Background:Autophagy dysfunction plays a crucial role in tau accumulation and neurodegeneration in Alzheimer’s disease(AD).This study aimed to investigate whether and how the accumulating tau may in turn affect autop...Background:Autophagy dysfunction plays a crucial role in tau accumulation and neurodegeneration in Alzheimer’s disease(AD).This study aimed to investigate whether and how the accumulating tau may in turn affect autophagy.Methods:The primary hippocampal neurons,N2a and HEK293T cells with tau overexpression were respectively starved and treated with vinblastine to study the effects of tau on the initiating steps of autophagy,which was analysed by Student’s two-tailed t-test.The rapamycin and concanamycin A were employed to inhibit the mammalian target of rapamycin kinase complex 1(mTORC1)activity and the vacuolar H+-ATPase(v-ATPase)activity,respectively,which were analysed by One-way ANOVA with post hoc tests.The Western blotting,co-immunoprecipitation and immunofuorescence staining were conducted to gain insight into the mechanisms underlying the tau effects of mTORC1 signaling alterations,as analysed by Student’s two-tailed t-test or One-way ANOVA with post hoc tests.The autophagosome formation was detected by immunofuorescence staining and transmission electron microscopy.The amino acids(AA)levels were detected by high performance liquid chromatography(HPLC).Results:We observed that overexpressing human full-length wild-type tau to mimic AD-like tau accumulation induced autophagy deficits.Further studies revealed that the increased tau could bind to the prion-related domain of T cell intracellular antigen 1(PRD-TIA1)and this association significantly increased the intercellular level of amino acids(Leucine,P=0.0038;Glutamic acid,P=0.0348;Alanine,P=0.0037;Glycine,P=0.0104),with concordant upregulation of mTORC1 activity[phosphorylated eukaryotic translation initiation factor 4E-binding protein 1(p-4EBP1),P<0.0001;phosphorylated 70 kD ribosomal protein S6 kinase 1(p-p70S6K1),P=0.0001,phosphorylated unc-51-like autophagyactivating kinase 1(p-ULK1),P=0.0015]and inhibition of autophagosome formation[microtubuleassociated protein light chain 3 II(LC3 II),P=0.0073;LC3 puncta,P<0.0001].As expected,this tau-induced deficit of autophagosome formation in turn aggravated tau accumulation.Importantly,we also found that blocking TIA1 and tau interaction by overexpressing PRD-TIA1,downregulating the endogenous TIA1 expression by shRNA,or downregulating tau protein level by a small proteolysis targeting chimera(PROTAC)could remarkably attenuate tau-induced autophagy impairment.Conclusions:Our findings reveal that AD-like tau accumulation inhibits autophagosome formation and induces autophagy deficits by activating the TIA1/amino acid/mTORC1 pathway,and thus this work reveals new insight into tau-associated neurodegeneration and provides evidence supporting the use of new therapeutic targets for AD treat-ment and that of related tauopathies.展开更多
目的探究T细胞内抗原1(T-cell intracellular antigen 1,TIA1)在乳腺癌组织中的表达和TIA1启动子区甲基化状态,并分析乳腺癌多层螺旋CT征象与TIA1甲基化状态之间的关系。方法选取2019年2月至2020年3月浙江省台州市第一人民医院收治的50...目的探究T细胞内抗原1(T-cell intracellular antigen 1,TIA1)在乳腺癌组织中的表达和TIA1启动子区甲基化状态,并分析乳腺癌多层螺旋CT征象与TIA1甲基化状态之间的关系。方法选取2019年2月至2020年3月浙江省台州市第一人民医院收治的50例乳腺癌患者,荧光定量PCR法检测所有患者肿瘤组织和癌旁组织TIA1表达水平。通过生物信息学软件MethPrimer预测TIA1启动子区域发现存在cPG岛。采用甲基化特异性聚合酶链反应(Methylmion Specific PCR,MSP)检测肿瘤组织和癌旁组织中TIA1甲基化状态,采用多层螺旋CT对患者进行检查,观察肿块大小、形态、钙化区域、淋巴结转移及边缘,分析TIA1甲基化状态与CT征象之间的相关性。结果乳腺癌组织TIA1表达水平低于癌旁组织(0.50±0.12 vs 0.95±0.10,P=0.00),但TIA1基因启动子甲基化率高于癌旁组织(64% vs 42%,χ2=4.86,P<0.05)。乳腺癌患者TIA1基因启动子甲基化率在肿块形态和有无钙化方面差异无统计学意义(P>0.05)。肿块直径≥2 cm患者TIA1基因启动子甲基化率显著高于肿块直径<2 cm患者(78.57% vs 45.45%,P<0.05),且淋巴结转移患者TIA1基因启动子甲基化率显著高于无淋巴结转移患者(79.17% vs 50.00%,P<0.05)。肿块边缘有毛刺的患者TIA1基因启动子甲基化率显著高于无毛刺的患者(80.77% vs 45.83%,P<0.05)。结论乳腺癌CT显像与TIA1基因启动子甲基化率之间存在相关性,这可为乳腺癌恶变程度和预后的判断提供依据。展开更多
