Background:Despite significant advancements in the development of anticancer therapies over the past few decades,the clinical management of colorectal cancer remains a challenging task.This study aims to investigate t...Background:Despite significant advancements in the development of anticancer therapies over the past few decades,the clinical management of colorectal cancer remains a challenging task.This study aims to investigate the inhibitory effects of cancer-targeting liposomes against colorectal cancer.Materials and Methods:Liposomes consisting of 3β-[N-(N′,N′-dimethylamino ethane)carbamoyl]-cholesterol(DC-CHOL),cholesterol(CHOL),and dioleoylphosphatidylethanolamine(DOPE)at a molar ratio of 1:1:0.5 were created and used as carriers to deliver an apoptosis-inducing plasmid encoding the tumor necrosis factor-related apoptosis-inducing ligand(pTRAIL)gene,along with the toll-like receptor(TLR7)agonist Rsiquimod(R848).The rationale behind this design is that pTRAIL can trigger cancer cell apoptosis by activating the DR4/5 receptor,while R848 can stimulate the immune microenvironment.Results:Experimental results demonstrated the synergistic effects of R848 and pTRAIL encapsulated by liposomes(RTL)in suppressing the proliferation of colorectal cancer cells.Moreover,further in vivo investigations revealed the strong anti-tumor efficacy of RTL in xenograft and orthotropic in situ models of colorectal cancer.Conclusions:These findings collectively highlight the therapeutic potential of R848/pTRAIL-loaded liposomes in the treatment of colorectal cancer.展开更多
【目的】探讨咪喹莫特类似物R848对胸腺基质淋巴细胞生成素(TSLP)介导Naive T细胞分化的影响。【方法】从健康志愿者外周血中提取单个核细胞(PBMC),免疫磁珠分选出CD14+PBMC,随后用不同因素诱导分化为成熟树突状细胞(DC);另用免疫磁珠...【目的】探讨咪喹莫特类似物R848对胸腺基质淋巴细胞生成素(TSLP)介导Naive T细胞分化的影响。【方法】从健康志愿者外周血中提取单个核细胞(PBMC),免疫磁珠分选出CD14+PBMC,随后用不同因素诱导分化为成熟树突状细胞(DC);另用免疫磁珠阴性分选出Na觙ve T细胞后,将其与诱导分化后的DC共培养,并分组:A组为Naive T+PBS/DC;B组为Naive T+TSLP/DC;C组为Naive T+(TSLP+R848)/DC;D组为Na觙veT+(TSLP+anti-OX40L)/DC。ELISA检测上清液细胞因子IFN-γ、IL-4和IL-13水平,荧光定量RT-PCR测定转录因子GATA-3及T-bet m RNA的表达。【结果】(1)TSLP诱导后DC表达OX40L增加,而R848可抑制OX40L表达(P<0.05)。(2)B、C、D三组T细胞表达细胞因子IL-4和IL-13水平均高于A组;B组IL-4和IL-13均较C组和D组高,而D组高于C组(均P<0.05);C组表达IFN-γ水平明显高于其余三组(均P<0.05);A、B、D三组之间IFN-γ无明显差异(P>0.05)。(3)C组和D组GATA-3 m RNA水平均低于B组,但高于空白对照A组,其中D组GATA-3 m RNA水平高于C组;C组表达T-bet m RNA水平高于其余三组(均P<0.05);A、B、D三组之间T-bet水平无明显差异(P>0.05)。【结论】TSLP通过上调人DC表达OX40L,在转录因子水平介导Na觙ve T细胞向Th2细胞分化;这种TSLP介导的Th2细胞优势反应可被R848所抑制。展开更多
Ultrasound-generated antigens combined with TLR7/8 agonists as adjuvants have demonstrated significant anti-tumor efficacy as an in-situ vaccine.However,the use of TLR7/8 agonists can cause severe inflammatory responses....Ultrasound-generated antigens combined with TLR7/8 agonists as adjuvants have demonstrated significant anti-tumor efficacy as an in-situ vaccine.However,the use of TLR7/8 agonists can cause severe inflammatory responses.In this study,we present a novel tumor-targeting nano-adjuvant termed aPDL1-PLG/R848 NPs,which are composed of aPDL1 antibody,Fc-III-4C peptide linker(Fc-linker)and poly(L-glutamic acid)-grafted-R848.Under ultrasound irradiation,antigen-presenting cells activate immune mechanisms in vivo under dual stimulation of in situ antigens and immune adjuvants.The strategy inhibits primary tumor growth and induces a strong antigen-specific immune memory effect to prevent tumor recurrence in vivo.This work offers a safe and potent platform for an in situ cancer vaccine based on ultrasound therapy.展开更多
