Purpose This study was conducted to assess the clinical significance of programmed cell death-ligand 1 (PD-L1)-positive circulating tumor cells (CTCs) as predictive biomarkers for the efficacy of PD-(L)1 inhibitor-based treatment in advanced hepatocellular carcinoma (HCC).Experimental design We enrolled 59 patients with unresectable HCC who received immunotherapy-based treatment and analyzed CTCs, PD-L1+CTCs and molecules in peripheral blood. An innovative telomerase reverse transcriptase-based method was applied to detect specific CTCs. Kaplan-Meier analysis and Cox regression were used to evaluate clinical outcomes, such as progression-free survival (PFS) and overall survival (OS).Results CTCs were detected in 86.4% (51/59) of patients, with a PD-L1-positive rate of 83.7% (41/49). Compared with the “PD-L1+CTC Low” group, the “PD-L1+CTC High” group had significantly shorter PFS (median PFS: 7.72 vs 16.1 months, p=0.037) and shorter OS (median OS: 13.89 vs 36.97 months, p=0.031). The “PD-L1+CTC Fewer & Low” group had the longest survival outcomes (median PFS: 17.03 months, median OS: 36.97 months), whereas the “PD-L1+CTC More & High” group had the poorest outcomes (median PFS: 5.3 months, median OS: 10.77 months) (p<0.05). Multivariate analysis revealed that the PD-L1+CTC count and ratio were an independent predictor of PFS. Significant correlations were found between PD-L1+CTC and several immune-related molecules, such as immune checkpoint molecules (CD28, TIM-3, and CD80) and regulatory factors (BDNF, and BLTA).Conclusions PD-L1+CTCs are potential indicators correlating with the shorter PFS and OS of immunotherapy-based treatment in patients with advanced HCC. The “PD-L1+CTC Low & Fewer” classification was associated with better clinical outcomes and immune-related molecules.

Hepatocellular carcinoma (HCC) was characterized by a highly complex genome, with structural variations (SVs) playing a significant role in its development. In this study, we employed Oxford Nanopore Technology long-read sequencing in paired tumor and adjacent normal liver tissues from 74 Chinese HCC patients to thoroughly characterize the landscape of somatic SVs. Our analysis revealed that somatic SVs were more prevalent in hepatitis B virus (HBV)-related HCC, with chromosome 1 emerging as a major hotspot, and several members of the chromosome 1 open reading frame (C1orf) family genes expression level exhibited significant age-related difference. Notably, HBV-related HCC cases exhibited a higher frequency of deletions, particularly among younger ones (≤ 35 years old). In addition, we observed an increased burden of HBV integration events in younger ones. Remarkably, the divergent-paired related homeobox ( DPRX ) loci was identified as a novel gene for HBV integration in younger patients. Together, these findings delineated the somatic SV landscape in HCC and underscored age-associated HBV-related genomic alterations as key pathological features of hepatocarcinogenesis.

