The biliary system is crucial for liver function, regulating bile production, secretion, and transport. Dysfunctions within this system can lead to various diseases, such as cholangiopathies and biliary fibrosis, which may progress from benign to malignant states like cholangiocarcinoma. While liver organoid research is well-established and technologically advanced, bile duct organoids (BDOs) offer significant potential. BDOs can accurately simulate the physiological structure and function of bile ducts, making them valuable tools for in-vitro biliary disease research. Here, we review the development of BDO models, focusing on stem cell-derived organoids and tissue-derived organoids. We also illustrate the role of cultivation strategies and extracellular scaffolds in supporting organoid growth and stability, including the influence of cellular components of the microenvironment and physicochemical factors. Furthermore, we discuss the applications of BDOs in biliary development, disease modeling, regenerative medicine, and drug screening. Additionally, we emphasize the transformative potential in BDO biobanks and personalized medicine, which helps to pave the way for innovative therapeutic strategies and personalized medicine. Finally, we summarize the current and prospective advancements in BDO technologies, highlighting the integration of emerging technologies such as artificial intelligence, 3D bioprinting, and organoid-on-chip systems. These technologies hold great promise for significantly enhancing both clinical and research applications in the field of biliary diseases.

Primary liver cancer is one of the most common fatal malignancies worldwide. Observational studies have shown that immune cells are closely linked to primary liver cancer, however, due to issues like reverse causality and confounding variables, the causal direction and extent of this association remain unclear. Thus, this study aimed to explore the potential causal association between immune cells and primary liver cancer, encompassing hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (ICC).

A two-sample mendelian randomization (MR) analysis was performed using summary statistics from genome-wide association studies (GWAS) of the 731 immune traits and primary liver cancer. The primary liver cancer dataset consisted of a total of 456,348 subjects, with 123 cases of HCC and 456,225 controls, as well as 104 cases of ICC and 456,244 controls, all of European ancestry. The primary method for assessing causality was inverse variance weighting (IVW), with sensitivity analysis utilized for testing heterogeneity and pleiotropy.

Two immunophenotypes were significantly associated with HCC risk: CD3 on CD45RA + CD4+ (OR [95% CI]: 1.334 [1.077 to 1.651], p = 0.008), CD80 on monocyte (OR [95% CI]: 0.578 [0.397 to 0.844], p = 0.004). Additionally, six immunophenotypes were identified to be significantly associated with the risk of ICC: SSC-A on NK (OR [95% CI]: 1.685 [1.166 to 2.436], p = 0.006); CD3 on CD28- CD8br: (OR [95% CI]: 1.826 [1.206 to 2.766], p = 0.004); CD45RA on naive CD4+: (OR [95% CI]: 1.391 [1.119 to 1.729], p = 0.003); Resting Treg %CD4: (OR [95% CI]: 1.290 [1.069 to 1.558], p = 0.008); HLA DR on HSC: (OR [95% CI]: 0.539 [0.343 to 0.846], p = 0.007); Plasmacytoid DC %DC: (OR [95% CI]: 0.610 [0.462 to 0.806], p < 0.001). And sensitivity analyses confirmed the robustness of the main findings.

MR analysis has revealed the causal relationship between immune cells and primary liver cancer through genetic methods. These findings could assist in clinical decision-making and provide new directions for the treatment and research of primary liver cancer.

Cyclin-dependent kinases (CDKs) are pivotal in regulating cell cycle progression and transcription, making them crucial targets in cancer research. The two types of CDKs that regulate different biological activities are transcription-associated CDKs (e.g., CDK7, 8, 9, 12, and 13) and cell cycle-associated CDKs (e.g., CDK1, 2, 4, and 6). One characteristic of cancer is the dysregulation of CDK activity, which results in unchecked cell division and tumor expansion. Targeting transcriptional CDKs, which control RNA polymerase II activity and gene expression essential for cancer cell survival, has shown promise as a therapeutic approach in recent research. While research into selective inhibitors for transcriptional CDKs is ongoing, inhibitors that target CDK4/6, such as palbociclib and ribociclib, have demonstrated encouraging outcomes in treating breast cancer. CDK7, CDK8, and CDK9 are desirable targets for therapy since they have shown oncogenic roles in a variety of cancer types, such as colorectal, ovarian, and breast malignancies. Even with significant advancements, creating selective inhibitors with negligible off-target effects is still difficult. This review highlights the need for more research to optimize therapeutic strategies and improve patient outcomes by giving a thorough overview of the non-transcriptional roles of CDKs in cancer biology, their therapeutic potential, and the difficulties in targeting these kinases for cancer treatment.

Intrahepatic cholangiocarcinoma (ICC) is a highly invasive liver tumor with a poor prognosis, arises from the intrahepatic bile ducts. It is the second most common type of liver cancer. Understanding the mechanisms driving ICC progression is crucial for identification of biomarkers and therapeutic targets. Galectin-1 (Gal-1), encoded by the LGALS1 gene, is known to be upregulated in various malignancies and plays a significant role in cancer progression. However, its underlying mechanisms in ICC have yet to be fully elucidated. The study employed RNA-seq analysis, western blot, cell migration, colony forming, EdU assay, qRT-PCR, luciferase assay and mIHC to investigate the expression pattern of Gal-1 in ICC and its role in the progression of the disease. Our findings revealed a significant upregulation of Gal-1 in ICC tissues. Notably, downregulation of Gal-1inhibited ICC cell proliferation and migration. Further, Gal-1 appears to promote ICC progression through miR-21/STAT3-related pathways, playing a critical role to the tumor microenvironment. These results suggest that Gal-1 may serve as a promising molecular diagnostic marker and therapeutic target for ICC.

Intrahepatic cholangiocarcinoma (iCCA) is the second most prevalent primary liver cancer, and there are limited treatment options when resection is not eligible. Systemic chemotherapy (SYS) offers modest survival benefits, highlighting the need for more effective approaches. This study aimed to evaluate and compare hepatic arterial infusion chemotherapy (HAIC) with SYS in patients with unresectable iCCA in terms of efficacy and safety.

A propensity score-matched analysis was conducted on 111 patients with unresectable iCCA from March 2019 to October 2023. The cohort comprised 37 HAIC-treated and 74 SYS-treated patients. The primary endpoint was overall survival (OS), while the secondary endpoints included progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and adverse events (AEs).

