Background and aims:Radiofrequency ablation(RFA)is the first-line treatment for early-stage hepatocellular carcinoma(HCC).However,recurrence after curative RFA remains a significant challenge for HCC patients.Although...Background and aims:Radiofrequency ablation(RFA)is the first-line treatment for early-stage hepatocellular carcinoma(HCC).However,recurrence after curative RFA remains a significant challenge for HCC patients.Although RFA induces an immune response,the anti-tumor effect is often limited by the immunosuppressive tumor microenvironment.Enhancing anti-tumor immunity is essential to improve treatment efficacy and prevent recurrence.In this study,we explore the efficacy and underlying mechanisms of the combination of RFA and anti-PD-1 in suppressing abscopal and recurrent tumors.Methods:We established a bilateral subcutaneous HCC mouse model and performed complete RFA on the right-flank tumor.Anti-PD-1 or anti-IgG was administered post-RFA.Tumor growth,immune cell profiles,and molecular pathways were assessed using flow cytometry,immunohistochemistry staining,RNA-sequencing,and Western blot.Chemokines released by the tumor were detected by ELISA.An in vivo tumor rechallenge experiment was performed after a complete tumor regression to evaluate the immune memory induced by the RFA+anti-PD-1 treatment.Results:RFA combined with anti-PD-1 significantly suppressed abscopal tumor growth and prolonged survival.Compared with RFA monotherapy,the infiltration of CD8^(+)T cells and dendritic cells was significantly increased in the combined treatment group,while PMN-MDSCs were markedly reduced.Mechanistically,the chemokine signaling pathway and JAK-STAT signaling pathway were activated in the tumor of the RFA+anti-PD-1 group with upregulation of CXCL10 to recruit CD8^(+)T cells.In addition,the combination therapy induced durable immune memory that inhibited rechallenge tumor outgrowth.Conclusions:Our study discovered that RFA combined with anti-PD-1 induced anti-tumor immunity to inhibit abscopal tumors and durable immune memory to prevent recurrence,suggesting RFA+anti-PD-1 as a potential therapeutic strategy for multifocal HCC and preventing recurrence.展开更多
Recent studies have suggested that the anti-tumour effect of the programmed cell death protein 1 monoclonal antibody(aPD-1)depends on the expression of interleukin-12(IL-12)by dendritic cells(DCs).Since DCs are abunda...Recent studies have suggested that the anti-tumour effect of the programmed cell death protein 1 monoclonal antibody(aPD-1)depends on the expression of interleukin-12(IL-12)by dendritic cells(DCs).Since DCs are abundant in skin tissues,transdermal delivery of IL-12 targeting DCs may significantly improve the anti-tumour effect of aPD-1.In this study,a novel mannosylated chitosan(MC)-modified ethosome(Eth-MC)was obtained through electrostatic adsorption.The Eth-MC loaded with plasmid containing the IL-12 gene(pIL-12@Eth-MC)stimulated DCs to express mature-related molecular markers such as CD86,CD80,and major histocompatibility complex-II in a targeted manner.The pIL-12@Eth-MC was then mixed with polyvinyl pyrrolidone solution to make microspheres using the electrospray technique,and sprayed onto the surface of electrospun silk fibroin-polyvinyl alcohol nanofibres to obtain a PVP-pIL-12@Eth-MC/silk fibroin-polyvinyl alcohol composite nanofibrous patch(termed a transcutaneous immunization(TCI)patch).The TCI patch showed a good performance on transdermal drug release.Animal experiments on melanoma-bearing mice showed that topical application of the TCI patches promoted the expression of IL-12 and inhibited the growth of tumour.Furthermore,combined application of the TCI patch and aPD-1 showed a stronger anti-tumour effect than aPD-1 monotherapy.The combination therapy significantly promoted the expression of IL-12,interferon-γand tumour necrosis factor-α,the infiltration of CD4+and CD8+T cells into tumour tissues,and thus promoted the apoptosis of tumour cells.The present study provides a convenient and non-invasive strategy for improving the efficacy of immune checkpoint inhibitor therapy.This study was approved by the Institutional Animal Care and Use Committee at Donghua University(approval No.DHUEC-NSFC-2020-11)on March 31,2020.展开更多
基金funded by the National Natural Science Foundation of China to Xuezhen Zeng(No.82372777)the Kelin Foundation for Distinguished Young Scholars to Xuezhen Zeng(No.R07014).