This study aimed to investigate the impact of hepatic segment distribution on the prognosis of patients with primary hepatocellular carcinoma (HCC). A retrospective analysis was conducted on 456 eligible patients with primary HCC admitted to the Hepatobiliary Surgery Department of Cancer Hospital, Chinese Academy of Medical Sciences, from January 2013 to December 2017. Tumor hepatic segment location was confirmed by postoperative pathology or imaging examinations, and clinical data (e.g. age, surgical method, operation time) were collected from the HIS electronic medical record system. Follow-up ended on May 5, 2019, with overall survival (OS) as the primary endpoint. Results showed that most tumors (70.39%, 321/456) were located in the right hemi-liver, with the highest incidence in Segment VI (53.73%, 245/456) and Segment VII (46.05%, 210/456). Univariate analysis identified tumor diameter, tumor characteristics, surgical method, operation time, intraoperative blood loss, and hepatic segment location as factors associated with OS. Multivariate COX analysis revealed that tumor diameter, open surgery, and intraoperative blood loss were independent predictors of poor long-term prognosis. Notably, hepatic segment location exerted contrasting effects: Segment VI tumors independently associated with poorer prognosis (HR=2.04, 95% CI=1.11-3.74, p=0.021), while Segment VII tumors served as a significant survival benefit (HR=0.47, 95% CI=0.25-0.88, p=0.018). Further subgroup analysis indicated that the protective effect of Segment VII was more pronounced in tumors ≥6 cm (p=0.03), whereas for tumors <6 cm, Segment VII involvement had no significant prognostic impact (p=0.32). For Segment VI tumors, laparotomy was associated with poorer prognosis regardless of tumor size (<6 cm: p=0.029; ≥6 cm: p=0.039), a trend not observed in non-Segment VI tumors ≥6 cm (p=0.9). Hepatic segment distribution is an independent prognostic indicator and may serve as a surrogate marker reflecting tumor biology, surgical complexity, and treatment selection, rather than an independent causal determinant, with Segment VI and VII showing distinct prognostic roles: Segment VI involvement is associated with worse survival, while Segment VII involvement confers a survival benefit. These findings provide valuable insights for precision surgical decision-making in HCC, such as prioritizing minimally invasive approaches or neoadjuvant therapy for Segment VI tumors and considering de-escalated treatment for large Segment VII tumors. Future studies should explore the molecular and immune mechanisms underlying these segment-specific differences and validate the results through multicenter prospective trials.

BackgroundThis study systematically reviews the efficacy and safety of the single or combined use of programmed factor 1 (PD-1)/programmed factor 1 ligand (PD-L1) inhibitors for treating metastatic or advanced renal cell carcinoma (RCC).MethodsRelevant articles were collected for meta-analysis through searches on PubMed, Web of Science, Embase, Cochrane Library, and Clinical Trials, as well as for relevant randomized controlled experiments.ResultsBased on eleven studies, the effectiveness of the experimental group was found to be significantly better than the control in terms of overall survival (OS) [R=0.74, 95%CI: 0.69~0.80, P<0.00001], progression-free survival (PFS) [HR=0.68, 95%CI: 0.57~0.81, P<0.0001], objective response rate (ORR) [RR=1.71, 95%CI: 1.39~2.12, P<0.00001], complete response rate (CR) [RR=2.99 95%CI: 2.34~3.83, P<0.0001], partial response rate (PR) [RR=1.56, 95%CI: 1.20~2.01, P=0.001], and disease control rate (DCR) [RR=1.13, 95%CI: 1.06~1.20, P<0.0001]. No statistical significance was observed between the experimental and control groups in overall adverse reactions (AEs) [RR=1.01, 95%CI: 0.98~1.04, P=0.598], the incidence of stage I~II adverse reactions [RR=1.02, 95%CI: 0.88~1.17, P=0.818], or stage III~V adverse reactions [RR=0.98, 95%CI: 0.81~1.18, P=0.817]. Regarding subgroup analysis, the incidence of dysphonia, rash, hypothyroidism, arthralgia, and pruritus in the experimental group was significantly higher than in the control. Compared with the control group, the incidence of diarrhea, nausea, indigestion, and fatigue in the experimental group was not statistically significant.ConclusionA good efficacy was found in treating metastatic or advanced RCC using PD-1/PD-L1 inhibitors alone or in combination, which significantly improved and enhanced OS, PFS, ORR, CR, PR, and DCR in patients with RCC. The incidence of adverse reactions in patients was not increased, and adverse reactions were controllable. These findings indicate that the single or combined use of PD-1/PD-L1 inhibitors shows good efficacy and safety in the treatment of metastatic or advanced RCC.