The HAIC group demonstrated comparable survival outcomes to those of the SYS group, with a median OS of 23.7 and 19.3 months [hazard ratio (HR) =0.84, 95% confidence interval (CI): 0.53-1.35; P=0.487] and median PFS of 10.7 vs. 10.3 months (HR =0.75, 95% CI: 0.46-1.22; P=0.246), respectively. However, HAIC showed superior tumor control, achieving a significantly higher ORR (35.13% vs. 12.16%, P<0.05) and DCR (83.78% vs. 64.86%, P<0.05) as compared to SYS. Safety analysis revealed markedly lower grade 3-4 AEs in the HAIC group.

This study demonstrated that HAIC can achieve comparable survival outcomes with superior local tumor control and reduced systemic toxicity as compared to SYS, suggesting its potential as an alternative treatment option for select patients.

Extracellular vesicles (EVs) are nanoscale structures involved in intercellular communication and play a key role in cancer pathology. Intrahepatic cholangiocarcinoma (ICC) is a highly invasive malignancy marked by abnormal sialylated glycosylation. Analyzing proteins and glycans in EVs provides insights into ICC molecular subtyping and mechanisms. Optimizing EV isolation methods for ICC-derived EVs enables comprehensive proteomic and glycomic analysis.

We systematically evaluated five EV isolation methods-Ultracentrifugation (UC), exoEasy, Total Exosome Isolation (TEI), EVtrap, and ÄKTA-by analyzing the biophysical properties, proteomic profiles, and glycomic structures of EVs. Subsequently, we applied TMT-based quantitative proteome and light/heavy methylamine labeling for the quantification of sialylated N-glycan linkage isomers to investigate alterations in proteins and N-glycans within EVs secreted by HuCCT1 and HCCC-9810 cells with overexpressing ST6 β‑galactoside α2,6‑sialyltransferase 1 (ST6GAL1).

By evaluating the biophysical properties, proteome, and N-glycome of EVs extracted using five different methods, UC was identified as the optimal approach for this study, as it offered a balance between operational complexity, cost-effectiveness, and the preservation of EVs activity. In this study, a total of 1,928 high-confidence proteins and over 84 high-confidence glycans were quantified. EVs secreted by HuCCT1 and HCCC-9810 cells overexpressing ST6GAL1 exhibited consistent upregulation of 16 proteins, consistent downregulation of 10 proteins, as well as consistent upregulation of 3 glycans and consistent downregulation of 3 glycans.

Quantitative proteomic and glycomic analysis of ICC-derived EVs revealed that ST6GAL1 overexpression led to significant alterations in proteins involved in cancer cell adhesion and glycosylation pathways, along with specific changes in N-glycan structures. Notably, these modifications extended beyond α2,6-sialylation, suggesting that interactions between glycosyltransferases and glycans may drive these alterations.

The tumor microenvironment has shown abilities to influence the progression and prognosis of intrahepatic cholangiocarcinoma (iCCA). However, little is known about the effect of IgG4+ plasma cells in iCCA.

We stained IgG4+plasma cells by immunohistochemistry and performed Kaplan-Meier survival analysis to detect the prognostic value. We also stained CD3, CD4, CD8 and Foxp3 positive T cells to explore its associations with IgG4+plasma cells.

We found that tumor infiltrated IgG4+plasma cells were associated with poor prognosis of iCCA, rather than IgG4+plasma cells in adjacent liver tissue. The number of IgG4+plasma cells was associated with CD8+ T cells. We divided the iCCA patients into 3 groups by IgG4+plasma cells to CD8+ T cells ratio. The group I (IgG4-/CD8+) had the best prognosis. The group II (IgG4-/CD8- or IgG4+/CD8+) and group III (IgG4+/CD8-) were independent risk factors of recurrence-free survival (HR = 1.69, group II; HR = 2.11, group III) and overall survival (HR = 1.76, group II; HR = 2.38, group III) compared with group I.

We examined the distribution of IgG4+ plasma cells in iCCA and discovered its prognostic utility when combined with CD8+ T cell distribution in iCCA.

Intrahepatic cholangiocarcinoma (ICC) is the second most common primary hepatocellular cancer. This study investigated whether ETV4, ALYREF, and PKM2 affect glycolytic metabolism and ferroptosis, thereby potentially influencing ICC.

Bioinformatic analysis was used to explore the expression levels and prognosis of ETV4, ALYREF, and PKM2 in ICC and their regulatory relationships were confirmed using in vitro experiments. Glycolytic metabolism and ferroptosis were examined, and chromatin immunoprecipitation and RNA immunoprecipitation experiments were performed to verify whether the ETV4, PKM2, and ALYREF could bind. The effect of ETV4/ALYREF on ICC was further confirmed by in vivo experiments.

ETV4, ALYREF, and PKM2 were highly expressed in ICC. Overexpressed (oe)-ETV4 and oe-PKM2 promoted cell migration and increased glucose (GLU) utilization and lactate and intracellular adenosine triphosphate (ATP) production. Addition of the ferroptosis inducer Erastin to the above groups revealed that sh-ETV4 and sh-ALYREF increased lipid reactive oxygen species (ROS), malondialdehyde (MDA), and Fe2+ levels, and oe-PKM2 reversed these effects in the sh-ETV4 and sh-ALYREF groups. Oe-ETV4 promoted the expression of PKM2, whereas sh-ALYREF inhibited the same. ETV4 could bind to ALYREF and PKM2 promoter, and ALYREF could promote the stability of PKM2 in an m5C-dependent manner. In vivo, ETV4 promotes tumor growth and the expression of proteins related to glycolytic metabolism by regulating ALYREF.

ETV4 promotes ICC development and ferroptosis resistance by facilitating glycolytic metabolism, and regulating PKM2 transcription by directly binding to the PKM2 promoter. Additionally, it mediates m5C-dependent PKM2 stabilization by directly binding to ALYREF. This study identified a new potential therapeutic target for ICC.

