Background The anatomical TNM staging system inadequately reflects the profound survival heterogeneity in gallbladder cancer (GBC), thereby limiting accurate risk stratification and contributing to suboptimal clinical decision-making. To overcome this limitation, we developed a multidimensional and dynamic prognostic framework integrating clinicopathological features, systemic inflammatory markers, and tumor biomarkers, leveraging large-scale multicenter real-world data. Methods A total of 1,354 patients with GBC from 44 medical centers were retrospectively analyzed and randomly assigned to training (n = 947) and validation (n = 407) cohorts. Independent prognostic factors were identified using LASSO and multivariable Cox regression to construct three risk scores: clinicopathological (C-score), blood-based inflammatory (B-score), and tumor marker (T-score). An integrated prognostic model was subsequently developed and evaluated through machine-learning–based benchmarking. Model performance was assessed using time-dependent ROC analysis (6–48 months), calibration curves, and decision curve analysis (DCA). Gene Set Enrichment Analysis (GSEA) was performed to explore the biological relevance of the scoring systems. Results Age, R0 resection status, TNM stage, CA19-9 (log1p), neutrophil-to-lymphocyte ratio (NLR), and lymphocyte-to-CRP ratio (LCR) were identified as independent prognostic factors. The integrated model consistently outperformed individual C-, B-, and T-scores across all follow-up intervals, demonstrating strong discriminative ability with a 12-month AUC of 0.857 in both cohorts. Calibration and decision curve analyses confirmed good model reliability and clinical utility. GSEA revealed distinct molecular associations underlying the three scores, including ECM–receptor interaction, immune-inflammatory signaling (JAK–STAT/NF-κB), and metabolic stress pathways (HIF-1/p53). A web-based dynamic prediction platform was further developed to enable individualized survival estimation. Conclusion This multidimensional framework provides a biologically interpretable and dynamically adaptable tool for prognostic stratification in gallbladder cancer. Implemented through a web-based platform, the model facilitates individualized risk assessment and supports data-driven clinical decision-making.

Liver invasion is one of the most frequent events in the progression of gallbladder cancer (GBC). However, the cellular and pathological role of the tumor-liver-interface microenvironment in liver invasion is still enigmatic. Here, we applied single-cell and spatial transcriptomics to systematically investigate the cellular component and gene expression regulation of the microenvironment from the tumor to the liver, specifically the invasive boundary. Our analyses revealed that CXCL9+ macrophage-rich immune cell niches were accumulated in the tumor-liver invasive margin, where 2 subclasses of the CXCL9+ immune cell niches, CXCL9+TRAC+ (CT) and CXCL9+C1QB+ (CC) niches, were identified. CD8+ T cells were recruited by CXCL9+ macrophages through CXCL9-CXCR3 interaction in the CT niche, which was located adjacent to the liver. Moreover, the CC niche was proximal to the tumor core, where tumor cells induced CD8+ T cell exhaustion via LGALS4 expression. In addition, our cohort study showed that high CXCL9 and low LGALS4 in the liver invasion margin demonstrated a favorable prognosis and better responses to anti-PD-1 immunotherapy for patients with gallbladder cancer. Altogether, these findings demonstrate novel cellular and molecular mechanisms underlying liver invasion and offer clinical value for immunotherapies.

