Search Results (5,711)

Gallbladder cancer (GBC) is an uncommon but highly lethal malignancy, characterized by striking geographic clustering, late diagnosis, and a prevention profile that differs from other gastrointestinal malignancies. This narrative review synthesizes epidemiologic, mechanistic, clinical, and public-health evidence on GBC determinants with a prevention-oriented lens. Rather than merely cataloguing risk factors, we propose a pragmatic framework that combines epidemiologic credibility, population-attributable burden, baseline risk, biologic plausibility, feasibility, and the benefit-risk balance of possible interventions. The strongest opportunities for prevention arise from risk-stratified management of gallbladder disease and congenital biliary abnormalities, reduction of excess and dysfunctional adiposity, promotion of physical activity, control of chronic Salmonella carriage in endemic settings, and mitigation of selected food- and water-borne toxic exposures. Several other determinants, including diet, alcohol consumption, smoking, reproductive factors, and socioeconomic disadvantage, may inform risk stratification or broader cancer-prevention policy, but currently offer less specific actionability. Effective prevention will require matching interventions to context: population-wide metabolic and lifestyle prevention, targeted clinical management of high-risk gallbladder conditions, and endemic-area strategies addressing infection, environmental exposure, and access to surgery. GBC prevention is therefore possible through carefully evaluated risk-adapted strategies. Full article

Chronic hepatitis B virus (HBV) infection remains a leading cause of hepatocellular carcinoma (HCC) worldwide, particularly in Asia. Although nucleos(t)ide analogs suppress viral replication and reduce HCC incidence, residual risk persists, highlighting the need to better understand HBV-driven hepatocarcinogenesis. HBV alters host transcription, DNA repair, lipid metabolism, and the hepatic immune microenvironment. These changes help sustain liver injury, fibrogenesis, and malignant transformation. Ferroptosis, an iron-dependent form of programmed cell death driven by excessive lipid peroxidation, has emerged as an important mechanism in this context, because the liver is central to iron and lipid metabolism. Here, we propose that HBV-driven lipid remodeling shapes ferroptosis susceptibility in a stage-dependent, double-edged manner. During chronic infection, oxidative stress and enrichment of peroxidation-prone lipid substrates may promote ferroptosis-associated hepatocyte injury, immune dysregulation, and fibrosis. After malignant transformation, HBV-related oncogenic programs may instead favor ferroptosis evasion through altered lipid composition and strengthened antioxidant defenses, thereby supporting tumor survival and progression. In this review, we summarize mechanisms linking HBV, lipid metabolic remodeling, and ferroptosis while distinguishing direct HBV-related evidence from findings inferred from other liver disease or cancer models. This HBV-ferroptosis framework may help refine risk stratification and suggest new therapeutic opportunities in HBV-related HCC. Full article

Human papillomavirus (HPV) is the most common sexually transmitted infection globally. While most infections resolve spontaneously, persistent infection with high-risk genotypes (HPV-16 and HPV-18) is causally linked not only to cervical cancer but to a broader spectrum of malignancies, including anal, oral, and oropharyngeal cancers, affecting both women and men. Since its introduction in 2006, the HPV vaccine has proven near-complete efficacy against cervical cancer and approximately 82.7% efficacy against oral and oropharyngeal cancers, establishing vaccination as a cornerstone of cancer prevention. In 2021, Europe’s Beating Cancer Plan set the target of 90% HPV vaccination coverage among girls by 2030, alongside the elimination of cervical cancer (fourth most common cancer in women). All EU/EEA member states now recommend the two-dose HPV vaccination for both adolescent girls and boys within national immunisation programmes, with three countries following the WHO 2022 one-dose regimen guideline (off-label use as the EMA still did not approve it). Five countries have already reached the 90% coverage threshold among girls by age 15. However, despite universal public funding of HPV vaccination across the European Union, the variation of vaccination rates between member states is striking. Uptake ranges from 4% in Bulgaria to 97% in Portugal (2024), with major economies such as France falling below 50%. This paper will try to investigate the current structural, cultural, and programmatic determinants of HPV vaccination differences across EU member states. It will mainly focus on the potential enablers and inhibitors of vaccination: vaccine hesitancy, cervical cancer prevalence, immigration, single-dose regimen adoption, parent support, physician recommendation practices, and school-based vaccination policy. Full article

