Lenvatinib, a first-line therapy for hepatocellular carcinoma (HCC), is frequently compromised by the development of acquired resistance. Although aberrant protein ubiquitination contributes to drug resistance, the underlying mechanisms remain incompletely understood. In this study, we identify ubiquitin-specific protease 32 (USP32) as a clinically significant oncogene that is overexpressed in HCC and closely associated with aggressive tumor progression and lenvatinib resistance. USP32 knockdown markedly inhibits tumor growth, metastasis, and restores lenvatinib sensitivity both in vitro and in vivo, whereas USP32 overexpression aggravates malignant behaviors. Mechanistically, USP32 directly interacts with and deubiquitinates DEAD-box helicase 17 (DDX17) by cleaving K48-linked polyubiquitin chains, thereby stabilizing DDX17. Furthermore, DDX17 transcriptionally represses the tumor-suppressive microRNA miR-3163, leading to hyperactivation of the MAPK signaling pathway involved in lenvatinib resistance. Notably, combined treatment with the MAPK inhibitor selumetinib and lenvatinib synergistically overcomes drug resistance in lenvatinib-resistant xenograft models. Collectively, our findings identify USP32 as an important regulator of HCC malignancy and lenvatinib resistance via the DDX17/miR-3163/MAPK axis, highlighting its potential as a therapeutic target in refractory HCC.

BACKGROUND:Although several postoperative adjuvant therapies for hepatocellular carcinoma (HCC) have shown efficacy in reducing recurrence, no globally accepted guidelines exist for patients with vascular invasion after curative resection. This study aimed to evaluate the efficacy and safety of lenvatinib (Len) as adjuvant therapy for patients with HCC with vascular invasion after curative liver resection (LR). METHODS:Patients with HCC with vascular invasion, including microvascular invasion (MVI) and portal vein tumor thrombus (PVTT), who underwent curative resection at 2 hospitals were retrospectively reviewed. Overall survival (OS), recurrence-free survival (RFS), and recurrence patterns were compared between patients treated with LR alone and those receiving postoperative Len (LR + Len) using propensity score matching (PSM). RESULTS:A total of 326 patients with HCC with vascular invasion were enrolled. Among them, 248 (76.1%) had MVI; 79 patients were allocated to the LR + Len group, and the others to the LR group. After PSM (1:3), 73 patients were included in the LR + Len group and 172 patients in the LR group. In the matched cohort, patients in the LR + Len group had significantly longer RFS (hazard ratio [HR], 1.54; 95% CI, 1.03-2.29; P =.032) and OS (HR, 2.11; 95% CI, 1.08-4.10; P =.025) than those undergoing LR alone. Subgroup analysis revealed that adjuvant Len significantly improved both RFS and OS in patients with MVI, but it conferred no benefit to those with PVTT. CONCLUSION:Postoperative adjuvant Len therapy was associated with longer OS and RFS for patients with HCC with vascular invasion, particularly for those with MVI.

The prevailing notion is that effector T cell activation mediates anti-PD-1 efficacy in cancer. Here, we conducted a mechanistic study parallel to our phase 2 trial of perioperative anti-PD-1 therapy in patients with resectable recurrent hepatocellular carcinoma (HCC) (NCT04615143) to study its mechanism of action. Late-recurrence patients present two distinct subtypes characterized by T cell or B cell dominant responses in the tumor microenvironment by dynamic single-cell multi-omics analysis. Clonal antibody repertoire analysis and spatially paired scRNA-seq/BCR-seq reveal somatic hypermutation promoting antibody binding against hepatitis B virus core antigen (HBcAg) within tumor tertiary lymphoid structures (TLSs) in these type B-late recurrence patients. Mechanistically, HBcAg is exported into the extracellular space, triggering local B cell and antibody responses and complement activation. In mice, these high-affinity HBcAg-reactive antibodies lead to complement-mediated antitumor activity with enhanced anti-PD-1 efficacy. Thus, we uncover enhanced anti-virus B cell immunity within the TLS as a mechanism to anti-PD-1 in HCC.