Checkpoint blockade based immune therapy has shown to be effective but benefit only the minority of patients whose tumors have been pre-infiltrated by T cells. To overcome this obstacles, a PEG-modified Bi2Se3 nanocag...Checkpoint blockade based immune therapy has shown to be effective but benefit only the minority of patients whose tumors have been pre-infiltrated by T cells. To overcome this obstacles, a PEG-modified Bi2Se3 nanocage (NC) loaded with imiquimod (R848), which could efficiently destroy the tumors thus producing enough tumor-associated antigens (TAA) and with the existence of R848, a toll-like-receptor-7 agonist, could generate strong anti-cancer immune responses is reported in this study. Moreover, immunogenic Bi2Se3 NC-PEG/R848 mediated photothermal therapy (PTT) sensitizes tumors to checkpoint inhibition mediated by a PD-L1 antibody, not only ablating cancer cells upon NIR laser but also causing strong anti-cancer immunity to suppress distant tumor growth post PTT. Both in vitro and in vivo experiments demonstrate that the Bi2Se3 NC-PEG/R848 could effectively activate a PTT-induced immune response as well as silence immune resistance based on PD-L1 checkpoint blockade to ablate the primary tumor and further inhibit the tumor metastasis. Bi2Se3 NC reported here exhibits high photothermal conversion efficiency and stability, as well as competent drug loading capacity with large hollow structures and high surface area. Our study not only provides a facial way to synthesize Bi2Se3 NC, but also offers an alternative strategy for tumor metastasis.展开更多
目的利用甘露糖(Mannase)修饰瑞喹莫德(R848)脂质体,赋予其对肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)的主动靶向能力,以提高其对小鼠移植瘤的免疫治疗作用。方法用二硬脂酰磷脂酰乙醇胺-聚乙二醇-甘露糖(Mannase-PEG_(200...目的利用甘露糖(Mannase)修饰瑞喹莫德(R848)脂质体,赋予其对肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)的主动靶向能力,以提高其对小鼠移植瘤的免疫治疗作用。方法用二硬脂酰磷脂酰乙醇胺-聚乙二醇-甘露糖(Mannase-PEG_(2000)-DSPE)脂材掺入脂质体,并采用pH梯度主动载药法包封R848,制备获得甘露糖修饰的R848脂质体(M-LS/R848)。利用粒度仪、透射电镜等表征其理化性质,透析法分析其释药稳定性。体外考察其对小鼠巨噬细胞RAW264.7的靶向性,体内评估其对小鼠结肠癌细胞MC38移植瘤的靶向性和免疫治疗效果,并利用免疫荧光染色探讨其免疫治疗机制。结果本研究制备的M-LS/R848大小均一,平均粒径为(123.43±2.43)nm,Zeta电位为(-0.33±0.24)mV,为球形或类球形粒子,R848包封率达(85.59±0.37)%,48 h R848释放率为(46.73±0.54)%,稳定性较高。体外靶向实验表明RAW264.7细胞对M-LS/R848的摄取率为(5.29±0.16)%,显著高于LS/R848的摄取率[(1.38±0.15)%]。体内靶向实验表明甘露糖修饰脂质体在MC38肿瘤中的累积显著高于非修饰脂质体。体内抑瘤实验表明M-LS/R848治疗组肿瘤平均体积和重量显著小于LS/R848组,甚至有两只小鼠肿瘤治愈并产生对MC38肿瘤的免疫记忆效应。免疫荧光分析显示M-LS/R848给药组肿瘤组织中M1型巨噬细胞和CD8阳性细胞较LS/R848组明显增多。结论利用甘露糖修饰R848脂质体可有效提高其对肿瘤的靶向性并增强免疫治疗效果。展开更多
基金approved by the Institutional Animal Care and Use Committee(IACUC)of the First Affiliated Hospital of Fujian Medical University(No.2023048).