INTRODUCTION:Subsegment IVb (S4b) is a common site of direct invasion in gallbladder cancer, and its complete resection is critical to achieve negative surgical margins. However, the portal venous anatomy of S4b is highly complex and exhibits significant interindividual variation, which the conventional Couinaud classification fails to adequately delineate. This study utilized three-dimensional visualization to systematically characterize the anatomical features and branching patterns of the portal veins supplying S4b, with the goal of establishing a practical anatomical classification to inform preoperative planning for radical gallbladder cancer surgery. METHODS:This retrospective analysis utilized thin-slice computed tomography data from 115 patients who underwent abdominal contrast-enhanced computed tomography for a spectrum of clinical indications (e.g., evaluation of focal liver lesions, trauma assessment, or other abdominal pathologies). Three-dimensional reconstruction of the portal venous system was performed using dedicated visualization software. We focused on analyzing the origin, number, course, and common trunk patterns of the internal branches of the left portal vein supplying S4b. RESULTS:S4b was supplied by 1 to 6 portal vein branches, with 2 to 6 branches observed in 87.1% of cases. The overwhelming majority (>90%) of these branches originated from the umbilical portion of the left portal vein. Based on the common trunks supplying both S4a and S4b, the vascular anatomy was classified into four types: type A (no common trunk, 23.4%), type B (one common trunk 53%), type C (two common trunks 20%), and type D (more than two common trunks or a branch derived from the right portal vein 3.4%). Overall, 73% of patients exhibited 1-2 common trunks. The diameter of these common trunks correlated with their number, and their trajectories consistently coursed toward the S4a segment. CONCLUSIONS:The proposed four-type classification systematizes the broad spectrum of S4b portal venous anatomy. This framework facilitates a preoperatively planned, targeted resection in radical gallbladder cancer surgery, which is instrumental in optimizing procedural safety and oncologic outcomes.

PURPOSE:The purpose of this study is to evaluate the efficacy and safety of nab-paclitaxel combined with gemcitabine (AG regimen) as a first-line chemotherapy for patients with unresectable gallbladder cancer (GBC). MATERIALS AND METHODS:A retrospective analysis was conducted on patients diagnosed with GBC via histological examination. Inclusion criteria included locally advanced or metastatic gallbladder cancer deemed unresectable by multidisciplinary team (MDT) consensus; age 18-75 years; ECOG performance status ≤ 2; and AG as first-line chemotherapy. Treatment response was assessed using RECIST 1.1 criteria. Primary endpoints were overall response rate (ORR) and progression-free survival (PFS); secondary endpoints included disease control rate (DCR), overall survival (OS), and adverse events incidence. RESULTS:Thirty-one patients were enrolled. The ORR was 48.4%, and DCR was 90.3%. The median PFS was 8.4 months (95% CI, 5.0-11.8), and the median OS was 20.9 months (95% CI, 16.8-24.9). Among the 11 patients with locally advanced disease, 7 patients (63.6%) successfully converted to a resectable state and underwent surgery. Grade 3/4 treatment-related adverse events occurred in 11 patients (35.5%), predominantly hematologic toxicities, and no treatment-related deaths or treatment discontinuations due to adverse events were observed. CONCLUSION:First-line AG chemotherapy demonstrated promising efficacy and manageable safety for patients with unresectable GBC, with a substantial portion achieving disease control and some converting to resectable status.

INTRODUCTION:In the AJCC 8th edition, N2M0 and M1 gallbladder cancer (GBC) are classified into Stage IVB, leading clinicians to treat them identically. This study separates N2M0 from M1 disease, re-evaluating chemotherapy, surgical quality, and establishing a clinical risk stratification system (CRSS). MATERIALS AND METHODS:Stage IVB GBC patients (2000-2022) were identified from the SEER database. A 1:3 propensity score matching (PSM) balanced N2M0 and M1 baseline characteristics. Overall survival was compared, optimal lymph node ratio (LNR) was identified, and the prediction model internally verified using 1000 bootstrap resamples. RESULTS:From 7270 initial IVB patients, PSM matched 125 N2M0 and 343 M1 patients. N2M0 patients demonstrated superior median overall survival versus M1 patients (17 vs. 9 months, P = 0.001). However, this survival advantage depended entirely on adjuvant chemotherapy. Untreated N2M0 patients had a worse prognosis than treated M1 patients (P = 0.002). For N2M0 patients, extended multi-organ resection offered no survival benefit over standard radical cholecystectomy (HR = 0.724, P = 0.225). A lower LNR (<0.875) predicted longer survival. We developed a three-tier CRSS separating median survival: 22 months (Tier 1: Low LNR + Chemotherapy), 12 months (Tier 2: High LNR or No Chemotherapy), and 9 months (Tier 3: M1 Systemic Disease). The nomogram C-index was 0.676. CONCLUSIONS:N2M0 GBC is biologically different from M1 disease, and should not be grouped together in future staging. For N2M0 patients, the best strategy is to focus on thorough lymph node dissection and mandatory adjuvant chemotherapy, not extended organ resection.

