Antibody-drug conjugates (ADCs) are one of the most significant advancements in modern cancer therapeutics. Combining the target selectivity of monoclonal antibodies with the cytotoxic potential of payloads, ADCs effectively kill cancer cells and offer hope to patients with even refractory cancer types. Beyond simply increasing the number of therapeutic options available for cancer patients, ADCs have become a powerful frontline agent in overcoming multidrug resistance (MDR). As one of the most challenging obstacles to effective cancer care, MDR is mediated by ATP-binding cassette (ABC) transporter-mediated drug efflux, target-based mutations, and dysregulated apoptosis. The clinical success of ADCs specifically engineered to overcome MDR, including in heterogeneous tumors and cancer cells that exhibit bypass signaling, is well established. This is especially evident with trastuzumab deruxtecan (T-DXd) in HER2-low, HER2-positive, and HER2-mutant cancers; sacituzumab govitecan (SG) in TROP2-expressing triple-negative breast cancer (TNBC) and urothelial carcinoma; and enfortumab vedotin in Nectin-4-positive bladder cancer. By overcoming MDR, ADCs have enabled more effective treatment algorithms across multiple malignancies. Most importantly, the clinical application of ADCs has become inextricably linked to cancer genomics. HER2 testing has evolved from a two-tiered system to a continuous spectrum including HER2-ultralow, HER2-low, HER2-positive, and ERBB2-mutant categories. Each of these categories exhibits different eligibility guidelines for ADC patient selection. As cancer cells continue to evolve and develop resistance to even ADCs through mutations and variants, researchers and clinicians have used pharmacogenomics to predict ADC response and resistance. To define the genomic architecture of ADC-resistant tumor subpopulations, single-cell transcriptomic studies and liquid biopsy approaches are being used to enable real-time examination of the tumor genome during ADC therapy, thereby optimizing treatment and circumventing resistance driven by emerging mutations and variants. This review provides a comprehensive analysis of the molecular structure of ADCs, the pharmacological principles underlying their potent cytotoxic activity against MDR cancer cells, the genomic and transcriptomic biomarkers that guide ADC patient selection, and the emerging resistance mechanisms that will shape the next generation of promising ADC development.
Metabolic reprogramming is a substantial obstacle for anticancer drug screening, as targeted therapeutics often lose efficiency due to the dynamic adaption of cancer cells. Glutamine metabolism in cancer profoundly impacts tumor initiation, progression and metastasis. The existing agents are compromised by resistance and off-target toxicity. In this study, a real-time NMR tracking method for intracellular glutamine metabolic flux was established. This method enables comprehensive profiling of nitrogen metabolism and serves as a valuable tool for characterizing specific cancer metabolic phenotypes and screening drugs against targeted cancer cells. Applying this approach to traditional Chinese medicine (TCM) discovery, we identified Astragalus membranaceus as a potent regulator of glutamine metabolism. Through virtual screening via molecular docking, 12 potential compounds from Astragalus membranaceus were initially flagged as candidate binders toward the allosteric pocket of glutaminase 1 (GLS1). Crucially, subsequent in vitro recombinant human GLS1 enzyme activity assays successfully ruled out computational false positives and demonstrated that Compound 2 (quercetin) acts as the exclusive, direct enzymatic inhibitor among the tested monomers, capable of effectively suppressing GLS1 activity. Overall, this work provides a robust platform for real-time metabolic profiling of glutamine metabolism and drug screening at the living cell level, and offers new insights into the mechanisms of TCMs in anticancer therapy.