Objectives:The main objectives of this research are to evaluate the outcomes of patients with initially unresectable hepatocellular carcinoma (HCC) who received transcatheter arterial chemoembolization (TACE)/hepatic artery infusion chemotherapy (HAIC)-based combination therapy and to investigate the effects of liver resection following comprehensive conversion therapy on the short-term benefits and long-term survival of these patients. Materials and Methods:A total of 301 initially unresectable HCC patients who received TACE/HAIC-based combination therapy between January 2019 and December 2021 were retrospectively reviewed. The study analyzed the conversion rate to resection, changes in tumor burden after treatment, and the survival outcomes. Results:The study found that 20.9% (63/301) of initially unresectable HCC patients were able to undergo liver resection. The conversion resection rate among all patients was 38.2% (29/76) and 17.3% (23/132) for those with Barcelona Clinic Liver Cancer (BCLC) stage A and C. Patients who underwent surgery achieved promising outcomes with a pathological complete response (pCR) rate of 31.7% (20/63) and a 100% R0 resection rate. Kaplan-Meier survival analysis showed that patients who had successful surgery after conversion therapy had significantly longer median overall survival (OS) (not reached vs. 58.5 months) and progression-free survival (PFS) (42.83 months vs. 9.7 months) compared to those who did not (both p < 0.05). Additionally, patients achieving radiographic complete response (CR) had significantly better OS and PFS than those who did not. Multivariable logistic regression analysis showed that age (OR = 0.95, p < 0.001), positive HBsAg expression (OR = 0.34, p = 0.011), and alpha-fetoprotein levels ≥400 (OR = 0.49, p = 0.045), ECOG PS score of 1 (OR = 0.43, p = 0.038), BCLC stage B (OR = 0.23, p < 0.001) and stage C (OR = 0.44, p = 0.045), systemic inflammation response index (OR = 0.73, p = 0.018) were independent predictors for successful conversion surgery (all p < 0.05). Conclusion:Patients with initially unresectable HCC can achieve promising curative effects and conversion resection rates with TACE/HAIC-based comprehensive therapy. More importantly, patients who undergo liver resection following conversion resection had significantly better long-term survival.

e16304 Background: Prognoses for patients with hepatocellular carcinoma (HCC) remain suboptimal due to the high recurrence rates following curative intended hepatectomy. In this study, we evaluated the safety and efficacy of neoadjuvant Cadonilimab (a bi-specific antibody targeting PD-1 and CTLA-4) in combination with transarterial chemoembolization (TACE) for patients at high risk of recurrence. Methods: In this phase II, single-arm trial, resectable HCC (BCLC stage A/B) participants were enrolled. Patients should meet at least one of the following high-risk criteria: a) Multiple tumors or satellite lesions (≥2); b) tumor diameter > 5cm (single tumor); c) AFP ≥ 400 ug/L; d) Predicted to be positive by the Microvascular invasion prediction model of our center according to preoperative imaging. All patients received two cycles of Cadonilimab(10mg/kg, Q3W) following TACE treatment. After resection, patients received additional 12 months of Cadonilimab (10mg/kg, Q3W). The primary endpoint was the major pathological response(MPR, defined as ≥90% necrosis of the resected tumour),while the secondary endpoints include objective response rate (ORR), one-year recurrence rate and safety. Results: Until December 13th, 2024, a total of 30 patients were enrolled and underwent successful resection. Among the patients, 25(83.3%) were male, with the mean age of 54 years. Fourteen (46.7%) patients had multiple tumors, and 22(73.3%) had tumors larger than 5cm. Nineteen (63.3%) achieved MPR, while26(86.7%) exhibited 70% or greater tumour necrosis. According to mRECIST , the ORR was 83.3% (25/30) and DCR was 100%(30/30). Of these patients, 2 patients achieved complete response (CR), and 23 patients achieved partial response (PR). Additionally, 27 patients maintained stable decease, while only 3 patients received PR as assessed by RECIST1.1. The 1-year recurrence-free survival rate of entire cohort was 90%. Notably, patients who did not achieve MPR had a higher recurrence rate(27.3% vs. 5.3%). Simultaneously, this subgroup also demonstrated a lower ORR (54.6 vs. 100%) per mRECIST. The most common TRAEs of any grade were ALT/AST increase (90%), abdominal pain (86.7%) and bilirubin increase (56.7%). Four patients experienced grade 3 adverse events, including infusion reaction (n = 2), and white blood cell decreased (n = 2). Notably, abdominal pain and fever were more frequently associated with TACE. No grade 4 or 5 events were observed. The incidence of immune-related adverse event (irAE) was 23.3%. Furthermore, it was observed that patients who experienced irAEs tended to demonstrate a better pathological response. Conclusions: Cadonilimab combined with TACE as a neoadjuvant therapy demonstrated acceptable safety profile and promising efficacy for high risk HCC patients. Clinical trial information: NCT05578430 .

