北京大学学报(医学版) ›› 2022, Vol. 54 ›› Issue (5): 884-895. doi: 10.19723/j.issn.1671-167X.2022.05.016
程晓静1,蒋栋2,张连海1,王江华2,李雅真2,翟佳慧2,闫宝琪2,张露露2,谢兴旺2,*(),李子禹1,*(),季加孚1,*()
Xiao-jing CHENG1,Dong JIANG2,Lian-hai ZHANG1,Jiang-hua WANG2,Ya-zhen LI2,Jia-hui ZHAI2,Bao-qi YAN2,Lu-lu ZHANG2,Xing-wang XIE2,*(),Zi-yu LI1,*(),Jia-fu JI1,*()
摘要:
目的: KRAS G12V是最为常见的KRAS突变类型之一,是一个T细胞表位抗原,目前尚无针对该位点的靶向药物,本研究旨在克隆能够特异性识别该表位抗原的T细胞受体(T cell receptor, TCR),通过体内外实验对该TCR基因修饰T细胞(TCR engineered T cells, TCR-T)靶向KRAS G12V突变肿瘤的安全性和有效性进行评估。方法: 从1例结直肠癌患者的肿瘤浸润淋巴细胞中获得靶向KRAS G12V8-16表位的高亲和力TCR序列,构建该TCR慢病毒载体并感染人源T细胞,获得TCR-T。采用抗原肽激活、γ-干扰素(interferon-γ, IFN-γ) 酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)、T细胞体外增殖等实验,体外评价该TCR-T的免疫杀伤活性和脱靶-交叉反应性;通过体内实验评价该TCR-T的抑瘤效果、安全性等指标。结果: 获得了能特异性识别HLA-A*11:01限制性KRAS G12V8-16表位的高亲和力TCR序列KVA11-01。KVA11-01 TCR-T能够显著杀伤体外过表达HLA-A*11:01和KRAS G12V的多种肿瘤细胞。非特异杀伤实验显示,KVA11-01仅杀伤同时表达HLA-A*11:01和KRAS G12V的肿瘤细胞。体内抑瘤实验显示,KVA11-01 TCR-T可以显著抑制PANC-1和HeLa(体外过表达HLA-A*11:01和KRAS G12V)细胞裸鼠皮下移植瘤的生长。TCR-T细胞可以显著浸润至肿瘤组织内部,有良好的实体肿瘤归巢能力。结论: KVA11-01 TCR-T能够在体内外有效靶向并杀伤携带KRAS G12V突变的多种恶性肿瘤细胞,具有良好的实体瘤组织归巢能力,KVA11-01 TCR-T有望成为携带KRAS G12V突变的实体恶性肿瘤患者的有效治疗手段。
中图分类号:
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