Mechanical stress profoundly influences tumor-bone interaction. However, the underlying mechanism is largely elusive. Here, we reveal that mechanical loading activates osteocyte connexin 43 (Cx43) hemichannels (HCs) and promotes the release of soluble factors, thus establishing a tumor-suppressive bone microenvironment that limits the growth of metastatic breast cancer in bone. Conditioned media from fluid flow shear stress (FFSS)-stimulated osteocyte MLO-Y4 cells significantly inhibited tumor spheroid expansion and migration of both triple-negative and ER-positive breast cancer cells. This inhibitory effect was abolished by blockade of Cx43 HCs. FFSS-activated osteocytes also reduced filopodial extensions and epithelial-mesenchymal transition (EMT) marker expression in cancer cells. Mechanistically, FFSS stimulation induced Cx43 HC-mediated release of prostaglandin E2 (PGE2) and ATP, which exerted opposing effects: PGE2 promoted, while ATP suppressed, breast cancer cells. The relative ATP:PGE2 ratio served as a key determinant of the extent of tumor inhibition. In vivo tibial loading for two weeks prior to tumor inoculation further demonstrated the anti-metastatic function of Cx43 HCs. Mechanical loading suppressed breast cancer proliferation, growth, and osteolytic damage in wild-type (WT) and R76W transgenic mice (defective in gap junctions (GJs) but retaining HCs), but not in Δ130-136 mice (defective in both HCs and GJs). Consistently, Cx43(M1)-mediated HC blockade attenuated the tumor-suppressive effects of mechanical loading. ATP and PGE2 levels were elevated in loaded tibiae of WT and R76W, but not Δ130-136 mice. Collectively, these findings identify Cx43 HC activation by mechanical stress as a key mechanism driving the bone's transition from a metastasis-permissive to a metastasis-resistant microenvironment, mediated by ATP-dominant paracrine signaling from osteocytes that suppresses breast cancer progression.
Cuproptosis is a type of recently reported cell death characterized by aberrant accumulation of copper ions within cells, leading to mitochondrial stress and protein aggregation. Recent studies suggest that certain cancer cells are particularly susceptible to cuproptosis-inducing agents. However, the genetic determinants of cellular sensitivity to cuproptosis and the therapeutic potential of cuproptosis inducers in cancer treatment remain unclear. Here, we report the discovery of a small molecule, N1,N1-dimethyl-N4-(4-(pyridin-2-yl)thiazol-2-yl)benzene-1,4-diamine (dPTBD), that targets KRAS-driven cancer via a tetracycline-inducible cell-based drug screening. dPTBD exhibited significant antitumor efficacy in KRAS-driven cancers both in vitro and in vivo. Mechanistic studies revealed that dPTBD acted as a copper ionophore, promoting intracellular copper accumulation particularly in the mitochondria, leading to metabolic disruption and cuproptotic cell death. Adding trace amount of copper massively enhanced the cytotoxic effect of dPTBD, resulting in an immediate mitochondrial dysfunction and cuproptosis. In preclinical models, dPTBD, either alone or combined with physiologically tolerable amount of copper, significantly suppressed tumor growth in KRAS-mutant pancreatic and colon cancer xenografts. Taken together, our study reveals that induction of cuproptosis is a new therapeutic strategy for KRAS-driven cancer and identifies dPTBD as a lead compound for future development.
Resistance to cyclin-dependent kinase (CDK) 4/6 inhibitors, such as ribociclib (RB), limits breast cancer therapy. Although PEG10 siRNA (siPEG10) can counteract this resistance, achieving targeted, synchronized co-delivery to reshape the immunosuppressive tumor microenvironment (TME) remains a formidable challenge. By loading RB and siPEG10 into PEG-β-CD-modified mesoporous silica nanoparticles (PMSNs) and coating them with breast cancer cell membranes (CM), the nanocomposite RB/siPEG10@PMSNs-CM was synthesized. The nanomaterials were characterized for their size, charge, drug loading, and pH-responsive release. Therapeutic efficacy, immunotherapy-related potentials, and underlying mechanisms were evaluated using in vitro functional assays, an anoikis resistance model, and in vivo breast cancer models. Toxicity was assessed in liver and kidney tissue. The nanodrug RB/siPEG10@PMSNs-CM (<200 nm) exhibited enhanced cellular uptake and pH-dependent drug release. The zeta potential of the RB/siPEG10@PMSNs-CM was negative. The nanodrug significantly inhibited tumor cell growth, migration, invasion, and metastasis, while inducing apoptosis. Crucially, the nanodrug overcame anoikis resistance, upregulated cleaved caspase-3 and cleaved PARP, reversed epithelial-mesenchymal transition (EMT) markers (increased E-cadherin and decreased Vimentin), and inhibited p-STAT3 and p-ERK signaling. Furthermore, the nanodrug exhibited lower toxicity to liver and kidney than RB. In Conclusion, the developed biomimetic nano-delivery system RB/siPEG10@PMSNs-CM successfully co-delivers RB and siPEG10 and exhibits a favorable safety profile. This platform exhibits anti-primary tumor and anti-metastatic efficacy and effectively overcomes anoikis resistance in breast cancer, which correlated with the suppression of the MEK/ERK and STAT3 signaling pathways, offering a promising combination strategy for advanced breast cancer immunotherapy and targeted treatment.
