The Study Of GPC3 Targeted FLI/MPI Dual-modality Imaging In The Differential Diagnosis Of Hepatocellular Carcinoma And Liver Fibrosis

BackgroundMost(80%-90%)hepatocellular carcinomas(HCCs)occur in cirrhotic liver,and the early symptoms are inconspicuous.Traditional imaging techniques can only detect morphological and anatomical lesions of the tissues,and it is difficult to detect early-stage HCC without typical imaging features.Molecular imaging technology can visualize abnormal features at the cellular and molecular levels in the early stage of HCC formation,which is of great value for exploring the evolution of HCC and identifying early-stage HCC.However,the excessive deposition of collagen in liver fibrotic tissues leads to the accumulation of HCC targeted imaging probes in liver fibrotic tissues,resulting in a decrease in the targeted delivery efficiency of HCC,and it is difficult to realize accurate detection of HCC lesions and liver cirrhosis.Highly specific HCC targeted imaging probe,imaging techniques with high sensitivity and good penetration depth,a clinically relevant animal model of HCC coexisting with hepatic fibrosis,and low non-specific accumulation in non-tumor tissue,are major concerns in the differential diagnosis of HCC and liver cirrhosis.In this study,we predegraded hepatic fibrotic collagen by HSA-C to reduce the non-specific retention of imaging probes in hepatic fibrotic tissues,followed by enhanced targeting visualization of HCC with GPC3-targeted FLI/MPI dual-modality imaging probe TSI to enhanced the differential diagnosis of early-stage HCC.ObjectiveTo explore the relationship between the progression of HCC and liver cirrhosis/fibrosis,and to evaluate the correlation between the intrahepatic accumulation of the probe and the degree ofliver fibrosis.Enhanced glypican-3-targeted identification of hepatocellular carcinoma with liver fibrosis by pre-degrading excess fibrotic collagen.MethodsSynthesizing HSA-C for degrading collagen,and synthesizing TSI nanoprobes for targeted imaging of HCC.Used transmission electron microscopy,nanoparticle size and potentiometric analyzers,etc.to detect chemical characterization of nanoparticles.Cell experiments were perfonned to verify the ability of HSA-C to degrade collagen in vitro and to evaluate its cytotoxicity.Cell experiments were performed to verify the specific recognition ability of TJ2 polypeptide to HCC cells and GPC3 protein in vitro,and to evaluate the cytotoxicity of TSI nanoprobes.Different degrees of liver fibrosis or cirrhosis were induced in mice,not only the correlation between the accumulation of non-HCC-targeting probes in the liver and collagen deposition was verified in vivo,but also the ability of HSA-C to degrade liver fibrosis in vivo was verified Animal model of liver fibrosis coexisting with HCC was established to explore the targeting detection accuracy of TSI probe for HCC with or without HSA-C pretreatmentResults1.HSA-C was spherical with an average diameter of 183.7 nm;the average size of TSI nanoprobes was about 20.15± 1.83 with good fluorescence imaging and magnetic particle imaging properties.2.HSA-C could effectively degrade collagen Ⅰ in activated hepatic stellate cells,HSA-C reduced the type Ⅰ collagen of activated LX-2 cells by 61.5%.The TJ2 polypeptide targeting probe specifically targeted HCC cells,and its targeting properties were blocked by anti-GPC3 antibodies.Cytotoxicity experiments proved that HSA-C and TSI nanoprobes had good biological safety.3.Induced different degrees of liver fibrosis or cirrhosis,which proved that the accumulation of imaging probes in liver fibrosis tissue was linearly correlated with the deposition of collagen fibers.HSA-C reduced the content of collagen fibers by 24.7%and the non-specific retention of nanoprobes by 27.2%in the 8-week group of liver fibrosis.4.The-TBRs of TSI HCC-targeted nanoprobes detected by FLI and MPI imaging were 5.43 times and 1.34 times higher than that of non-targeted probes,respectively.5.HSA-C pretreatment could improve the detection accuracy of TSI nanoprobes in HCC coexisting with liver fibrosis mice,TBR was 2.61 times that of TSI alone group,and the TBR was 2.61 times higher than that of TSI alone,and could distinguish the border between HCC and liver fibrosis.ConclusionCombined with HSA-C pretreatment,TSI nanoprobes preferentially aggregated in HCC lesions with higher TBR,and clearly delineated the boundary between HCC and liver fibrosis.This strategy is expected to alleviate advanced liver fibrosis and improve the differential diagnosis accuracy of HCC and liver cirrhosis.