Wakayama Medical University

C-Mannosyl tryptophan (C-Man-Trp), a unique monomeric glycosyl amino acid, is up-regulated in the blood of ovarian cancer patients; however, the underlying mechanisms remain unclear. In the present study, C-Man-Trp production and its dynamics were investigated in female B6C3F1 mice transplanted with mouse ovarian cancer OV2944-HM-1 (HM-1) cells. After transplantation, C-Man-Trp levels increased in the plasma, urine, ascites, peritoneal exudate cells (PECs), and tumor masses of mice. Furthermore, changes in the transcriptional expression of C-Man-Trp metabolism-related genes, C-mannosyltransferases (Dpy19l1 and Dpy19l3), and thrombospondin type I repeat superfamily genes (Thbs1, Spon1, and cellular communication network factor 1) were noted in tumor-associated cells and tissues. A cell-sorting analysis revealed that PECs mainly comprised myeloid-derived immune cells, such as macrophages and myeloid-derived suppressor cells, in addition to a small population of HM-1 tumor cells. C-Man-Trp levels were high in the macrophage fraction, but lower in the myeloid-derived suppressor cell fraction. C-Man-Trp was also produced in the ex vivo culture medium of macrophages isolated from PECs. Under macrophage depletion using clodronate liposomes, the ovarian cancer-stimulated up-regulation of C-Man-Trp was significantly suppressed in the plasma, ascites, PECs, and tumor masses of HM-1 cell-transplanted mice. C-Man-Trp levels in the plasma and peritoneal cavity cells of normal healthy mice were also suppressed by clodronate liposomes, whereas the expression of C-Man-Trp metabolism-related genes showed different changes from those in mice transplanted with HM-1 cells. Collectively, these results demonstrate that tumor-stimulated macrophages play a pivotal role in the dynamics of C-Man-Trp in mice with ovarian cancer.

Visceral fat (VF), particularly epicardial adipose tissue (EAT), plays a crucial role in the development of coronary artery disease (CAD). Cathelicidin (LL37) is an antimicrobial peptide involved in innate immunity and has been implicated in inflammatory processes. However, the relationship between VF accumulation, cathelicidin, and atherosclerosis remains unclear. Seventy-eight subjects without CAD were enrolled and classified by obesity type: normal-weight (normal; n=20), subcutaneous fat (SF; n=19), and VF (n=39). Plasma LL37 concentrations were compared across groups. LL37 expression in EAT was assessed in 9 patients undergoing open-heart surgery, stratified by CAD status. In animal experiments, angiotensin II-infused wild-type and Apoe-/-mice fed a Western-type diet were treated with cathelicidin-related antimicrobial peptide (CRAMP) small interfering RNA (siRNA), scrambled siRNA, or vehicle. Atherosclerotic lesions were evaluated by enface Sudan IV staining. Plasma LL37 concentrations were higher in the VF than in the normal-weight and SF groups (median [IQR] 99.11 [91.70-129.86] vs. 58.63 [50.03-70.05] and 69.81 [54.64-87.39] ng/mL, respectively; P<0.001) and were correlated with VF area and EAT volume. LL37 expression in EAT was higher in patients with than without CAD (2.34 [2.06-3.90] % vs. 1.34 [0.81-1.49] %; P=0.034). Treatment with CRAMP siRNA significantly reduced atherosclerotic lesion area in Apoe-/-mice without affecting blood pressure or lipid profiles. LL37 is associated with VF accumulation and CAD. However, these findings are exploratory and warrant prospective validation to determine its potential utility as a biomarker.

Objectives: This study aimed to prospectively investigate the associations among obesity, physical activity, and short-term high medical expenditures in Japanese employees and their dependents. Methods: Participants were 245,044 employees and their dependents aged 40-74 years who underwent the Specific Health Checkup in fiscal year 2008. Health checkup and medical expenditure data for 2008-2010 were provided by health insurers. They were divided into 12 groups according to the combination of body mass index categories (normal, overweight, and obesity) and engagement in exercise and/or daily physical activity (inactive, daily physical activity only, exercise only, and active). The multivariable-adjusted odds ratios of the groups for high total medical expenditures in the next year compared to the inactive normal body mass index group were estimated. High medical expenditures were defined as the top 5% of total annual expenditures, consistent with prior literature identifying high-cost users. Similar analyses were performed by sex and age (<65 years, ≥65 years). Results: Of the participants, 61.8% were men (mean age, 52.1 years). Multivariable-adjusted odds ratios were significantly high only in the inactive groups with overweight or obesity in men and younger individuals. In women and older individuals, the odds ratios were significantly high only in inactive women with obesity; however, the odds ratios were high in women who exercised only and in active older individuals, both with obesity. Conclusions: Exercise or daily physical activity might attenuate the possibility of incurring high short-term medical expenditures in men and younger individuals with overweight or obesity. These findings suggest that physical activity recommendations may need to be tailored for women and older individuals with obesity, and further longitudinal research is warranted.