Bronchobiliary fistula (BBF) is a rare but fatal disease. Due to its rarity, only a limited number of cases have been reported, leading to a lack of consensus on appropriate treatment strategies. We conducted a retrospective analysis of clinical data of 17 patients diagnosed with BBF between January 1, 2012, and May 30, 2025, focusing on the the presenting symptoms, diagnostic approaches, treatment modalities, and outcomes. All 17 patients had cough and pathognomonic biliptysis. Sputum analysis confirmed bile components in all samples from 3 patients, and fiber bronchoscopy revealed yellow-green bilious sputum in 6 patients. Computed tomography (CT) or magnetic resonance imaging (MRI) demonstrated communication between the bile duct and the bronchial tree in 9 patients. Cholangiography showed contrast medium passing through a fistulous tract into the bronchi in 10 patients. Surgical intervention was performed in 6 patients, 3 of whom achieved long-term survival (> 24 months). Minimally invasive interventions provided effective symptomatic palliation in 10 of 11 patients. Among these 10 patients, 3 achieved long-term survival (> 24 months), with an additional 3 remaining alive and under ongoing follow-up (though not yet reaching the 24-month threshold). Of the 8 fatalities, 2 were directly attributed to uncontrolled BBF and its complications (sepsis or hepatic failure). The remaining deaths resulted from progressive malignancy (n = 3), postoperative complications (pneumothorax/respiratory failure, n = 1; hemorrhagic shock/disseminated intravascular coagulation, n = 1), and post-transplant septic shock (n = 1). BBF is associated with poor prognosis. Minimally invasive therapies offer effective palliation in malignant cases, whereas surgical intervention may provide curative potential in selected benign cases. Individualized, multidisciplinary management is essential for optimizing outcomes.

Lenvatinib is the first-line therapy for inoperable HCC. However, intrinsic and acquired drug resistance occurs during the treatment period. Autophagy is an adaptive response that favors tumor survival under stress. In the present study, we aim to reveal the unknown autophagic engagement in lenvatinib resistance. Lenvatinib-resistant HCC cell lines and xenograft mouse HCC models were established to identify the key regulator of lenvatinib resistance in HCC. By in vitro functional restoration assays and autophagic flux detection, we demonstrated that the Syntaxin-6 (STX6) -mediated autophagy induced lenvatinib resistance of HCC cells. Mechanistically, Co-immunoprecipitation assay and mass spectrometry indicated that the interactions of STX6 with Beclin1, VTI1A, and VAMP3 facilitated autophagy, leading to the lenvatinib resistance. Additionally, STX6 enhanced the ability of proliferation, migration, and invasion of HCC in vitro and in vivo. Clinically, STX6 expression was significantly elevated in HCC tissues compared to it in para-tumor tissues. High STX6 expression predicted poor outcomes for patients following resection. Moreover, high expression of STX6 displayed low preventive efficacy of lenvatinib as a postoperative adjuvant treatment for HCC patients with a high risk of recurrence. Collectively, we identified that STX6-mediated autophagy plays a crucial role in lenvatinib resistance in HCC, providing a potential therapeutic target to overcome lenvatinib resistance for HCC patients.