BACKGROUND:While the inflammation-to-carcinoma transition in gallbladder carcinogenesis is well recognized, the molecular pathogenesis of gallbladder adenomyomatosis (GBA), which is clinically associated with both chronic cholecystitis (CCS) and gallbladder carcinoma (GBC), remains poorly understood. Hence, we aimed to evaluate the premalignant potential of GBA and to identify previously unrecognized molecular pathways involved in gallbladder carcinogenesis. METHODS:We first conducted single-cell RNA sequencing and downstream bioinformatics analyses on samples from nine patients with CCS, GBA, and GBC. Key molecular findings were subsequently validated using publicly available bulk RNA-seq datasets (n = 80) and multiplex immunohistochemistry in an independent cohort (n = 62). Functional validation of tumor microenvironment-associated molecules was performed through a series of in vitro and in vivo assays. RESULTS:We profiled the transcriptomic landscape of 89 428 single cells and uncovered distinct epithelial and immune ecosystems across different disease stages. A transitional epithelial subset (SPP1-CCL20⁺ EpiC3) and PTGER4+CCL5⁺ CD8⁺T cells were identified in GBA tissues, indicating malignant potential. Notably, PRDX1 was selectively upregulated in the GBC-derived Macro04 macrophages, which displayed strong interactions with exhausted T cells and malignant epithelial cells. Functionally, PRDX1 knockdown in macrophages disrupted mitochondrial oxidative metabolism, leading to metabolic reprogramming and diminished M2 polarization, thereby attenuating tumor cell proliferation, migration, and invasion both in vitro and in vivo. Mechanistically, PRDX1 loss downregulated PPAR-δ and CPT1A, whereas PPAR-δ agonist GW1516 restored CPT1A expression as well as the associated metabolic and polarization defects, supporting a PRDX1/PPAR-δ/CPT1A regulatory axis governing macrophage immunometabolism in GBC. CONCLUSIONS:Our study reveals that GBA harbors malignant potential at both epithelial and immune microenvironmental levels. The expression of CCL20, CCL5, and PRDX1 may serve as molecular markers for stratifying high-risk GBA, while PRDX1 represents a promising therapeutic target for reprogramming the tumor immune microenvironment in GBC.

BACKGROUND:The accurate early-stage diagnosis of gallbladder cancer (GBC) is regarded as one of the major challenges in the field of oncology. However, few studies have focused on the comprehensive classification of GBC based on multiple modalities. This study aims to develop a comprehensive diagnostic framework for GBC based on both imaging and non-imaging medical data. METHODS:This retrospective study reviewed 298 clinical patients with gallbladder disease or volunteers from two devices. A novel end-to-end interpretable diagnostic framework for GBC is proposed to handle multiple medical modalities, including CT imaging, demographics, tumor markers, coagulation function tests, and routine blood tests. To achieve better feature extraction and fusion of the imaging modality, a novel global-hybrid-local network, namely GHL-Net, has also been developed. The ensemble learning strategy is employed to fuse multi-modality data and obtain the final classification result. In addition, two interpretable methods are applied to help clinicians understand the model-based decisions. Model performance was evaluated through accuracy, precision, specificity, sensitivity, F1-score, area under the curve (AUC), and matthews correlation coefficient (MCC). RESULTS:In both binary and multi-class classification scenarios, the proposed method showed better performance compared to other comparison methods in both datasets. Especially in the binary classification scenario, the proposed method achieved the highest accuracy, sensitivity, specificity, precision, F1-score, ROC-AUC, PR-AUC, and MCC of 95.24%, 93.55%, 96.87%, 96.67%, 95.08%, 0.9591, 0.9636, and 0.9051, respectively. The visualization results obtained based on the interpretable methods also demonstrated a high clinical relevance of the intermediate decision-making processes. Ablation studies then provided an in-depth understanding of our methodology. CONCLUSION:The machine learning-based framework can effectively improve the accuracy of GBC diagnosis and is expected to have a more significant impact in other cancer diagnosis scenarios.

