Screening and identification of tumor-specific T cell receptors from tumor infiltrating lymphocytes using colorectal cancer organoid model
10.13431/j.cnki.immunol.j.20240036
- VernacularTitle:结直肠癌类器官的反应性T细胞受体筛选及功能鉴定
- Author:
Yaoyao MEI
1
,
2
;
Yingming WANG
;
Xiaojian HAN
;
Meiying SHEN
;
Ya LI
;
Zhengqiang WEI
;
Aishun JIN
Author Information
1. 400016,重庆医科大学基础医学院免疫学教研室
2. 400016,肿瘤免疫调节与免疫干预重庆市重点实验室
- Keywords:
Colorectal cancer organoids;
Tumor-reactive T cell;
T cell receptor;
Anti-tumor
- From:
Immunological Journal
2024;40(3):279-285
- CountryChina
- Language:Chinese
-
Abstract:
This study intends to establish a colorectal cancer(CRC)organoid model,expand and isolate CRC-reactive tumor-infiltrating lymphocytes(TILs),screen tumor-specific T cell receptors(TCRs)and perform functional verification,in order to provide a technological platform and research foundation for the clinical transformation of individualized adoptive T-cell immunotherapy for colorectal cancer.An organoid model derived from colon cancer patient tissues was constructed using in vitro 3D culture techniques,which then subjected to HE staining and immunohistochemistry for detecting morphological characteristics and representative molecular expression.Subsequently,CRC organoids were co-cultured with TILs for sorting reactive TILs using flow cytometry,and the characteristics of reactive TCR clones was analyzed through single T cell receptor gene cloning technology.Furthermore,the function of TCRs was verified through cytotoxicity experiments.Morphological analysis and representative molecules(CK20 and CDX2)expression indicated that there is high similarity between colorectal cancer organoids and patient tumors.In the in vitro expanded and cultured TILs,colorectal cancer-reactive T cells with upregulated CD137 expression and increased IFN-γ secretion were screened out successfully,among which TCR2-T cells demonstrated superior tumor reactivity and in vitro tumor killing function.In conclusion,a platform for screening and function validation of reactive TCRs based on CRC-Org has been established,providing a technological platform for the translational application of individualized T-cell therapy for colorectal cancer.