Colorectal cancer (CRC) exhibits substantial molecular heterogeneity, yet detailed single-cell transcriptomic data from Japanese patients remain limited. Here, we performed single-cell RNA sequencing (scRNA-seq) on tumor tissues and adjacent normal tissues from four Japanese CRC patients to characterize malignant cell populations. We identified a common cancer-associated cell cluster observed across all four cases, characterized by high expression of the long noncoding RNA KCNQ1 opposite strand/antisense transcript 1 (KCNQ1OT1). Further analysis revealed a marked decrease in Ras Association Domain Family Member 6 (RASSF6) expression within this cluster, despite clinical datasets reporting that elevated RASSF6 expression correlates with poor overall survival in Western cohorts. Reanalysis of publicly available bulk RNA-seq data from independent East Asian cohorts confirmed that RASSF6 expression was consistently reduced in tumor tissues compared with adjacent normal colon tissues, while KCNQ1OT1 expression showed no apparent differences across cohorts. Collectively, our single-cell analysis delineates a recurrent cancer-associated transcriptional state observed within this cohort and highlights distinct gene expression features that may be underappreciated by bulk or population-averaged analyses, underscoring the value of single-cell profiling for refining molecular interpretations of CRC.

Antigen heterogeneity remains a major obstacle to the effective application of chimeric antigen receptor (CAR)-T cell therapy in solid tumors. We investigated the potential of epitope spreading as a strategy to overcome this limitation and examined whether CAR-T cells concomitantly producing IL-7 and CCL19 (7 × 19 CAR-T cells) could act as potent inducers of epitope spreading, as well as the underlying mechanisms. We used a murine model inoculated with a mixture of cancer cell lines—genetically modified to express the CAR target antigen—and their respective parental lines lacking target expression. Following administration of 7 × 19 CAR-T cells, flow cytometry and peptide stimulation assays were performed to evaluate the induction of tumor antigen-specific T cells and dendritic cells within tumor-draining lymph nodes and tumor tissues. We also evaluated the antitumor efficacy of 7 × 19 CAR-T cells derived from an allogeneic donor and their capacity to induce epitope spreading in vivo and ex vivo. In multiple tumor mixture models, 7 × 19 CAR-T cells demonstrated marked antitumor activity and promoted epitope spreading in murine solid tumor models, enabling endogenous T cells to recognize and target tumor-associated antigens. This response was dependent on cross-presentation by defined dendritic cell subsets in both the tumor microenvironment and tumor-draining lymph nodes within 2 weeks of 7 × 19 CAR-T cell administration. Notably, 7 × 19 CAR-T cells derived from allogeneic donors also induced epitope spreading in tumor-bearing hosts, thereby increasing survival. The 7 × 19 CAR system is a promising strategy for overcoming antigen heterogeneity in solid tumors by promoting epitope spreading.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00262-026-04316-z.

Concurrent chemoradiotherapy (CCRT) is the standard treatment for unresectable stage III non-small-cell lung cancer (NSCLC). Completion of CCRT affects subsequent therapy and impacts overall survival. The aim of this study was to develop a model for predicting completion of CCRT and thus guide individualized treatment strategies. We conducted a pooled analysis of data from the Japan Lung Cancer Society-Integrated Database, which includes data from eight randomized phase II/III trials of CCRT for unresectable stage III NSCLC in Japan. Clinical factors associated with CCRT completion were identified through multivariable logistic regression analysis and used to develop a scoring model. The scoring model includes neutrophil count ≥4500/μL = two points, C-reactive protein concentration <1.00 mg/dL = two points, cisplatin-based therapy = one point, and PaO 2 ≥83.0 mmHg = one point for a total of six points (NCCP score). NCCP score categories were defined as low (0–3) or high (4–6). Completion rates of CCRT were 39.7% (95% CI = 34.1–45.5%) in the low and 63.9% (95% CI = 56.6–70.7%) in the high NCCP score groups (p < 0.001). High NCCP scores were also significantly associated with better objective response rate, disease control rate, progression-free survival, time-to-treatment failure, and overall survival) ( p <0.001, 0.003, 0.003, <0.001, and <0.001, respectively). The NCCP score, composed of neutrophil count, use of cisplatin, CRP concentration, and PaO 2 , predicts completion of CCRT and may contribute to optimizing CCRT treatment as a supplementary risk stratification in patients with stage III NSCLC.

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