Tumor fibrosis is recognized as a malignant hallmark in various solid tumors; however, the clinical importance and associated molecular characteristics of tumor fibrosis in liver metastases (LM) from colorectal cancer (CRLM) remain poorly understood. Here we show that patients with CRLM whose liver metastases (LM) exhibited tumor fibrosis (Fibrosis+ LM) had significantly worse progression-free survival (P = 0.025) and overall survival (P = 0.008). Single-cell RNA sequencing revealed that the tumor microenvironment of the Fibrosis+ LM was characterized by T cells with an exhausted phenotype, macrophages displaying a profibrotic and suppressive phenotype and fibrosis-promoting fibroblasts. Further investigation highlighted the pivotal role of VCAN_eCAF in remodeling the tumor fibrosis in the tumor microenvironment of Fibrosis+ LM, emphasizing potential targetable interactions such as FGF23 or FGF3-FGFR1. Validation through multiplex immunohistochemistry/immunofluorescence and spatial transcriptomics supported these findings. Here we present a comprehensive single-cell atlas of tumor fibrosis in LM, revealing the intricate multicellular environment and molecular features associated with it. These insights deepen our understanding of tumor fibrosis mechanisms and inform improved clinical diagnosis and treatment strategies.

BACKGROUND:Patients with synchronous colorectal cancer liver metastasis (CRLM) often face sub-optimal outcomes from systemic therapy or resection. This study investigates the prognostic value of the pre-treatment S-index, a reliable non-invasive marker for liver fibrosis, for outcomes in synchronous CRLM patients. METHODS:This study included two populations of patients with synchronous CRLM: one population undergoing resection and another population receiving systemic therapy for unresectable CRLM. Pre-treatment S-index levels were assessed from blood samples. Patients were categorized into high and low S-index groups, and comparisons were made regarding outcomes: progression-free survival (PFS), early post-operative recurrence and fibrosis in liver metastases in the resection population, and treatment response in the systemic therapy population. Multiplex immunohistochemistry/immunofluorescence (mIHC/IF) was used to investigate the distribution of immunosuppressive T-cell subsets in liver metastases. RESULTS:For synchronous CRLM patients receiving resection (n = 1000), patients with high preoperative S-index demonstrated significantly worse PFS both before (HR = 1.556, 95 % CI: 1.255-1.929; P < 0.001) and after IPTW-adjusted Cox proportional hazards regression analysis (IPTW-adjusted HR = 1.439, 95 % CI: 1.094-1.894; P = 0.036). High S-index patients also exhibited an elevated risk of early recurrence, both before and after adjustment (OR = 1.556, 95 % CI: 1.255-1.929, P < 0.001; IPTW-adjusted OR = 1.439, 95 % CI: 1.094-1.894, P = 0.009). For synchronous CRLM patients receiving system therapy (n = 123), a high pre-treatment S-index (OR = 34.691, P = 0.005) was a significant predictor of progression disease in multivariate analyses. Further, the S-index showed an AUC of 0.814 (95 %CI: 0.762-0.866, P < 0.001) for detecting fibrosis in liver metastases, with a specificity of 0.928. mIHC/IF analysis revealed that inhibitory T cells, especially CD4+PD1+ T cells and CD4+FOXP3+ T cells, were significantly elevated in the liver metastases of the high S-index group. CONCLUSION:This study contributed valuable evidence regarding pre-treatment S-index for association with outcomes among synchronous CRLM patients receiving resection or system therapy. Furthermore, it underscores a significant association between a high S-index and the presence of fibrosis in liver metastases, as well as more infiltration of immunosuppressive T cells in the tumor.

Hepatocellular carcinoma (HCC) is a major cause of cancer-related mortality, it ranks as the second most common cause of cancer deaths in China. Most HCC patients are first diagnosed at an advanced stage. In recent years, targeted therapy combined with immunotherapy has become the preferred regimen for systemic treatment of intermediate-advanced HCC, while targeted therapy combined with immunotherapy plus local treatment could further improve the efficacy in many clinical studies. To better guide the clinical treatment for effective and safe combination therapy, our interdisciplinary panel on the treatment of intermediate-advanced HCC comprising hepatologists, hepatobiliary surgeons, oncologists, radiologists, interventional radiologists and traditional Chinese medicine physicians have formulated this consensus based on current clinical studies and clinical medication experience for reference. The consensus contained 15 recommendations, including the applicable population and management, local treatment selection, conversion strategy, treatment strategy after tumor progression and management of common adverse reactions.

