Ovarian cancer (OV) is a highly lethal gynecological malignancy with limited effective biomarkers for prognosis and precision immunotherapy. B and T lymphocyte attenuator (BTLA) is an immune checkpoint molecule involved in tumor immune regulation, but its role in OV remains unclear.

Transcriptomic data from The Cancer Genome Atlas (TCGA) were analyzed to evaluate BTLA expression and its potential clinical relevance in OV. Exploratory multivariable Cox regression analysis was performed to examine the association between BTLA expression and patient prognosis. Functional characteristics were investigated through gene set enrichment and variation analyses, protein-protein interaction analysis, and regulatory network construction. Single-cell RNA sequencing (ScRNA-seq) was applied to explore cellular heterogeneity and BTLA-related cell-cell communication. Drug sensitivity analyses and molecular docking were conducted to assess therapeutic relevance. Experimental assays were performed to validate BTLA expression.

BTLA was significantly upregulated in OV tissues compared with normal controls. Higher BTLA expression was associated with patient prognosis in exploratory Cox regression and Kaplan-Meier (KM) analyses. Functional enrichment revealed associations with immune- and tumor-related pathways including epithelial mesenchymal transition (EMT), interleukin-10 signaling, and interferon responses. Single-cell analysis demonstrated BTLA-related transcriptional heterogeneity across cell populations, with epithelial cells showing the highest activity. Drug sensitivity profiling identified differential responses between BTLA expression groups, and molecular docking suggested moderate binding affinity between BTLA and genistein (Vina score = - 6.3 kcal/mol). Experimental validation confirmed increased BTLA expression at both mRNA and protein levels in OV tissues.

These findings suggest that BTLA may be involved in the tumor immune microenvironment of OV and may provide preliminary clues as a potential biomarker and therapeutic target.

Antigen loss remains a major barrier to chimeric antigen receptor (CAR) T cell efficacy in solid tumors. In aggressive thyroid cancers, dedifferentiation is accompanied by coordinated loss of lineage-restricted surface antigens, limiting immune recognition.

Thyroid-stimulating hormone receptor (TSHR) expression was assessed on multiple histotypes of thyroid cancer via immunohistochemistry. TSHR-directed chimeric antigen receptor T (CART) cell therapy was assessed against differentiated thyroid cancer subtypes which highly expressed TSHR and dedifferentiated thyroid cancer subtypes which downregulated TSHR in vitro and in xenograft and patient-derived xenograft (PDX) mouse models. MAPK inhibitors were assessed in combination with TSHR-CART cell therapy in dedifferentiated thyroid cancers which downregulated TSHR in vitro and in PDX mouse models.

Using TSHR as a clinically relevant antigen target, we demonstrated that pharmacologic tumor redifferentiation can restore target expression and sensitize tumors to CART cell therapy. TSHR-CART cells mediate durable antigen-specific cytotoxicity in TSHRhigh differentiated thyroid cancer models but are limited in TSHRlow dedifferentiated tumors. In patient-derived anaplastic thyroid cancer xenografts, MAPK inhibition restores TSHR expression and converts tumors from CAR-resistant to CAR-responsive. Concurrent redifferentiation therapy and CART cell treatment yields superior tumor control and survival versus monotherapy, without impairing CART cell function.

These findings establish tumor redifferentiation as a generalizable strategy to overcome antigen loss and enhance CART cell therapy in thyroid cancer and potentially other solid tumors.

Radical prostatectomy (RP) often leads to urinary incontinence (UI). There is sufficient evidence from randomized controlled trials (RCTs) that pelvic floor muscle and sphincter training (PFMT) are effective interventions to reduce UI.

To compare the effects of high-effort resistance training (HERT) combined with a non-invasive biofeedback-assisted device PFMT (dPFMT) versus HERT combined with conventional physiotherapeutic PFMT (pPFMT) on UI in prostate cancer patients undergoing RP.

Men with a history of RP for localized prostate cancer were randomly assigned to dPFMT (n = 55) or pPFMT group (n = 54). Both PFMT intervention groups underwent a 12-week whole body HERT program performed at moderate to high loads (> 60% 1-repetition maximum, performed to momentary failure).