Extrahepatic cholangiocarcinoma (EHC) and gallbladder cancer (GBC), which make up most biliary tract cancers, have distinct molecular and clinical features compared with intrahepatic cholangiocarcinoma. However, effective adjuvant treatments specifically for patients with resectable EHC and GBC are scarce. To evaluate the safety and efficacy of immunotherapy combining with chemoradiotherapy. In the ACCORD randomized clinical trial, from April 2020 to June 2022, patients with EHC and GBC after curative resection were assessed for eligibility. Patients were randomized 1:1 into to the combination camrelizumab plus concurrent capecitabine and radiotherapy group or the observation group. Data were analyzed from June to August 2024. The intervention group received camrelizumab every 3 weeks after surgery. After 2 courses of camrelizumab treatment, patients received capecitabine with concurrent radiotherapy. Patients in the observation group received no anticancer treatment unless relapse was detected. The primary end point was overall survival (OS) and the secondary end points included recurrence-free survival (RFS) and safety. Of 93 included patients, 48 (52%) were female, and the median (range) age was 62 (31-70) years. Patients' baseline characteristics were comparable in the 2 groups. With a median (IQR) follow up of 36 (32-39) months, patients in the combination treatment group significantly better OS and RFS. The 1-year, 2-year and 3-year OS rates were 95.7% (95% CI, 83.7-98.9), 71.4% (95% CI, 56.4-82.5), and 58.2% (95% CI, 40.4-72.4), respectively, in the combination treatment group and 80.9% (95% CI, 66.4-89.5), 52.9% (95% CI, 37.7-65.9), and 30.5% (95% CI, 16.5-45.7), respectively, in the observation group (hazard ratio, 0.43; 95% CI, 0.24-0.79; P = .004). The 1-year, 2-year and 3-year RFS rates were 78.3% (95% CI, 63.4-87.7), 54.0% (95% CI, 38.6-67.1), and 40.3% (95% CI, 25.3-54.8), respectively, in the combination treatment group and 55.3% (95% CI, 40.1-68.1), 27.0% (95% CI, 15.2-40.3), and 17.2% (95% CI, 7.7-29.8), respectively, in the observation group (hazard ratio, 0.46; 95% CI, 0.28-0.76; P < .001). In the combination treatment group, only 6 patients (13%) experienced treatment delay for camrelizumab, and all patients completed the chemoradiation treatment, with no treatment-related deaths. In this randomized clinical trial, camrelizumab plus concurrent capecitabine and radiotherapy as an adjuvant therapy demonstrated superior survival outcomes over observation in patients with resectable EHC/GBC with a well-tolerable safety profile. The observed camrelizumab plus concurrent capecitabine and radiotherapy efficacy warrants further study with active treatment (chemotherapy or chemoradiation therapy) as the control group. ClinicalTrials.gov Identifier: NCT04333927.

Hilar cholangiocarcinoma (hCCA) is a malignant tumor originating from the epithelial cells of the bile ducts, and it is characterized by an aggressive nature, complex surgical management, high mortality, and poor prognosis. Despite recent advances in surgical techniques, medical devices, and related technologies, there remains a pressing need to standardize diagnostic and therapeutic pathways to improve treatment outcomes and extend long-term patient survival. To better integrate and refine these standards, this consensus was reached through a national conference held in Changsha, Hunan Province, involving multidisciplinary experts from various regions across China. This collaborative effort, drawing from various medical facilities and academic organizations nationwide, resulted in the reaching of the "Chinese Multicenter Expert Consensus on the Diagnosis and Treatment of Hilar Cholangiocarcinoma: 2025 Edition" based on current clinical studies and over 40 years of clinical practice experience in managing hCCA. The consensus provides a comprehensive overview of hCCA, including its epidemiological characteristics, diagnostic and screening methods, pathological features, staging and classification systems, and various treatment modalities, while offering specific and actionable recommendations for clinical practice that highlight well-defined indications for surgical, local, and systemic therapies and that emphasize the importance of multidisciplinary approaches to both diagnostic and therapeutic workflows.

Purpose: The patterns and risk factors of postsurgical recurrence of patient with hepatocellular carcinoma (HCC) with microvascular invasion (MVI) are not clarified. This study aimed to decipher and compare the postoperative recurrent patterns and the risk factors contributing to recurrence between MVI positive (MVI ((+))) and MVI negative (MVI ((-))) HCC after hepatectomy. Patients and methods: Patients with HCC who underwent hepatectomy in three Chinese academic hospitals between January 1, 2009, and December 31, 2018, were enrolled. Recurrent patterns included early (<= 2 years) or late (>2 years) recurrence, recurrent sites and number, and risk factors of recurrence were compared between the MVI ((+) )and MVI ((-)) groups by propensity score-matching (PSM). Results: Of 1756 patients included, 581 (33.1%) were MVI(+), and 875 (49.8%) patients developed early recurrence. Compared with the MVI ((-)) group, the MVI((+) )group had a higher 2-year recurrence rate in the PSM cohort (hazard ratio [HR], 1.82; 95% confidence interval [CI], 1.59-2.10; P < 0.001), and more patients with multiple tumor recurrence. Patients with early recurrence in the MVI((+) )group had a worse overall survival (OS) than those in the MVI((-) )group (HR, 1.24; 95% CI, 1.02-1.50; P = 0.034). Resection margin (RM) <= 1.0 cm is a surgical predictor of early recurrence for the MVI(+) group (HR, 0.68; 95% CI, 0.54-0.87; P = 0.002), but not for the MVI((-) )group. Conclusion: Compared to MVI(-) HCC, MVI(+) HCC tends to be early, multiple recurrence and lung and lymph node metastasis after resection. RM <= 1.0 cm is a surgical risk factor of early recurrence for patient with MVI.

