Search Results (894)

Background: Pancreatoblastoma (PB) is an exceptionally rare malignant epithelial neoplasm of the pancreas that recapitulates the developing pancreatic anlage. It is defined histologically by acinar-predominant differentiation with characteristic squamoid nests, and it may also show ductal and endocrine differentiation within the same tumor, features that distinguish it from acinar cell carcinoma and solid pseudopapillary neoplasm but that are readily overlooked. It occurs predominantly in young children, although adult-onset disease is well documented. We provide a population-based characterization of PB across the full age spectrum and benchmark it against pancreatic ductal adenocarcinoma (PDAC). Methods: Cases diagnosed between 2000 and 2021 were identified in the Surveillance, Epidemiology, and End Results (SEER) 17-registry database using site and histology codes. Cancer-specific survival (CSS) was estimated and compared, and Cox proportional hazards regression was used to explore associations with cancer-specific mortality. Results: Thirty-nine cases of PB were identified, compared with 155,924 cases of PDAC. The median age at diagnosis was 17 years (range, under 1 to 78 years); 12.8% (n = 5) were younger than 1 year. Males accounted for 69.2% (n = 27) of cases. The cohort was divided at the conventional pediatric-to-adult threshold of 18 years into a pediatric subgroup (age < 18 years; n = 20) and an adult subgroup (age ≥ 18 years; n = 19). CSS at 1 and 5 years was 95.0% and 83.5% in the pediatric subgroup, versus 67.7% and 24.6% in the adult subgroup (log-rank p < 0.001). Five-year CSS was 44.8% in males and 75.0% in females. In an exploratory multivariable model, older age was associated with higher cancer-specific mortality both as a dichotomous variable (adjusted hazard ratio [HR] for age ≥ 18 years 10.7, 95% confidence interval [CI] 2.4–48.2; p = 0.002) and, in a parallel model, as a continuous variable (adjusted HR 1.4 per 10-year increment, 95% CI 1.1–1.8; p = 0.002), indicating an age–mortality gradient. Male sex showed an association in the same direction that did not reach statistical significance (adjusted HR 3.3, 95% CI 0.96–11.6; p = 0.06). Relative to PDAC, PB was more frequently diagnosed in males and was associated with markedly superior survival (1- and 5-year CSS 81.8% and 54.9%, versus 28.8% and 4.0%). Conclusions: Pancreatoblastoma is predominantly a malignancy of young males and carries a substantially more favorable prognosis than PDAC, but outcomes differ markedly across the age spectrum, with adult-onset disease showing considerably poorer survival. Translationally, these population-level estimates support age-stratified prognostic counseling, argue for the referral of adults to centers experienced in rare pancreatic tumors, and provide a rationale for prospective molecular profiling to determine whether adult and pediatric PBs are biologically distinct and whether Wnt/beta-catenin pathway activation is therapeutically actionable. Full article

The immune checkpoint blockade (ICB) has changed the way many solid tumors are treated; yet, only a minority of patients respond durably. The biomarkers that guide therapy (the PD-L1 expression, tumor mutational burden, and microsatellite status) read only fixed molecular features and miss the shifting tumor–immune exchange. Patient-derived tumor organoids reproduce the tumor epithelium and predict the chemotherapy responses, but, in their usual form, they omit the immune compartment on which the ICB acts. This review asks how immune-competent organoids can close that gap, and how well they predict the response. One idea organizes the field: how a model is built determines what it can predict. Two complementary construction routes have been developed. One preserves endogenous immunity through an air–liquid interface culture; the other rebuilds it by a co-culture with defined effector populations. Both now incorporate microenvironmental reconstruction and quantitative functional readouts. The most developed examples are in colorectal cancer, with breast cancer applications only beginning to appear. Evidence for the ICB response prediction, graded on a four-level scale, remains at the proof-of-concept stage. Organoid functional assays complement rather than replace molecular biomarkers. Their clinical translation requires progress on four fronts: standardized protocols, longer immune cell viability, spatial and multi-omic integration, and prospective paired validation. Full article