Dyspnea is one of the most distressing symptoms in patients with advanced cancer. Although systemic opioids are recommended as first-line pharmacologic treatment, 30-50% of patients do not achieve adequate relief. Midazolam is often used for persistent dyspnea despite opioid administration; however, robust evidence supporting its efficacy as a second-line treatment remains limited. To evaluate the feasibility of a randomized controlled trial assessing the efficacy and safety of continuous subcutaneous midazolam infusion as second-line treatment for persistent dyspnea despite morphine administration in hospitalized patients with advanced cancer. A protocol for a multicenter, randomized, double-blind, placebo-controlled feasibility trial (J-SUPPORT2201/JORTC-PAL22) is described. The trial is being conducted at nine sites in Japan. Participants are hospitalized adult patients with advanced cancer who are not receiving active anticancer treatment and who experience dyspnea at rest (Integrated Palliative Care Outcome Scale dyspnea score ≥2) despite continuous morphine infusion. Protocol treatment includes standardized morphine escalation plus continuous subcutaneous infusion of midazolam (9 or 6 mg/day for vulnerable patients) or placebo for 24 hours. The primary endpoint is feasibility, defined as completion of protocol treatment at 24 hours. Secondary endpoints include dyspnea intensity, anxiety, rescue morphine use, communication, adverse events, and 30-day survival. This feasibility trial will provide methodological insights and preliminary clinical data regarding midazolam as a second-line option for persistent dyspnea despite morphine infusion in patients with advanced cancer and help design future confirmatory trials.
This review aims to summarize and critically evaluate published clinical trials investigating the role of [18F]FDG PET in bladder cancer (BC) management, and to provide an overview of ongoing trials in this field. A literature search was conducted in PubMed up to 31 August 2025 and on ClinicalTrials.gov to identify clinical trials assessing the role of [18F]FDG PET/CT in patients with BC. The search strategy used the terms "FDG PET," "bladder cancer," "PET/CT," "clinical trials," and "PET/MRI." Review articles, editorials, letters, comments, and case reports were excluded. Two investigators independently screened the literature, and additional trial information was retrieved from ClinicalTrials.gov. Five eligible full-text publications and eight registered clinical trials were identified. Among the publications, two reported study protocols, two were pilot studies, and one was an ancillary analysis from a larger trial. In addition, eight ongoing clinical trials were retrieved from ClinicalTrials.gov. Available data suggest that [18F]FDG PET/MRI may be more suitable than PET/CT for evaluating primary bladder lesions, while PET/CT is particularly effective in detecting distant metastases. Several ongoing studies are also investigating delayed acquisition protocols to improve detection of nodal metastases. [18F]FDG PET/CT and PET/MRI show promise for enhancing local and distant staging in muscle-invasive bladder cancer, although their role in nodal evaluation remains uncertain. Current evidence is limited by heterogeneous study designs, small patient cohorts, and the lack of standardized imaging protocols.