Pancreatic ductal adenocarcinoma (PDAC) remains one of the most lethal malignancies, with a subset occurring in the context of hereditary cancer predisposition. BRIP1 is involved in homologous recombination repair, and pathogenic germline variants are established risk factors for ovarian cancer. However, its role in pancreatic cancer susceptibility and treatment response remains poorly defined. We report PDAC in a father and son whose tumors independently harbored the same BRIP1 p.K159E variant of uncertain significance (VUS), with clinical features suggestive of potential homologous recombination deficiency (HRD). A father and son developed PDAC at 70 and 39 years, respectively. Tumor profiling identified the same BRIP1 p.K159E VUS in both, alongside canonical PDAC-associated alterations. Germline testing was not performed. Both patients showed clinically meaningful sensitivity to platinum-based chemotherapy. The father achieved a partial response to FOLFIRINOX for early metastatic recurrence but subsequently deteriorated and died within one year of diagnosis. The son achieved prolonged disease control with platinum-based chemotherapy and multimodal treatment, including surgery and local therapies for oligoprogression, surviving approximately five years. Maintenance olaparib was briefly administered but discontinued because of hematological toxicity and subsequent disease progression. The occurrence of PDAC in two first-degree relatives, including early-onset disease in the son, together with the same BRIP1 p.K159E VUS in both tumors, represents an unusual familial and molecular observation. Although germline status and pathogenicity cannot be established, the shared alteration and platinum sensitivity raise the hypothesis that this BRIP1 variant may warrant further investigation in the context of homologous recombination repair. However, platinum sensitivity alone cannot be considered evidence of homologous recombination deficiency. The uncertain benefit from PARP inhibition further highlights the limitations of inferring therapeutic actionability from an individual HRR gene VUS. Further germline and functional investigation of BRIP1 alterations in PDAC is warranted. Full article

Background/Objectives: Oral potentially malignant diseases (OPMDs) with signs of epithelial dysplasia are characterized by a high risk of malignancy. However, there are still no algorithms for their early diagnosis and prediction, due to the many possible predisposing factors, including human papillomavirus (HPV) infection and decreased immunity. There is a need to assess the relationship between the manifestations of oral epithelial dysplasia (OED), HPV invasion in the oral cavity, and interferon status to improve the identification and characterization of patients with different degrees of dysplasia. Methods: This study was designed and reported with reference to the STROBE checklist. The study aimed to assess the association between interferon status and HPV invasion in patients with OED. As per the inclusion and exclusion criteria, this observational study included 180 patients with histologically confirmed oral epithelial dysplasia associated with leukoplakia or oral lichen planus. Interferon (IFN) status was assessed by measuring serum interferon activity, spontaneous interferon production, IFN-α/β, and IFN-γ activity in peripheral blood. HPV DNA was detected and genotyped in biopsy specimens using polymerase chain reaction. Results: HPV-positive patients demonstrated significantly lower IFN-α/β and IFN-γ activity compared with HPV-negative patients (p < 0.001). Severe OED was more frequently observed in HPV-positive individuals, whereas mild dysplasia predominated among HPV-negative patients (p < 0.05). Interferon activity progressively decreased with increasing dysplasia severity, with the lowest IFN-α/β and IFN-γ activity detected in patients with severe dysplasia (p < 0.001 and p < 0.05, respectively). A significant positive correlation between IFN-α/β and IFN-γ activity was identified in patients with moderate dysplasia (p = 0.003). Multivariate binary logistic regression analysis demonstrated that HPV positivity (adjusted OR = 4.62, 95% CI: 2.18–9.79; p < 0.001) and reduced IFN-α/β activity (adjusted OR = 1.34 per 10-unit decrease, 95% CI: 1.14–1.58; p < 0.001) were independently associated with severe dysplasia. The multivariable model demonstrated acceptable discriminatory ability for identifying patients with severe dysplasia (AUC = 0.736), with a sensitivity of 89.4% and a specificity of 56.1%. Conclusions: The findings suggest that HPV-associated OED is linked to reduced interferon status, particularly lower IFN-α/β and IFN-γ activity in more severe dysplastic lesions. Combined assessment of interferon status and HPV detection may help identify patients at higher risk of severe OED. Full article

