2508 Background: Advanced hepatocellular carcinoma (HCC) remains a high unmet-medical-need malignancy with limited therapeutic options. Ori-C101 is a novel, armored, autologous GPC3-directed CAR-T cell therapy. Following promising results from early-phase trials (ChiCTR190028121; NCT05652920, BEACON study), we shall herein update outcomes from the BEACON study with focusing on long-term safety, durability of response, and survival after more than 2 years of follow-up. Methods: This is an open-label, multi-center, phase Ib dose-escalation and expansion study enrolled patients (pts) with GPC3 + advanced HCC who had progressed on ≥2 prior lines of systemic therapy (including ICIs and TKIs). A single dose of Ori-C101 was administered via hepatic arterial infusion to enhance regional cell delivery. Integrated analyses assessed safety, tolerability, PK, and efficacy (per RECIST v1.1)，aiming to determine the RP2D. Results: As of Dec 24, 2025, 19 pts received Ori-C101 infusion across 4 dose levels (DLs). All pts had BCLC stage B/C disease and 31.6% (6/19) had extrahepatic metastases. Pts were previously treated with a median of 3 lines (range 2–8) therapies. Safety: Safety remained manageable; no late-onset nor cumulative toxicities were observed through the extended follow-up period. The most common ≥G3 TEAEs (≥10.0%) were transient hematologic toxicities and hepatic laboratory abnormalities. CRS occurred in 100.0% (19/19) of pts; while ≥G3 CRS observed in 42.1% (8/19). No ICANS occurred. One pt at DL4 experienced a DLT of G4 CRS complicated by secondary DIC. Efficacy: In 18 efficacy-evaluable pts, Ori-C101 demonstrated a robust dose-dependent response. Confirmed ORR was 50.0% (9/18); DCR was 77.8% (14/18). At RP2D (DL3), the confirmed ORR and DCR were 66.7% (6/9) and 88.9% (8/9), respectively. Critically, responses were not only rapid but also remarkably durable. 88.9% (8/9) of responders achieved objective response within 1.1 months; at M3, 83.3% (5/6) of responders at the RP2D remained PR. Notably, 1 pt at DL4 achieved CR with a duration exceeding 20 months. Preliminary overall survival data indicate a substantial long-term survival benefit with a median OS of 14.4 months (range 2.6–22.0). In addition, Dose-Exposure-Responses analysis showed dose-dependent CAR-T cells expansion, pharmacodynamic effects and improved tumor response. Conclusions: Ori-C101 demonstrates a manageable safety profile and compelling, durable anti-tumor activity in GPC3 + advanced HCC. The combination of high ORR and prolonged survival benefit distinguishes Ori-C101 as a potential paradigm-shifting therapy for patients who have failed standard-of-care treatments. A phase II/III study is currently underway to further confirm the efficacy and asses the safety of Ori-C101. Clinical trial information: NCT05652920 .

Disordered cholesterol-oxysterol profiles are observed in the tumor microenvironment, yet their roles in tumor-associated macrophages (TAMs) remain underexplored. This study reveals that TAMs across human pan-cancers exhibit elevated phosphotyrosine interaction domain-containing protein 1 (PID1) expression and prominent immunosuppressive gene signatures. PID1 deficiency in myeloid cells upregulates low-density lipoprotein (LDL) receptor expression, thereby promoting LDL uptake and intracellular accumulation of free cholesterol and reactive oxygen species (ROS). Increased ROS drives cholesterol oxidation to generate the oxysterols 5α,6α-epoxycholesterol (5α,6α-EC) and 7β-hydroxycholesterol (7β-OHC), which inhibit mTOR-STAT6 signaling in macrophages. Pid1 deletion switches immunosuppressive macrophages toward an antitumor subtype that downregulates arginase 1 expression while upregulating proinflammatory cytokines, thereby potentiating CD8+ T cell-mediated immunosurveillance across multiple tumor types. Moreover, combination treatment with the oxysterol and chemotherapeutic agent 5-fluorouracil produces synergistically enhanced antitumor effects. Previous studies showed that cholesterol-derived oxysterols differentially regulate macrophage cell fates, with 25-OHC promoting the protumor and immunosuppressive phenotype of TAMs. Targeting PID1 reroutes cholesterol and ROS metabolism toward the production of 5α,6α-EC and 7β-OHC in TAMs, representing a promising immunometabolic strategy to restore antitumor immunosurveillance.