Background: Neoadjuvant chemotherapy followed by interval debulking surgery is an established treatment strategy for selected patients with advanced epithelial ovarian cancer. However, simple postoperative prognostic tools integrating pre-treatment disease extent and post-treatment pathological stage remain limited. The neoadjuvant rectal (NAR) score was originally developed as a composite stage-migration endpoint in rectal cancer and has subsequently been explored in other solid tumors. We evaluated the feasibility and prognostic value of an ovarian NAR-like score (oNAR) in patients with advanced ovarian cancer treated with neoadjuvant chemotherapy and interval debulking surgery. Methods: This single-center retrospective cohort included patients with advanced ovarian cancer treated with platinum-taxane-based neoadjuvant chemotherapy followed by interval debulking surgery. The oNAR score was calculated using clinical T category before neoadjuvant chemotherapy and pathological T and N categories after interval surgery: oNAR = [5 × ypN − 3 × (cT − ypT) + 12]²/9.61. The primary analysis evaluated oNAR as a continuous score. Low, intermediate, and high oNAR categories based on the original rectal NAR thresholds were used only for exploratory visualization. The primary endpoint was progression-free survival (PFS). Cox regression, Kaplan–Meier analysis, log-rank testing, Harrell concordance index, and exploratory ROC analysis were used. Results: Among 74 reviewed records, 73 patients had sufficient cT, ypT, and ypN data for oNAR calculation and were included in the primary analysis. Thirty-one PFS events and 10 deaths occurred. The median oNAR was 14.98 (IQR, 14.98–30.07). As a continuous variable, higher oNAR was associated with shorter PFS (HR per 1-point increase, 1.036; 95% CI, 1.010–1.061; p = 0.0055), corresponding to an HR of 1.42 per 10-point increase. The Harrell C-index for PFS was 0.669. In a parsimonious comparison, oNAR showed higher discrimination than ypN alone and discrimination comparable to a combined ypT/ypN model. PFS differed significantly across low-, intermediate-, and high-oNAR groups (log-rank p = 0.0053). Median PFS was not reached in the low-oNAR group, compared with 13.47 months in the intermediate group and 12.09 months in the high group. The association persisted in sensitivity analyses restricted to patients with complete/optimal cytoreduction, maximal cytoreduction, and high-grade serous histology. Conclusions: The oNAR score was feasible to calculate using routinely available clinical and pathological staging variables and was significantly associated with PFS after neoadjuvant chemotherapy and interval debulking surgery. These findings support oNAR as a feasible stage-migration-based prognostic summary that may help capture post-NACT pathological T/N information and warrants external validation in larger ovarian cancer cohorts. Full article

Background/Objective: Niclosamide exhibits considerable potential for the treatment of drug-resistant cancers. However, its poor aqueous solubility substantially limits its oral bioavailability and therapeutic efficacy. To overcome this limitation, we developed a novel pharmaceutical supramolecular crystalline phase comprising niclosamide and nafamostat. Methods: In this study, we developed a niclosamide–nafamostat pharmaceutical supramolecular crystalline phase using a conventional solvent evaporation technique. Comprehensive solid-state characterization was carried out using X-ray powder diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and differential scanning calorimetry (DSC). In vitro drug transport kinetics and barrier transport efficiency were evaluated across artificial transpermeable membranes using a diffusion cell setup. In vitro anticancer efficacy was systematically screened against human lung, breast, and pancreatic cancer cell lines. Finally, in vivo translation was established using female NOD/SCID mice tumor-bearing animal models; therapeutic efficacy and total tumor burden were evaluated. Results: Niclosamide–nafamostat cocrystal (NNC) was found to form a distinct crystalline phase, with characterization results supporting a unique crystal lattice stabilized through strong intermolecular hydrogen-bonding interactions and exhibiting high thermal purity. Crucially, the supramolecular crystalline phase considerably enhances membrane transport, yielding superior cumulative permeation and enhanced apparent permeability (Papp) values compared to those of pure niclosamide. In vitro evaluation across multiple cancer cell lines demonstrated a tenfold increase in antiproliferative potency compared to the parent compounds. Furthermore, in vivo studies revealed a twofold increase in tumor growth inhibition and a tenfold reduction in total tumor burden (p < 0.05). Conclusions: These findings demonstrate that the NNC supramolecular crystalline phase platform successfully optimizes membrane permeability and antitumor efficacy, highlighting its potential for treating advanced cancers. Full article