Persistent infection with high-risk human papillomavirus, particularly HPV16, is a major cause of cervical cancer and orogenital malignancies. Therapeutic DNA vaccines targeting viral oncoproteins represent a promising strategy to induce antitumor immune responses. However, the impact of antigen combinations in bicistronic vectors and the immunization sequence on therapeutic efficacy remains poorly understood. Thus, this study aimed to characterize in silico previously described E5CP and E7CP vaccines, develop a new bicistronic construct, and evaluate its antitumor efficacy under different immunization regimens in vivo. The constructs were initially characterized regarding predicted physicochemical, immunological, and structural properties, followed by molecular confirmation of plasmid construction and qualitative assessment of transcript detection after transient transfection. In vivo, C57BL/6 mice bearing C3 tumors were immunized with five different regimens combining the monocistronic or bicistronic constructs. At the endpoint, tumors and spleens were collected for histopathological and immunological analyses. The results showed that regimens using the bicistronic construct or including E5 in the first dose were associated with lower mean tumor volumes or delayed tumor growth compared to the control groups. Histopathological and immunophenotypic changes were variable among regimens, and an exploratory analysis suggested an association between lower nitric oxide levels and smaller tumor volumes. Therefore, the order of antigen administration and the use of bicistronic constructs warrant further investigation as strategies that could be optimized to improve the therapeutic efficacy of HPV DNA vaccines. Full article

Background: Oral squamous cell carcinoma (OSCC) has a relatively poor survival, and TNM staging alone offers limited prognostic discrimination. C-reactive protein (CRP) is a non-specific inflammatory marker and is one of the most studied candidate prognostic biomarkers of OSCC, while CYFRA 21-1 and squamous cell carcinoma antigen (SCC-Ag) are tumor-derived keratin proteins reflecting malignant cell turnover, often examined alongside CRP. However, no prior review has synthesized the value of these three markers together in OSCC. Methods: Following the PRISMA 2020 guidelines, PubMed, Embase, and Web of Science were searched for studies reporting an extractable hazard ratio (HR), odds ratio, or comparable association of serum and/or salivary CYFRA 21-1, SCC-Ag, and/or CRP with disease-free survival (DFS), recurrence, or overall survival (OS) in OSCC. HRs were pooled using Knapp–Hartung random-effects models, wherever at least two independent cohorts contributed; salivary CYFRA 21-1 and recurrence were pooled as a standardized mean difference (SMD). Studies reporting only cross-sectional associations with tumor stage, grade, or metastasis and non-independent (same-registry) studies were synthesized descriptively following Synthesis without Meta-analysis (SWiM) guidance. Risk of bias was assessed with QUIPS and certainty of evidence with the GRADE framework. Results: Twenty-eight studies were included (21 quantitative, 7 narrative), representing ~3500 study participants across published cohorts, with some overlap between studies. For OS, two independently pooled single-marker estimates were both non-significant: serum SCC-Ag alone (n = 2, HR 1.43, 95% CI: 0.09–23.14, p = 0.35, I² = 66%) and serum CRP alone (n = 5, HR 1.33, 95% CI: 0.89–1.99, I² = 52%). A further seven non-independent, same-registry studies reporting combined SCC-Ag + CRP or single-marker models ranged from HR 1.82 to 11.11, directionally consistent with the pooled estimates. For DFS, pooled serum CYFRA 21-1 alone gave HR 1.61 (95% CI: 0.39–6.61; p = 0.15, I² = 0%), with six further non-independent or single-study estimates (HR 1.38–4.05) in the same direction. For recurrence, pooled salivary CYFRA 21-1 gave an SMD of Hedges’ g = 0.79 (95% CI: 0.04–1.55; p = 0.048, I² = 0%). All outcomes were rated very low certainty by GRADE. Conclusions: Within the limitations of the study and the small number of contributing studies, salivary CYFRA 21-1 showed some prognostic value for recurrence, while pooled estimates for serum CRP alone, serum SCC-Ag alone, and serum CYFRA 21-1 alone for DFS all trended toward increased risk but did not reach statistical significance. Given the very low evidence certainty, these findings are hypothesis-generating and require further validation in independent, multicenter, prospective cohorts before clinical application. Full article