文摘Background:Despite significant advancements in the development of anticancer therapies over the past few decades,the clinical management of colorectal cancer remains a challenging task.This study aims to investigate the inhibitory effects of cancer-targeting liposomes against colorectal cancer.Materials and Methods:Liposomes consisting of 3β-[N-(N′,N′-dimethylamino ethane)carbamoyl]-cholesterol(DC-CHOL),cholesterol(CHOL),and dioleoylphosphatidylethanolamine(DOPE)at a molar ratio of 1:1:0.5 were created and used as carriers to deliver an apoptosis-inducing plasmid encoding the tumor necrosis factor-related apoptosis-inducing ligand(pTRAIL)gene,along with the toll-like receptor(TLR7)agonist Rsiquimod(R848).The rationale behind this design is that pTRAIL can trigger cancer cell apoptosis by activating the DR4/5 receptor,while R848 can stimulate the immune microenvironment.Results:Experimental results demonstrated the synergistic effects of R848 and pTRAIL encapsulated by liposomes(RTL)in suppressing the proliferation of colorectal cancer cells.Moreover,further in vivo investigations revealed the strong anti-tumor efficacy of RTL in xenograft and orthotropic in situ models of colorectal cancer.Conclusions:These findings collectively highlight the therapeutic potential of R848/pTRAIL-loaded liposomes in the treatment of colorectal cancer.
文摘【目的】探讨咪喹莫特类似物R848对胸腺基质淋巴细胞生成素(TSLP)介导Naive T细胞分化的影响。【方法】从健康志愿者外周血中提取单个核细胞(PBMC),免疫磁珠分选出CD14+PBMC,随后用不同因素诱导分化为成熟树突状细胞(DC);另用免疫磁珠阴性分选出Na觙ve T细胞后,将其与诱导分化后的DC共培养,并分组:A组为Naive T+PBS/DC;B组为Naive T+TSLP/DC;C组为Naive T+(TSLP+R848)/DC;D组为Na觙veT+(TSLP+anti-OX40L)/DC。ELISA检测上清液细胞因子IFN-γ、IL-4和IL-13水平,荧光定量RT-PCR测定转录因子GATA-3及T-bet m RNA的表达。【结果】(1)TSLP诱导后DC表达OX40L增加,而R848可抑制OX40L表达(P<0.05)。(2)B、C、D三组T细胞表达细胞因子IL-4和IL-13水平均高于A组;B组IL-4和IL-13均较C组和D组高,而D组高于C组(均P<0.05);C组表达IFN-γ水平明显高于其余三组(均P<0.05);A、B、D三组之间IFN-γ无明显差异(P>0.05)。(3)C组和D组GATA-3 m RNA水平均低于B组,但高于空白对照A组,其中D组GATA-3 m RNA水平高于C组;C组表达T-bet m RNA水平高于其余三组(均P<0.05);A、B、D三组之间T-bet水平无明显差异(P>0.05)。【结论】TSLP通过上调人DC表达OX40L,在转录因子水平介导Na觙ve T细胞向Th2细胞分化;这种TSLP介导的Th2细胞优势反应可被R848所抑制。
基金Ministry of Science and Technology of China,Grant/Award Number:2022YFE0110200Natural Science Foundation of Jilin Province,Grant/Award Number:20230101037JCNational Natural Science Foundation of China,Grant/Award Numbers:52203198,52025035。
文摘Ultrasound-generated antigens combined with TLR7/8 agonists as adjuvants have demonstrated significant anti-tumor efficacy as an in-situ vaccine.However,the use of TLR7/8 agonists can cause severe inflammatory responses.In this study,we present a novel tumor-targeting nano-adjuvant termed aPDL1-PLG/R848 NPs,which are composed of aPDL1 antibody,Fc-III-4C peptide linker(Fc-linker)and poly(L-glutamic acid)-grafted-R848.Under ultrasound irradiation,antigen-presenting cells activate immune mechanisms in vivo under dual stimulation of in situ antigens and immune adjuvants.The strategy inhibits primary tumor growth and induces a strong antigen-specific immune memory effect to prevent tumor recurrence in vivo.This work offers a safe and potent platform for an in situ cancer vaccine based on ultrasound therapy.
