The therapeutic efficacy of whole tumor vaccine is limited by the relatively low immunogenicity,which determines subsequent activation of the immune response.In this study,we developed a whole tumor cell vaccine with ...The therapeutic efficacy of whole tumor vaccine is limited by the relatively low immunogenicity,which determines subsequent activation of the immune response.In this study,we developed a whole tumor cell vaccine with multiple immunogenicity enhancement strategies,including CD47 knockout by CRISPR/Cas9 gene editing based on non-viral polymer gene carriers,membrane insertion of immune adjuvant DSPE-PEG-Mannose and heat-induced immunogenic cell death.Specifically,CD47 knockout blocks the CD47/SIRPα“don't eat me”signal between tumor cells and APCs,thereby enhancing APC-mediated phagocytosis of tumor cells.Next,the adjuvant DSPE-PEG-Mannose onto the surface of CD47^(KO)B16F10 cells using a simple membrane insertion method for stimulating the maturation of bone marrow-derived dendritic cells(BMDCs)and triggering a stronger immune response.Finally,heat treatment via a simple water bath induced immunogenic cell death(ICD),leading to cell inactivation while releasing specific damage-associated molecular patterns(DAMPs)to activate both the innate and adaptive immune systems.After the programmable process of CD47 gene editing,insertion of immune adjuvants and heat-induced death,the tumor cells transformed into a highly immunogenic whole tumor cell vaccine,achieving significant tumor prevention and therapeutic effects.This work provided a novel,simple,and effective strategy for the construction of a whole tumor cell vaccine.展开更多
Tumor vaccines trigger tumor-specific immune responses to prevent or treat tumors by activating the hosts’immune systems,and therefore,these vaccines have potential clinical applications.However,the low immunogenicit...Tumor vaccines trigger tumor-specific immune responses to prevent or treat tumors by activating the hosts’immune systems,and therefore,these vaccines have potential clinical applications.However,the low immunogenicity of the tumor antigen itself and the low efficiency of the vaccine delivery system hinder the efficacy of tumor vaccines that cannot produce high-efficiency and long-lasting antitumor immune effects.Here,we constructed a nanovaccine by integrating CD47KO/CRT dual-bioengineered B16F10 cancer cell membranes and the unmethylated cytosine-phosphate-guanine(CpG)adjuvant.Hyperbranched PEI25k was used to load unmethylated cytosine-phosphate-guanine(CpG)through electrostatic adsorption to prepare PEI25k/CpG nanoparticles(PEI25k/CpG-NPs).CD47KO/CRT dual-bioengineered cells were obtained by CRISPR-Cas9 gene editing technology,followed by the cell surface translocation of calreticulin(CRT)to induce immunogenic cell death(ICD)in vitro.Finally,the extracted cell membranes were coextruded with PEI25k/CpG-NPs to construct the CD47KO/CRT dual-bioengineered cancer cell membrane-coated nanoparticles(DBE@CCNPs).DBE@CCNPs could promote endocytosis of antigens and adjuvants in murine bone marrow derived dendritic cells(BMDCs)and induce their maturation and antigen cross-presentation.To avoid immune checkpoint molecule-induced T cell dysfunction,the immune checkpoint inhibitor,the anti-PD-L1 antibody,was introduced to boost tumor immunotherapy through a combination with the DBE@CCNPs nanovaccine.This combination therapy strategy can significantly alleviate tumor growth and may open up a potential strategy for clinical tumor immunotherapy.展开更多
Tumor nanovaccines have potential applications in the prevention and treatment of malignant tumors.However,it remains a longstanding challenge in exploiting efficient nanocarriers for inducing potent specifically cell...Tumor nanovaccines have potential applications in the prevention and treatment of malignant tumors.However,it remains a longstanding challenge in exploiting efficient nanocarriers for inducing potent specifically cellular immune responses.Toward this objective,we herein explore an intensive tumor immunotherapeutic strategy by combining mannosylated nanovaccines and gene regulated PD-L1 blockade for immune stimulation and killing activity.Here,we fabricate a mannose modified PLL-RT(Man-PLL-RT)mediated nanovaccines with dendritic cells(DCs)targeting capacity.Man-PLL-RT is capable of co-encapsulating with antigen(ovalbumin,OVA)and adjuvant(unmethylated cytosine-phosphate-guanine,CpG)by electrostatic interaction.This positively charged Man-PLL-RT/OVA/CpG nanovaccines can facilitate the endocytosis,maturation and cross presentation in DCs.However,the nanovaccines arouse limited inhibition of tumor growth,which is mainly due to the immunosuppressed microenvironment of tumors.Combining tumor nanovaccines with gene regulated PD-L1 blockade leads to an obvious tumor remission in B16F10 melanoma bearing mice.The collaborative strategy provides essential insights to boost the benefits of tumor vaccines by regulating the checkpoint blockade with gene therapy.展开更多
基金supported by the National Key Research and Development Program of China(2021YFB3800900)the National Natural Science Foundation of China(52433006,52495010,52495015)+1 种基金the Talent Cultivation Project Funds for the Innovation Laboratory for Sciences and Technologies of Energy Materials of Fujian Province(HRTP-[2022]52)supported by the General Plan Project of Quanzhou,Fujian Province,China(2024NS009[to Nuo Dong])。
文摘The therapeutic efficacy of whole tumor vaccine is limited by the relatively low immunogenicity,which determines subsequent activation of the immune response.In this study,we developed a whole tumor cell vaccine with multiple immunogenicity enhancement strategies,including CD47 knockout by CRISPR/Cas9 gene editing based on non-viral polymer gene carriers,membrane insertion of immune adjuvant DSPE-PEG-Mannose and heat-induced immunogenic cell death.Specifically,CD47 knockout blocks the CD47/SIRPα“don't eat me”signal between tumor cells and APCs,thereby enhancing APC-mediated phagocytosis of tumor cells.Next,the adjuvant DSPE-PEG-Mannose onto the surface of CD47^(KO)B16F10 cells using a simple membrane insertion method for stimulating the maturation of bone marrow-derived dendritic cells(BMDCs)and triggering a stronger immune response.Finally,heat treatment via a simple water bath induced immunogenic cell death(ICD),leading to cell inactivation while releasing specific damage-associated molecular patterns(DAMPs)to activate both the innate and adaptive immune systems.After the programmable process of CD47 gene editing,insertion of immune adjuvants and heat-induced death,the tumor cells transformed into a highly immunogenic whole tumor cell vaccine,achieving significant tumor prevention and therapeutic effects.This work provided a novel,simple,and effective strategy for the construction of a whole tumor cell vaccine.