Background:Lenvatinib or Sorafenib combined with programmed cell death protein-1 (PD-1) inhibitor as recommend treatment of advanced hepatocellular carcinoma (HCC) with extrahepatic metastasis (EHM). We aimed to compared the prognosis of Lenvatinib plus PD-1 inhibitor (Len+PD-1) versus Sorafenib plus PD-1 (Sora+PD-1) as an initial therapy for HCC with EHM.Methods:Incorporating a sum of 229 HCC patients with EHM were encompassed within this study, with 127 in the Sora+PD-1 group and 102 in the Len+PD-1 group. Through propensity score matching (PSM), we compared overall survival (OS), progression-free survival (PFS), and patient safety between these two groups.Results:The median OS were 13.0 months and 14.2 months in the Sora+PD-1 group and Len+PD-1 group. The 6-, 12-, and 24-month OS rates were 92.9%, 58.9% and 5.6% in Sora+PD-1 group and 93.1%, 61.8% and 22.6% in Len+PD-1 group, respectively. The Len+PD-1 group had obviously better OS than the Sora+PD-1 group (P = 0.002). The 3-, 6-, and 12-month PFS rates were 76.4%, 27.6% and 1.6% in Sora+PD-1 group and 86.2%, 50.5% and 12.2% in Len+PD-1 group, respectively. Compared with Sora+PD-1 group, the Len+PD-1 group had obviously better PFS (P < 0.001). Analysis within subgroups showed that OS was significant in patients receiving TACE in Len+PD-1 group than Sora+PD-1 group (p = 0.003).Conclusion:Len+PD-1 group had longer OS and PFS than Sora+PD-1 group for patient with EHM. In addition, OS in patients received TACE was improved with Len+PD-1 treatment. For patients without TACE, there was no significance between Sora+PD-1 and Len+PD-1 groups.

BackgroundMicrovascular invasion (MVI) stands as a pivotal pathological hallmark of hepatocellular carcinoma (HCC), closely linked to unfavorable prognosis, early recurrence, and metastatic progression. However, the precise mechanistic underpinnings governing its onset and advancement remain elusive.MethodsIn this research, we downloaded bulk RNA-seq data from the TCGA and HCCDB repositories, single-cell RNA-seq data from the GEO database, and spatial transcriptomics data from the CNCB database. Leveraging the Scissor algorithm, we delineated prognosis-related cell subpopulations and discerned a distinct MVI-related malignant cell subtype. A comprehensive exploration of these malignant cell subpopulations was undertaken through pseudotime analysis and cell-cell communication scrutiny. Furthermore, we engineered a prognostic model grounded in MVI-related genes, employing 101 algorithm combinations integrated by 10 machine-learning algorithms on the TCGA training set. Rigorous evaluation ensued on internal testing sets and external validation sets, employing C-index, calibration curves, and decision curve analysis (DCA).ResultsPseudotime analysis indicated that malignant cells, showing a positive correlation with MVI, were primarily concentrated in the early to middle stages of differentiation, correlating with an unfavorable prognosis. Importantly, these cells showed significant enrichment in the MYC pathway and were involved in extensive interactions with diverse cell types via the MIF signaling pathway. The association of malignant cells with the MVI phenotype was corroborated through validation in spatial transcriptomics data. The prognostic model we devised demonstrated exceptional sensitivity and specificity, surpassing the performance of most previously published models. Calibration curves and DCA underscored the clinical utility of this model.ConclusionsThrough integrated multi-transcriptomics analysis, we delineated MVI-related malignant cells and elucidated their biological functions. This study provided novel insights for managing HCC, with the constructed prognostic model offering valuable support for clinical decision-making.