OBJECTIVE:Hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), mostly characterised by HBV integrations, is prevalent worldwide. Previous HBV studies mainly focused on a few hotspot integrations. However, the oncogenic role of the other HBV integrations remains unclear. This study aimed to elucidate HBV integration-induced tumourigenesis further. DESIGN:Here, we illuminated the genomic structures encompassing HBV integrations in 124 HCCs across ages using whole genome sequencing and Nanopore long reads. We classified a repertoire of integration patterns featured by complex genomic rearrangement. We also conducted a clustered regularly interspaced short palindromic repeat (CRISPR)-based gain-of-function genetic screen in mouse hepatocytes. We individually activated each candidate gene in the mouse model to uncover HBV integration-mediated oncogenic aberration that elicits tumourigenesis in mice. RESULTS:These HBV-mediated rearrangements are significantly enriched in a bridge-fusion-bridge pattern and interchromosomal translocations, and frequently led to a wide range of aberrations including driver copy number variations in chr 4q, 5p (TERT), 6q, 8p, 16q, 9p (CDKN2A/B), 17p (TP53) and 13q (RB1), and particularly, ultra-early amplifications in chr8q. Integrated HBV frequently contains complex structures correlated with the translocation distance. Paired breakpoints within each integration event usually exhibit different microhomology, likely mediated by different DNA repair mechanisms. HBV-mediated rearrangements significantly correlated with young age, higher HBV DNA level and TP53 mutations but were less prevalent in the patients subjected to prior antiviral therapies. Finally, we recapitulated the TONSL and TMEM65 amplification in chr8q led by HBV integration using CRISPR/Cas9 editing and demonstrated their tumourigenic potentials. CONCLUSION:HBV integrations extensively reshape genomic structures and promote hepatocarcinogenesis (graphical abstract), which may occur early in a patient's life.

Background: Scarce evidence exists for clinical target volume (CTV) definitions of regional lymph nodes (LNs) in intrahepatic cholangiocarcinoma (iCCA) or combined hepatocellular-cholangiocarcinoma (cHCC-CCA). We investigated the mapping pattern of nodal recurrence after surgery for iCCA and cHCC-CCA and provided evidence for the nodal CTV definition. Methods: We retrospectively reviewed the medical records of patients with iCCA or cHCC-CCA who underwent surgery between 2010 and 2020. Eligibility criteria included patients pathologically diagnosed with iCCA or cHCC-CCA after surgery and a first recurrent event in regional LNs during follow-up. All recurrent LNs were registered onto reference computed tomography images based on the vascular structures to reconstruct the node mapping. Fifty-three patients were eligible. LN regions were classified into four risk groups. Results: Hepatic hilar and portal vein-vena cava were the most common recurrent regions, with recurrence rates of 62.3 % and 39.6 % (high-risk regions), respectively. Recurrence rates in the left gastric, diaphragmatic, common hepatic, superior mesenteric vessels, celiac trunk, and paracardial regions ranged from 15.1 % to 30.2 % (intermediate-risk regions). There were fewer recurrences in the para-aortic (16a1, a2, b1) and splenic artery and hilum regions, with rates <10 % (low-risk regions). No LN recurrence was observed in the para-oesophageal or para-aortic region (16b2) (very low-risk regions). Based on node mapping, the CTV should include high- and intermediate-risk regions for pathologically negative LN patients during postoperative radiotherapy. Low-risk regions should be included for pathologically positive LN patients. Conclusion: We provide evidence for CTV delineation in patients with iCCA and cHCC-CCA based on recurrent LN mapping.