Background: Testicular germ cell tumors (TGCTs) are the most common solid malignancies in young men. In real-world clinical practice, non-random treatment allocation may result in confounding by indication, as patients with a greater disease burden are more likely to receive chemotherapy. We evaluated the association between treatment strategy and recurrence-free survival using propensity score–based inverse probability of treatment weighting (IPTW). Methods: We retrospectively analyzed 114 patients who underwent radical orchiectomy for TGCT between 2015 and 2024. Patients were classified into active surveillance and chemotherapy groups. Stabilized IPTW was used to balance baseline clinicopathological characteristics. Recurrence-free survival was evaluated using Kaplan–Meier analysis and Cox proportional hazards regression. Residual post-weighting imbalance was addressed by additional covariate adjustment. Results: During follow-up, 29 of 114 patients (25.4%) experienced recurrence. Unadjusted Kaplan–Meier analysis demonstrated no statistically significant difference in recurrence-free survival between treatment groups (log-rank p = 0.150). Consistently, the unadjusted Cox proportional hazards model showed no statistically significant association between treatment strategy and recurrence-free survival (HR = 1.85, 95% CI 0.79–4.34; p = 0.158). After IPTW, treatment strategy remained not significantly associated with recurrence-free survival (HR = 0.96, 95% CI 0.43–2.13; p = 0.911). Additional adjustment for residual post-weighting imbalance yielded similar results (HR = 0.94, 95% CI 0.43–2.03; p = 0.876). Conclusions: After adjustment for measured baseline differences using IPTW, treatment strategy was not statistically significantly associated with recurrence-free survival. Given the limited number of recurrence events, wide confidence intervals, residual covariate imbalance, incomplete propensity-score overlap, and evidence of non-proportional hazards, these findings should be considered exploratory and should not be interpreted as evidence of equivalence. Full article

Chimeric antigen receptor (CAR)-T therapy has achieved partial therapeutic efficacy in solid tumors, but its overall effectiveness remains limited. IL-21-armored CD276.CAR-T (IL21.CD276.CAR-T) represents a promising strategy to enhance anti-tumor activity against esophageal squamous cell carcinoma (ESCC). However, resistance mechanisms remain unclear. We generated IL21.CD276.CAR-T cells and evaluated their cytotoxicity in vitro and in B-NDG xenograft models. Poliovirus receptor (PVR) expression on ESCC cells was analyzed, and shRNA-mediated PVR knockdown was performed to validate its role in resistance. IL-21R expression on ESCC cells was examined to exclude direct IL-21 signaling. IL-21 enhances the cytotoxicity of CD276.CAR-T cells against ESCC. Although IL21.CD276.CAR-T exhibited potent cytotoxicity in vitro, it failed to achieve complete tumor regression, and resistant cells still emerged. Mechanistically, resistance was not caused by CD276 antigen loss or IL-21R expression, but by upregulation of PVR on cancer cell membrane after IL21.CAR-T exposure. PVR knockdown restored CAR-T sensitivity in vitro, enhanced the anti-tumor capacity of IL21.CD276.CAR-T cells, and partially improved anti-tumor efficacy in vivo without systemic toxicity. PVR may act as a mediator of resistance to IL21.CD276.CAR-T in ESCC. Targeting PVR represents a novel strategy to overcome IL21.CAR-T resistance and improve therapeutic outcomes in ESCC. Full article