Conventional cancer therapy has largely been organized around maximal cytotoxic elimination, yet intense tumor killing can impose evolutionary selection pressures that promote resistance, clonal escape, and relapse. Here, we propose therapeutic super-competitor cells (TSCs) as a programmable, evolution-aware extension of educative cancer care rather than as an established clinical platform. In this framework, TSCs are engineered cells designed to home to or be locally positioned within tumor-associated niches, sense malignant ecological cues, transiently acquire bounded competitive fitness, restrict malignant access to space, metabolites, and stromal support, and then undergo monitored clearance or benign differentiation. TSCs are therefore not defined by direct target-cell cytolysis, although competitive pressure, metabolic restriction, or immune remodeling may secondarily reduce tumor viability. This Review integrates concepts from cancer ecology, tumor microenvironment biology, developmental cell competition, immunotherapy, and synthetic biology while distinguishing established principles from speculative engineering applications, evaluating feasible early disease contexts, and outlining measurable endpoints for ecological replacement, including spatial occupancy, clonal displacement, resource gradients, persistence kinetics, and responsiveness to safety triggers. We further discuss organoid and tumor-on-chip competition assays, spatial and lineage tracking, response biomarkers, therapeutic-window definition, layered containment, and ethical/regulatory safeguards. By reframing engineered cell therapy as a controllable ecological intervention rather than simply a vehicle for maximal killing, the TSC framework provides a testable but still hypothetical direction for educative cancer care and evolution-aware oncology.
Breast cancer is the most frequently diagnosed malignancy among women globally, accounting for approximately 2.3 million new cases and 670,000 deaths in 2022. Trace element imbalance, heavy metal exposure, and oxidative stress play critical roles in the pathogenesis of breast cancer (BC). This study aimed to determine the serum concentrations of total sulfhydryl (TSH) groups-a marker of non-enzymatic antioxidant capacity distinct from thyroid-stimulating hormone-ischemia-modified albumin (IMA), essential trace elements (Zn, Mg, Cu, Mn, Fe), and toxic heavy metals (Cd, Pb) in newly diagnosed BC patients before initiating chemotherapy, and to compare these parameters with healthy individuals. The study was conducted at Van Yüzüncü Yıl University Dursun Odabaş Medical Center and included 30 newly diagnosed BC patients and 30 healthy volunteers. Blood samples were collected into trace element-grade anticoagulant-free serum tubes. Serum levels of trace elements and heavy metals were analyzed using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES, Thermo Scientific iCAP 6900). TSH levels were measured using the DTNB (Ellman) method, and IMA levels were assessed colorimetrically. Statistical analyses were performed using SPSS 19.0; normality was assessed by the Shapiro-Wilk test; group comparisons employed the Mann-Whitney U test, with a significance level set at p < 0.05. The results demonstrated that serum concentrations of Cu, Fe, Mg, Mn, and Zn were significantly lower in the patient group compared to controls (p < 0.001). Conversely, levels of Cd and Pb were significantly higher in BC patients (p < 0.001). IMA levels were significantly increased (44.71 ± 4.03 vs. 22.52 ± 0.27 ABSU; p < 0.001), while TSH levels were significantly decreased (1.58 ± 0.45 vs. 3.26 ± 0.05 mmol/L; p < 0.001). Spearman correlation analysis identified a significant positive association between serum Zn and IMA within the patient group (r = 0.493, p = 0.007). These findings indicate that breast cancer is associated with disrupted mineral homeostasis, increased toxic metal accumulation, and impaired antioxidant defense, leading to oxidative stress. However, the cross-sectional design limits causal inference; these results represent associations that require validation in larger, prospective cohorts. The combined evaluation of trace elements, IMA, and TSH may serve as potential supportive biomarkers in breast cancer.
Although immune checkpoint blockade has offered new hope to patients with bladder cancer, its overall response rate remains modest. In this setting, the quantity and functional status of CD8+T cells largely dictate therapeutic efficacy. Integrative bioinformatics analyses, validated in our institutional tissue cohort, we demonstrated that histone deacetylase 10 (HDAC10) was markedly up-regulated in bladder cancer and inversely correlated with CD8+T cells infiltration. In vivo experiments, using Hdac10-KO cell line, we revealed that Hdac10-KO significantly inhibited tumour growth and promoted the infiltration and activity of CD8+T cells. Mechanically, HDAC10 deacetylated the K342 site of interleukin enhancer-binding factor 3 (ILF3), thereby inducing the ubiquitination and degradation of it by WWP2. This reduced the stability regulation of CXCL9 mRNA by ILF3, decreasing CXCL9 expression and weakening CD8+T cell infiltration and activation. Notably, we found that the probiotic Clostridium butyricum (C.B.) can inhibit the expression of HDAC10 by increasing the short-chain fatty acid butyric acid, thereby sensitizing immunotherapy for bladder cancer in mouse models. This study revealed the specific mechanism by which HDAC10 led to immunotherapy resistance in bladder cancer and proposed potential combined treatment strategies.