ImportanceCertain patients with hepatocellular carcinoma with portal vein tumor thrombus could benefit from surgical resection, and postoperative adjuvant therapy may lower the incidence of tumor recurrence.ObjectiveTo compare the efficacy and safety of sorafenib plus transarterial chemoembolization vs sorafenib alone as postoperative adjuvant therapy for patients with hepatocellular carcinoma with portal vein tumor thrombus.Design, Setting, and ParticipantsThis was a phase 3, multicenter, randomized clinical trial conducted in 5 hospitals in China. A total of 158 patients were enrolled and randomized from October 2019 to March 2022, with a median follow-up of 28.4 months. Portal vein tumor thrombus was graded by the Cheng classification. Eligible patients with hepatocellular carcinoma with Cheng grade I to III portal vein tumor thrombus (ie, involving segmental or sectoral branches, right- or left-side branch, or main trunk of portal vein) were included.InterventionsPatients were randomly assigned 1:1 to receive transarterial chemoembolization with sorafenib or sorafenib alone as postoperative adjuvant therapy. Sorafenib treatment was started within 3 days after randomization, with an initial dose of 400 mg orally twice a day. In the transarterial chemoembolization with sorafenib group, transarterial chemoembolization was performed 1 day after the first administration of sorafenib.Main Outcomes and MeasuresThe primary end point was recurrence-free survival. Efficacy was assessed in the intention-to-treat population and safety was assessed in patients who received at least 1 dose of study treatment.ResultsOf 158 patients included, the median (IQR) age was 54 (43-61) years, and 140 (88.6%) patients were male. The median (IQR) recurrence-free survival was significantly longer in the transarterial chemoembolization with sorafenib group (16.8 [12.0-NA] vs 12.6 [7.8-18.1] months; hazard ratio [HR], 0.57; 95% CI, 0.39-0.83; P = .002). The median (IQR) overall survival was also significantly longer with transarterial chemoembolization with sorafenib than with sorafenib alone (30.4 [20.6-NA] vs 22.5 [15.4-NA] months; HR, 0.57; 95% CI, 0.36-0.91; P = .02). The most common grade 3/4 adverse event was hand-foot syndrome (23 of 79 patients in the transarterial chemoembolization with sorafenib group [29.1%] vs 24 of 79 patients in the sorafenib alone group [30.4%]). There were no treatment-related deaths in either group. The transarterial chemoembolization with sorafenib group did not show additional toxicity compared with the sorafenib monotherapy group.Conclusion and RelevanceIn this study, the combination of sorafenib and transarterial chemoembolization as postoperative adjuvant therapy in patients with hepatocellular carcinoma with portal vein tumor thrombus resulted in longer recurrence-free survival and overall survival than sorafenib alone and was well tolerated.Trial RegistrationClinicalTrials.gov Identifier: NCT04143191

Background & AimsIntrahepatic cholangiocarcinoma (ICC) is the second most common primary liver cancer with high lethality. Clonorchis sinensis (C. sinensis) infection is an important risk factor for ICC. Here we investigated the clinical impact and underlying molecular characteristics of C. sinensis-infected ICC.MethodsWe performed single-cell RNA sequencing, whole exome sequencing, RNA-sequencing, metabolomics and spatial transcriptomics in 251 ICC patients from three medical centers. The alterations of metabolic and immune microenvironment of C. sinensis-infected ICCs were validated through in vitro co-culture system and hydrodynamic injection ICC mouse model.ResultsWe revealed that C. sinensis infection was significantly associated with ICC patients’ overall survival and immunotherapy response. Fatty acid biosynthesis and the expression of FASN, a key enzyme catalyzing long-chain fatty acid synthesis, were significantly enriched in C. sinensis-infected ICCs. ICC cell lines treated with C. sinensis-produced excretory/secretory products (ESPs) displayed an elevation of FASN and free fatty acid. The metabolic alteration of tumor cells was closely correlated with the enrichment of tumor-associated macrophage-like (TAM-like) macrophages and the impairment function of T cells, which led to the immunosuppressive microenvironment formation and tumor progression. Spatial transcriptomics analysis revealed that malignant cells were in closer juxtaposition with TAM-like macrophages in C. sinensis-infected ICCs than non-C. sinensis-infected ICCs. Importantly, FASN inhibitor significantly reversed immunosuppressive microenvironment and enhanced anti-PD-1 efficacy in ICC mouse models treated with ESPs from C. sinensis.ConclusionsWe uncover the metabolic signature and immune microenvironment of C. sinensis-infected ICCs and highlight the combination of FASN inhibitors with immunotherapy as a promising strategy for treating C. sinensis-infected ICCs.Impact and implicationsC. sinensis-infected ICC patients have a poorer prognosis and worse response to immunotherapy than non-C. sinensis-infected ICCs. The underlying molecular characteristics of C. sinensis-infected ICCs remains unclear. Herein, we demonstrate that up-regulation of FASN and free fatty acids in C. sinensis-infected ICCs leads to immunosuppressive microenvironment formation and tumor progression. Thus, administration of FASN inhibitors could significantly reverse immunosuppressive environment and further enhance anti-PD-1 efficacy in combating C. sinensis-infected ICCs.

