Limited clinical application of antibody-drug conjugates(ADCs)targeting tumor associated antigens(TAAs)is usually caused by on-target off-tumor side effect.Tumor-specific mutant antigens(TSMAs)only expressed in tumor ...Limited clinical application of antibody-drug conjugates(ADCs)targeting tumor associated antigens(TAAs)is usually caused by on-target off-tumor side effect.Tumor-specific mutant antigens(TSMAs)only expressed in tumor cells which are ideal targets for ADCs.In addition,intracellular somatic mutant proteins can be presented on the cell surface by human leukocyte antigen class I(HLA I)molecules forming tumor-specific peptide/HLA I complexes.KRAS G12 V mutation frequently occurred in varied cancer and was verified as a promising target for cancer therapy.In this study,we generated two TCR-mimic antibodydrug conjugates(TCRm-ADCs),2E8-MMAE and 2 A5-MMAE,targeting KRAS G12 V/HLAA*0201 complex,which mediated specific antitumor activity in vitro and in vivo without obvious toxicity.Our findings are the first time validate the strategy of TCRm-ADCs targeting intracellular TSMAs,which improves the safety of antibody-based drugs and provides novel strategy for precision medicine in cancer therapy.展开更多
目的肝细胞癌(hepatocellular carcinoma,HCC)的发生具有显著的性别差异,但其分子机制尚不清楚。本研究通过检测Hras12V转基因小鼠雌雄肝肿瘤组织的蛋白质组学表达差异,探讨肝肿瘤发生的性别差异机制。方法采用双向荧光差异凝胶电泳(two...目的肝细胞癌(hepatocellular carcinoma,HCC)的发生具有显著的性别差异,但其分子机制尚不清楚。本研究通过检测Hras12V转基因小鼠雌雄肝肿瘤组织的蛋白质组学表达差异,探讨肝肿瘤发生的性别差异机制。方法采用双向荧光差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)和基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)技术检测Hras12V雌雄小鼠肝肿瘤组织的蛋白质组学差异表达谱,分离并鉴定差异表达蛋白,应用蛋白质印迹法验证2D-DIGE结果,并对差异表达蛋白进行生物信息学分析,包括蛋白功能注释、分类(gene ontology,GO)分析、京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路分析。结果经双向荧光差异凝胶电泳-图像分析得到1 313个蛋白点,其中差异倍数≥1.5(P<0.05)的蛋白点553个。从中选择94个差异蛋白点进行MALDI-TOF-MS鉴定,得到56种差异性表达的蛋白质。其中,与雌鼠相比,雄鼠肝肿瘤组织中高表达的蛋白有43种,低表达的蛋白有13种。选取4个差异蛋白FABP1、Albumin、Prdx6和CALR进行了蛋白质印迹法验证,其中FABP1在雄鼠肝癌中显著上调,Albumin和Prdx6在雄鼠肝癌中显著下调,CALR在雌雄小鼠肝癌中差异无统计学意义,结果与质谱数据基本一致,证明了2D-DIGE结果的可靠性。对雌雄鼠肝肿瘤组织中的差异蛋白进行GO分析,细胞组分分析显示,差异蛋白主要分布在细胞质、细胞溶质、线粒体和细胞核中;分子功能分析显示,差异蛋白主要承担poly(A)RNA结合、ATP结合和酶结合等功能;生物过程分析显示,差异蛋白主要参与运输、脂代谢过程和凋亡过程的负调控等。KEGG通路分析结果表明,差异蛋白涉及9条信号通路,主要通路包括代谢途径、内质网加工过程和细胞色素P450的异物代谢等通路。结论 Hras12V转基因�展开更多
基金supported by the National Key Research and Development Program of China‘Precision Medicine Research’(Grant No.2017YFC0908602)the State Key Program of National Natural Science of China(Grant No.81430081)National Key R&D Program of China(No.2017YFE0102200)。
文摘Limited clinical application of antibody-drug conjugates(ADCs)targeting tumor associated antigens(TAAs)is usually caused by on-target off-tumor side effect.Tumor-specific mutant antigens(TSMAs)only expressed in tumor cells which are ideal targets for ADCs.In addition,intracellular somatic mutant proteins can be presented on the cell surface by human leukocyte antigen class I(HLA I)molecules forming tumor-specific peptide/HLA I complexes.KRAS G12 V mutation frequently occurred in varied cancer and was verified as a promising target for cancer therapy.In this study,we generated two TCR-mimic antibodydrug conjugates(TCRm-ADCs),2E8-MMAE and 2 A5-MMAE,targeting KRAS G12 V/HLAA*0201 complex,which mediated specific antitumor activity in vitro and in vivo without obvious toxicity.Our findings are the first time validate the strategy of TCRm-ADCs targeting intracellular TSMAs,which improves the safety of antibody-based drugs and provides novel strategy for precision medicine in cancer therapy.
文摘目的肝细胞癌(hepatocellular carcinoma,HCC)的发生具有显著的性别差异,但其分子机制尚不清楚。本研究通过检测Hras12V转基因小鼠雌雄肝肿瘤组织的蛋白质组学表达差异,探讨肝肿瘤发生的性别差异机制。方法采用双向荧光差异凝胶电泳(two dimension difference gel electrophoresis,2D-DIGE)和基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)技术检测Hras12V雌雄小鼠肝肿瘤组织的蛋白质组学差异表达谱,分离并鉴定差异表达蛋白,应用蛋白质印迹法验证2D-DIGE结果,并对差异表达蛋白进行生物信息学分析,包括蛋白功能注释、分类(gene ontology,GO)分析、京都基因与基因组百科全书(kyoto encyclopedia of genes and genomes,KEGG)通路分析。结果经双向荧光差异凝胶电泳-图像分析得到1 313个蛋白点,其中差异倍数≥1.5(P<0.05)的蛋白点553个。从中选择94个差异蛋白点进行MALDI-TOF-MS鉴定,得到56种差异性表达的蛋白质。其中,与雌鼠相比,雄鼠肝肿瘤组织中高表达的蛋白有43种,低表达的蛋白有13种。选取4个差异蛋白FABP1、Albumin、Prdx6和CALR进行了蛋白质印迹法验证,其中FABP1在雄鼠肝癌中显著上调,Albumin和Prdx6在雄鼠肝癌中显著下调,CALR在雌雄小鼠肝癌中差异无统计学意义,结果与质谱数据基本一致,证明了2D-DIGE结果的可靠性。对雌雄鼠肝肿瘤组织中的差异蛋白进行GO分析,细胞组分分析显示,差异蛋白主要分布在细胞质、细胞溶质、线粒体和细胞核中;分子功能分析显示,差异蛋白主要承担poly(A)RNA结合、ATP结合和酶结合等功能;生物过程分析显示,差异蛋白主要参与运输、脂代谢过程和凋亡过程的负调控等。KEGG通路分析结果表明,差异蛋白涉及9条信号通路,主要通路包括代谢途径、内质网加工过程和细胞色素P450的异物代谢等通路。结论 Hras12V转基因�
