Efficacy of hepatic arterial infusion chemotherapy and its multimodality therapeutic regimens in treatment of patients with advanced hepatocellular carcinoma and related prognostic factors

ObjectiveTo investigate the efficacy of continuous hepatic arterial infusion chemotherapy (HAIC) with the FOLFOX regimen and its multimodality therapeutic regimen in the treatment of patients with advanced hepatocellular carcinoma, as well as the influencing factors for prognosis. MethodsA retrospective analysis was performed for the clinical data of 66 patients with advanced hepatocellular carcinoma who received continuous HAIC with FOLFOX regimen in Nanfang Hospital, Southern Medical University, from September 2018 to November 2021. The patients were observed in terms of objective response rate (ORR), disease control rate (DCR), median progression-free survival (mPFS), and median overall survival (mOS) after treatment, and treatment-related adverse reactions were recorded. For the patients with portal vein tumor thrombus, the effect of the treatment on portal vein tumor thrombus was assessed. The Kaplan-Meier method was used for survival analysis, and the Cox regression analysis was used to investigate the influencing factors for prognosis. ResultsAccording to the RECIST1.1 criteria, FOLFOX-HAIC and its multimodality therapeutic regimen achieved an ORR of 33.3% (22/66) and a DCR of 86.4% (57/66) in the treatment of 66 patients with advanced hepatocellular carcinoma, with an mPFS time of 8.2 months and an mOS time of 22.1 months. Among the 39 patients with portal vein tumor thrombus, 2 achieved complete remission, 8 achieved partial remission, 24 achieved stable disease, and 5 had disease progression, with an ORR of 25.6% (10/39) and a DCR of 87.2% (34/39). The main adverse reactions included gastrointestinal reactions (167%, 11/66), pyrexia (12.1%, 8/66), liver area pain (10.6%, 7/66), bone marrow suppression (3.0%, 2/66), and contrast agent allergy (3.0%, 2/66), and there were no grade ＞Ⅳ toxic or side effects or deaths caused by such complications. The Cox regression analysis showed that extrahepatic metastasis (hazard ratio ［HR］=2.668, 95% confidence interval ［CI］: 1.357-5.245, P＜0.05) and prothrombin time (PT) (HR=1.282, 95%CI: 1.080-1.630, P＜0.05) were independent risk factors for PFS, and aspartate aminotransferase level (HR=1.008, 95%CI: 1.002-1.013, P＜005) and PT (HR=1.303, 95%CI: 1.046-1.630, P＜0.05) were independent risk factors for OS. ConclusionFOLFOX-HAIC and its multimodality therapeutic regimen has a certain clinical effect with controllable adverse reactions in the treatment of advanced hepatocellular carcinoma

Establishment of a zebrafish model of alcohol-induced acute hepatic steatosis

ObjectiveTo establish a zebrafish model of hepatic steatosis induced by acute alcohol exposure as a model system that will be useful for investigations of disease pathogenesis and high-throughput screenings. MethodsAt 5 days post fertilization (dpf), 288 juvenile zebrafish (wild-type strain AB) with normal liver development were selected and randomly divided into four groups (n=72 each) for culturing in water and Hank′s balanced salt solution alone (controls) or supplemented with ethanol at 2.0%, 2.5% and 3.0% concentrations. At 4, 24 and 32 h of culturing, the physical activity (swimming, circling) was observed, the survival rate was recorded, and the morphology was assessed by whole-mount and liver section microscopy. Oil Red O staining was used to determine the percent of steatosis and pathological features were assessed by hematoxylin-eosin staining. Differences between two groups were assessed by independent samples t-test. ResultsStarting at 24 h of ethanol exposure, the survival rates showed a decreasing trend that corresponded to increased alcohol concentration, so that by 32 h of exposure there were remarkable differences among the three alcohol-treated groups: 2.0% ethanol: 100% survival; 2.5% ethanol: (91.21±1.61)% survival; and 3.0% ethanol: 0% survival. Abnormalities in liver morphology and physical activity were also observed at the 24 h time point and showed concentration-dependence: 2.0% ethanol: normal morphology, increased activity, abnormal circling; 2.5% ethanol: some deformities; and 3.0% ethanol: bent spinal cord, pericardial and yolk sac edema, stationary behavior. Significant steatosis was observed in the 2.0% ethanol group at 32 h of exposure, but not at 24 h, as evidenced by the percent of steatosis［(71.25±0.15)% vs. controls: (31.25±0.05)%, P=0.002］ and pathological features (swollen size, lipid droplet-induced nucleus displacement). ConclusionA model of acute alcohol-induced hepatic steatosis was successfully established in 5 dpf zebrafish by treating with 2.0% alcohol for 32 h. This model may represent a useful tool for investigating the disease pathogenesis and performing high-throughput screenings of potentially therapeutic drugs