Patients with intrahepatic cholangiocarcinoma (iCCA) harboring FGFR2 fusion/rearrangement benefit from targeted therapies, highlighting the need for reliable testing strategies to identify FGFR2 alterations. We assessed 226 iCCA cases using RNA-based NGS, DNA-based NGS, and break-apart FISH to evaluate the effectiveness of these methods in detecting FGFR2 fusion/rearrangement. The detection rates for FGFR2 fusion/rearrangement were 9.7% (22/226) for RNA-based NGS, 7.1% (16/226) for DNA-based NGS, and 10.2% (23/226) for FISH. Among the 26 FGFR2 fusion/rearrangement-positive cases identified by any method, only 15 (57.7%) were positive by all three techniques, yielding a concordance rate of 95.1% (215/226). RNA-based NGS confirmed oncogenic FGFR2 fusion in 81% (21/26) of positive cases and identified five novel oncogenic fusions. Thirty-five percent (6/17) of the partner genes were located on chromosome 10, with BICC1 being the most common fusion partner, while the rest were distributed across the other 9 chromosomes. FISH demonstrated a sensitivity of 95.2% and specificity of 98.5%, compared to oncogenic FGFR2 fusions confirmed by RNA-based NGS, while DNA-based NGS exhibited a sensitivity of 71.4% and specificity of 99.5%, identifying FGFR2 mutations in 4 cases. FGFR2-FISH positive cases displayed no significant heterogeneity in positive cell distribution. Oncogenic FGFR2 fusion/rearrangement was associated with small duct type iCCA, especially in cases with positive serum HBsAg and absent cholangiolocarcinoma components and peripheral liver steatosis. This study provides a comprehensive comparison of three assays for detecting FGFR2 fusion/rearrangement, along with clinicopathologic characterization of oncogenic FGFR2 fusion in iCCA.

Intrahepatic cholangiocarcinoma (ICC) is a rare malignant tumor with limited treatment options. Celastrol (Cela) shows potential treatment for ICC, but its clinical use is hindered by poor water solubility and toxic side effects. To address these challenges and enhance its anti-tumor efficacy, we developed hyaluronic acid (HA)-coated triphenylphosphine complex-modified liposomes (HCTL) for accurate delivery of Cela to tumor cell mitochondria.HCTL enhances Cela's water solubility and demonstrates a high rate of encapsulation, stability, and sustained drug release behavior. Moreover, HCTL exhibits outstanding anti-ICC efficacy by efficiently inducing apoptosis in ICC cells via the mitochondrial pathway due to its precise targeting capabilities. In an in-situ ICC mouse model activated by hydrodynamic transfection of AKT and Yap, HCTL downregulates tumor-associated proliferative indices, attenuates the severity of liver injury and modulates the tumor microenvironment. Importantly, HCTL overcomes systemic toxicity associated with Cela. To sum up, HCTL is a potentially effective drug delivery system for ICC treatment.

Intrahepatic cholangiocarcinoma (ICC), an aggressive liver cancer, lacks simple and accurate clinical tests, which poses challenges to postoperative diagnosis and treatment. Recent studies have indicated that platelet levels might be relevant to the postoperative prognosis of ICC. However, their prognostic significance in ICC remains unclarified. This study included 218 ICC patients who underwent hepatic resection. Comprehensive analyses of patients' postoperative prognosis were conducted primarily focusing on their platelet levels associated with prognostic traits. To further investigate the underlying mechanism between platelet levels and patients' postoperative prognosis, we elucidated the association between platelets and tumor metastasis using HCCC-9810 and HUCC-T1 cells as well as mouse models. In the retrospective cohort study, elevated serum platelet levels (≥300 × 109/L) or tumoral platelet levels (≥0.23) individually indicated an unfavorable postoperative prognosis in individuals with ICC. Multivariate analysis showed that tumoral platelet levels can be an independent prognostic factor, while the loss of prognostic superiority of serum platelet levels in the analysis may be attributed to the influence of confounding inclusion variables. Epithelial/mesenchymal transition (EMT) marker expression changes in HCCC-9810 and HUCC-T1 cells with platelet treatment were analyzed to understand how platelets contribute to ICC malignant recurring progression. The significant role of the TGF-β/Smad2 pathway in ICC metastasis was identified. In addition, aspirin was found to have the potential to reduce ICC metastasis by inhibiting platelet function. In conclusion, this study indicated that ICC patients with postoperative serum platelet levels ≥300 × 109/L or tumoral platelet levels ≥0.23 have significantly higher risk of poor postoperative prognosis. This is due to platelet-derived TGFβ1 leading to EMT in ICC cells, thus promoting tumor metastasis.

Clonorchis sinensis infection is an important risk factor for intrahepatic cholangiocarcinoma (ICC). C. sinensis positive (C.s+) ICC patients had much shorter overall survival (OS) compared with C. sinensis negative (C.s-) group. This study aims to explore the impact and underlying mechanism of C. sinensis infection on ICC progression.

In this study, ICC patients underwent surgery from two medical centers enrolled. RNA sequencing was used to determine the downstream activated pathways and genes. Furthermore, we demonstrated the potential mechanism of C. sinensis infection in promoting ICC progression through in vitro co culture systems and two animal models.

Through RNA sequencing, we found fatty acid metabolism and the expression of fatty acid synthase (FASN), a key enzyme catalyzing long-chain fatty acid synthesis, were significantly elevated in C.s+ ICCs. Then, we found excretory/secretory products (ESPs) secreted by C. sinensis could significantly upregulate the expression of transcription factor E2F1, thereby promoting FASN expression and fatty acid synthesis in tumor cells, which ultimately accelerating tumor progression. However, the promotive effect disappeared when FASN was knocked down. Meanwhile, ESPs could promote tumor growth, increasing FASN expression and free fatty acid level in both subcutaneous and orthotopic mouse models.

This study indicates that C. sinensis infection could upregulate the level of FASN and activate fatty acid synthesis pathway, thereby accelerating ICC progression. This provides a new insight for the clinical treatment of ICC with C. sinensis infection.

The precise impact of LI-RADS-defined risk factors on the diagnosis and prognosis of intrahepatic cholangiocarcinoma (iCCA) remains unclear.

To assess the value of LI-RADS categories and features for iCCA diagnosis, focusing on the diagnostic and prognostic implications of LI-RADS-defined risk factors.

Totally 214 high risk patients, including 107 surgically-confirmed solitary iCCAs and 107 hepatocellular carcinomas (HCC) from two centers were retrospectively enrolled. Clinical and MRI features based on LI-RADS v2018 were compared, and the performance of targetoid features for discriminating iCCA was evaluated. Recurrence-free survival (RFS) was compared across different pathologic diagnoses and LI-RADS categories. Multivariate Cox analysis was performed to identify the independent risk factors for RFS.