文摘Background and aims:Radiofrequency ablation(RFA)is the first-line treatment for early-stage hepatocellular carcinoma(HCC).However,recurrence after curative RFA remains a significant challenge for HCC patients.Although RFA induces an immune response,the anti-tumor effect is often limited by the immunosuppressive tumor microenvironment.Enhancing anti-tumor immunity is essential to improve treatment efficacy and prevent recurrence.In this study,we explore the efficacy and underlying mechanisms of the combination of RFA and anti-PD-1 in suppressing abscopal and recurrent tumors.Methods:We established a bilateral subcutaneous HCC mouse model and performed complete RFA on the right-flank tumor.Anti-PD-1 or anti-IgG was administered post-RFA.Tumor growth,immune cell profiles,and molecular pathways were assessed using flow cytometry,immunohistochemistry staining,RNA-sequencing,and Western blot.Chemokines released by the tumor were detected by ELISA.An in vivo tumor rechallenge experiment was performed after a complete tumor regression to evaluate the immune memory induced by the RFA+anti-PD-1 treatment.Results:RFA combined with anti-PD-1 significantly suppressed abscopal tumor growth and prolonged survival.Compared with RFA monotherapy,the infiltration of CD8^(+)T cells and dendritic cells was significantly increased in the combined treatment group,while PMN-MDSCs were markedly reduced.Mechanistically,the chemokine signaling pathway and JAK-STAT signaling pathway were activated in the tumor of the RFA+anti-PD-1 group with upregulation of CXCL10 to recruit CD8^(+)T cells.In addition,the combination therapy induced durable immune memory that inhibited rechallenge tumor outgrowth.Conclusions:Our study discovered that RFA combined with anti-PD-1 induced anti-tumor immunity to inhibit abscopal tumors and durable immune memory to prevent recurrence,suggesting RFA+anti-PD-1 as a potential therapeutic strategy for multifocal HCC and preventing recurrence.
基金supported by the Science&Technology Commission of Shanghai Municipality of China,Nos.18490740400,20DZ2254900the National Key Research&Development Program of China,No.2018YFC1706200.
文摘Recent studies have suggested that the anti-tumour effect of the programmed cell death protein 1 monoclonal antibody(aPD-1)depends on the expression of interleukin-12(IL-12)by dendritic cells(DCs).Since DCs are abundant in skin tissues,transdermal delivery of IL-12 targeting DCs may significantly improve the anti-tumour effect of aPD-1.In this study,a novel mannosylated chitosan(MC)-modified ethosome(Eth-MC)was obtained through electrostatic adsorption.The Eth-MC loaded with plasmid containing the IL-12 gene(pIL-12@Eth-MC)stimulated DCs to express mature-related molecular markers such as CD86,CD80,and major histocompatibility complex-II in a targeted manner.The pIL-12@Eth-MC was then mixed with polyvinyl pyrrolidone solution to make microspheres using the electrospray technique,and sprayed onto the surface of electrospun silk fibroin-polyvinyl alcohol nanofibres to obtain a PVP-pIL-12@Eth-MC/silk fibroin-polyvinyl alcohol composite nanofibrous patch(termed a transcutaneous immunization(TCI)patch).The TCI patch showed a good performance on transdermal drug release.Animal experiments on melanoma-bearing mice showed that topical application of the TCI patches promoted the expression of IL-12 and inhibited the growth of tumour.Furthermore,combined application of the TCI patch and aPD-1 showed a stronger anti-tumour effect than aPD-1 monotherapy.The combination therapy significantly promoted the expression of IL-12,interferon-γand tumour necrosis factor-α,the infiltration of CD4+and CD8+T cells into tumour tissues,and thus promoted the apoptosis of tumour cells.The present study provides a convenient and non-invasive strategy for improving the efficacy of immune checkpoint inhibitor therapy.This study was approved by the Institutional Animal Care and Use Committee at Donghua University(approval No.DHUEC-NSFC-2020-11)on March 31,2020.