BACKGROUND:Gallbladder cancer (GBC) is a leading cause of cancer-related death worldwide, and its prognosis remains poor, with 5-year survival of approximately 5%. In this study, we analyzed the involvement of a novel proteoglycan, Sparc/osteonectin, cwcv, and kazal-like domains proteoglycan 1 (SPOCK1), in the tumor progression and prognosis of human GBC. METHODS:SPOCK1 expression levels were measured in fresh samples and stored specimens of GBC and adjacent nontumor tissues. The effect of SPOCK1 on cell growth, DNA replication, migration and invasion were explored by Cell Counting Kit-8, colony formation, EdU retention assay, wound healing, and transwell migration assays, flow cytometric analysis, western blotting, and in vivo tumorigenesis and metastasis in nude mice. RESULTS:SPOCK1 mRNA and protein levels were increased in human GBC tissues compared with those in nontumor tissues. Immunohistochemical analysis indicated that SPOCK1 levels were increased in tumors that became metastatic, compared with those that did not, which was significantly associated with histological differentiation and patients with shorter overall survival periods. Knockdown of SPOCK1 expression by lentivirus-mediated shRNA transduction resulted in significant inhibition of GBC cell growth, colony formation, DNA replication, and invasion in vitro. The knockdown cells also formed smaller xenografted tumors than control GBC cells in nude mice. Overexpression of SPOCK1 had the opposite effects. In addition, SPOCK1 promoted cancer cell migration and epithelial-mesenchymal transition by regulating the expression of relevant genes. We found that activation of the PI3K/Akt pathway was involved in the oncogenic functions of SPOCK1 in GBC. CONCLUSIONS:SPOCK1 activates PI3K/Akt signaling to block apoptosis and promote proliferation and metastasis by GBC cells in vitro and in vivo. Levels of SPOCK1 increase with the progression of human GBC. SPOCK1 acts as an oncogene and may be a prognostic factor or therapeutic target for patients with GBC.

BACKGROUND:Neoadjuvant treatment has demonstrated clinical benefits in advanced gallbladder cancer (GBC). However, the overall response rate remains suboptimal, and the underlying mechanisms driving treatment efficacy are not fully understood. MATERIALS AND METHODS:This study aimed to evaluate the therapeutic effects of neoadjuvant chemo-immunotherapy (NAT) combining gemcitabine, nab-paclitaxel, and anti-PD-1 immunotherapy in advanced GBC, and to investigate the associated tumor microenvironment (TME) alterations. Single-cell RNA sequencing and multiplex immunohistochemistry were utilized to analyze the cellular composition of the TME in patients who successfully underwent downstaging. RESULTS:NAT significantly reshaped the tumor-immune landscape, characterized by an expansion of follicular helper T (Tfh) cells and the formation of tertiary lymphoid structures (TLSs) in 50% of treated tumors. Inflammatory cancer-associated fibroblasts (CAFs) increased and exhibited upregulation of CCL19 and CXCL12 , potentially promoting Tfh cell recruitment and TLS formation. Additionally, NAT led to an expansion of GZMB + cytotoxic CD8 + T cells with an exhausted phenotype in GBC, but not in adjacent normal tissues. The treatment also increased the number and effector functions of natural killer cells while reducing tumor-promoting macrophages and angiogenesis-related CAFs. Furthermore, NAT decreased the cancer stem cell-like subpopulation while increasing a cancer cell subset with enhanced antigen-presenting capacity. CONCLUSION:This study suggested the potential efficacy of NAT in advanced GBC and revealed alterations in the TME following treatment. The findings provided insights into the mechanisms underlying NAT responses and offered valuable directions for optimizing therapeutic strategies in GBC.