基金supported by the National Basic Research Project (973 Program) of China (No.2014CB932200)the National Natural Science Foundation of China (Nos.81503016, 81771880, and 81401453)the Application Foundation and Cutting-edge Technologies Research Project of Tianjin (Young Program)(No.15JCQNJC13800).
文摘Checkpoint blockade based immune therapy has shown to be effective but benefit only the minority of patients whose tumors have been pre-infiltrated by T cells. To overcome this obstacles, a PEG-modified Bi2Se3 nanocage (NC) loaded with imiquimod (R848), which could efficiently destroy the tumors thus producing enough tumor-associated antigens (TAA) and with the existence of R848, a toll-like-receptor-7 agonist, could generate strong anti-cancer immune responses is reported in this study. Moreover, immunogenic Bi2Se3 NC-PEG/R848 mediated photothermal therapy (PTT) sensitizes tumors to checkpoint inhibition mediated by a PD-L1 antibody, not only ablating cancer cells upon NIR laser but also causing strong anti-cancer immunity to suppress distant tumor growth post PTT. Both in vitro and in vivo experiments demonstrate that the Bi2Se3 NC-PEG/R848 could effectively activate a PTT-induced immune response as well as silence immune resistance based on PD-L1 checkpoint blockade to ablate the primary tumor and further inhibit the tumor metastasis. Bi2Se3 NC reported here exhibits high photothermal conversion efficiency and stability, as well as competent drug loading capacity with large hollow structures and high surface area. Our study not only provides a facial way to synthesize Bi2Se3 NC, but also offers an alternative strategy for tumor metastasis.
文摘目的利用甘露糖(Mannase)修饰瑞喹莫德(R848)脂质体,赋予其对肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)的主动靶向能力,以提高其对小鼠移植瘤的免疫治疗作用。方法用二硬脂酰磷脂酰乙醇胺-聚乙二醇-甘露糖(Mannase-PEG_(2000)-DSPE)脂材掺入脂质体,并采用pH梯度主动载药法包封R848,制备获得甘露糖修饰的R848脂质体(M-LS/R848)。利用粒度仪、透射电镜等表征其理化性质,透析法分析其释药稳定性。体外考察其对小鼠巨噬细胞RAW264.7的靶向性,体内评估其对小鼠结肠癌细胞MC38移植瘤的靶向性和免疫治疗效果,并利用免疫荧光染色探讨其免疫治疗机制。结果本研究制备的M-LS/R848大小均一,平均粒径为(123.43±2.43)nm,Zeta电位为(-0.33±0.24)mV,为球形或类球形粒子,R848包封率达(85.59±0.37)%,48 h R848释放率为(46.73±0.54)%,稳定性较高。体外靶向实验表明RAW264.7细胞对M-LS/R848的摄取率为(5.29±0.16)%,显著高于LS/R848的摄取率[(1.38±0.15)%]。体内靶向实验表明甘露糖修饰脂质体在MC38肿瘤中的累积显著高于非修饰脂质体。体内抑瘤实验表明M-LS/R848治疗组肿瘤平均体积和重量显著小于LS/R848组,甚至有两只小鼠肿瘤治愈并产生对MC38肿瘤的免疫记忆效应。免疫荧光分析显示M-LS/R848给药组肿瘤组织中M1型巨噬细胞和CD8阳性细胞较LS/R848组明显增多。结论利用甘露糖修饰R848脂质体可有效提高其对肿瘤的靶向性并增强免疫治疗效果。