Background/Objectives: Postoperative hypocalcemia is the most common complication following thyroidectomy; however, previous studies have largely relied on biochemical definitions without distinguishing symptomatic hypocalcemia from asymptomatic biochemical abnormalities. This study aimed to identify the independent predictors of symptomatic postoperative hypocalcemia. Methods: This retrospective single-center study included 245 adult patients who underwent thyroidectomy between January 2017 and December 2022. Patients were classified as normocalcemic (n = 88), asymptomatic biochemical hypocalcemic (n = 87), or symptomatic hypocalcemic (n = 70) according to calcium status within the first 72 postoperative hours. Demographic, surgical, biochemical, ultrasonographic, cytological, and histopathological variables were analyzed. Independent predictors were identified using multivariable logistic regression. Results: Symptomatic hypocalcemia occurred more frequently after bilateral total thyroidectomy with central neck dissection (p = 0.001). Patients with symptomatic hypocalcemia had lower preoperative serum calcium and lower postoperative albumin and parathyroid hormone levels than the other groups (all p < 0.05). Although high-risk FNAB cytology and malignant histopathology were associated with hypocalcemia in univariate analyses, neither remained independently predictive after adjustment. Bilateral total thyroidectomy with central neck dissection (adjusted OR 6.63, 95% CI 1.68–26.11; p = 0.007) and lower preoperative serum calcium (adjusted OR 0.46, 95% CI 0.22–0.94; p = 0.036) were independent predictors of symptomatic hypocalcemia after thyroidectomy. Conclusions: Symptomatic postoperative hypocalcemia should be considered a distinct clinical outcome rather than merely a biochemical abnormality. Bilateral total thyroidectomy with central neck dissection and lower preoperative serum calcium independently predicted symptomatic hypocalcemia, whereas ultrasonographic findings, FNAB cytology, histopathological diagnosis, and incidental parathyroidectomy did not. Although the absence of perioperative vitamin D and magnesium measurements is an important limitation, the identified predictors may provide clinically useful information for perioperative risk stratification. Full article

Background/Objectives: Sarcoma comprises mesenchymal malignancies for which PD-1/PD-L1 checkpoint inhibitors have produced few durable responses. The CD47–SIRPα innate checkpoint suppresses macrophage phagocytosis through a “don’t eat me” signal and is the most clinically advanced myeloid checkpoint, but its evidence base in sarcoma has not been systematically assembled or appraised. Methods: A PRISMA 2020-compliant systematic review (PROSPERO CRD420261452040) was undertaken; three bibliographic databases and three trial registers, without language restriction, were searched on 14 September 2026. Expression, preclinical and clinical studies were appraised and graded separately. Synthesis followed SWiM guidelines and no pooled effect estimate was calculated. Results: Of 391 records identified, 50 studies met the eligibility criteria as separate analysis units (45 journal articles, 4 conference abstracts, 1 registry record), appraised across expression (20), preclinical (38) and clinical (18, including 2 interventional trials) domains. CD47 expression was subtype-dependent and bimodal, consistent in chordoma and angiosarcoma, frequently negative in leiomyosarcoma and Ewing sarcoma, and contested in undifferentiated pleomorphic sarcoma; reported positivity in osteosarcoma varied six-fold between cohorts, and the assay was often incompletely described. In preclinical models, CD47 blockade consistently increased phagocytosis and combination regimens outperformed single-agent use, but few studies used patient-derived or humanized models. Only two interventional trials enrolled sarcoma patients; both were single-arm, one was terminated without posted results, and nine of ten pharmacological strategies had no sarcoma-specific clinical data. Certainty was low for expression and patient-cohort evidence, and very low for preclinical and interventional evidence. Conclusions: The CD47–SIRPα axis is biologically well characterized and expressed in a definable subset of sarcomas, but has not been tested in these tumors in a design capable of detecting a treatment effect. Four of the fifty records were conference abstracts, which carry a higher risk of reporting bias and non-publication. The rate-limiting step lies in the assay and the trial architecture rather than in the biology: a harmonized immunohistochemical standard and a biomarker-selected trial with an internal comparator are prerequisites for an informative next study. Full article

Background and Objectives: Bethesda Category IV thyroid nodules present a diagnostic challenge due to their variable risk of malignancy and uncertain cytological features. Therefore, determining the risk of malignancy before surgery is important. This study aims to identify clinical, biochemical, and ultrasonographic factors associated with malignancy and to explore the performance of a multivariable analysis based on routinely available preoperative features in thyroid nodules. Methods: This retrospective, single-center study included 105 patients who underwent thyroidectomy for Bethesda Category IV cytology. The mean age was 50.13 ± 11.78 years. 87 patients (82.9%) were female, and 18 (17.1%) were male. Patients were classified into benign/borderline (n = 79) and malignant (n = 26) groups based on their postoperative histopathological findings. Demographic characteristics, serum thyroid function tests, thyroid autoantibody levels, and ultrasonographic findings were compared. Variables significantly associated with malignancy were included in a Firth bias-reduced penalized multivariable logistic regression analysis. The discriminative performance of the multivariable analysis was evaluated using ROC curve analysis, and internal validation was performed using bootstrap resampling. The AUC was compared with that of ACR TI-RADS using DeLong’s test. Results: The malignant group showed a significantly higher prevalence of male sex, irregular margins, a taller-than-wide shape, and microcalcification (all p < 0.05). In the multivariable logistic regression analysis, only irregular margins (OR = 9.46, 95% CI: 1.88–47.59; p = 0.006) were independently associated with malignancy. The model demonstrated acceptable discrimination in this study population (n = 105), with an AUC of 0.717. The AUC of the multivariable analysis was 0.717, compared with 0.704 for ACR TI-RADS, and the difference was not statistically significant (ΔAUC = 0.013; DeLong p = 0.803). Conclusions: In Bethesda Category IV thyroid nodules, irregular margins remained independently associated with malignancy. The multivariable analysis did not demonstrate significantly better discrimination than ACR TI-RADS. These findings should be considered exploratory and require external validation in larger cohorts. Full article