In the LI-RADS defined high-risk patients, iCCAs differed from HCCs in MRI manifestation. The LR-M category enabled the accurate classification of most iCCAs (89/107, 83.2%), achieving high sensitivity (83.2%), specificity (85.1%), and accuracy (84.1%). The optimal diagnostic performance for iCCA was achieved when at least one targetoid appearance was required for LR-M categorization (AUC = 0.828). Although 26.2% iCCAs presented at least one major feature and 15.0% iCCAs were miscategorized as probably or definitely HCC, only one iCCA case was categorized as LR-5. RFS varied according to both pathologic diagnosis (P = 0.030) and LI-RADS category (P = 0.028), with LI-RADS category demonstrating an independent association with RFS (HR = 1.736, P = 0.033).

In high-risk patients, iCCAs frequently exhibit HCC major features, leading to miscategorization as probable HCC. However, the LR-5 category remains highly specific for ruling out iCCA. Furthermore, in high-risk patients with solitary resected iCCA or HCC, LI-RADS category enables the prediction of postsurgical prognosis independently from pathological diagnosis.

Intrahepatic cholangiocarcinoma (iCCA) denotes a rare, highly malignant, and heterogeneous class of primary liver adenocarcinomas exhibiting phenotypic characteristics of cholangiocyte differentiation. Among the distinctive pathological features of iCCA, one that differentiates the most common macroscopic subtype (eg, mass-forming type) of this hepatic tumor from conventional hepatocellular carcinoma is a prominent desmoplastic reaction manifested as a dense fibro-collagenous-enriched tumor stroma. Cancer-associated fibroblasts (CAFs) represent the most abundant mesenchymal cell type in the desmoplastic reaction. Although the protumor effects of CAFs in iCCA have been increasingly recognized, more recent cell lineage tracing studies, advanced single-cell RNA sequencing, and expanded biomarker analyses have provided new awareness into their ontogeny, as well as underscored their biological complexity as reflected by the presence of multiple subtypes. In addition, evidence supports CAFs' potential to display cancer-restrictive roles, including immunosuppression. However, CAFs also play important roles in facilitating metastasis, as exemplified by lymph node metastasis and peritoneal carcinomatosis, which are common in iCCA. Herein, the authors provide a timely appraisal of the origins and phenotypic and functional complexity of CAFs in iCCA, together with providing mechanistic insights into lymphangiogenesis and peritoneal metastasis relevant to this lethal human cancer.

The biliary tract is a ductal network comprising the intrahepatic (IHBDs) and extrahepatic bile duct (EHBDs). Biliary duct disorders include cholangitis, neoplasms, and injury. However, the underlying mechanisms are not fully understood. With advancements in 3D culture technology, cholangiocyte organoids (COs) derived from primary tissues or induced pluripotent stem cells (iPSCs) can accurately replicate the structural and functional properties of biliary tissues. These organoids have become powerful tools for studying the pathogenesis of biliary diseases, such as cystic fibrosis and primary sclerosing cholangitis, and for developing new therapeutic strategies for cholangiocarcinoma. Additionally, COs have the potential to repair bile duct injuries and facilitate transplantation therapies. This review also discusses the use of organoids in genetically engineered mouse models to provide mechanistic insights into tumorigenesis and cancer progression. Continued innovation and standardization of organoid technology are crucial for advancing precision medicine for biliary diseases and cancer.

Intrahepatic cholangiocarcinoma (ICC) is a highly invasive bile duct cancer with poor prognosis due to frequent recurrence and limited effective treatments. Cancer stem cells (CSCs) contribute to ICC's therapeutic resistance and recurrence, driven by distinct cellular subpopulations with variable tumorigenic properties. Recent advances in single-cell RNA sequencing (scRNA-seq) have enabled a deeper exploration of cellular heterogeneity in tumors, offering insights into unique CSC subgroups that impact ICC progression and patient outcomes. This study aimed to investigate the effect of CSC heterogeneity on the prognosis of ICC.

The scRNA-seq dataset GSE142784 was retrieved from the Gene Expression Omnibus (GEO) database, and Bulk RNA-seq data were obtained from The Cancer Genome Atlas (TCGA) databases. Hallmarks and AUCell R package were adopted for analyzing the signaling pathway activity, CellChat for observing cell communication between subgroups, and SCENIC for analyzing transcription factors expression. The immune cell infiltration and drug sensitivity of the model were analyzed using the CIBERSORT algorithm and the "pRRophetic" R packages, respectively. And immunohistochemistry (IHC) tests were used to evaluate expression of transcription factors in ICC patients.

Based on scRNA-seq data, five clusters (DLK+, CD13+, CD90+, CD133+, and other cholangiocarcinoma cells) were observed in ICC, which presented different signaling pathway activities, such as HSF1 and STAT1 were highly expressed in the CD133 cluster, and consistent with the results of IHC tests. Pathways like Notch and Wnt/β-catenin signaling transferred among above subgroups. Further, subgroups favored varied immune response and drug sensitivity, and CD133+ subgroup patients showed significantly shortened recurrence-free survival (RFS).

Configuring the subgroup of ICC is helpful for predicting the prognosis and drug resistance in ICC and can provide new strategies for cancer treatment.

This study aims to introduce the concept of habitat subregions and construct an accurate prediction model by analyzing refined medical images, to predict lymph node metastasis (LNM) in patients with intrahepatic cholangiocarcinoma (ICC) before surgery, and to provide personalized support for clinical decision-making.

Clinical, radiological, and pathological data from ICC patients were retrospectively collected. Using information from the arterial and venous phases of multisequence CT images, tumor habitat subregions were delineated through the K-means clustering algorithm. Radiomic features were extracted and screened, and prediction models based on different subregions were constructed and compared with traditional intratumoral models. Finally, a lymph node metastasis prediction model was established by integrating the features of several subregional models, and its performance was evaluated.

A total of 164 ICC patients were included in this study, 103 of whom underwent lymph node dissection. The patients were divided into LNM- and LNM + groups on the basis of lymph node status, and significant differences in white blood cell indicators were found between the two groups. Survival analysis revealed that patients with positive lymph nodes had significantly worse prognoses. Through cluster analysis, the optimal number of habitat subregions was determined to be 5, and prediction models based on different subregions were constructed. A comparison of the performance of each model revealed that the Habitat1 and Habitat5 models had excellent performance. The optimal model obtained by fusing the features of the Habitat1 and Habitat5 models had AUC values of 0.923 and 0.913 in the training set and validation set, respectively, demonstrating good predictive ability. Calibration curves and decision curve analysis further validated the superiority and clinical application value of the model.