Gallbladder cancer (GBC), the most common malignancy of the bile duct, is highly aggressive and has an extremely poor prognosis, which is a result of early metastasis. As it is regulated being at multiple levels, the metastatic cascade in GBC is complex. Recent evidence suggests that microRNAs (miRNAs) are involved in cancer metastasis and are promising therapeutic targets. In this study, miR-101 was significantly downregulated in tumor tissues, particularly in metastatic tissues. In GBC patients, low miR-101 expression was correlated with tumor size, tumor invasion, lymph node metastasis, TNM stage, and poor survival. Moreover, miR-101 was an independent prognostic marker for GBC. Additionally, miR-101 inhibited GBC cell proliferation, migration, invasion, and TGF-β-induced epithelial-mesenchymal transition (EMT) in vitro and in vivo. Mechanistically, the gene encoding the zinc finger protein X-linked (ZFX) was identified as a direct target of miR-101. More importantly, miR-101 significantly reduced activation of the MAPK/Erk and Smad signaling pathways, resulting in inhibition of TGF-β-mediated induction of EMT. Altogether, our findings demonstrate a novel mechanism by which miR-101 attenuates the EMT and metastasis in GBC cells and suggest that miR-101 can serve as a potential biomarker and therapeutic target for GBC management.

Reprogrammed cellular metabolism plays a critical role in the development and progression of various cancers. However, the mechanisms by which these metabolic changes drive malignancy in gallbladder cancer (GBC) remain unclear. In this study, we identified significant alterations in branched-chain amino acid (BCAA) metabolism in GBC through comprehensive transcriptomic and metabolic analyses. Reduced activity of the BCAA catabolic enzymes ACADS, ACADSB, and BCKDHA was associated with poor prognosis in GBC patients. Dietary reduction of BCAAs in mouse model of GBC significantly slowed tumor growth. The lncRNA LPAL2 was found to correlate with the expression of BCAA catabolic enzymes. In vivo and in vitro assays demonstrated that LPAL2 inhibited GBC cell proliferation, downregulated intracellular BCAA levels, and suppressed mTORC1 activation. Furthermore, LPAL2 and ACADS were identified as independent prognostic factors for survival. Mechanically, IGF2BP1 maintaining LPAL2 stability through methylation. Additionally, LPAL2 decreased YBX1 stability by promoting its ubiquitination-mediated degradation, while YBX1 inhibited the transcription of certain BCAA catabolic enzymes through binding to their promoters. In summary, the LPAL2-YBX1 interaction regulates BCAA metabolism to influence GBC cell proliferation, which could be targeted for therapeutic interventions in GBC treatment.

Gallbladder cancer (GBC) frequently mimics gallbladder benign lesions (GBBLs) in radiological images, leading to preoperative misdiagnoses. To address this challenge, we initiated a prospective, multicenter clinical trial (ChicCTR2100049249) and proposed a multimodal, non-invasive diagnostic model to distinguish GBC from GBBLs. A total of 301 patients diagnosed with gallbladder-occupying lesions (GBOLs) from 11 medical centers across 7 provinces in China were enrolled and divided into a discovery cohort and an independent external validation cohort. An artificial intelligence (AI)-based integrated model, GBCseeker, is created using cell-free DNA (cfDNA) genetic signatures, radiomic features, and clinical information. It achieves high accuracy in distinguishing GBC from GBBL patients (93.33% in the discovery cohort and 87.76% in the external validation cohort), reduces surgeons' diagnostic errors by 56.24%, and reclassifies GBOL patients into three categories to guide surgical options. Overall, our study establishes a tool for the preoperative diagnosis of GBC, facilitating surgical decision-making.

The journal retracts the article, titled “Schisandrin B induces apoptosis and cell cycle arrest of gallbladder cancer cells” [...]

[This corrects the article DOI: 10.7150/jca.46351.].