Peripheral nerves contribute to tumor progression, but the mechanisms by which neural signals regulate cancer cell plasticity and immune resistance remain unclear. Using a paired murine model of intraneural and nonintraneural tumor growth with single-cell RNA sequencing, we identified a nerve-associated cancer cell state marked by increased expression of secretory leukocyte protease inhibitor (SLPI). SLPI was elevated in intraneural tumors and in human tumors with perineural invasion. Sensory neuron-derived substance P (SP) induced SLPI secretion through tumor cell TACR1, whereas sensory denervation or TACR1 blockade reduced SLPI production. SLPIhigh cancer cells were enriched for WNT/β-catenin and stemness programs. Consistent with this, SLPI increased β-catenin and c-MYC expression, expanded ALDH+ stem-like cells, and enhanced mammosphere formation, whereas Slpi deletion reduced stemness and impaired tumor growth in vivo. SLPI also limited immune-mediated tumor cell killing. Proteomic and biochemical analyses identified granzyme B as a direct SLPI-binding partner, and SLPI inhibited granzyme B-dependent cleavage of caspase-3 and gasdermin E, thereby reducing cytotoxic lymphocyte-induced cell death. In immunocompetent mouse models, pharmacologic inhibition of the upstream SP-TACR1 pathway with aprepitant synergized with anti-PD-1 therapy. In clinical datasets, high SLPI expression was associated with residual disease after immunotherapy, poor response, and adverse outcome. Together, these findings identify the SP-TACR1-SLPI axis as a neural pathway linking cancer stemness to immune escape and nominate this pathway as a target for combination immunotherapy.
Methionine dependence represents a well-known metabolic vulnerability in cancer. Despite promising preclinical results, methionine restriction is impaired by the presence of methionine-independent cancer cells. After confirming both inter- and intra-tumoral heterogeneity in methionine dependence, we demonstrate that "methionine-independent" cells are rather "methionine self-sufficient," relying on the vitamin B12 (B12)-dependent methionine synthase (MTR) to sustain growth without exogenous methionine, which renders them highly vulnerable to B12 deprivation. Dual methionine and B12 deprivation produced synergistic cytotoxicity, inhibiting proliferation and inducing apoptosis across multiple cancer types and primary tumor cells, while sparing fibroblasts. This synergy persisted under moderate nutrient restriction, supporting translational potential. Moreover, dual therapy prevented the adaptive metabolic shift seen with methionine deprivation alone, avoiding rebound proliferation and resistance. In vivo, a methionine-restricted diet plus a synthesized B12 antagonist significantly suppressed growth of methionine-independent pancreatic xenografts without hematologic toxicity. These findings uncover a selective, synergistic anticancer strategy targeting methionine self-sufficiency.
Here, we examined the consequences of biologically relevant vitamin D deficiency, a known risk factor for aggressive prostate cancer, using ex vivo and in vivo models. Phenotypic and single-cell RNA sequencing of mouse prostate organoids showed that vitamin D deficiency stunted luminal cell differentiation more than androgen deficiency, which is a known driver of prostate development. Mice fed a vitamin D-deficient diet showed significantly altered expression of androgen-responsive genes in their prostate luminal cells, as determined by single-cell RNA sequencing. MDA-PCa-2b and 22Rv1 human prostate cancer cells, when maintained for 6 months in 1,25-dihydroxyvitamin D, had very different responses compared to short-term treatments. Notably, the MDA-PCa-2b cells lost the ability to form xenografts, despite normal proliferation in vitro. RNA sequencing showed that these cells also had disruptions in androgen signaling and in multiple cancer-related pathways, which differed by treatment duration. This study offers new insights and validation of vitamin D's role in both benign and malignant prostate biology, underscoring its essential hormonal functions and supporting strategies for vitamin D supplementation to reduce prostate cancer risk in vulnerable populations.