This study successfully constructed an accurate prediction model based on habitat subregions that can effectively predict the lymph node metastasis of ICC patients preoperatively. This model is expected to provide personalized decision support to clinicians and help to optimize treatment plans and improve patient outcomes.

Cholangiocarcinoma (CHOL) is the second most common primary liver malignancy, characterized by high aggressiveness and heterogeneity. It is typically diagnosed at an advanced stage, leading to a poor prognosis. Although Peptidyl Proline Isomerase H (PPIH) has been implicated in various tumors, its role in CHOL remains unexplored. This study aims to investigate the diagnostic value and potential function of PPIH in CHOL.

We analyzed the expression levels, prognostic significance, and diagnostic efficiency of PPIH in CHOL using data from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO) datasets, coupled with gene enrichment analyses. The CIBERSORT database was employed to assess the correlation between PPIH expression and immune cell infiltration in CHOL. Additionally, immunohistochemical experiments were conducted to validate PPIH expression levels in CHOL tissues and to explore its correlation with TP53 gene mutations.

Our findings indicate that overexpression of PPIH mRNA in CHOL is associated with poor prognosis, with increased PPIH protein levels observed in CHOL tissues. Furthermore, PPIH expression showed a positive correlation with TP53 mutations. PPIH demonstrated strong diagnostic value for CHOL. Moreover, PPIH may influence tumor progression through its involvement in cell cycle regulation and spliceosome pathways, and is associated with immune cell infiltration levels.

The results of this study suggest that PPIH is a potential novel biomarker with significant diagnostic value for patients with CHOL. PPIH may also play a role in modulating the immune microenvironment, contributing to poor prognosis.

This was the first phase 1 study conducted in the United States. It consisted of dose-escalation (part A) and multiple indication-specific cohort expansion (part B), investigating the safety and preliminary efficacy of toripalimab (anti-programmed cell death-1 inhibitor) in patients with advanced malignancies.

Patients with advanced malignancies that progressed after treatment with at least one prior line of standard systemic therapy, including the patients with advanced/recurrent cholangiocarcinoma (CCA), received toripalimab 240 mg every 3 weeks in part B. The primary endpoint was safety assessment. Efficacy endpoints included objective response rate (ORR), disease control rate (DCR), duration of response (DoR), progression-free survival (PFS) as assessed by the investigators according to Response Evaluation Criteria in Solid Tumors (version 1.1) and overall survival (OS).

In part B, 166 patients, including the 42 patients with CCA, were enrolled and received toripalimab. Among the 166 patients, treatment-emergent adverse events (TEAEs) of any grade occurred in 158 (95.2%) patients, and 97 (58.4%) patients experienced TEAEs of Grade 3 or greater. The most common TEAE was fatigue (42.2%). Seven (4.2%) patients experienced TEAEs with a fatal outcome, none of which were identified by investigators as related to toripalimab. Investigator-assessed immune-related adverse events (irAE) of Grade 3 or higher occurred in 7 (4.2%) patients. In the CCA cohort, with the median follow-up of 4.4 months, the ORR and DCR were 4.8% (95% CI: 0.58, 16.16) and 40.5% (95% CI: 25.63, 56.72), respectively; median DoR was 7.8 (range 4.4+ to 7.8) months; median PFS was 2.1 (95% CI: 1.91, 3.88) months; median OS was not estimable.

Toripalimab had manageable side effects in patients with refractory cholangiocarcinoma and exhibited preliminary evidence of anti-tumor activity. However, further information regarding biomarkers is needed. ClinicalTrials.gov ID: NCT03474640.

Targeted therapy for intrahepatic cholangiocarcinoma (ICC) shows superior survival outcomes but patients with certain targetable alterations are no more than 20%. Genetic alteration screening for all ICC patients is of high cost and not routinely performed. This study intends to develop a histopathology-based artificial intelligence (AI)-assisted system for predicting genetic alteration of ICC.

We constructed a Genetic Alteration Prediction (GAP) system based on multi-instance learning and self-supervised learning to predict genetic alterations using whole-slide images (WSIs) of H&E-stained slides. A total of 2069 WSIs from 232 ICC patients underwent surgery of the FAH-SYSU dataset were used for model construction and adjustment by five-fold cross-validation. Another 150 patients from three medical centres were used as independent external validations. We also compared the cost-effectiveness of GAP-assisted precise treatment and all-sequencing strategy to non-sequencing strategy.

The GAP was able to predict actionable genetic alterations of ICC, including FGFR2 and IDH. The area under the receiver operating characteristic curves (AUC) for FGFR2 and IDH were 0.754 and 0.713 in the internal dataset, and 0.724 and 0.656 in the external dataset, respectively. Furthermore, compared to giving chemotherapy without sequencing for every patient, GAP-assisted precise treatment could increase 1 progression-free quality-adjusted life month with a cost of $13871.72, the co-responding figure for all-sequencing strategy is $44538.93. Decision curve analysis showed that AI-assisted strategy provides better clinical benefits.

We constructed an AI-assisted genetic alteration screening system which is predictable to ICC actionable targets and has potential to assist precise targeted treatment of advanced ICC.

In the setting of hepatocellular carcinoma (HCC) surveillance, a range of observations can be identified. Often, imaging findings and biochemical results are consistent with the diagnosis of HCC. However, challenges in HCC surveillance can arise in different clinical contexts, particularly when imaging results and tumor biomarkers are discordant. In this article, the authors describe 5 clinical scenarios based on our experiences in which additional evaluation was necessary to determine whether HCC was present or not.

Microvascular invasion (MVI) serves as a significant predictor of poor prognosis in intrahepatic cholangiocarcinoma (ICC). This study aims to establish a comprehensive model utilizing MR radiomics for preoperative MVI status stratification and outcome prediction in ICC patients.

A total of 249 ICC patients were randomly assigned to training and validation cohorts (174:75), along with a time-independent test cohort consisting of 47 ICC patients. Independent clinical and imaging predictors were identified by univariate and multivariate logistic regression analyses. The radiomic model was developed based on robust radiomic features extracted using a logistic regression classifier. The predictive efficacy of the models was evaluated by receiver operating characteristic curves, calibration curves and decision curves. Multivariate Cox analysis identified the independent risk factors for recurrence-free survival and overall survival, Kaplan-Meier curves were plotted, and a nomogram was used to visualize the predictive model.