Background and Objectives: Hereditary cancer predisposition is associated with pathogenic germline variants in numerous genes. Identification of inherited cancer susceptibility enables personalized screening, preventive interventions, and targeted therapies. It also facilitates cascade testing of relatives who may be at increased risk and benefit from tailored clinical management. We describe the first results of hereditary cancer patient identification, selection, and genetic testing at the University Hospital of Split. Materials and Methods: Next-generation sequencing and hereditary cancer gene panel analysis were performed in individuals who met the National Comprehensive Cancer Network clinical criteria for genetic testing at the University Hospital of Split between May 2025 and July 2026. A comprehensive cancer gene panel covering 101 genes was used. Results: A total of 297 individuals underwent genetic testing, including 268 (90.24%) females and 29 (9.76%) males. Of these, 138 (46.46%) were unaffected individuals who met testing criteria based on a family history of cancer. Among individuals with a personal history of malignancy, breast cancer was the most common diagnosis, occurring in 130 patients (43.77%). Pathogenic or likely pathogenic (P/LP) variants, as well as risk variants, were identified in 80 individuals (26.94%), including seven individuals carrying two P/LP variants. Variants were detected in 29 different genes, most frequently BRCA1 (n = 14), BRCA2 (n = 11), CHEK2 (n = 9) and PALB2 (n = 7). The detected variant types included frameshift (n = 30), missense (n = 22), nonsense (n = 21), splice-site variants (n = 4), start loss variants (n = 4), large deletions (n = 4), and in-frame deletions (n = 2). Conclusions: To our knowledge, this is the first study to report the results of panel germline testing for hereditary cancer syndromes in a Croatian population. These findings provide initial epidemiological data on the spectrum and distribution of P/LP variants. The relatively high detection rate may reflect the recent implementation of genetic testing and the careful selection of individuals with a strong clinical suspicion of hereditary cancer predisposition. Full article

Human papillomavirus (HPV) drives cervical carcinogenesis, but viral infection alone is insufficient for malignant transformation. The vaginal microbiome may modulate HPV persistence and cervical intraepithelial neoplasia (CIN) progression, yet existing reviews remain inconclusive. This systematic review identifies specific vaginal microbiome risk factors associated with HPV and CIN and proposes plausible mechanisms linking dysbiosis to disease progression. Following (PRISMA) guidelines, databases were searched for studies published through October 2025. Thirty-three studies from 2013 to 2025 across 10 countries met the inclusion criteria, with 87.9% exhibiting low risk of bias. Most studies demonstrated marked Lactobacillus depletion alongside Prevotella and Gardnerella enrichment in HPV/CIN cases. Disease severity correlated with shifts toward high-diversity anaerobe communities, though alpha and beta diversity metrics varied across cohorts. Importantly, longitudinal data suggests a temporal sequence where baseline dysbiosis precedes clinical progression. Crucially, although the included cohorts directly observed and quantified these taxonomic shifts, the underlying mechanisms of disease progression, such as enzymatic degradation of mucosal barriers mediated by Gardnerella and immunosuppression driven by Fusobacterium, remain largely inferred from external in vitro literature. Lactobacillus iners emerged as a notable species-level risk factor in HPV persistence. Vaginal dysbiosis, characterised by Lactobacillus depletion and anaerobic expansion, is strongly correlated with HPV persistence and CIN progression, potentially facilitated through convergent barrier degradation and immune subversion. These findings highlight targets for microbiome-based risk stratification and therapeutic intervention. Full article