Urinary leakage was assessed with the 24-hour pad weight test (24-h pad test) at the baseline (T0) and at the end of the 12-week lasting programs (T1). Individual changes of cumulative pad weight in gram (g) from T0 to T1 were calculated and compared between groups. An UI reduction of ≥ 50% was considered as treatment success and compared between groups (primary endpoint).

Ninety-six participants (88%) completed the intervention. All participants demonstrated a statistically significant reduction in 24-h pad test values from T0 to T1 (Mean (SD): -86.39 g (336.25 g); 𝑝 < 0.001). Treatment success did not differ between groups (OR = 1.19, 95% CI 0.53-2.66; 𝑝 = 0.670) and was achieved by 52.1% of all participants. In a post hoc exploratory analysis restricted to participants achieving treatment success, the relative reduction in urinary leakage from T0 to T1 was greater in the dPFMT group than in the pPFMT group (mean (SD) = -83.97% (13.88%) vs. -74.68% (12.93%); 𝑝 = 0.017). This exploratory finding should be interpreted with caution, particularly because no adjustment for multiple testing was applied. Time since surgery was a significant negative predictor of treatment success (p < 0.001), whereas age (p = 0.313) and follow-up rehabilitation treatment (AHB; p = 0.994) were not. None of the examined variables predicted the magnitude of change in urinary leakage.

Both device-based and physiotherapist-guided PFMT were associated with significant reductions in objectively measured urinary leakage after RP. No statistically significant between-group differences were observed under conditions evaluated in this study.

ClinicalTrials.gov Identifier NCT06206993, retrospectively registered on 15 December 2023.

Peptide receptor radionuclide therapy with [¹⁷⁷Lu]Lu-DOTATATE is an established treatment for somatostatin receptor-expressing neuroendocrine tumors. Although strong anti-tumor effects have been demonstrated in experimental models, curative responses in patients remain limited. Improved understanding of the molecular responses induced by [¹⁷⁷Lu]Lu-DOTATATE may help identify strategies for treatment optimization. This study aimed to characterize proteomic alterations in GOT1 small intestinal neuroendocrine tumor xenografts following [¹⁷⁷Lu]Lu-DOTATATE therapy.

GOT1 tumor-bearing BALB/c nude mice received a non-curative intravenous administration of 15 MBq [¹⁷⁷Lu]Lu-DOTATATE or saline control. Tumors were collected 1 or 13 days after treatment to represent early response and regrowth phases. Tumor volumes were monitored using magnetic resonance imaging. Proteomic profiling was performed using liquid chromatography tandem mass spectrometry with tandem mass tag labeling. Differential protein expression was analyzed using Welch's t-test with significance defined as fold change ≥ 1.5 and p < 0.01. Functional enrichment and pathway analyses were conducted using Gene Ontology annotation and Ingenuity Pathway Analysis.

Treatment induced a transient reduction in tumor volume followed by regrowth. In total, 3861 proteins were quantified, of which 155 showed significantly altered expression after treatment. Affected proteins were associated with cytoskeletal organization, oxidative stress response, protein metabolism, and systemic regulation. Pathway analyses identified inhibition of several signaling pathways linked to cell migration and invasiveness, including RhoA, Rac, integrin, and CXCR4 signaling. Upstream regulator analysis suggested activation of RABL6 and inhibition of p53 signaling during tumor regrowth. Proteins related to endoplasmic reticulum stress and ubiquitin-proteasome activity were also altered. Selected findings were validated by ELISA.

[¹⁷⁷Lu]Lu-DOTATATE therapy induced extensive proteomic changes in GOT1 tumors, including suppression of pathways associated with invasiveness and modulation of p53- and stress-related signaling. These findings suggest potential therapeutic benefits of combining [¹⁷⁷Lu]Lu-DOTATATE with agents targeting p53 regulation or endoplasmic reticulum stress pathways to improve treatment efficacy in neuroendocrine tumors.

CRISPR activation (CRISPRa) enables precise up-regulation of gene expression for ex vivo and in vivo applications. However, a lack of scalable, high-coverage tools has limited comprehensive genetic screening in murine models. Here, we introduce Partita, a next-generation mouse whole-genome CRISPRa sgRNA platform, designed for unparalleled efficiency in gene activation studies. Partita uses a high-density targeting strategy, deploying 10 sgRNAs per transcriptional start site, structured into five gene class-specific sublibraries to maximize transcriptional induction. To demonstrate the capabilities of Partita, we performed a series of large-scale screens: an in vitro enrichment/depletion screen, whole-genome CRISPRa screens in a double-hit lymphoma model to uncover resistance factors to proapoptotic drugs (venetoclax, nutlin-3a, and etoposide) and an in vivo screen to identify accelerators of MYC-driven lymphomagenesis. Each experiment revealed both expected and unexpected regulators, with high validation rates. By enabling robust gain-of-function screening, Partita unlocks new avenues for functional genomics and expands the toolkit for discovering key drivers of biological processes across diverse research fields.