Re-recurrent rectal cancer (RRRC) represents a highly complex disease following curative-intent treatment of recurrent rectal cancer (RRC). While management principles for primary and locally recurrent rectal cancer (RRC) have been defined by expert collaborations, no specific guidelines currently outline the perioperative management of RRRC. This systematic review aimed to appraise the reported perioperative strategies and oncological outcomes of patients undergoing curative-intent treatment for RRRC. Eligibility criteria, Studies reporting perioperative management and outcomes of adult patients undergoing curative-intent treatment for RRRC were included. Non-English articles, letters, abstracts, and studies lacking surgical or oncological data were excluded. Information sources, The review was conducted according to PRISMA guidelines and registered in PROSPERO (CRD420251244390). MEDLINE (PubMed), Cochrane Library, Web of Science, and Scopus were searched for relevant articles. Risk of bias, Methodological quality was assessed using the Newcastle-Ottawa Scale for cohort studies. Synthesis of results, Given heterogeneity in treatment strategies and outcome reporting, results were synthesized narratively. Included studies, Three retrospective cohort studies comprising 169 patients treated with curative intent surgery for RRRC were included. Synthesis of results, Neoadjuvant therapy was administered in 20-92% of included patients, depending on the previous cumulative radiation dose. Pelvic exenteration was frequently required, with total exenteration performed in 6-20% and sacrectomy in up to 15% of cases; reconstructive procedures were reported in less than 16%. IORT was used in 44-77% of patients in centers where it was available. R0 resection rates ranged from 33% to 62%, with oncological outcomes directly associated with margin status. Limitations of evidence, Evidence was limited to retrospective observational studies with small sample sizes, heterogeneous management, and varied institutional resources, precluding meta-analysis. Interpretation, Curative-intent surgery for RRRC is feasible in highly selected patients, with R0 being the principal prognostic determinant of oncological outcome. However, significant variability in perioperative pathways, margin definition, MRI-based classification, and reconstructive strategies underlines the necessity for the development of standardized, consensus-based recommendations to optimize multidisciplinary treatment. https://www.crd.york.ac.uk/PROSPERO/view/CRD420251244390, identifier CRD420251244390.
Although there are several ongoing clinical trials using neoantigen peptide-based cancer vaccines, challenges still exist to implement in clinical approval such as poor antigen stability, inefficient delivery, and inadequate immune activation. To address these limitations, we developed a polymer-based polyvalent peptide and adjuvant (SPPA) that co-delivers lipid-conjugated tumor-specific peptides with Toll-like receptor 7/8 (3M - 052) and a STING (2'3'-cGAMP) agonist. This nanoplatform enables efficient peptide encapsulation, sustained release, and targeted delivery to antigen-presenting cells (APCs), thereby enhancing both innate and adaptive immune responses. We synthesized and characterized a library of lipid-conjugated tumor-associated and neoantigenic peptides. In vitro, SPPA significantly upregulated pro-inflammatory genes and cytokine secretion, confirming robust innate immune activation and demonstrated effective cellular uptake and lymphatic trafficking. In vivo, SPPA alone or in combination with anti-PD-1 antibody (αPD-1) elicited strong cytotoxic T lymphocyte (CTL) responses and inhibited tumor growth in four aggressive syngeneic mouse models: Triple-negative breast cancer (4T1), HER2+ breast cancer (TUBO), lung carcinoma (LLC1), and renal cell carcinoma (RENCA). The combination therapy led to pronounced tumor growth inhibition, survival benefit, and immune cell infiltration, including elevated CD8+IFN-γ+ T cells and M1 macrophages, and reduced regulatory T cells and MDSCs. Spatial transcriptomics revealed localized transcriptional reprogramming, with downregulation of extracellular matrix genes and activation of inflammatory pathways. Collectively, these findings establish SPPA as a potent and versatile nanovaccine platform capable of inducing durable antitumor immunity, especially when combined with immune checkpoint blockade. This approach offers strong translational potential for personalized immunotherapy across diverse solid tumor types.
