Objectives:The Kirsten rat sarcoma virus(KRAS)G12D oncogenic mutation poses a significant challenge in treating solid tumors due to the lack of specific and effective therapeutic interventions.This study aims to explore...Objectives:The Kirsten rat sarcoma virus(KRAS)G12D oncogenic mutation poses a significant challenge in treating solid tumors due to the lack of specific and effective therapeutic interventions.This study aims to explore innovative approaches in T cell receptor(TCR)engineering and characterization to target the KRAS G12D7-16 mutation,providing potential strategies for overcoming this therapeutic challenge.Methods:In this innovative study,we engineered and characterized two T cell receptors(TCRs),KDA11-01 and KDA11-02 with high affinity for the KRAS G12D7-16 mutation.These TCRs were isolated from tumor-infiltrating lymphocytes(TILs)derived from tumor tissues of patients with the KRAS G12D mutation.We assessed their specificity and anti-tumor activity in vitro using various cancer cell lines.Results:KDA11-01 and KDA11-02 demonstrated exceptional specificity for the HLA-A*11:01-restricted KRAS G12D7-16 epitope,significantly inducing IFN-γrelease and eliminating tumor cells without cross-reactivity or alloreactivity.Conclusions:The successful development of KDA11-01 and KDA11-02 introduces a novel and precise TCR-based therapeutic strategy against KRAS G12D mutation,showing potential for significant advancements in cancer immunotherapy.展开更多
目的采用FQ-PCR溶解曲线技术初步分析肾综合征出血热(Hemorrhagic fever with renal syndrome,HFRS)患者外周血T细胞TCRα链各可变区基因家族(TRAV)CDR3谱系特征。方法提取4例健康志愿者和15例HFRS患者外周血单个核细胞(PBMC)的总RNA,...目的采用FQ-PCR溶解曲线技术初步分析肾综合征出血热(Hemorrhagic fever with renal syndrome,HFRS)患者外周血T细胞TCRα链各可变区基因家族(TRAV)CDR3谱系特征。方法提取4例健康志愿者和15例HFRS患者外周血单个核细胞(PBMC)的总RNA,逆转录成c DNA,荧光定量PCR(FQ-PCR)扩增34条TRAV基因家族CDR3受体库,采用溶解曲线技术分析CDR3谱系特征,并测序分析部分单克隆家族CDR3区基因和氨基酸序列的组成和特征。结果健康志愿者外周血T细胞TCRα链34个TRAV家族CDR3的溶解曲线谱型图呈现CDR3多克隆增生的高斯分布;15例HFRS患者多数TRAV家族CDR3的溶解曲线谱型图为多克隆增生的高斯分布,但出现数量不等的单克隆、寡克隆、极低水平表达的TRAV家族。部分测序结果显示TCRα链优势取用TRAV家族的CDR3区有着不同的基因序列和氨基酸组成。结论HFRS患者外周血中TCRα链各TRAV家族CDR3溶解曲线谱型图出现数量不等,形态不一的单峰、寡峰/偏峰、极低水平的现象,表现出不同的异常频率,TRAV优势取用家族的CDR3区有着不同的基因序列和氨基酸组成,实验结果可为进一步研究HFRS个体特异性T细胞应答提供基础。展开更多
基金funded by the key R&D Project of Hubei Province(Social Development),China(2022BCA018)the Cooperative Innovation Center of Industrial Fermentation(Ministry of Education&Hubei Province),China(2022KF16)to Kanghong Hu.
文摘Objectives:The Kirsten rat sarcoma virus(KRAS)G12D oncogenic mutation poses a significant challenge in treating solid tumors due to the lack of specific and effective therapeutic interventions.This study aims to explore innovative approaches in T cell receptor(TCR)engineering and characterization to target the KRAS G12D7-16 mutation,providing potential strategies for overcoming this therapeutic challenge.Methods:In this innovative study,we engineered and characterized two T cell receptors(TCRs),KDA11-01 and KDA11-02 with high affinity for the KRAS G12D7-16 mutation.These TCRs were isolated from tumor-infiltrating lymphocytes(TILs)derived from tumor tissues of patients with the KRAS G12D mutation.We assessed their specificity and anti-tumor activity in vitro using various cancer cell lines.Results:KDA11-01 and KDA11-02 demonstrated exceptional specificity for the HLA-A*11:01-restricted KRAS G12D7-16 epitope,significantly inducing IFN-γrelease and eliminating tumor cells without cross-reactivity or alloreactivity.Conclusions:The successful development of KDA11-01 and KDA11-02 introduces a novel and precise TCR-based therapeutic strategy against KRAS G12D mutation,showing potential for significant advancements in cancer immunotherapy.