Background/Objectives: Cardiovascular comorbidities are frequent in patients with solid malignancies, but their distribution across tumor types and their association with recorded all-cause mortality remain insufficiently defined. This study assessed clinically defined cardiovascular phenotypes across major cancer groups and examined their association with recorded all-cause mortality. Methods: We performed a retrospective cohort study of 2020 consecutive adult patients hospitalized with malignancies at University Hospital “Tsaritsa Yoanna—ISUL” between September 2023 and December 2025. Patients were classified as having respiratory, gastrointestinal, genitourinary, breast, or other solid malignancies. Four non-mutually exclusive cardiovascular phenotypes were predefined: atherosclerotic, cardiometabolic, structural heart disease, and thromboembolic. Mortality status was ascertained through the National Health Insurance Fund database and hospital electronic records. As complete time-to-event data were unavailable, mortality was analyzed as a binary outcome using multivariable logistic regression. Results: Recorded all-cause mortality occurred in 471 patients (23.3%) and differed significantly across solid tumor types (p < 0.001). Mortality was highest in respiratory cancers (42.3%), followed by gastrointestinal cancers (30.2%), other malignancies (23.5%), genitourinary cancers (14.9%), and breast cancer (9.1%). Cardiovascular phenotypes also differed across tumor groups. In the fully adjusted model, structural heart disease was associated with recorded all-cause mortality status after multivariable adjustment (OR 1.61, 95% CI 1.16–2.23, p = 0.004). The atherosclerotic phenotype showed a borderline association (OR 1.42, 95% CI 1.00–2.02, p = 0.051), whereas cardiometabolic and thromboembolic phenotypes were not associated with recorded mortality after multivariable adjustment. Model AUC was 0.771. When cardiovascular burden was modeled continuously, each additional cardiovascular factor was associated with higher odds of recorded all-cause mortality status (OR 1.12, 95% CI 1.03–1.21, p = 0.005). Conclusions: Cardiovascular profiles differed across cancer types. Structural heart disease was the most consistent cardiovascular phenotype associated with recorded all-cause mortality, while overall cardiovascular burden provided additional but modest mortality-related information. In our cohort, routinely recorded cardiovascular diagnoses helped characterize mortality-related clinical profiles. However, these findings should be viewed as observational and hypothesis-generating, and further longitudinal studies are needed before any clinical use can be recommended. Full article

Backgrounds: Astragalus membranaceus (AM), a traditional Chinese medicinal herb, has garnered interest for its potential in anticancer therapy. Preclinical evidence suggests that AM inhibits tumor growth and could mitigate the adverse effects associated with conventional treatments. However, a comprehensive review of its efficacy across various cancer types is lacking. Objectives: This systematic review evaluates the preclinical evidence for the antitumor efficacy of AM and its active components to inform future clinical translation. Methods: A systematic literature search was conducted in PubMed, EMBASE, and Web of Science from their earliest records to February 2026 to identify relevant studies. Studies reporting on the antitumor effects of AM or its constituents in murine models were included. Results: Seventy-six studies met the inclusion criteria. The most frequently investigated compounds were formononetin (21 studies), Astragaloside IV (20 studies), and Astragalus polysaccharide (16 studies). The evidence indicates that these compounds, both as monotherapies and in combination with standard chemotherapy, can inhibit tumor growth and potentiate antitumor efficacy. Conclusions: AM and its bioactive constituents exhibit promising anticancer effects against solid tumors, including breast, lung, hepatocellular, and colorectal carcinomas in preclinical models. Clinical investigation is warranted to confirm the utility of AM for cancer therapy. Full article