Intrahepatic cholangiocarcinoma (ICC) is a deadly cancer with rapid tumor progression. While hyperactive mRNA translation caused by mis-regulated mRNA or tRNA modifications promotes ICC development, the role of rRNA modifications remains elusive. Here, we found that 18S rRNA m(6)A modification and its methyltransferase METTL5 were aberrantly upregulated in ICC and associated with poorer survival (log rank test, p < 0.05). We further revealed the critical role of METTL5-mediated 18S rRNA m(6)A modification in regulation of ICC cell growth and metastasis using loss- and gain-of function assays in vitro and in vivo. The oncogenic function of METTL5 is corroborated using liver-specific knockout and overexpression ICC mouse models. Mechanistically, METTL5 depletion impairs 18S rRNA m(6)A modification that hampers ribosome synthesis and inhibits translation of G-quadruplex-containing mRNAs that are enriched in the transforming growth factor (TGF)-beta pathway. Our study uncovers the important role of METTL5-mediated 18S rRNA m(6)A modification in ICC and unravels the mechanism of rRNA m(6)A modification-mediated oncogenic mRNA translation control.

Objective Revealing the single-cell immune ecosystems in true versus de novo hepatocellular carcinoma (HCC) recurrences could help the optimal development of immunotherapies. Design We performed 5’and VDJ single-cell RNA-sequencing on 34 samples from 20 recurrent HCC patients. Bulk RNA-sequencing, flow cytometry, multiplexed immunofluorescence, and in vitro functional analyses were performed on samples from two validation cohorts. Results Analyses of mutational profiles and evolutionary trajectories in paired primary and recurrent HCC samples using whole-exome sequencing identified de novo versus true recurrences, some of which occurred before clinical diagnosis. The tumour immune microenvironment (TIME) of truly recurrent HCCs was characterised by an increased abundance in KLRB1+CD8+ T cells with memory phenotype and low cytotoxicity. In contrast, we found an enrichment in cytotoxic and exhausted CD8+ T cells in the TIME of de novo recurrent HCCs. Transcriptomic and interaction analyses showed elevated GDF15 expression on HCC cells in proximity to dendritic cells, which may have dampened antigen presentation and inhibited antitumour immunity in truly recurrent lesions. In contrast, myeloid cells’ cross talk with T cells-mediated T cell exhaustion and immunosuppression in the TIME of de novo recurrent HCCs. Consistent with these findings, a phase 2 trial of neoadjuvant anti-PD-1 immunotherapy showed more responses in de novo recurrent HCC patients. Conclusion True and de novo HCC recurrences occur early, have distinct TIME and may require different immunotherapy strategies. Our study provides a source for genomic diagnosis and immune profiling for guiding immunotherapy based on the type of HCC recurrence and the specific TIME.