Gallbladder cancer (GBC) is among the most common malignancies of biliary tract system due to its limited treatments. The immunotherapeutic targets for T cells are appealing, however, heterogeneity of T cells hinds its further development. We systematically construct T cell atlas by single-cell RNA sequencing; and utilized the identified gene signatures of high_CNV_T cells to predict molecular subtyping towards personalized therapeutic treatments for GBC. We identified 12 T cell subtypes, where exhausted CD8+ T cells, activated/exhausted CD8+ T cells, and regulatory T cells were predominant in tumors. There appeared to be an inverse relationship between Th17 and Treg populations with Th17 levels significantly reduced, whereas Tregs were concomitantly increased. Furthermore, we first established subtyping criterion to identify three subtypes of GBC based on their pro-tumorigenic microenvironments, e.g., the type 1 group shows more M2 macrophages infiltration, while the type 2 group is infiltrated by highly exhausted CD8+ T cells, B cells and Tregs with suppressive activities. Our study provides valuable insights into T cell heterogeneity and suggests that molecular subtyping based on T cells might provide a potential immunotherapeutic strategy to improve GBC treatment.

Gallbladder cancer (GBC) is a highly aggressive malignancy lacking clinically available targeted therapeutic agents. Super-enhancers (SEs) are crucial epigenetic cis-regulatory elements whose extensive reprogramming drives aberrant transcription in cancers. To study SE in GBC, the genomic distribution of H3K27ac is profiled in multiple GBC tissue and cell line samples to establish the SE landscape and its associated core regulatory circuitry (CRC). The biliary lineage factor SOX9 and Wnt pathway effector TCF7L2, two master transcription factor (TF) candidates identified by CRC analysis, are verified to co-occupy each other's SE region, forming a mutually autoregulatory loop to drive oncogenic SE reprogramming in a subset of GBC. The SOX9/TCF7L2 double-high GBC cells are highly dependent on the two TFs and enriched of SE-associated gene signatures related to stemness, ErbB and Wnt pathways. Patients with more such GBC cells exhibited significantly worse prognosis. Furthermore, SOX9/TCF7L2 double-high GBC preclinical models are found to be susceptible to SE-targeted CDK7 inhibition therapy in vitro and in vivo. Together, this study provides novel insights into the epigenetic mechanisms underlying the oncogenesis of a subset of GBCs with poorer prognosis and illustrates promising prognostic stratification and therapeutic strategies for treating those GBC patients in future clinical trials.

Background The effect of neoadjuvant chemotherapy (NACT) in gallbladder cancer (GBC) patients remains controversial. The aim of this study was to assess the impact of NACT on overall survival (OS) and cancer specific survival (CSS) in patients with localized or locoregionally advanced GBC, and to explore possible protective predictors for prognosis. Methods Data for patients with localized or locoregionally advanced GBC (i.e., categories cTx-cT4, cN0-2, and cM0) from 2004 to 2020 were collected from the Surveillance, Epidemiology, and End Results (SEER) database. Patients in the NACT and non-NACT groups were propensity score matched (PSM) 1:3, and the Kaplan-Meier method and log-rank test were performed to analyze the impact of NACT on OS and CSS. Univariable and multivariable Cox regression models were applied to identify the possible prognostic factors. Subgroup analysis was conducted to identify patients who would benefit from NACT. Results Of the 2676 cases included, 78 NACT and 234 non-NACT patients remained after PSM. In localized or locoregionally advanced GBC patients, the median OS of the NACT and non-NACT was 31 and 16 months (log-rank P < 0.01), and the median CSS of NACT and non-NACT was 32 and 17 months (log-rank P < 0.01), respectively. Longer median OS (31 vs 17 months, log-rank P < 0.01) and CSS (32 vs 20 months, log-rank P < 0.01) was associated with NACT compared with surgery alone. Multivariable Cox regression analysis showed that NACT, stage, and surgery type were prognostic factors for OS and CSS in GBC patients. Subgroup analysis revealed that the survival hazard ratios (HRs) of NACT vs non-NACT for localized or locoregionally advanced GBC patients were significant in most subgroups. Conclusions NACT may provide therapeutic benefits for localized or locoregionally advanced GBC patients, especially for those with advanced stage, node-positive, poorly differentiated or undifferentiated disease. NACT combined with radical surgery was associated with a survival advantage. Therefore, NACT combined with surgery may provide a better treatment option for resectable GBC patients.