BackgroundTo predict clinical important outcomes for colorectal liver metastases (CRLM) patients receiving colorectal resection with simultaneous liver resection by integrating demographic, clinical, laboratory, and genetic data.MethodsRandom forest (RF) models were developed to predict postoperative complications and major complications (binary outcomes), as well as progression-free survival (PFS) and overall survival (OS) (time-to-event outcomes) of the CRLM patients based on data from two hospitals. The models were validated on an external dataset from an independent hospital. The clinical utility of the models was assessed via decision curve analyses (DCA).ResultsThere were 1067 patients included in survival prediction analyses and 1070 patients included in postoperative complication prediction analyses. The RF models provided an assessment of the model contributions of features for outcomes and suggested KRAS, BRAF, and MMR status were salient for the PFS or OS predictions. RF model of PFS showed that the Brier scores at 1-, 3-, and 5-year PFS were 0.213, 0.202 and 0.188; and the AUCs of 1-, 3- and 5-year PFS were 0.702, 0.720 and 0.743. RF model of OS revealed that Brier scores of 1-,3-, and 5-year OS were 0.040, 0.183 and 0.211; and the AUCs of 1-, 3- and 5-year OS were 0.737, 0.706 and 0.719. RF model for postoperative complication resulted in an AUC of 0.716 and a Brier score of 0.196. DCA curves clearly demonstrated that the RF models for these outcomes exhibited a superior net benefit across a wide range of threshold probabilities, signifying their favorable clinical utility. The RF models consistently exhibited robust performance in both internal cross-validation and external validation. The individualized risk profile predicted by the models closely aligned with the actual survival outcomes observed for the patients. A web-based tool (https://kanli.shinyapps.io/CRLMRF/) was provided to demonstrate the practical use of the prediction models for new patients in the clinical setting.ConclusionThe predictive models and a web-based tool for personalized prediction demonstrated a moderate predictive performance and favorable clinical utilities on several key clinical outcomes of CRLM patients receiving simultaneous resection, which could facilitate the clinical decision-making and inform future interventions for CRLM patients.

Background: Literature is lacking on the impact of advancements in minimally invasive surgery (MIS) on outcomes for patients with gastroenteropancreatic neuroendocrine carcinomas (GEP-NECs). Herein, we compared perioperative and oncologic outcomes among patients with GEP-NECs undergoing open, laparoscopic, and robotic resection.Methods: Patients with GEP-NECs diagnosed 2010-2019 were identified from the National Cancer Database (NCDB). We used the inverse probability of treatment weighting method to account for selection bias. Patients were stratified by surgical approach; and pairwise comparisons were conducted by analyzing short- and long-term outcomes.Results: Receipt of MIS increased from 34.2% in 2010 to 67.5 % in 2019. Altogether, 6560 patients met and 333 (5.1%) underwent robotic resection. Compared with open resection, laparoscopic or robotic resection were associated with shorter post-operative length of stay, reduced 30-day and 90-day postoperative mortality, and prolonged overall survival (OS). Compared with laparoscopic resection, robotic resection was associated with reduced 90-day post-operative mortality, however, there was no significant difference in OS.Conclusion: This NCDB analysis demonstrates that MIS approaches for treating GEP-NECs have become more common, with improved perioperative mortality, shorter post-operative length of stay and favorable OS, compared with open resection.

This study aimed to investigate the prognostic value of the preoperative alkaline phosphatase-to-albumin ratio (APAR) in patients with hepatocellular carcinoma (HCC) who underwent radical hepatectomy. The clinicopathological data from 330 patients was retrospectively analyzed. Receiver operating characteristic curves of APAR for diagnostic tumor recurrence were plotted with a cut-off value of 1.74. A high preoperative APAR value was significantly associated with hepatitis B surface antigen level, tumor diameter, and tumor-node-metastasis stage. The disease-free survival (DFS) and overall survival (OS) of patients with a high preoperative APAR were shorter than those with a low APAR. The independent risk factors for DFS were an APAR ≥1.74, and macrovascular invasion or tumor thrombus. The independent risk factors for OS were an APAR ≥1.74, existing clinical symptoms, α-fetoprotein level ≥20 ng/ml, macrovascular invasion or tumor thrombus, and family history of cancer. In conclusion, a preoperative APAR (≥1.74) is an independent risk factor influencing the poor prognosis of patients with HCC after curative hepatectomy, and patients with such a result should be closely monitored.

Table S1 shows the patients information in CRC.

Figure S6. siRNA-mediated RIPK1 or MCU Silencing Inhibites the OCR. (A, B) Oxygen consumption rate of HT29 transfected with si-RIPK1 (200 pM) and si-MCU (200 pM, n=5). *p Ë, 0.05, **p Ë, 0.01, control vs treatment.