The imaging model included tumor size and intrahepatic duct dilatation. The radiomics model comprised 25 stable radiomics features. The Imaging-Radiomics (IR) model, which integrates independent predictors and robust radiomics features, demonstrates desirable performance for MVI (AUCtraining= 0.890, AUCvalidation= 0.885 and AUCtest= 0.815). The calibration curve and decision curve validate the clinical utility. Preoperative MVI prediction based on IR model demonstrated comparable accuracy in MVI stratification and outcome prediction when compared to histological MVI.

The IR model and the nomogram based on IR model-predicted MVI status may serve as potential tools for MVI status stratification and outcome prediction in ICC patients preoperatively.

Accurately differentiating hepatocellular carcinoma (HCC) from intrahepatic cholangiocarcinoma (ICC) is essential for therapeutic decision-making. This study aimed to explore the value of Fluor 18 (18F)-conjugated fibroblast-activation protein inhibitor (FAPI-42) positron emission tomography-computed tomography (PET/CT) in distinguishing HCC from ICC preoperatively.

Patients with suspected intrahepatic lesions who underwent 18F-FAPI-42 PET/CT were retrospectively assessed and placed into an HCC group and an ICC group based on postoperative pathology. Clinical indicators and PET/CT metabolic parameters were documented. Univariate and multivariate logistic regression analyses identified the independent predictive factors. The receiver operating characteristic curve and the area under the receiver operating curve (AUC) were used to determine the diagnostic efficacy.

A total of 48 patients (age 55.4±10.8 years; 35 males and 13 females), including 28 in the HCC and 20 in the ICC group, were included. Univariate analysis revealed significant differences in clinical indicators such as gender, platelet count, and hepatitis B surface antigen, carbohydrate antigen 19-9 (CA19-9), and alpha fetoprotein (AFP) levels between the groups (P<0.05). Metabolic parameters including the maximum standardized uptake value (SUVmax), the total lesion-FAPI (TL-FAPI), and the target-to-background ratio (TBR) were statistically different between groups (P<0.01), and patients with ICC had higher values than those with HCC, while the FAPI tumor volume (FAPI-TV) was not statistically different (P>0.05). Multivariate binary logistic regression analysis identified the SUVmax (P=0.003) as an independent predictor for ICC, and the AUC of the regression-based diagnostic model for predicting ICC was 0.914 at an optimal cutoff value of 12.13, with a sensitivity of 85.0% and a specificity of 89.3%.

SUVmax is an independent predictor for ICC. With the optimal cutoff value of 12.13, 18F-FAPI-42 PET/CT indicates good diagnostic efficacy in distinguishing between HCC and ICC.

Intrahepatic cholangiocarcinoma (iCCA) is a highly aggressive cancer of the peripheral bile ducts and is recognized by the abundance of cancer stem-like cells (CSCs) within the tumor mass. While CSC markers in iCCA are well-defined, the molecular vulnerabilities of this subpopulation remain elusive.

The 96-well, three dimensional (3D) tumorsphere culture was adapted from a well-established CSC model, validated for CSC markers through gene expression analysis. Kinase library screening was then conducted to reveal potential oncogenic vulnerable pathways. RNA interference was utilized to stably silence the candidate gene in three iCCA cell lines and its impact on iCCA cell proliferation and tumorsphere formation efficiency (TFE) was evaluated.

Kinase inhibitor library screening identified the top 50 kinase inhibitors crucial for tumorsphere viability, with 11 inhibitors targeting the IGF-1R/PI3K/AKT axis. Further dose-dependent analysis of the top 'hit' inhibitors confirmed IGF-1R as the candidate molecule. Upon stably silencing of IGF-1R, all three iCCA cell lines exhibited decreased AKT activation, impeded proliferation and reduced TFE, indicating a decline in CSC subpopulations.

IGF-1R plays a critical role in maintaining iCCA-stem like cell populations.

Our data highlight the potential utility of IGF-1R as a prognostic marker of iCCA and a therapeutic target for eliminating its CSC subpopulation.

Cholangiocarcinoma (CCA) is a dreaded complication of primary sclerosing cholangitis (PSC) that is difficult to diagnose and associated with high mortality. A lack of animal models of CCA recapitulating the hepatic microenvironment of sclerosing cholangitis has hindered the development of novel treatments. Herein, we sought to develop a mouse model of PSC-associated CCA.

Ten-week-old Mdr2-/- mice with congenital PSC-like disease, and healthy wild-type littermates were subjected to either modified retrograde biliary instillation or hydrodynamic tail vein injection of a sleeping beauty transposon-transposase plasmid system with activated AKT (myr-AKT) and Yap (YapS127A) proto-oncogenes (SB AKT/YAP1). The role of TGFβ was interrogated via ALK5 inhibitor (SB-525334) administration. Tumor phenotype, burden and desmoplastic reaction were analyzed histologically and via RNA sequencing.

While SB AKT/YAP1 plasmids administered via retrograde biliary injection caused tumors in Mdr2-/-, only 26.67% (4/15) of these tumors were CCA. Alternatively, hydrodynamic tail vein injection of SB AKT/YAP1 resulted in robust tumorigenesis in all fibrotic Mdr2-/- mice with high CCA burden compared to healthy mice. Tumors phenotypically resembled human CCA, expressed multiple CCA (but not hepatocellular carcinoma) markers, and exhibited a profound desmoplastic reaction. RNA sequencing analysis revealed profound transcriptional changes in CCA evolving in a PSC-like context, with specific alterations in multiple immune pathways. Pharmacological TGFβ inhibition led to enhanced immune cell tumor infiltration, reduced tumor burden and suppressed desmoplastic collagen accumulation compared to placebo.

We established a new high-fidelity cholangiocarcinoma model in mice, termed SB CCA.Mdr2-/-, which recapitulates the increased susceptibility to CCA in the setting of biliary injury and fibrosis observed in PSC. Through transcriptomics and pharmacological studies, we show dysregulation of multiple immune pathways and TGFβ signaling as potential drivers of CCA in a PSC-like microenvironment.

Animal models for primary sclerosing cholangitis (PSC)-related cholangiocarcinoma (PSC-CCA) are lacking. Thus, we have developed and characterized a new mouse model of PSC-CCA, termed SB CCA.Mdr2-/-, which features reliable tumor induction on a PSC-like background of biliary injury and fibrosis. Global gene expression alterations were identified and standardized tools, including automated whole slide image analysis methodology for tumor burden and feature analysis, were established to enable systematic research into PSC-CCA biology and formal preclinical drug testing.

To establish an MRI-based model for microvascular invasion (MVI) prediction in mass-forming intrahepatic cholangiocarcinoma (MF-iCCA) and further evaluate its potential survival and therapeutic benefit.