Background The evidence of radiofrequency ablation (RFA) following transarterial chemoembolisation (TACE) combined with sorafenib for intermediate-stage recurrent hepatocellular carcinoma (RHCC) is limited. Patient responses to this treatment vary because of the heterogeneous nature of RHCC, making it important to identify patients who are most likely to benefit from this combination therapy. The aim of this study was to evaluate the efficacy of RFA following TACE and sorafenib for the intermediate-stage RHCC.Methods This retrospective, multicentre, cohort study included 363 patients with intermediate-stage RHCC underwent TACE combined with sorafenib (TACE-sorafenib group) or RFA following TACE and sorafenib (TACE-sorafenib + RFA group) between January 01, 2009 to December 31, 2015 from four institutions in China. Overall survival (OS), progression-free survival (PFS) and efficacy of patients were compared between the two groups by propensity score-matching (PSM).Findings The 1-, 3-, and 5-year OS rates were 97.7%, 83.7%, 54.7% in TACE-sorafenib + RFA group, and 93.3%, 57.0%, 32.7% in TACE-sorafenib group. The 1-, 2-, and 3-year PFS rates were 85.3%, 58.0%, 26.9% in TACE-sorafenib + RFA group, and 55.3%, 30.7%, 15.3% in TACE-sorafenib group. Compared with the TACE-sorafenib group, the TACE-sorafenib + RFA group had significantly longer OS (HR, 0.54; 95%CI, 0.40-0.73; P < 0.001) and PFS (HR, 0.52; 95% CI, 0.41-0.66; P < 0.001). Subgroup analysis was conducted to precisely screen out the beneficial population from RFA treatment.Interpretation Our findings suggest that addition of RFA following TACE and sorafenib combination was superior to TACE combined with sorafenib for intermediate-stage RHCC, resulting in longer OS and PFS. Patients who had good response to TACE and achieved downstaging successfully could not benefit from the RFA therapy.Funding This research was funded by National Natural Science Foundation of China (No. 81627803), Chen Xiao-Ping Science and Technology Development Fund (CXPJJH1200009-06).Copyright (c) 2022 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Purpose:Reports showed that some of intrahepatic cholangiocarcinoma (ICC) patients with lymph node metastasis (LNM) may also gain survival benefit undergone resection. However, the effect of the extent of LNM on prognosis and surgical indication is barely discussed.Methods:From September 1994 to November 2018, primary ICC patients undergone initial curable surgery were enrolled. Based on the extent of LNM, we divided these patients into 4 groups, including patients with no LNM (group N0), LNM to hepatoduodenal ligament or common hepatic artery (region A, group A), LNM to gastrohepatic lymph nodes for left liver ICC and periduodenal and peripancreatic lymph node for right liver ICC (region B, group B), or LNM beyond these regions (region C, group C). Multivariable Cox regression analysis was performed to identify the prognostic factors for recurrence-free survival (RFS) and overall survival (OS) in all groups.Results:A total of 133 patients were enrolled. There were 56, 21, 17, and 39 patients in groups N0, A, B, and C, respectively. There was significant difference between groups N0 and C in RFS (P < 0.001) and OS (P = 0.002). When we compared group N0 + A + B with group C, we also found that RFS (P < 0.001) and OS (P = 0.007) were significantly different. In multivariable analysis, the extent of LNM was an independent risk factor for RFS (P < 0.050).Conclusion:ICC patients with the LNM to regions A and B could still achieve good prognosis with resection. Surgery should be carefully considered when LNM to region C.

UNLABELLED The objective of this study was to test the hypothesis that the integrity of the large bowel wall in AIDS patients is compromised in a manner that favours the chronic translocation of bacteria and/or products of bacterial metabolism into the bloodstream. When such translocation occurs, it induces a characteristic stress/inflammatory response in the body. Urinary butyrate, a unique product of colonic microbial metabolism, was used to assess gut wall permeability. Excretion of the pro-inflammatory cytokine IL-6 in the urine was used as a marker for the stress/inflammatory response. Four groups of subjects were studied, controls (n = 12), HIV + (n = 35) and AIDS patients with (n = 14) and without (n = 17) weight loss. RESULTS measurable amounts of interleukin 6 (IL-6) and butyrate were found in the urine of all subjects. There were no significant differences in IL-6 excretion between the controls (0.68 +/- 0.64 pg/ml), asymptomatic HIV + subjects (0.59 +/- 0.37 pg/ml) and AIDS patients without weight loss (1.18 +/- 0.33 pg/ml) but IL-6 levels were significantly higher in the AIDS group with weight loss (4.02 +/- 1.26 pg/ml, P < 0.05). A similar pattern of results was found with interleukin 1 receptor antagonist (IL-1ra). Like IL-6 and (IL-1ra), urinary butyrate levels were increased in the AIDS patients with weight loss (2.83 +/- 0.67 mumol/l) relative to the controls (1.31 +/- 0.13 mumol/l, P < 0.05), with the HIV + patients (1.65 +/- 0.18 mumol/l) and AIDS patients without weight loss (1.90 +/- 0.22 mumol/l) falling in between. The data are consistent with a low, but chronic rate of bacteria and/or bacterial products seeping across a compromised colonic wall causing a chronic low stress response in AIDS patients.