γδT cells are a kind of innate immune T cell.They have not attracted sufficient attention because they account for only a small proportion of all immune cells,and many basic factors related to these cells remain unc...γδT cells are a kind of innate immune T cell.They have not attracted sufficient attention because they account for only a small proportion of all immune cells,and many basic factors related to these cells remain unclear.However,in recent years,with the rapid development of tumor immunotherapy,γδT cells have attracted increasing attention because of their ability to exert cytotoxic effects on most tumor cells without major histocompatibility complex(MHC)restriction.An increasing number of basic studies have focused on the development,antigen recognition,activation,and antitumor immune response ofγδT cells.Additionally,γδT cell-based immunotherapeutic strategies are being developed,and the number of clinical trials investigating such strategies is increasing.This review mainly summarizes the progress of basic research and the clinical application ofγδT cells in tumor immunotherapy to provide a theoretical basis for further the development ofγδT cell-based strategies in the future.展开更多
基金supported by grants from the Natural Science Foundation of China(91949205,31730035,81721005)the Science and Technology Committee of China(2016YFC1305800)+1 种基金the Special Project of Technological Innovation of Hubei Province(2018ACA142)Guangdong Provincial Key S&T Program(2018B030336001)。
文摘Background:Autophagy dysfunction plays a crucial role in tau accumulation and neurodegeneration in Alzheimer’s disease(AD).This study aimed to investigate whether and how the accumulating tau may in turn affect autophagy.Methods:The primary hippocampal neurons,N2a and HEK293T cells with tau overexpression were respectively starved and treated with vinblastine to study the effects of tau on the initiating steps of autophagy,which was analysed by Student’s two-tailed t-test.The rapamycin and concanamycin A were employed to inhibit the mammalian target of rapamycin kinase complex 1(mTORC1)activity and the vacuolar H+-ATPase(v-ATPase)activity,respectively,which were analysed by One-way ANOVA with post hoc tests.The Western blotting,co-immunoprecipitation and immunofuorescence staining were conducted to gain insight into the mechanisms underlying the tau effects of mTORC1 signaling alterations,as analysed by Student’s two-tailed t-test or One-way ANOVA with post hoc tests.The autophagosome formation was detected by immunofuorescence staining and transmission electron microscopy.The amino acids(AA)levels were detected by high performance liquid chromatography(HPLC).Results:We observed that overexpressing human full-length wild-type tau to mimic AD-like tau accumulation induced autophagy deficits.Further studies revealed that the increased tau could bind to the prion-related domain of T cell intracellular antigen 1(PRD-TIA1)and this association significantly increased the intercellular level of amino acids(Leucine,P=0.0038;Glutamic acid,P=0.0348;Alanine,P=0.0037;Glycine,P=0.0104),with concordant upregulation of mTORC1 activity[phosphorylated eukaryotic translation initiation factor 4E-binding protein 1(p-4EBP1),P<0.0001;phosphorylated 70 kD ribosomal protein S6 kinase 1(p-p70S6K1),P=0.0001,phosphorylated unc-51-like autophagyactivating kinase 1(p-ULK1),P=0.0015]and inhibition of autophagosome formation[microtubuleassociated protein light chain 3 II(LC3 II),P=0.0073;LC3 puncta,P<0.0001].As expected,this tau-induced deficit of autophagosome formation in turn aggravated tau accumulation.Importantly,we also found that blocking TIA1 and tau interaction by overexpressing PRD-TIA1,downregulating the endogenous TIA1 expression by shRNA,or downregulating tau protein level by a small proteolysis targeting chimera(PROTAC)could remarkably attenuate tau-induced autophagy impairment.Conclusions:Our findings reveal that AD-like tau accumulation inhibits autophagosome formation and induces autophagy deficits by activating the TIA1/amino acid/mTORC1 pathway,and thus this work reveals new insight into tau-associated neurodegeneration and provides evidence supporting the use of new therapeutic targets for AD treat-ment and that of related tauopathies.