One hundred and fifty-six pathologically confirmed MF-iCCAs with traditional surgery (121 in training and 35 in validation cohorts), 33 with neoadjuvant treatment and 57 with first-line systemic therapy were retrospectively included. Univariate and multivariate regression analyses were performed to identify the independent predictors for MVI in the traditional surgery group, and an MVI-predictive model was constructed. Survival analyses were conducted and compared between MRI-predicted MVI-positive and MVI-negative MF-iCCAs in different treatment groups.

Tumor multinodularity (odds ratio = 4.498, p < 0.001) and peri-tumor diffusion-weighted hyperintensity (odds ratio = 4.163, p < 0.001) were independently significant variables associated with MVI. AUC values for the predictive model were 0.760 [95% CI 0.674, 0.833] in the training cohort and 0.757 [95% CI 0.583, 0.885] in the validation cohort. Recurrence-free survival or progression-free survival of the MRI-predicted MVI-positive patients was significantly shorter than the MVI-negative patients in all three treatment groups (log-rank p < 0.001 to 0.046). The use of neoadjuvant therapy was not associated with improved postoperative recurrence-free survival for high-risk MF-iCCA patients in both MRI-predicted MVI-positive and MVI-negative groups (log-rank p = 0.79 and 0.27). Advanced MF-iCCA patients of the MRI-predicted MVI-positive group had significantly worse objective response rate than the MVI-negative group with systemic therapy (40.91% vs 76.92%, χ2 = 5.208, p = 0.022).

The MRI-based MVI-predictive model could be a potential biomarker for personalized risk stratification and survival prediction in MF-iCCA patients with varied therapies and may aid in candidate selection for systemic therapy.

Question Identifying intrahepatic cholangiocarcinoma (iCCA) patients at high risk for microvascular invasion (MVI) may inform prognostic risk stratification and guide clinical treatment decision. Findings We established an MRI-based predictive model for MVI in mass-forming-iCCA, integrating imaging features of tumor multinodularity and peri-tumor diffusion-weighted hyperintensity. Clinical relevance The MRI-based MVI-predictive model could be a potential biomarker for personalized risk stratification and survival prediction across varied therapies and may aid in therapeutic candidate selection for systemic therapy.

Intrahepatic cholangiocarcinoma is a biliary neoplasm usually showing a dismal prognosis. In early stages, surgical resection is the best treatment option, significantly increasing the overall survival. This approach is also recommended in the case of relapsing disease. In this study, we report the case of a patient affected by intrahepatic cholangiocarcinoma with multiple relapses and still alive for over 18 years. We also provide a systematic review regarding long-survivor (> 60 months) of intrahepatic cholangiocarcinoma.

A 41-year-old woman with no pathological history was diagnosed with localized intrahepatic cholangiocarcinoma and surgically treated with left hepatectomy. After the first intervention, the patients underwent three further surgical resections because of locoregional recurrences. Histologically, there were some significant similarities among all neoplasms, including the tubule-glandular architecture, but also morphological heterogeneity. The tumor immune microenvironment remained stable across the different lesions. The molecular analysis with next-generation sequencing demonstrated that all neoplasms shared the same genomic profile, including NBN and NOTCH3 mutations and chromosomes 1 and 3 alterations.

This case study highlights the essential role of a stringent follow-up after resection of intrahepatic cholangiocarcinoma for detecting early relapsing tumors. Moreover, it shows the importance of the molecular characterization of multiple tumors for understanding their real nature. The accurate study of long-surviving patients highlights the features that are critical for outcome improvement.

CCA has a poor prognosis. Different anatomical subtypes are characterized by distinct clinical features, surgical options, and prognoses, which can potentially impact survival outcomes following radical resection. In addition to the malignancy of CCA itself, clinical staging and treatment methods are the main factors that can affect survival. This study aims to update a more reliable prediction model for the prognosis of CCA based on different anatomical locations.

A total of 1172 CCA patients (305 iCCA, 467 pCCA, and 400 dCCA) who underwent surgical resection between 2015 and 2022 were included in the analysis. The covariates included in the analysis were age, sex, tumor diameter, differentiation grade, T stage, N stage, M stage, neural invasion, cancer thrombus, history of hepatitis B or biliary calculi, and receipt of adjuvant chemotherapy. The data were randomly divided into training (80 %) and validation cohort (20 %).

We developed a nomogram of the sensitive model and calculated concordance indices of different constructed prognostic survival models. Meanwhile, we validated the effectiveness of the nomogram model and compared it with the TNM system through decision curve analysis (DCA) and internal cohort validation. The nomogram model had a better net benefit than the TNM system at any given threshold for iCCA, pCCA, and dCCA, regardless of their location.

We have updated the prognostic model for OS in CCA patients who underwent radical resection according to the different tumor locations. This model can effectively predict OS and has the potential to facilitate individual clinical decision-making.

Liver fluke infection is a major risk for cholangiocarcinoma (CCA). It has been established that the Asian liver flukes, Clonorchis sinensis and Opisthorchis viverrini secrete growth factors, digestive enzymes, and extracellular vesicles (EVs) which contribute to abnormal cell development in the bile ducts where the worms reside. These secretions - combined with aberrant inflammation and repeated cycles of chronic wounding at the site of parasite attachment and grazing on the epithelium - promote biliary hyperplasia and fibrosis and ultimately malignant transformation. Application of post-genomic and gene-editing tools to the study of liver fluke immunobiology and pathogenesis has accelerated the discovery of essential virulence factors to which targeted therapies and diagnostics can be directed.

Systemic chemotherapy (SC) stands the only first-line treatment for advanced intrahepatic cholangiocarcinoma (iCCA) for the past few decades. Immune checkpoint inhibitors (ICIs) have been proved to provide additional benefit in disease control. However, oncological outcome of iCCA remains poor and awaits further improvement with new treatment modalities. Promising results have been observed in lenvatinib plus pembrolizumab (Len-P) as a second-line therapy in iCCA. This study aimed to explore the safety and efficacy of Len-P as a first-line therapy for iCCA patients in real-world clinical practice.

We retrospectively enrolled 133 patients with advanced iCCA who received Len-P or SC between May 2019 and May 2023. Overall survival (OS), progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and adverse events (AEs) were compared between the two groups.