BACKGROUND:No neoadjuvant treatment has been considered to be standard therapy for patients with resectable intrahepatic cholangiocarcinoma with high-risk factors for recurrence. The GOLP regimen (gemcitabine-oxaliplatin, lenvatinib, and an anti-programmed death 1 antibody) has shown promising efficacy with a manageable safety profile in advanced intrahepatic cholangiocarcinoma and biliary tract cancer. METHODS:In a phase 2-3 trial, we randomly assigned, in a 1:1 ratio, patients with resectable high-risk intrahepatic cholangiocarcinoma to the neoadjuvant group (intravenous gemcitabine-oxaliplatin plus toripalimab every 3 weeks for three cycles and oral lenvatinib once daily for 9 weeks, followed by curative resection) or the control group (curative resection and no neoadjuvant treatment). All patients received adjuvant capecitabine for eight cycles after surgery. The primary end point was event-free survival. Secondary end points included overall survival and safety. RESULTS:A total of 178 patients underwent randomization (88 patients to the neoadjuvant group and 90 to the control group). At the interim analysis at a median follow-up of 16.9 months, the median event-free survival was significantly longer in the neoadjuvant group (18.0 months; 95% confidence interval [CI], 13.8 to 27.6) than in the control group (8.7 months; 95% CI, 7.2 to 12.4) (P<0.001). Overall survival at 24 months was 79% (95% CI, 70 to 90) in the neoadjuvant group and 61% (95% CI, 50 to 75) in the control group (hazard ratio for death, 0.43; 95% CI, 0.23 to 0.79; P = 0.005, which did not meet the significance criterion [two-sided alpha, 0.0019]). Across all treatment phases, adverse events occurred in 97% of the patients in the neoadjuvant group and in 70% of those in the control group. During the neoadjuvant phase, adverse events of grade 3 or higher occurred in 28% of the patients, and treatment-related adverse events of grade 3 or higher in 26%. No treatment-related adverse event led to death. CONCLUSIONS:Neoadjuvant GOLP led to significantly longer event-free survival than control therapy, with mainly low-grade adverse events, among patients with resectable high-risk intrahepatic cholangiocarcinoma. (Funded by the Clinical Research Plan of Shanghai Hospital Development Center and others; ZSAB-neoGOLP ClinicalTrials.gov number, NCT04669496.).

3028 Background: Pts with advanced HCC face limited treatment options beyond immunotherapy and anti-angiogenic agents. GPC3, a surface antigen overexpressed in HCC and associated with poor prognosis, represents a promising therapeutic target due to its tumor-specific expression. MRG006A, a potential first-in-class GPC3-targeted ADC, demonstrated potent pre-clinical anti-tumor activity. Here we report the preliminary safety and efficacy of MRG006A in advanced HCC with intermediate/high GPC3 expression from a phase I/II trial. Methods: MRG006A-001 (NCT07093970) is an ongoing first-in-human, open-label, multi-center phase I/II study. The phase I study comprises dose escalation (Ia) and dose optimization/expansion (Ib) stages. The dose-escalation employed an accelerated titration plus 3+3 design. Eligible pts who had failed standard treatment received MRG006A intravenously at 1.6-6.4 mg/kg every three weeks (Q3W). GPC3 expression was only required for dose expansion cohorts. The primary endpoints were safety and tolerability. Secondary endpoints included objective response rate (ORR), disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), clinical benefit rate (CBR), etc. Results: As of Dec 19, 2025, the maximum administered dose was established as 6.4 mg/kg Q3W, with ≥ G3 treatment-related adverse event (TRAE) of platelet count decreased reported in all six pts at this dose level and dose-limiting toxicity (G4 platelet count decreased) observed in one patient. During dose escalation, tumor response was observed at 3.2 mg/kg and 4.8 mg/kg. Accordingly, dose optimization in phase Ib proceeded with three levels (3.2, 4.0, and 4.8 mg/kg Q3W). Twenty-six HCC pts with intermediate or high GPC3 expression were enrolled across three cohorts; 61.5% had BCLC stage C disease; median 2 (range 1-4) prior lines of therapy, and 25 (96.2%) had received immune checkpoint inhibitors and anti-angiogenic agents. Among 25 efficacy evaluable pts, ORR, DCR and CBR were 23.1%, 68.0% and 32.0%, respectively. In pts with high GPC3 expression (n=12), ORR, DCR and CBR increased to 33.3%, 75.0% and 50.0%, respectively; median PFS and DOR were 7.0 and 4.2 months (median follow-up 5.7 months). Most pts (24 [92.3%]) experienced TRAE of any Grade. TRAEs of ≥ G3 were reported in 11 (42.3%) pts . The most common TRAEs were platelet count decreased (88.5%), blood bilirubin increased (50.0%), AST increased (46.2%), white blood cell count decreased (38.5%), and nausea (30.8%). No treatment-related permanent discontinuations or deaths occurred. Conclusions: MRG006A demonstrated manageable safety and promising anti-tumor activity in heavily pretreated pts with GPC3-expressing advanced HCC. Our findings support the therapeutic potential of GPC3-directed ADC and merits further clinical investigations. Clinical trial information: NCT07093970 .