Figure S2. Nuclear localization of NF-kB is Increased in CRC (A) Representative photomicrographs of immunostaining for NF-κB and phosphorylated NF-κB (P65 and p-P65) in normal and CRC tissues. Scale bar = 50μm.

Background:Although immunotherapy combined with targeted therapy can be effective for hepatocellular carcinoma (HCC), not all HCC patients respond to this treatment. Models for predicting tumour response in HCC patients receiving immunotherapy combined with targeted therapy are lacking.Methods:A total of 221 HCC patients from two independent prospective cohorts were retrospectively reviewed. The patients were randomly divided into training and validation cohorts at a ratio of 7:3. Standard clinical data were collected from each patient, including age, sex, hepatitis B infection status, laboratory tests, and immune target-related adverse events (itrAEs). Tumour responses were evaluated using the Response Evaluation Criteria in Solid Tumours (RECIST) v1.1 guidelines. ItrAEs were assessed based on the Common Terminology Criteria for Adverse Events version 4.0. The nomogram for tumour response prediction was constructed based on the results of the multivariate logistic regression analysis, areas under the receiver operating characteristic curves (AUROCs) were used to determine the sensitivity and specificity of the model, and calibration plots and Hosmer-Lemeshow chi-square tests were performed to assess the calibration of the model.Results:In the multivariate logistic regression analysis, a solitary tumour (P=0.006), neutropenia (P=0.003) and hypertension (P=0.042) independently predicted objective response (OR). A nomogram for OR was established with AUROCs of 0.734, 0.675, 0.730, and 0.707 in the training, validation, first-line and second-line treatment sets, respectively. Tumour sizes less than 5 cm (P=0.005), a solitary tumour (P=0.037), prognostic nutritional indices greater than or equal to 54.3 (P=0.037), neutropenia (P=0.004) and fatigue (P=0.041) independently predicted disease control (DC). A nomogram for DC was established with AUROCs of 0.804, 0.667, and 0.768 in the training, first-line and second-line treatment sets, respectively. All the Hosmer-Lemeshow tests and calibration curves showed acceptable calibration.Conclusions:The current provides clinicians with new insights into selecting patients for immunotherapy combined with targeted therapy and contributes to the development of immunotherapy for HCC. It is necessary to expand the scale of our research and perform prospective studies to verify our findings.

由于手术切除的难度较大和术后出现肝功能衰竭的风险较高,因此巨大肝细胞癌(hepatocellular carcinoma,HCC)患者的治疗是比较困难的.供体来源较少、费用较高和术后复发率较高等问题导致肝移植术不是常规的治疗方式.其余的治疗方案,例如经肝动脉化疗栓塞(transhepatic arterial chemoembolization,TACE)和射频消融术(radiofrequency ablation,RFA)不能根治肿瘤,所以也不是巨大HCC患者的首选治疗方案.肝切除术是治疗巨大HCC的最佳方式. 本例患者肿瘤巨大,且肿瘤位于位置特殊的肝尾状叶,大大提升了手术难度.而且患者术前实验室检查肝炎指标和肿瘤标志物全部为阴性,联合影像学检查也无法明确诊断,通过多学科诊疗(multi-disciplinary team,MDT)团队专家讨论,选择手术切除治疗.术后病理结果为HCC,随访显示该例患者疗效确切,尚未复发.

肝脏是结直肠癌患者最常见的转移部位.随着系统治疗药物的研发,结直肠癌肝转移(colorectal liver metastases,CRLM)患者生存率显著提高,良好的系统治疗联合局部治疗是目前可以长期控制疾病甚至治愈的治疗策略.热消融或立体定向放射治疗(stereotactic radiotherapy,SRT)作为一种局部治疗方式,在特殊部位的转移病灶、老年患者或有合并症的患者中均可以安全实施.本文基于现有的研究证据对热消融和SRT在CRLM中的临床应用和研究进展进行综述.

Figure S5. MICU1 is Down-regulated in CRC. (A, B) Representative western blots and averaged data for MICU1 from normal and CRC tissue, n = 7. Data are mean {plus minus} SEM. of three independent experiments. *p Ë, 0.05, normal vs CRC.