Background/Objectives: Systemic inflammation is common in patients with cancer and may reflect tumor activity, disease extent, and the patient’s overall clinical condition. The systemic immune-inflammation index (SII), calculated from neutrophil, platelet, and lymphocyte counts, is a simple marker, but its relationship with tumor type, cardiovascular burden, and mortality is not fully clarified. To compare patients with respiratory system malignancies (RSM) and gastrointestinal tract malignancies (GITM) in terms of cardiovascular comorbidity, inflammatory profile, tumor-related characteristics, and recorded all-cause mortality, and to assess the association between SII and mortality. Methods: We analyzed 879 patients with solid malignancies, including 350 with RSM and 529 with GITM. SII was calculated as platelet count × neutrophil count/lymphocyte count. Cardiometabolic and cardiovascular burden was assessed according to the number of recorded cardiometabolic risk factors and cardiovascular diseases. The main outcome was recorded all-cause mortality. Results: Patients with RSM were younger and more often male, and more frequently had stage IV disease, whereas patients with GITM had a more pronounced cardiometabolic risk profile. Mortality was higher in RSM than in GITM (42.3% vs. 29.9%), and SII was also higher in RSM (median 1117.8 vs. 716.8). In the overall cohort, higher SII was associated with mortality after adjustment for age, sex, cancer type, stage, and metastatic disease (OR 1.11 per 1000-unit increase, 95% CI 1.02–1.21; p = 0.013). Mortality increased across SII tertiles from 22.5% to 34.5% and 47.4%. Adding SII to the clinical model led to only a small increase in AUC, from 0.719 to 0.729. Conclusions: SII was higher in patients with RSM and was associated with recorded all-cause mortality. However, its added prognostic value was modest. SII should therefore be interpreted as a supportive inflammatory marker, together with tumor stage, metastatic disease, and the broader clinical condition of the patient. Full article

Immune checkpoint inhibitors (ICIs) are now being introduced into perioperative treatment for several solid tumors. This strategy is usually explained by tumor reduction before surgery or by the elimination of minimal residual disease (MRD) after surgery. However, these explanations may not be sufficient to understand why the timing of ICI treatment, especially before lymph node (LN) removal, is important. In this review, we discuss tumor-draining lymph nodes (tdLNs) from two different aspects. tdLNs are anatomical routes for regional and distant metastasis, but they are also sites where tumor antigens are presented and tumor-specific T cell responses are generated. In particular, preclinical and translational studies suggest that tdLNs may maintain stem-like or progenitor-exhausted T cells (TPEX) that can respond to PD-1 blockade and supply more differentiated exhausted T cells to tumor sites. However, current clinical trials of perioperative ICIs demonstrate therapeutic benefit in specific diseases and regimens, but do not directly establish tdLN preservation or tdLN-resident TPEX maintenance as the decisive mechanism of efficacy. We therefore present the tdLN-reservoir model as a hypothesis-generating framework rather than as a clinically validated basis for modifying lymph node management. We also discuss the possible roles of neoadjuvant and adjuvant ICI in relation to antigen flow, minimal residual disease, metastatic-site draining LNs, postoperative lymphatic dysfunction, and future immune-guided clinical trials. Importantly, current evidence does not support altering standard lymph node surgery or radiotherapy solely to preserve putative tdLN immune-reservoir function. Full article

Hypoxia and the evolving concept of pseudohypoxia are critical in driving tumor angiogenesis, contributing to malignancy progression and therapeutic resistance. Angiogenesis, a common feature of many solid tumors, is promoted by hypoxia-induced overexpression of pro-angiogenic factors (e.g., VEGF, FGF) and genetic mutations (e.g., VHL, SDH) that stabilize hypoxia-inducible factors (HIF) even in normal oxygen conditions, a phenomenon known as pseudohypoxia. Recent experimental studies challenge the view that hypoxia universally enhances vessel growth. In certain models, severe oxygen deprivation impairs angiogenesis. Furthermore, tumor-mediated metabolic reprogramming can drive immune evasion via HIF stabilization in immune cells. These paradoxes, together with persistent therapy resistance and the limited effectiveness of current anti-angiogenic treatments, reveal critical gaps in our understanding of how hypoxic signaling modulates vascular and immune dynamics within the tumor microenvironment. These complexities demand more detailed exploration of underlying processes and the development of innovative therapeutic strategies. Here, we review recent mechanistic studies on tumor angiogenesis, summarizing therapeutic and diagnostic advances from both preclinical and clinical studies. We further discuss strategies to exploit hypoxic vulnerabilities, including HIF inhibitors, hypoxia-activated prodrugs, vascular normalization, combination regimens to restore immunity, biomarker-guided patient selection, and advanced hypoxia-targeted imaging to improve outcomes in angiogenesis-driven cancers. Full article

