemotherapy remain crucial whenever clinically feasible.

The pan-immune-inflammation value (PIV) derived from four blood cell counts warrants cautious interpretation in rectal-cancer immunotherapy, as CRT cannot be disentangled from anti-PD-1 effects within the small n = 16 subgroup.

Bacteria in tumors are increasingly recognized to be a part of the tumor microenvironment and their biological function depends on the location and relative abundance of a given microorganism or microbial community. Genetic sequencing data directly from tissue samples to detection of active metabolic signals from microorganisms, cell-microbiome interactions, ultrastructural localization of microorganisms; these are all different kinds of inferences about the functional role of the microbiome, and the evidence for "microbial presence and function" grows in a stepwise fashion. Furthermore, most current microbial detection methods do not, alone, predict the viability, biological activity, or functional properties of microorganisms. In this review, we compare different existing microbial detection techniques and distinguish three spatial paradigms: organized communities in tissue, intracellular bacteria, and extracellular bacteria. We summarize how each paradigm influences signal transduction, metabolism, metastatic spread, treatment resistance, antitumor immunity, and immunotherapy responses. We focus on low-biomass contamination, batch effects, negative controls, absolute quantification, and orthogonal validation. Finally, we classify results according to localization, biological role, clinical evidence, application, and applications, while distinguishing observations of human patients from mechanistic and therapeutic preclinical studies. Spatial microbiology provides useful information about tumor-microbial interactions, but clinically actionable conclusions require standard analysis, reproducible localization, viability or activity, and functional disruption experiments to demonstrate microbial effects and independent clinical validation.

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, with limited therapeutic options and poor clinical outcomes. Although PDAC has traditionally been thought to arise from ductal epithelial cells, recent studies using genetically engineered mouse models have demonstrated that acinar cells can also serve as a cell of origin, giving rise to tumors with distinct biological properties. Here, we investigated whether such lineage-specific differences are preserved in human PDAC. RNA sequencing was performed on resected specimens from 40 treatment-naïve patients with PDAC, and tumors were classified into Acinar- and Ductal-subtypes using lineage-specific gene expression signatures derived from mouse models. This classification strategy was further applied to human PDAC cell lines, followed by in vitro drug sensitivity assays. Patients with Acinar-subtype tumors had significantly longer overall survival than those with Ductal-subtype tumors (median, 54.0 vs. 26.5 months; p < 0.01), and multivariate analysis identified the ductal cell-derived tumor signature as an independent predictor of poor prognosis (HR, 3.74 [95% CI, 1.41-9.97]; p = 0.01). Among patients who developed postoperative recurrence, Acinar-subtype tumors were associated with more durable disease control (≥ 10 months) following gemcitabine-based chemotherapy than Ductal-subtype tumors (63.6% vs. 16.7%; p = 0.02). Consistently, pancreatic cancer cell lines classified as Acinar-subtype showed greater sensitivity to gemcitabine (p < 0.01) and paclitaxel (p < 0.05). These findings extend the clinical relevance of cell-of-origin signatures beyond prognostic stratification, highlighting the potential of cell-of-origin-based classification for treatment stratification and precision medicine in PDAC.