Cyclin E1 (CCNE1), a critical regulator of cell cycle progression, has been implicated in various cancers; however, its prognostic significance and functional role across breast cancer (BC) subtypes remain inadequately defined. This study aims to comprehensively analyze CCNE1 mRNA expression and evaluate its potential as both a prognostic biomarker and a therapeutic guide. We analyzed CCNE1 mRNA expression using large-scale BC cohorts (TCGA, METABRIC, GEO) and assessed associations with tumour grade, overall survival, and mutation status. The muTarget platform was employed to investigate associations between CCNE1 expression and key genetic alterations. Furthermore, gene set enrichment analysis was performed to identify enriched biological pathways, and functional studies were conducted using mTORC1 inhibition in TP53-mutant, RB1-deficient TNBC cells to evaluate cell viability and CCNE1 expression levels. CCNE1 expression was significantly associated with higher tumour grade, overall survival, and distinct mutation statuses. Gene set enrichment analysis revealed enrichment of proliferation-related pathways, including G2-M checkpoint, E2F targets, and mTORC1 signaling in CCNE1-high tumors. Moreover, functional studies showed that mTORC1 inhibition successfully reduced CCNE1 expression and impaired viability in TP53-mutant, RB1-deficient TNBC cells. CCNE1 overexpression characterises a clinically aggressive subset of breast cancers, particularly TNBC. Targeting mTORC1 may represent a promising therapeutic approach in CCNE1-high, TP53-mutated, and RB1-deficient tumours, supporting the clinical utility of CCNE1 in patient stratification and targeted therapy.
Despite the success in infectious diseases, mRNA vaccines have inherent limitations in stability and immunogenicity. Non-small cell lung cancer remains the leading cause of cancer-related death worldwide, with poor prognosis in advanced stages, underscoring the urgent need for novel therapeutic strategies. Previously, we have developed a scarless circular RNA synthesis platform (NeoAna) that exhibited enhanced stability and long-lasting antigen expression with minimal cytotoxicity compared to mRNA. In this study, we construct a circRNA vaccine using NY-ESO-1 as the antigen and deliver circRNA vaccine with lipid nanoparticles. The circRNA vaccine elicited robust innate and adaptive immune responses both in vitro and in vivo. It demonstrated superior antitumor efficacy relative to mRNA vaccine, achieving 93.1% tumor inhibition when combined with immune checkpoint blockade. Prophylactic vaccination prevented tumor establishment, while adjuvant therapy conferred protection against tumor re-challenge, highlighting durable immune memory. Mechanistically, circRNA vaccine promoted dendritic cell maturation and enhanced tumor infiltration of T cells, and mediated antitumor immunity in a CD8+ T cell-dependent manner. Additionally, single-nucleus RNA sequencing revealed that it remodeled the tumor microenvironment toward a pro-inflammatory and antitumor phenotype. Following circRNA vaccination, the proportions of cytotoxic T cells, pro-inflammatory macrophages, and antigen-presenting cancer-associated fibroblasts were all increased within the tumor microenvironment, accompanied by activation of immunity-related signaling pathways. In conclusion, our findings demonstrate the immunogenicity and therapeutic potential of the NeoAna circRNA-based therapeutic vaccine, supporting its further evaluation in clinical trials.
AR pathway-independent prostate cancer (ARIPC), particularly neuroendocrine prostate cancer (NEPC), represents one of the most lethal states of metastatic castration-resistant prostate cancer. However, how fatty acid synthesis (FAS) is organized in ARIPC and whether distinct lipogenic states shape neuroendocrine lineage transdifferentiation remain unclear. By integrating single-cell and bulk transcriptomic analyses of mCRPC cohorts, we identify NEPC as a fatty-acid-synthesis-low state associated with poor survival. Within this context, fatty acid synthase (FASN) emerges as a key indicator and functional contributor to lipogenic activity. FASN depletion suppresses lipogenesis while increasing NEPC-associated programs, migration, and metastatic colonization. We further identify FGFRL1 as the FGF family member most consistently associated with fatty acid synthesis activity in ARIPC. FGFRL1 depletion reduces FASN expression and relative free fatty-acid content, while targeted GC-MS supports broader fatty-acid remodeling and fluorescent uptake assays show increased exogenous fatty-acid uptake. FASN restoration partially restores relative free fatty-acid content and attenuates NEPC-associated and migratory phenotypes. Directional perturbation, rescue, AKT phosphorylation, and co-immunoprecipitation analyses further support the functional FGFRL1-FASN relationship. ONECUT2 is prioritized as a candidate downstream transcriptional regulator whose expression correlates with the neuroendocrine program. Together, these findings support an FGFRL1-FASN metabolic axis that regulates neuroendocrine lineage transdifferentiation and metastatic progression in ARIPC.