Transformative therapeutic innovation should not begin with a molecule—or even a molecular target. It should begin with a clearly defined unmet clinical need. Here, we present a seven-step Translational Therapeutic Development Roadmap that systematically connects an unmet medical need to a developable drug candidate through the disciplined sequence of (i) defining the need, (ii) understanding disease and resistance biology, (iii) building a mechanistic hypothesis, (iv) defining a target product profile (TPP), (v) molecular design and experimental validation, (vi) developability and manufacturability assessment, and (vii) clinical translation. A central conclusion emerging from this review is that resistance biology should be viewed not merely as a cause of therapeutic failure, but as a primary design input for next-generation therapeutic innovation. Our analysis identifies continuous alignment among unmet clinical needs, resistance biology, mechanistic hypothesis, molecular design, developability, and clinical translation as the defining characteristic of successful therapeutic development. We use dual-payload antibody–drug conjugates (ADCs) as a contemporary and highly illustrative case study of this resistance-informed therapeutic development approach. Single-payload ADCs such as trastuzumab deruxtecan and sacituzumab govitecan have transformed treatment across multiple solid tumors, yet most patients ultimately relapse through antigen loss, defective intracellular trafficking, drug efflux, payload-target alterations, and tumor heterogeneity, creating an emerging post-ADC treatment gap. Dual-payload ADCs, which deliver two mechanistically distinct warheads from a single antibody, represent a form of molecular combination therapy designed to increase the barrier to resistance and address multiple escape pathways simultaneously, as well as provide a clinically relevant model for resistance-informed therapeutic design. Using dual-payload ADCs as a worked example, we demonstrate how resistance biology directly informs payload pairing, molecular architecture, conjugation strategy, experimental validation, and developability. Our analysis indicates that successful dual-payload ADC design depends not simply on combining two cytotoxic payloads, but on selecting complementary mechanisms with non-overlapping resistance liabilities while satisfying predefined target product profiles and manufacturability requirements. We further summarize resistance-guided payload pairing strategies, including topoisomerase I plus tubulin inhibitors, topoisomerase I plus DNA-damage-response inhibitors, cytotoxic plus immunomodulatory payloads, and cell-permeable plus non-permeable combinations; the conjugation chemistries that enable defined dual-payload products; the preclinical validation, pharmacological optimization, and developability hurdles that separate promising biology from viable therapeutics; and the rapidly expanding clinical landscape, including the first-in-human program KH815 and emerging bispecific dual-payload constructs. Finally, we demonstrate that the same translational roadmap extends beyond ADCs to radiopharmaceutical conjugates, multispecific antibodies, targeted protein degraders, and cell and gene therapies, indicating that it represents a general framework for therapeutic innovation rather than an ADC-specific strategy. Collectively, this review supports the concept that therapeutic innovation is most successful when unmet clinical needs, resistance biology, molecular design, developability, and clinical translation are considered as an integrated continuum rather than as independent stages of drug discovery. This Translational Therapeutic Development Roadmap provides an organizing framework for guiding the rational development of next-generation targeted therapeutics across diverse therapeutic modalities. Full article

Background: The highly heterogeneous and dynamically evolving tumor microenvironment leads to the development of drug resistance and recurrence in traditional therapies. Although immunotherapy demonstrates unique advantages, its clinical utility remains constrained by the suboptimal immunogenicity and the limited effect of monotherapy. Herein, a hybrid drug delivery system based on a spatiotemporal hierarchical controlled-release strategy was proposed to achieve dual immunotherapy with immune checkpoint blockade (ICB) and immunogenic cell death (ICD) to promote synergistic anti-tumor therapy. Methods: A liposome–micelle hybrid drug delivery system (RUT-RPP-LP) was constructed using a lipid bilayer composed of dioleoyl phosphatidylethanolamine/hemisuccinyl cholesterol to encapsulate rutin (RUT) and to form an inner cavity-encapsulated resveratrol micelle (RPP). RUT-RPP-LP was characterized, and its pH sensitivity and release behavior were investigated. Subsequently, a colon cancer tumor-bearing mouse model was constructed to evaluate the in vivo targeted anti-tumor effect and biological safety. On this basis, the combined mechanism of ICB and ICD was preliminarily explored. Results: RUT-RPP-LP possessed excellent formulation characteristics, stability, and biocompatibility, achieving graded controlled release of drugs via responding to the TME and lysosomal acidity, respectively. Obviously, RUT-RPP-LP could specifically target the tumor site, induce the occurrence of ICD, and simultaneously block the PD-1/PD-L1 immune checkpoint signaling pathway, thereby enhancing the function of T cells and inducing apoptosis of tumor cells. Conclusions: The RUT-RPP-LP based on the hierarchical controlled-release strategy exerted a spatiotemporally coordinated enhancement of anti-tumor immunity, and may provide a novel combinatorial approach to overcome the low response of immunotherapy in solid tumors. Full article