There were 72 patients and 61 patients in the Len-P and SC groups, respectively. The median OS for the Len-P and SC groups was 16.3 and 17.8 months, respectively. The median PFS for the Len-P and SC groups was 8.9 and 11.4 months, respectively. There was no significant difference in ORR and DCR between the Len-P and SC groups (ORR: 22.2% vs. 23%; P=0.92; DCR: 69.4% vs. 77%; P=0.58). Additionally, the overall incidence of AEs was lower in the Len-P group than SC group. Low inflammation-based scores were indicative of favorable outcomes in patients undergoing Len-P therapy.

This study demonstrated that Len-P is promising for the treatment of advanced ICC, with highly improved safety. It emerges as a viable treatment alternative for advanced iCCA. Inflammation-based scores show potential utility in identifying individuals likely to benefit from Len-P therapy.

Background: Intrahepatic cholangiocarcinoma (ICC), one type of highly malignant tumor, has a poor prognosis. However, the specific role of the polycystic kidney and hepatic disease 1 (PKHD1) gene in ICC has not yet been evaluated. This study aimed to investigate the potential function and mechanism of the PKHD1 gene in ICC. Methods: Quantitative real-time PCR was applied to detect the expression of PKHD1 mRNA in human ICC and adjacent normal tissues. CRISPR/Cas9 technique was used to construct PKHD1 partially knockout (PKHD1-/+) ICC cell lines. In the vitro study, the effects of PKHD1 on the malignant biological behavior of ICC cells were examined by Edu, RTCA, migration, and invasion assays. The expression levels of proteins were detected using western blotting, immunohistochemistry, and flow cytometry. Furthermore, DAPT, an antagonist of the Notch1 signaling pathway, was used in the rescue experiment in vitro. Results: Compared with normal tissues, PKHD1 mRNA expression was significantly down-regulated in human cholangiocarcinoma tissues (P<0.001). At the same time, the expressions of Notch pathway-related proteins were dramatically increased in PKHD1(-/+) ICC cells (P<0.001). Moreover, tumor proliferation, migration, and invasion were promoted in loss-of-function experiments in vitro and in vivo, which was partially reversed by DAPT. Conclusions: PKHD1 inhibits the proliferation, migration, and invasion of ICC, and the Notch pathway may be the downstream mechanism of the negative regulatory effect of PKHD1 during the progression of ICC.

Intrahepatic cholangiocarcinoma (ICC) is a highly heterogeneous malignancy. The reasons behind the global rise in the incidence of ICC remain unclear, and there exists limited knowledge regarding the immune cells within the tumor microenvironment (TME). In this study, a more comprehensive analysis of multi-omics data was performed using machine learning methods. The study found that the immunoactivity of B cells, macrophages, and T cells in the infiltrating immune cells of ICC exhibits a significantly higher level of immunoactivity in comparison to other immune cells. During the immune sensing and response, the effect of antigen-presenting cells (APCs) such as B cells and macrophages on activating NK cells was weakened, while the effect of activating T cells became stronger. Simultaneously, four distinct subpopulations, namely BLp, MacrophagesLp, BHn, and THn, have been identified from the infiltrating immune cells, and their corresponding immune-related marker genes have been identified. The immune sensing and response model of ICC has been revised and constructed based on our current comprehension. This study not only helps to deepen the understanding the heterogeneity of infiltrating immune cells in ICC, but also may provide valuable insights into the diagnosis, evaluation, treatment, and prognosis of ICC.

Intrahepatic cholangiocarcinoma (ICC) is a highly desmoplastic tumor with poor prognosis even after curative resection. We investigated the associations between the composition of the ICC stroma and immune cell infiltration and aimed to develop a stromal-immune signature to predict prognosis in surgically treated ICC.

We recruited 359 ICC patients and performed immunohistochemistry to detect α-smooth muscle actin (α-SMA), CD3, CD4, CD8, Foxp3, CD68, and CD66b. Aniline was used to stain collagen deposition. Survival analyses were performed to detect prognostic values of these markers. Recursive partitioning for a discrete-time survival tree was applied to define a stromal-immune signature with distinct prognostic value. We delineated an integrated stromal-immune signature based on immune cell subpopulations and stromal composition to distinguish subgroups with different recurrence-free survival (RFS) and overall survival (OS) time.

We defined four major patterns of ICC stroma composition according to the distributions of α-SMA and collagen: dormant (α-SMAlow/collagenhigh), fibrogenic (α-SMAhigh/collagenhigh), inert (α-SMAlow/collagenlow), and fibrolytic (α-SMAhigh/collagenlow). The stroma types were characterized by distinct patterns of infiltration by immune cells. We divided patients into six classes. Class I, characterized by high CD8 expression and dormant stroma, displayed the longest RFS and OS, whereas Class VI, characterized by low CD8 expression and high CD66b expression, displayed the shortest RFS and OS. The integrated stromal-immune signature was consolidated in a validation cohort.

We developed and validated a stromal-immune signature to predict prognosis in surgically treated ICC. These findings provide new insights into the stromal-immune response to ICC.

Intrahepatic cholangiocarcinoma (ICCA) with hepatic hilus involvement is a more aggressive type of cholangiocarcinoma with worse outcomes.1,2 Surgical resection with negative margins is the only effective treatment for ICCA.3,4 Neoadjuvant therapy is considered to improve the possibility of surgery for patients;5,6 however, laparoscopic radical resection after neoadjuvant therapy for ICCA with hepatic hilus involvement remains at the exploratory stage due to technical challenges.7 METHODS: A 19-year-old man presented with an ICCA on the left side of the liver invading the blood vessels and bile ducts in the hepatic hilum. Five courses of neoadjuvant therapy were administered after a multidisciplinary team determined that the tumor was extremely difficult and risky to operate on. A laparoscopic left hepatectomy plus caudal lobectomy was performed to complete the resection of the negative margins. Three-dimensional visualization enabled precise preoperative planning and intraoperative guidance, including visualization of the tumor location, simulation of bile duct and vessel dissection steps, as well as determining the extent of liver resection. Vascular skeletonization, lymphadenectomy and biliary reconstruction were performed during operation.

The operation time was 415 min with a blood loss of 100 mL. Postoperative pathohistology confirmed cholangiocarcinoma with low to intermediate differentiation. The resection margin was negative (R0) and lymph node pathology was tumor-negative (0/10). The patient was discharged on postoperative day 10 without complications.

Laparoscopic radical resection after neoadjuvant therapy for ICCA with hepatic hilus involvement is safe and feasible in a large-throughput hepatic surgery center.