Background Hepatocellular carcinoma (HCC) is a global health challenge with high mortality rates, particularly in patients with advanced disease and lung metastasis. T-cell receptor (TCR)-T cell therapy based on specific neoantigens, is an emerging treatment with potential for HCC. However, the prognosis of patients remains poor, underscoring the need for novel targets and strategies.Methods We conducted a comprehensive study to investigate the role of C7orf50 and its neoantigens in HCC. We evaluated the functional impact on HCC progression and metastasis in vitro and in vivo, and further explored the mechanism by which C7orf50 promotes cancer metastasis and remodels tumor immune environment. Using exome and transcriptome sequencing, we identified neoantigens associated with C7orf50 and assessed their potential in TCR-T therapy.Results Our in vitro experiments revealed that C7orf50 overexpression enhances HCC cell proliferation, migration, and invasion, while knockdown inhibits these processes. In vivo, C7orf50 promoted tumor growth and lung metastasis, with a significant correlation between C7orf50 expression and poor clinical outcomes in patients with HCC. We further demonstrated that C7orf50 activates the NF-κB/PAI-1 pathway by binding to AEG-1 and facilitating its nuclear translocation, thereby promoting tumor-associated macrophage recruitment. Meanwhile, we found that TCR-T from C7orf50-related neoantigen could obviously realize the killing effect on HCC cells, revealing its great role in cell therapy.Conclusion C7orf50 is a critical mediator of HCC progression and lung metastasis, acting through the NF-κB/PAI-1 pathway and AEG-1. Its expression levels, along with those of PAI-1 and CD68, serve as independent prognostic markers. And C7orf50-related neoantigen shows great application potential in TCR-T therapy. These findings provide a foundation for developing C7orf50-targeted therapies and highlight its potential in precision medicine and immunotherapy for HCC.

BACKGROUND:Resection might offer additional benefit in patients with advanced-stage hepatocellular carcinoma treated with systemic therapy. However, high-quality evidence supporting the procedure is absent. We aimed to establish whether resection can provide survival benefit in patients with advanced hepatocellular carcinoma who respond to systemic therapy. METHODS:In this randomised, open-label, multicentre, phase 3 trial, we recruited treatment-naive patients with hepatocellular carcinoma, macrovascular invasion, and no extrahepatic metastasis from 24 hospitals in China. These patients were treated in the induction phase with three cycles of intravenous atezolizumab (1200 mg every 3 weeks) plus intravenous bevacizumab (15 mg/kg bodyweight every 3 weeks) and one cycle of atezolizumab monotherapy. Patients who completed the induction phase, had a partial response or stable disease according to Response Evaluation Criteria in Solid Tumors (RECIST) version 1.1, and were considered feasible for resection were randomly assigned (1:1) to undergo surgical resection followed by 12 months of atezolizumab plus bevacizumab initiated 4-6 weeks after surgery (ie, the surgery group), or to maintenance atezolizumab plus bevacizumab until loss of clinical benefit or intolerable toxicity (the maintenance therapy group). Randomisation was by computer-generated sequence with permuted blocks via a central interactive web response system, stratified by tumour response and Eastern Cooperative Oncology Group performance status. Treatment administered to patients was not masked. The primary endpoint was time to treatment failure, assessed by an independent review facility and analysed by intention to treat. Time to treatment failure was defined as the time from randomisation to the first documented treatment failure (ie, local recurrence or disease progression according to RECIST 1.1, emergence of extrahepatic spread, or death). This study is registered with ClinicalTrials.gov (NCT04649489) and is ongoing. FINDINGS:Between April 4, 2021, and July 18, 2024, a total of 489 patients were enrolled in the induction phase. Of them, 201 were randomly assigned to the surgery group (n=101; 93 male, eight female) or the maintenance therapy group (n=100; 87 male, 13 female). After a median follow-up of 18·4 months, the median time to treatment failure was 20·4 months in the surgery group and 11·8 months in the maintenance therapy group (hazard ratio 0·60, 95% CI 0·39-0·91; p=0·015). Grade 3 or 4 treatment-related adverse events occurred in 32 (39%) of 83 patients in the surgery group and 21 (21%) of 100 in the maintenance therapy group, the most common of which were increased alanine aminotransferase (seven patients [8%] in the surgery group vs one [1%] in the maintenance therapy group), reduced platelet count (seven [8%] vs three [3%]), and proteinuria (three [4%] vs seven [7%]). Two treatment-related deaths occurred in the surgery group due to abnormal liver function (considered related to atezolizumab) and liver failure (considered related to atezolizumab, bevacizumab, or surgery). INTERPRETATION:In patients with advanced hepatocellular carcinoma with macrovascular invasion after systemic therapy, time to treatment failure was longer in those who had liver resection than in those who received maintenance therapy. FUNDING:Shanghai Roche Pharmaceuticals and Ministry of Science and Technology of China.