Natural killer (NK) cells represent a promising tool for cancer immunotherapy; however, their efficacy against solid tumors is severely limited by the hostile tumor microenvironment (TME). This review provides a comprehensive overview of the physical, molecular, and metabolic barriers that drive NK cell dysfunction and immune evasion, emphasizing the physical challenge posed by extracellular matrix (ECM) density, which restricts infiltration. Beyond structural barriers, we examine the role of immunosuppressive cytokines (e.g., TGF-β) and immune checkpoint upregulation, both of which directly inhibit NK cell activation. Furthermore, NK cell signaling and cytotoxicity are profoundly affected by metabolic stressors such as hypoxia and acidosis, which act synergistically with the accumulation of immunosuppressive metabolites, including adenosine. These factors impair antitumor activity through multiple mechanisms, particularly the shedding of activating ligands. To investigate these complex interactions, we evaluate the advantages and disadvantages of different three-dimensional (3D) preclinical platforms, including tumor spheroids and Organ-on-Chip technologies, highlighting their distinct characteristics. Rather than advocating for a single technology, we emphasize that each model offers unique advantages for studying specific physical, chemical, and cellular components of the TME. Ultimately, leveraging the capabilities of these advanced 3D platforms is essential for deciphering microenvironmental barriers and unlocking the full therapeutic potential of NK cells against solid tumors. Full article

Objective: Current therapeutic regimens for pancreatic neuroendocrine neoplasms (pNENs) remain limited and fail to yield notable improvements in overall survival. Therefore, the development of novel agents is of paramount importance. Bruton’s tyrosine kinase inhibitors (BTKis) have demonstrated promising therapeutic potential in solid tumors; however, ibrutinib, a classic BTKi, exhibits unsatisfactory clinical efficacy against pNENs. In this study, we synthesized a novel pyrrolopyrimidine-based BTKi, QY21, and aimed to investigate its inhibitory effects on pNEN cell proliferation both in vitro and vivo and identify the core signaling pathways mediating its suppressive effects on pNENs. Methods: CCK-8, EdU, and colony formation assays were conducted to assess the effect of QY21 on pNENs in vitro. Transcriptome sequencing, quantitative real-time PCR, Western blotting, and flow cytometry were employed to explore the mechanisms. A xenograft tumor model in nude mice was established for in vivo validation. Results: QY21 significantly suppressed pNENs proliferation in vitro. Compared with the control and ibrutinib groups, QY21 exhibited stronger tumor growth inhibition in vivo. Histopathological analysis revealed a decreased Ki-67 index in the QY21 group, with no significant organ-toxic lesions observed. Transcriptome sequencing identified ATF3 as the core mediator responsible for the anti-proliferative effect of QY21. ATF3 was poorly expressed in pNENs, while QY21 markedly upregulated ATF3 expression. Mechanistically, QY21 induced ferroptosis by elevating ATF3 levels. The knockdown of ATF3 or administration of ferrostatin-1 significantly attenuated the anti-proliferative capacity of QY21, accompanied by reduced accumulation of reactive oxygen species and lipid peroxidation. Conclusions: This study demonstrates that the novel BTKi QY21 suppresses pNENs proliferation by triggering ATF3-mediated ferroptosis, providing a potential preclinical strategy for pNENs. Full article