Metabolic reprogramming is a defining hallmark of CRC. The Warburg effect is the principal metabolic feature of CRC cells, wherein glucose is preferentially catabolized into lactate to sustain accelerated proliferation. In parallel, CRC cells exhibit strong glutamine reliance to replenish tricarboxylic acid (TCA) cycle intermediates required for adenosine triphosphate (ATP) production, lipid biosynthesis and redox homeostasis. Consequently, mitochondria play a central role in supporting the augmented biosynthetic and energetic demands beyond basal energy homeostasis. In this regard, the mitochondrial pyruvate carrier (MPC), mitochondrial citrate carrier (CIC) and the mitochondrial glutamine carrier (SLC1A5_var) located in the inner mitochondrial membrane, are emerging areas of investigation in CRC metabolism. MPC is frequently lost or downregulated in CRC, whereas CIC was found to be upregulated and promote CRC growth and survival. In contrast, SLC1A5_var has been reported to exhibit elevated expression in colon cancer cells. Recent evidence indicates that its inhibition reduces CRC cell viability; however, its specific role in CRC progression remains to be elucidated. Notably, these transporters may influence the metabolic-epigenetic landscape of CRC through metabolite-dependent regulation of chromatin and transcriptional processes. This review highlights current insights into mitochondrial metabolite transporters in CRC and their potential metabolic and epigenetic implications. Thus, elucidating the roles of these transporters may provide novel therapeutic strategies for CRC management.

Drug‑tolerant persister cells represent a major barrier to durable responses in cases of epidermal growth factor receptor (EGFR)‑mutant lung adenocarcinoma treated with tyrosine kinase inhibitors. To define the molecular features and actionable vulnerabilities of this cell state, public transcriptomic datasets from tumour cell‑based models treated with EGFR inhibitors or other targeted agents within the receptor tyrosine kinase (RTK)/RAS/MAPK pathway were integrated with machine‑learning‑based feature selection, survival analysis and Cancer Dependency Map‑based pharmacogenomic screening. The present study also performed experimental validation of identified genes and drugs in PC‑9 and HCC827 cell models. A four‑gene persister‑associated gene signature comprising B‑cell translocation gene 1 protein, inhibitor of growth protein 4, proline‑rich nuclear receptor coactivator 1 and calcium‑binding and coiled‑coil domain‑containing protein 1 was identified, which defined a targeted therapy‑induced persister state and associated with poor patient survival. Functional validation using reverse transcription‑quantitative PCR, doxycycline‑inducible short hairpin RNA knockdown and β‑galactosidase staining assays demonstrated rapid induction of these genes during osimertinib treatment and showed that depletion of each gene markedly impaired drug‑tolerant persister cell formation. Pharmacogenomic analysis identified ZLN005, a peroxisome proliferator‑activated receptor γ coactivator‑1α agonist, as a candidate metabolic modulator that synergized with osimertinib to reduce persister cell viability. Mechanistically, ZLN005 enhanced mitochondrial oxidative metabolism and reactive oxygen species accumulation, leading to caspase‑3 activation and gasdermin E‑associated pyroptotic cell death. Collectively, these findings defined the transcriptional and metabolic features of targeted therapy‑induced persister cells and revealed a metabolic vulnerability to ZLN005 in EGFR‑mutant lung adenocarcinoma.