文摘目的探究T细胞内抗原1(T-cell intracellular antigen 1,TIA1)在乳腺癌组织中的表达和TIA1启动子区甲基化状态,并分析乳腺癌多层螺旋CT征象与TIA1甲基化状态之间的关系。方法选取2019年2月至2020年3月浙江省台州市第一人民医院收治的50例乳腺癌患者,荧光定量PCR法检测所有患者肿瘤组织和癌旁组织TIA1表达水平。通过生物信息学软件MethPrimer预测TIA1启动子区域发现存在cPG岛。采用甲基化特异性聚合酶链反应(Methylmion Specific PCR,MSP)检测肿瘤组织和癌旁组织中TIA1甲基化状态,采用多层螺旋CT对患者进行检查,观察肿块大小、形态、钙化区域、淋巴结转移及边缘,分析TIA1甲基化状态与CT征象之间的相关性。结果乳腺癌组织TIA1表达水平低于癌旁组织(0.50±0.12 vs 0.95±0.10,P=0.00),但TIA1基因启动子甲基化率高于癌旁组织(64% vs 42%,χ2=4.86,P<0.05)。乳腺癌患者TIA1基因启动子甲基化率在肿块形态和有无钙化方面差异无统计学意义(P>0.05)。肿块直径≥2 cm患者TIA1基因启动子甲基化率显著高于肿块直径<2 cm患者(78.57% vs 45.45%,P<0.05),且淋巴结转移患者TIA1基因启动子甲基化率显著高于无淋巴结转移患者(79.17% vs 50.00%,P<0.05)。肿块边缘有毛刺的患者TIA1基因启动子甲基化率显著高于无毛刺的患者(80.77% vs 45.83%,P<0.05)。结论乳腺癌CT显像与TIA1基因启动子甲基化率之间存在相关性,这可为乳腺癌恶变程度和预后的判断提供依据。
基金supported by grants from the National Natural Science Foundation of China(Nos.32270915,31970843,82071791,U20A20374,and 81972886)National Key Research and Development Program of China(No.2022YFC3602004)+4 种基金CAMS Initiative for Innovative Medicine(Nos.2021-I2M-1-005,2021-I2M-1-035,and 2021-I2M-1-053)Haihe Laboratory of Cell Ecosystem Innovation Fund(No.22HHXBSS00028)CAMS Central Public Welfare Scientific Research Institute Basal Research Expenses(No.3332020035)Changzhou Science and Technology Support Plan(No.CE20215008)Beijing Municipal Commission of Science and Technology Fund for Innovative Drug(No.Z221100007922040)
文摘γδT cells are a kind of innate immune T cell.They have not attracted sufficient attention because they account for only a small proportion of all immune cells,and many basic factors related to these cells remain unclear.However,in recent years,with the rapid development of tumor immunotherapy,γδT cells have attracted increasing attention because of their ability to exert cytotoxic effects on most tumor cells without major histocompatibility complex(MHC)restriction.An increasing number of basic studies have focused on the development,antigen recognition,activation,and antitumor immune response ofγδT cells.Additionally,γδT cell-based immunotherapeutic strategies are being developed,and the number of clinical trials investigating such strategies is increasing.This review mainly summarizes the progress of basic research and the clinical application ofγδT cells in tumor immunotherapy to provide a theoretical basis for further the development ofγδT cell-based strategies in the future.