Minimal residual disease (MRD) is proposed to drive early recurrence of hepatocellular carcinoma (HCC) after curative treatment, and patients with MRD may benefit from adjuvant treatment. We applied a customized targeted deep sequencing (TDS) to profile tumor genomic alterations in HCC patients undergoing liver transplantation (LT) or hepatectomy, and evaluated their associations with MRD status, recurrence risk, and adjuvant lenvatinib response across multiple independent cohorts. TDS revealed MRD-associated genomic patterns, including increased mutation burden and frequent alterations in TP53 and switch/sucrose non-fermentable (SWI/SNF) genes. Alterations in TP53 and SWI/SNF genes were consistently associated with higher early recurrence rates (48.5 % vs 30.0 %, P = 0.026 for TP53; 56.2 % vs 34.0 %, P = 0.024 for the SWI/SNF genes) and shorter recurrence-free survival (12.2 vs 34.6 months, P = 0.026 for TP53; 9.3 vs 18.6 months, P = 0.018 for the SWI/SNF genes) in the LT cohort, as well as the hepatectomy cohort. Multivariate analysis confirmed these alterations as independent risk factors for MRD. Integrative analysis demonstrated that TP53- and SWI/SNF-altered HCC represent clinically and biologically distinct phenotypes, differing in etiology, differentiation status, mutation burden, and transcriptomic subtypes: TP53 alterations correlated with an epithelial-mesenchymal transition-related subtype, while the SWI/SNF genes alterations related to a MYC-activated subtype. Notably, patients harboring TP53 alterations derived clinical benefit from adjuvant lenvatinib following curative surgery. In conclusion, TDS enables effective identification of MRD-associated genomic alterations and stratifies HCC patients who may benefit from adjuvant lenvatinib, providing a molecular basis for personalized postoperative management.