ATR inhibitor AZD6738 enhances the antitumor activity of radiotherapy and immune checkpoint inhibitors by potentiating the tumor immune microenvironment in hepatocellular carcinoma

Background Radioimmunotherapy has a promising antitumor effect in hepatocellular carcinoma (HCC), depending on the regulatory effect of radiotherapy on tumor immune microenvironment. Ionizing radiation (IR)-induced DNA damage repair (DDR) pathway activation leads to the inhibition of immune microenvironment, thus impairing the antitumor effect of radioimmunotherapy. However, it is unclear whether inhibition of the DDR pathway can enhance the effect of radioimmunotherapy. In this study, we aim to explore the role of DDR inhibitor AZD6738 on the combination of radiotherapy and immune checkpoint inhibitors (ICIs) in HCC.Methods C57BL/6 mouse subcutaneous tumor model was used to evaluate the ability of different treatment regimens in tumor growth control and tumor recurrence inhibition. Effects of each treatment regimen on the alterations of immunophenotypes including the quantification, activation, proliferating ability, exhaustion marker expression, and memory status were assessed by flow cytometry.Results AZD6738 further increased radiotherapy-stimulated CD8+ T cell infiltration and activation and reverted the immunosuppressive effect of radiation on the number of Tregs in mice xenografts. Moreover, compared with radioimmunotherapy (radiotherapy plus anti-PD-L1 (Programmed death ligand 1)), the addition of AZD6738 boosted the infiltration, increased cell proliferation, enhanced interferon (IFN)-γ production ability of TIL (tumor-infiltrating lymphocyte) CD8+ T cells, and caused a decreasing trend in the number of TIL Tregs and exhausted T cells in mice xenografts. Thus, the tumor immune microenvironment was significantly improved. Meanwhile, triple therapy (AZD6738 plus radiotherapy plus anti-PD-L1) also induced a better immunophenotype than radioimmunotherapy in mice spleens. As a consequence, triple therapy displayed greater benefit in antitumor efficacy and mice survival than radioimmunotherapy. Mechanism study revealed that the synergistic antitumor effect of AZD6738 with radioimmunotherapy relied on the activation of cyclic GMP–AMP synthase /stimulator of interferon genes (cGAS/STING) signaling pathway. Furthermore, triple therapy led to stronger immunologic memory and lasting antitumor immunity than radioimmunotherapy, thus preventing tumor recurrence in mouse models.Conclusions Our findings indicate that AZD6738 might be a potential synergistic treatment for radioimmunotherapy to control the proliferation of HCC cells, prolong survival, and prevent tumor recurrence in patients with HCC by improving the immune microenvironment

Additional file 1: Figure S1. of High fat plus high cholesterol diet lead to hepatic steatosis in zebrafish larvae: a novel model for screening anti-hepatic steatosis drugs

HF and HFC diets lead to hepatic steatosis in zebrafish larvae. Figure S2. Lipid accumulation in the livers and blood vessels of zebrafish larvae fed with HF and HFC diets. Figure S3. Genes changes in the livers of HF and HFC diets-fed zebrafish larvae. Table S1. Primer sequences used for quantitative RT-PCR. (DOCX 2678 kb