Glioblastoma (GBM) is the most malignant primary brain tumor, with limited therapeutic options. Dysregulation of FXYD5 has been reported in multiple malignancies, suggesting FXYD5 as a potential target for precision medicine. In the present study, FXYD5 expression and prognostic significance were analyzed using biological analysis on multiple databases and it was identified that FXYD5 was observably upregulated in GBM and inversely correlated with the prognosis of patients. Functional studies were investigated in LN229 and U251 GBM cell lines through FXYD5 knockdown and overexpression approaches, assessing proliferation (Cell Counting Kit‑8), apoptosis (flow cytometry), migration/invasion (Transwell assays) and lipid metabolism (free fatty acids, triglycerides, cholesterol, Nile Red staining). Knockdown of FXYD5 suppressed GBM cell proliferation, migration and invasion, while increasing apoptosis, indicating that FXYD5 may serve as a therapeutic target for GBM. Mechanistic studies examined the PI3K/AKT/long‑chain acyl‑coenzyme A (CoA) synthase 4 4 (ACSL4) pathway via western blotting and rescue experiments, finding that FXYD5 activated the PI3K/AKT signaling pathway, leading to upregulation of ACSL4 and enhanced lipid metabolism. In vivo, a subcutaneous xenograft mouse model was used to evaluate whether PI3K/AKT inhibition could antagonize FXYD5 overexpression‑induced tumor growth and PI3K/AKT inhibition reversed FXYD5 overexpression‑induced tumor growth in the subcutaneous mouse model. These findings revealed that FXYD5 promotes GBM progression via the PI3K/AKT/ACSL4 signaling axis and represents a potential therapeutic target for GBM.

Endocrine therapy has improved the prognosis of patients with hormone receptor-positive (HR⁺) breast cancer. However, acquired resistance remains a major cause of recurrence and progression, and the limited response of this subtype to immunotherapy highlights the need to better understand the mechanisms underlying immune escape. Recent studies indicate that, in addition to an immunosuppressive microenvironment, alterations in antigen processing and presentation may also contribute to immune escape, although systematic integration of these mechanisms remains lacking. Lipid metabolic reprogramming, as a key feature of tumor metabolic remodeling, supports tumor cell survival and adaptive evolution in endocrine-resistant HR⁺ breast cancer and may also regulate antitumor immune responses through multilevel mechanisms. At the tumor cell level, enhanced fatty acid oxidation, increased lipid synthesis, and altered cholesterol metabolism may affect tumor immune visibility by influencing antigen presentation-related processes, including MHC class I-associated pathways. Within the immune microenvironment, lipid accumulation may contribute to dendritic cell dysfunction and promote the polarization of tumor-associated macrophages toward immunosuppressive phenotypes, thereby impairing antigen presentation. Consequently, persistent and inefficient antigen stimulation may promote T-cell dysfunction and exhaustion. This article systematically integrates the role of lipid metabolism in antigen presentation and T-cell regulation, and proposes a dysfunctional multilevel mechanistic axis of "lipid metabolism-antigen presentation-T-cell function", providing a theoretical basis for understanding immune evasion associated with endocrine resistance and for developing combination therapeutic strategies.

Clear cell renal cell carcinoma (ccRCC) exhibits substantial molecular and immune heterogeneity. Mast cells participate in tumor-microenvironment remodeling, but the prognostic relevance of mast cell-associated apoptosis-related transcriptional features remains unclear.

Single-cell RNA sequencing was used to identify mast cells in ccRCC. Genes associated with the annotated mast-cell cluster were intersected with the HALLMARK_APOPTOSIS gene set to develop a mast cell-associated apoptosis score (MCAS) using machine-learning survival algorithms. MCAS was evaluated in TCGA-KIRC and two external cohorts. Associations with clinicopathological features, mutation profiles and model-derived genes were analyzed. YBX3 was functionally characterized using qRT-PCR, western blotting, CCK-8, colony formation, EdU, Transwell migration and flow cytometry assays.

A TPSAB1/CPA3-positive mast cell population was identified. The CoxBoost plus random survival forest model showed the best performance for MCAS construction. High MCAS scores predicted poor survival in the training and validation cohorts and were associated with advanced clinicopathological features and tumor mutation burden. SurvSHAP and SurvLIME jointly identified BNIP3, CX3CR1, HMOX1, and YBX3. High YBX3 expression was associated with unfavorable prognosis. Functionally, YBX3 knockdown inhibited proliferation, colony formation, DNA synthesis and migration, and promoted apoptosis with increased BAX and decreased Bcl-2, whereas YBX3 overexpression enhanced proliferative and migratory phenotypes.

MCAS is a mast cell-associated apoptosis-related transcriptomic signature for prognostic stratification in ccRCC. The in vitro findings suggest that YBX3 may contribute to malignant phenotypes and apoptosis resistance in ccRCC cells.