BACKGROUND:Patients with intrahepatic cholangiocarcinoma have a poor prognosis and high postoperative recurrence rates. Immunochemotherapy has been approved as a first-line treatment for advanced biliary tract cancer. We aimed to explore the activity and safety of camrelizumab, an anti-PD-1 immunotherapy, combined with capecitabine chemotherapy in the adjuvant treatment of patients with resected intrahepatic cholangiocarcinoma. METHODS:ACC was a single-arm, single-centre, open-label, phase 2 trial in adult patients (aged 18-75 years) with R0-resected, pathologically confirmed intrahepatic cholangiocarcinoma (staged as IA with G3 classification or IB-III per the American Joint Committee on Cancer staging system [8th edition, 2017]) done at Zhongshan Hospital, Fudan University (Shanghai, China). Eligible patients had no extrahepatic metastases, an Eastern Cooperative Oncology Group performance status of 0 or 1, and adequate organ function. 4-8 weeks after surgery, patients received eight 21-day cycles of intravenous camrelizumab (200 mg on day 1 of each cycle) plus oral capecitabine (1250 mg/m2 twice daily on days 1-14, followed by a 7-day rest period). The primary endpoint was recurrence-free survival, assessed in the full analysis set (FAS), which included patients who had received at least one dose of either drug. Safety was assessed in the safety set, which included patients who received at least one dose of either drug and completed at least one post-baseline safety assessment following enrolment. This study is registered with ClinicalTrials.gov (NCT04295317) and is currently ongoing but no longer recruiting new patients. FINDINGS:Between Sept 7, 2020, and Nov 18, 2022, 65 patients were enrolled (median age 64 years [IQR 54-70]) and included in the FAS and safety set. 40 (62%) patients were male and 25 (38%) were female. At the data cutoff of Nov 19, 2024, the median follow-up duration was 33·73 months (IQR 24·86-40·38) and 36 (55%) of 65 patients had recurrence (24 [67%] with intrahepatic recurrence only, six [17%] with both intrahepatic and extrahepatic recurrence, and six [17%] with extrahepatic recurrence only). Median recurrence-free survival was 24·29 months (95% CI 13·54-not reached). The most common treatment-related adverse events (occurring in ≥10% of patients) were reactive cutaneous capillary endothelial proliferation (45 [69%] patients), nausea (21 [32%] patients), hand-foot syndrome (19 [29%] patients), pruritus (11 [17%] patients), fatigue (11 [17%] patients), and dizziness (nine [14%] patients). Grade 3 treatment-related adverse events occurred in 15 (23%) patients, the most common of which was elevated bilirubin (two [3%] patients). Serious treatment-related adverse events were reported in four (6%) patients, including one case each of myocarditis, myalgia, type 1 diabetes, and hypothyroidism. No grade 4 treatment-related adverse events or treatment-related deaths occurred. INTERPRETATION:The combination of camrelizumab and capecitabine showed promising activity with an acceptable safety profile in the adjuvant setting for patients with resected intrahepatic cholangiocarcinoma. Further validation of this immunochemotherapy regimen is warranted in larger, multicentre trials. FUNDING:Clinical Research Special Fund, Zhongshan Hospital, Fudan University.

Hepatocellular carcinoma (HCC), which accounts for approximately 75–85% of primary liver cancers, ranks 4th in newly diagnosed cases among various types of cancer in China, and is the 2nd leading cause of cancer-related mortality, thereby posing a significant threat to the life and health of the Chinese population. Since the publication of the “Guidelines for Diagnosis and Treatment of Primary Liver Cancer in China” in June 2017, which were updated by the China’s National Health Commission in December 2019 and December 2021, additional high-quality evidence from researchers worldwide regarding the diagnosis, staging, and treatment of HCC has emerged, necessitating another update to the guidelines. The new edition (2024 Edition) was written by more than 120 multidisciplinary experts in the field of HCC in China, which not only reflects the real-world situation in China but also may reshape the nationwide diagnosis and treatment of HCC. The new guideline aims to encourage the implementation of evidence-based practice and improve the national average 5-year survival rate for patients with HCC, as proposed in the “Healthy China 2030: A Vision for Health Care.”

The aim of this study was to determine the expression of Glypican-3(GPC-3) and AXL in hepatocellular carcinomas (HCCs). A total of 140 patients diagnosed with HCC were included in this study. All patients had undergone radical surgery and had complete clinical information. Formalin-fixed paraffin-embedded tissue blocks of HCC from these patients were collected, and the expression levels of GPC-3 and AXL were detected by immunohistochemical (IHC) staining. Immunohistochemical analysis showed that GPC-3 and AXL were diffusely expressed in HCCs. The positive expression rate of GPC-3 was 77.1

The safety of immunotherapy in patients undergoing transplant remains unclear due to rejection risks. This study assessed the safety and efficacy of programmed death-1 (PD-1) inhibitors in patients with recurrent tumors who had undergone liver transplantation, emphasizing the value of using graft programmed death-ligand 1 expression as a predictor of rejection to guide patient selection. This single-center, open-label, prospective, single-arm study was conducted from July 2019 to May 2024 at Zhongshan Hospital, Fudan University (Shanghai, China). Eligible participants included patients with recurrent or metastatic liver cancer who had undergone liver transplantation and were unresponsive to locoregional or systemic therapies. The primary endpoints were the incidence and clinical outcomes of acute rejection. Secondary endpoints were overall survival and objective response rate. Twenty consecutive patients received PD-1 inhibitor therapy. Of these, 18 had HCC, and 2 had intrahepatic cholangiocarcinoma. Liver graft biopsies confirmed negative programmed death-ligand 1 expression in all participants before PD-1 inhibitor therapy. Three patients (15%) experienced acute rejection, with a 95% CI of 3.2%-37.9%. The 1-year and 2-year survival probabilities after PD-1 inhibitor treatment were 0.55 (95% CI: 0.33-0.77) and 0.24 (95% CI: 0.05-0.43), respectively. The median survival time after tumor recurrence was 24.6 months, exceeding the historically reported median survival time of 16.3 months. These exploratory findings suggest that, in selected recipients of liver transplant, PD-1 inhibitors may be associated with reduced rejection risk and potential survival benefit, although further validation is needed.