基金The authors are thankful to the National Key R&D Program of China(2021YFB3800900)National Natural Science Foundation of China(51925305,51873208,51973217)+2 种基金Taishan Scholar Foundation of Shandong Province(qnts20161035)Natural Science Foundation of Shandong Province(ZR2019ZD24,ZR2019YQ30)Jilin Province Science and Technology Development Program(20200201075JC,20210509005RQ).
文摘Tumor vaccines trigger tumor-specific immune responses to prevent or treat tumors by activating the hosts’immune systems,and therefore,these vaccines have potential clinical applications.However,the low immunogenicity of the tumor antigen itself and the low efficiency of the vaccine delivery system hinder the efficacy of tumor vaccines that cannot produce high-efficiency and long-lasting antitumor immune effects.Here,we constructed a nanovaccine by integrating CD47KO/CRT dual-bioengineered B16F10 cancer cell membranes and the unmethylated cytosine-phosphate-guanine(CpG)adjuvant.Hyperbranched PEI25k was used to load unmethylated cytosine-phosphate-guanine(CpG)through electrostatic adsorption to prepare PEI25k/CpG nanoparticles(PEI25k/CpG-NPs).CD47KO/CRT dual-bioengineered cells were obtained by CRISPR-Cas9 gene editing technology,followed by the cell surface translocation of calreticulin(CRT)to induce immunogenic cell death(ICD)in vitro.Finally,the extracted cell membranes were coextruded with PEI25k/CpG-NPs to construct the CD47KO/CRT dual-bioengineered cancer cell membrane-coated nanoparticles(DBE@CCNPs).DBE@CCNPs could promote endocytosis of antigens and adjuvants in murine bone marrow derived dendritic cells(BMDCs)and induce their maturation and antigen cross-presentation.To avoid immune checkpoint molecule-induced T cell dysfunction,the immune checkpoint inhibitor,the anti-PD-L1 antibody,was introduced to boost tumor immunotherapy through a combination with the DBE@CCNPs nanovaccine.This combination therapy strategy can significantly alleviate tumor growth and may open up a potential strategy for clinical tumor immunotherapy.
基金This work was supported by the National Natural Science Foundation of China(51925305,51873208,51973217,51520105004 and 51803210)National Science and Technology Major Projects for Major New Drugs Innovation and Development(2018ZX09711003-012)and Jilin Province Science and Technology Development Program(20180414027GH and 20200201075JC).
文摘Tumor nanovaccines have potential applications in the prevention and treatment of malignant tumors.However,it remains a longstanding challenge in exploiting efficient nanocarriers for inducing potent specifically cellular immune responses.Toward this objective,we herein explore an intensive tumor immunotherapeutic strategy by combining mannosylated nanovaccines and gene regulated PD-L1 blockade for immune stimulation and killing activity.Here,we fabricate a mannose modified PLL-RT(Man-PLL-RT)mediated nanovaccines with dendritic cells(DCs)targeting capacity.Man-PLL-RT is capable of co-encapsulating with antigen(ovalbumin,OVA)and adjuvant(unmethylated cytosine-phosphate-guanine,CpG)by electrostatic interaction.This positively charged Man-PLL-RT/OVA/CpG nanovaccines can facilitate the endocytosis,maturation and cross presentation in DCs.However,the nanovaccines arouse limited inhibition of tumor growth,which is mainly due to the immunosuppressed microenvironment of tumors.Combining tumor nanovaccines with gene regulated PD-L1 blockade leads to an obvious tumor remission in B16F10 melanoma bearing mice.The collaborative strategy provides essential insights to boost the benefits of tumor vaccines by regulating the checkpoint blockade with gene therapy.