Parasitic neglected tropical diseases (PNTDs) are primarily infections of poverty, impaired sanitation, and environmental exposure, but only a limited subset has recognized links to malignancy. The rationale for this Perspective is that these cancer-associated infections sit at the interface of communicable disease control, chronic inflammation, delayed organ damage, and non-communicable cancer prevention. At least 253.7 million people required preventive treatment for schistosomiasis in 2024, while infection-attributable cancer estimates suggest that liver flukes accounted for approximately 1300 to nearly 7000 new cholangiocarcinoma cases annually depending on modelling assumptions. The strongest recognized cancer associations involve Schistosoma haematobium with urinary bladder cancer and the food-borne liver flukes Opisthorchis viverrini and Clonorchis sinensis with cholangiocarcinoma. Importantly, infection with these parasites does not inevitably lead to cancer; malignant transformation is an uncommon, long-latency outcome shaped by parasite burden, reinfection, chronic inflammation, host susceptibility, co-infections, environmental exposures, nutrition, and access to care. This Perspective proposes a focused, risk-stratified framework that links parasite control, precision diagnostics, digital surveillance, targeted longitudinal follow-up, and implementation priorities for selected high-risk infections and settings. Rather than advocating broad cancer surveillance for all PNTDs, the article argues for staged, locally feasible approaches that strengthen existing control platforms while using advanced tools selectively for research, referral pathways, and populations at elevated risk. Full article

Background/Objectives: Peritoneal dissemination is a major pattern of failure in advanced digestive cancer and is poorly detected by cross-sectional imaging. Outside gastric cancer, peritoneal lavage cytology is rarely performed and carries no staging weight. We examined the association between cytological status and subsequent peritoneal carcinomatosis in patients without peritoneal disease on imaging or at operation. Methods: Retrospective prognostic study of consecutive patients with clinically staged T3–T4 digestive malignancy operated on by one surgical team (January 2019–December 2022). Lavage was performed before tumour manipulation and processed as cell blocks. The exposure was cytological status, positive (CY1) or negative (CY0); the outcome was radiologically evident peritoneal carcinomatosis within 12 months on computed tomography at 3, 6, 9 and 12 months, analysed as interval-censored data. Results: Of 62 patients, 10 (16.1%) had carcinomatosis at operation despite negative imaging. Of the 45 with digestive primaries and no peritoneal disease, 13 (28.9%) were CY1. Carcinomatosis developed in 10 of 13 CY1 and 5 of 32 CY0 patients (76.9% versus 15.6%; relative risk 4.92, 95% CI 2.09–11.62), an association preserved in patients without distant metastases and in those who underwent resection. The hazard ratio from a discrete-time proportional hazards model was 9.07 (95% CI 3.15–29.72; p = 0.00005). No clinicopathological variable examined was significantly associated with cytological status. Conclusions: In this exploratory series, positive lavage cytology was associated with early radiologically evident peritoneal carcinomatosis. The association is unadjusted and derives from a small, heterogeneous single-centre cohort; it does not establish independent prognostic value, nor does it support routine lavage cytology or changes to postoperative surveillance. It warrants prospective evaluation. Full article

Background/Objectives: Neuroblastoma is a high-risk pediatric malignancy in which cancer stem cells (CSCs) contribute to treatment resistance and disease recurrence. All-trans retinoic acid (ATRA) has established differentiation-inducing activity in neuroblastoma but is constrained by poor aqueous solubility and formulation challenges. Methods: ATRA nanocrystals (ATRA-NCs) stabilized with Poloxamer 407 were developed and characterized for their physicochemical properties. Their anticancer activity was evaluated in IMR-32 neuroblastoma cells and CD133+ CSCs, with unformulated ATRA used as the comparator, using multiple in vitro biological assays. Results: ATRA-NCs exhibited a mean hydrodynamic diameter of 194 ± 5.3 nm and a zeta potential of −60.1 mV. Compared with unformulated ATRA, ATRA-NCs produced greater concentration-dependent cytotoxicity and enhanced apoptotic responses in IMR-32 neuroblastoma cells. In CD133+ CSCs, ATRA-NCs reduced cell viability and suppressed the expression of stemness-associated genes, including SOX2, NANOG, LGR5, OCT4, BMI-1, and ALDH1A1. The formulation also reduced colony-forming capacity, impaired spheroid formation, and inhibited cellular migration. Conclusions: These findings demonstrate enhanced in vitro anticancer activity against neuroblastoma cancer cells and CD133+ CSCs following ATRA nanocrystallization and indicate modulation of multiple functional phenotypes associated with neuroblastoma CSCs. ATRA nanocrystallization may therefore provide a formulation approach for enhancing ATRA activity against neuroblastoma and warrants further preclinical evaluation.. Full article