Adding a PD-1/PD-L1 inhibitor to gemcitabine plus cisplatin (GemCis) has shown survival benefits in advanced biliary tract cancer (BTC). Dual inhibition of PD-1/PD-L1 and TIGIT may act synergistically, and further enhance antitumor effects. ZSAB-TOP was a single-arm, multicenter, phase 2 study (NCT05023109) evaluating efficacy and safety of first-line tislelizumab (a PD-1 inhibitor) plus ociperlimab (a TIGIT inhibitor) and GemCis in advanced BTC. Eligible patients received tislelizumab (200 mg) and ociperlimab (900 mg) on day 1 until unacceptable toxicity or disease progression, in combination with cisplatin (25 mg/m²) and gemcitabine (1000 mg/m²) on days 1 and 8 of a 21-day cycle for a maximum eight cycles. The primary endpoint was confirmed objective response rate (ORR) evaluated by the investigator, which was compared with a historical ORR of 25% with GemCis, with a statistical superiority setting at p ≤ 0.05. From March 8, 2022, to January 18, 2023, 45 patients were enrolled. Among the 41 patients in the efficacy analysis set, the confirmed ORR was 51.2% (95% CI 35.1–67.1), achieving the statistical superiority criteria (p = 0.0003). Patients who had TIGIT+/PD-L1+ (n = 16) tended to have a numerically greater confirmed ORR (75.0% [95% CI 47.6–92.7]). After a median follow-up of 14.6 months, median progression-free survival was 7.7 months (95% CI 6.0–9.4), with a median overall survival of 17.4 months (95% CI 11.7-not reached). Treatment-related adverse events of grade ≥3 occurred in 60.0% of patients; immune-mediated adverse events of any grade was observed in 42.2%, with the majority being grade 1 or 2. In conclusion, first-line tislelizumab and ociperlimab plus GemCis yielded clinically promising tumor response and survival outcomes in advanced BTC and were generally well tolerated without new safety signals.

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.

BACKGROUND:Despite comparable survival benefit has been obtained, the drug resistance remarkably reduced lenvatinib clinical efficacy. Here, we aimed to identify the potential mechanism by which cysteine and glycine-rich protein 2 (CSRP2) regulates the development of hepatocellular carcinoma (HCC) and participates in the resistance to lenvatinib. METHODS:We harnessed RNA sequencing, multiplex immunofluorescence staining, and hydrodynamic tail vein (HTV) injection HCC model to systematically explore the function of CSRP2 in HCC progression. To precisely delineate how immunosuppressive macrophages, influenced by CSRP2-regulated C-C motif chemokine ligand 28 (CCL28) signaling, respond to lenvatinib-induced cytotoxicity, we established an in vitro co-culture system and conducted functional cytotoxicity assays. RESULTS:Using RNA sequencing, multiplex immunofluorescence staining and HTV injection HCC mouse model, we identified CSRP2 as one of the most significantly upregulated genes in HCC tissues. CSRP2 overexpression drives anti-lenvatinib resistance by inducing high levels of tumor-associated macrophages (TAMs) infiltration and reshaping an immunosuppressive microenvironment. Then flow cytometry, mass spectrometry and chromatin immunoprecipitation were conducted to clarify the underlying mechanism of CSRP2. We showed CSRP2 promotes phosphorylation of activating transcription factor 2 (ATF2) at Thr69/71, leading to the transcriptional activation of CCL28 expression. HCC-derived CCL28 recruits TAMs to drive immunosuppression and anti-lenvatinib tolerance. BI6901, a potent and selective CCR10 antagonist, blocked TAMs recruitment and enhanced T-cell activation. Combining CCR10 inhibition improved the therapeutic benefit of anti-lenvatinib in HCC. CONCLUSIONS:These results illustrate that CSRP2 regulates the tumor microenvironment to promote HCC growth and drive lenvatinib tolerance via the CSRP2/ATF2/CCL28 axis. Targeting this pathway could synergize with lenvatinib to treat HCC more effectively.