The immunosuppressive tumor microenvironment is a major driver of tumor progression and therapeutic resistance. Basal-like breast cancer (BLBC), which largely overlaps with triple-negative breast cancer, generally displays an immunosuppressive tumor microenvironment enriched with tumor-associated macrophages and other immunosuppressive cell populations. However, the driving mechanisms that establish and maintain these tumor microenvironment features remain not fully understood. This study identifies tumor cell-intrinsic POU4F1 as a key regulator of the immunosuppressive tumor microenvironment in BLBC. Tumors with high POU4F1 expression exhibited a lower IFNγ-related signature and poor responses to immunotherapy. In immunocompetent 4T1 tumor-bearing mice, POU4F1 knockout led to decreased infiltration of M2-like macrophages and increased infiltration of proliferative and functional CD8+ T cells and NK cells. In vitro assays using human immune cells demonstrated that POU4F1 directly promoted monocyte recruitment and macrophage polarization, which in turn suppressed the proliferation and effector function of tumor-specific CD8+ T cells and NK cells. Mechanistically, POU4F1 upregulated CCL2 expression through NIK-mediated activation of the non-canonical NF-κB signaling pathway, thereby promoting monocyte recruitment and immunosuppressive phenotype polarization. Genetic ablation or pharmacological targeting of POU4F1 with Bobcat339 significantly inhibited tumor growth, remodeled the tumor immune microenvironment, and synergized with anti-PD-1 therapy in mouse models and patient-derived breast cancer organoids. These findings provide mechanistic insights into how POU4F1, a BLBC-specific transcription factor, orchestrates intercellular crosstalk to establish an immunosuppressive, tumor-supportive microenvironment, and indicate that targeting POU4F1 may represent a promising therapeutic strategy for BLBCs.
To evaluate the clinical applicability of previously established transcriptomic signatures (molecular subtypes, components and GemPred status) in metastatic pancreatic cancer, we conducted a retrospective pooled analysis of 178 patients from three phase 2 trials (PRODIGE35/37, AFUGEM; 2013-2016) testing first-line regimens (FOLFIRINOX, GemNab, FuNab, FOLFIRI3). RNA sequencing was performed on primary/metastatic tumors across French centers, with blinded assessment of subtypes (immune classical, pure basal-like, stroma-activated), quantitative components, and GemPred status. Primary endpoint: progression-free survival (PFS). Immune classical subtype showed superior median PFS (9.03 months) and OS (11.27 months) versus basal-like and stroma-activated subtypes (PFS: p=0.015; OS: p=0.010). Higher classical component correlated with improved PFS (HR=0.83, p=0.048) and OS (HR=0.71, p=0.001). Inactive stroma predicted better PFS (HR=0.64, p=0.001) and OS (HR=0.71, p=0.015). GemPred-negative patients treated with FOLFIRINOX versus GemNab had higher ORR (46.9% vs. 19.1%, p=0.046), longer PFS (8.2 vs. 2.3 months; HR=2.28, p=0.008), and OS (11.6 vs. 5.0 months; HR=2.04, p=0.021). No differences occurred in GemPred-positive patients. In a formal treatment-by-GemPred interaction analysis (FOLFIRINOX vs. GemNab), the interaction was significant for OS (adjusted p-interaction=0.050) but not for PFS (adjusted p-interaction=0.51). Transcriptomic signatures retain prognostic and predictive utility in metastatic pancreatic cancer, with GemPred representing a hypothesis-generating predictive signal for OS (e.g., a FOLFIRINOX OS benefit in GemPred-negative). Prospective validation is warranted for clinical implementation.