BACKGROUND & AIMS:Tumour-associated macrophages (TAMs) contribute to hepatocellular carcinoma (HCC) progression. However, while the pro-tumour and immunosuppressive roles of lipid-loaded macrophages are well established, the mechanisms by which lipid metabolism enhances the tumour-promoting effects of TAMs remain unclear. METHODS:Single-cell RNA sequencing was performed on mouse and human HCC tumour samples to elucidate the landscape of HCC TAMs. Macrophages were stimulated with various long-chain unsaturated fatty acids (UFAs) to assess immunosuppressive molecule expression in vitro. Additionally, in vivo and in vitro studies were conducted using mice with macrophage-specific deficiencies in fatty acid-binding protein 5 (FABP5) or peroxisome proliferator-activated receptor γ (PPARγ). RESULTS:Single-cell RNA sequencing identified a subpopulation of FABP5+ lipid-loaded TAMs characterized by enhanced immune checkpoint blocker ligands and immunosuppressive molecules in an oncogene-mutant HCC mouse model and human HCC tumours. Mechanistically, long-chain UFAs released by tumour cells activate PPARγ via FABP5, resulting in immunosuppressive properties in TAMs. FABP5 deficiency in macrophages decreases immunosuppressive molecule expression, enhances T cell-dependent antitumour immunity, diminishes HCC growth, and improves immunotherapy efficacy. CONCLUSIONS:This study demonstrates that UFAs promote tumourigenesis by enhancing the immunosuppressive tumour microenvironment via FABP5-PPARγ signalling and provides a proof-of-concept for targeting this pathway to improve the efficacy of tumour immunotherapy. IMPACT AND IMPLICATIONS:Despite the role of tumour-associated macrophages (TAMs) in promoting tumour progression being well established, the mechanisms by which lipid metabolism enhances the tumour-promoting effects of TAMs remain unclear. Our study reveals that FABP5-mediated unsaturated fatty acid metabolism in TAMs is crucial for modulating antitumour T-cell immunity and influencing the efficacy of immunotherapy. This finding provides novel insights into the immunomodulatory roles of FABP5+ lipid-loaded TAMs in hepatocellular carcinoma and suggests that targeting FABP5 could offer a new approach to liver cancer treatment.

4084 Background: Previously, we reported the results of Ori-C101 from investigator initiated trial in China (ChiCTR1900028121). The data demonstrated that Ori-C101 owned a favorable safety profile and promising efficacy. Among 10 GPC3 + HCC patients (pts) treated with Ori-C101, 9 pts (90%) achieved disease control and 6 pts (60%) met partial response per RECIST 1.1. Two pts with PR attained progression-free survival of one and two years respectively, with an overall survival close to 3 years. These results implied that Ori-C101 potentially held significant clinical benefits. Subsequently, extensive optimizations and improvements in the manufacturing process were implemented to enhance its clinical efficacy and persistency. Hence, a multicenter registration study was launched in China (the BEACON study), and herein, we will present the preliminary results. Methods: This is an open-label, multi-center, dose-escalation (3+3 design) study. GPC3 + advanced HCC pts who failed at least 2 lines of systemic treatments received a single hepatic arterial infusion with a total dose of 0.9 to 6×10 8 CAR-T cells. Primary endpoints are rate of dose-limiting toxicities (DLTs) and safety with the aim to determine a recommended phase II dose (RP2D). Secondary endpoints are cellular kinetics, overall response rate by investigator assessment, duration of response, overall survival and overall safety. Results: As of Dec 17th, 2024, a total of 10 eligible pts received Ori-C101 infusion at 3 dose levels (DLs). All pts had BCLC stage B or C, with 20% (2/10) had extrahepatic metastasis. The median number of prior lines of therapy was 4.5 (range 2-9), 100% pts received immune checkpoint inhibitors and tyrosine kinase inhibitors. All pts were evaluable for safety. All adverse events were reported regardless of study drug relationship. Of 10 pts evaluable for safety, the most common ≥ grade (G) 3 AEs were lymphocyte count decreased (100%), neutrophil count decreased (60.0%), blood fibrinogen decreased (40.0%), transaminases increased (40.0%), platelet count decreased (20.0%), blood bilirubin increased (20.0%). CRS was observed in 10 (100%) pts with 3 (30.0%) ≥ G3 CRS. No ICANS was observed. One pt developed DLT event due to CRS and secondary disseminated intravascular coagulation. 9 pts were evaluable for efficacy per RECIST 1.1. While 6 pts (66%) achieved disease control at DL2 or higher, all pts at the DL3 achieved objective response. Particularly, one pt who achieved CR showed encouraging durability and no signs of relapse at 9 months follow up evaluation, and follow up is ongoing. Conclusions: These preliminary data showed Ori-C101 has manageable safety profile and exciting efficacy with encouraging sign of good durability. Currently, more pts have been enrolled at dose expansion to confirm the DLs of RP2D. More information will be presented at coming ASCO conference. Clinical trial information: NCT05652920 .

e16312 Background: A prospective, single-arm, open-label exploratory, phase II study (ChiCTR2000035901) aims to explore the safety and efficacy of Sintilimab plus Anlotinib in unresectable ICC. Methods: Eligible unresectable ICC patients received Sintilimab (200 mg, iv, d1) and Anlotinib (12 mg, po, d1-14) every 3 weeks until disease progression or unacceptable toxicity. The primary endpoint was the overall response rate (ORR) assessed by RECIST v1.1 and secondary endpoints included progression-free survival (PFS), disease control rate (DCR), duration of response (DOR), overall survival (OS), surgical conversion rate, as well as safety and tolerability. Results: By the end of Dec. 2024, 28 patients were enrolled with a median age of 60.3 years (range 36-74), 57.1% were male, and all patients were classified as Child-Pugh A. 26 of 28 patients (92.9%) presented with TNM stage IIIb/IV and 11 (11/28, 39.3%) patients had distant metastasis. Confirmed ORR was 42.9% and DCR was 85.7%. Of these, 7.1% (2/28) patients achieved complete response (CR), and 35.7% (10/28) patients achieved partial response (PR). Median PFS was 8.70 months (95% CI: 4.01-13.39) and median OS was 18.33 (95% CI: 11.71-24.95). Among those, 7 (25.0%) patients further received surgical resection, and 3 of them remained disease-free survival at the last follow-up. Treatment-related AEs (TRAEs) occurred in 23/28 pts (82.1%), with the most common being hypertension in 57.1%. Grade 3 TRAEs occurred in 9/28 pts (32.1%), and there was no grade 4-5 TRAEs. Conclusions: Given its encouraging efficacy and safety profile, Sintilimab plus Anlotinib might represent a viable and safe chemotherapy-free regimen in unresectable ICC treatment. Baseline characteristics of patients. Clinical and pathologic indexes Responders Non-Responders P (CR+PR) (n=12) (SD+PD) (n=16) N % N % Age(years) ≤50 3 25.00% 2 12.50% 0.624 >50 9 75.00% 14 87.50% Gender Female 7 58.30% 5 31.25% 0.250 Male 5 41.70% 11 68.75% ALT, U/L ≤50 10 83.30% 13 81.25% 1.000 >50 2 16.67% 3 18.75% GGT, U/L ≤60 2 16.67% 2 12.50% 1.000 >60 10 83.30% 14 87.50% Albumin, g/L <35 12 100.00% 14 87.50% 0.492 ≥35 0 0.00% 2 12.50% PT, s ≤13 12 100.00% 13 81.25% 0.238 >13 0 0.00% 3 18.75% HBsAg Negative 11 91.67% 14 87.50% 1.000 Positive 1 8.33% 2 12.50% CA19-9, U/mL ≤34 3 25.00% 2 12.50% 0.624 >34 9 75.00% 14 87.50% Maximal primary tumor diameter, cm ≤5 2 16.67% 1 6.25% 0.560 >5 10 83.30% 15 93.25% Intrahepatic tumor number Single 8 66.67% 10 62.50% 1.000 Multiple 4 33.33% 6 37.50% Lymph gland metastasis No 3 25.00% 2 12.50% 0.624 Yes 9 75.00% 14 87.50% Distance metastasis No 10 83.30% 7 43.75% 0.054 Yes 2 16.67% 9 56.25% Abbreviation: CR, complete response; PR, partial response; SD, stable disease; PD, progressive disease; HBsAg, hepatitis B surface antigen; ALT, alanine aminotransferase; GGT, γ-glutamyl transpeptidase; PT, prothrombin time.