Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors.
- Author:
Xiao Jing CHENG
1
,
2
;
Dong JIANG
3
;
Lian Hai ZHANG
1
,
2
;
Jiang Hua WANG
3
;
Ya Zhen LI
3
;
Jia Hui ZHAI
3
;
Bao Qi YAN
3
;
Lu Lu ZHANG
3
;
Xing Wang XIE
3
;
Zi Yu LI
1
,
2
;
Jia Fu JI
1
,
2
Author Information
1. Department of Gastrointestinal Cancer Center, Key Laboratory of Carcinogenesis and Translational Research, Ministry of Education
2. Laboratory of Genetics, Peking University Cancer Hospital & Institute, Beijing 100142, China.
3. Beijing CorreGene Biotechnology Co., Ltd., Beijing 100094, China.
- Publication Type:Journal Article
- Keywords:
DNA mutational analysis;
Neoplasms;
T-cell receptor;
Tumor-infiltrating lymphocytes
- MeSH:
Animals;
DNA, Complementary;
Epitopes;
HLA-A Antigens;
Humans;
Interferon-gamma;
Mice;
Mice, Nude;
Mutation;
Neoplasms;
Proto-Oncogene Proteins p21(ras)/genetics*;
Receptors, Antigen, T-Cell/genetics*
- From:
Journal of Peking University(Health Sciences)
2022;54(5):884-895
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:KRAS gene is one of the most common mutations of proto-oncogenes in human tumors, G12V is one of the most common mutation types for KRAS. It's challenging to chemically acquire the targeted drug for this mutation. Recent studies reported that this mutation peptides can form a neoepitope for T cell recognition. Our study aims to clone the T cell receptor (TCR) which specifically recognizes the neoepitope for KRAS G12V mutation and constructs TCR engineered T cells (TCR-T), and to investigate if TCR-Ts have strong antitumor response ability.
METHODS:In this study, tumor infiltrating lymphocytes were obtained from one colorectal cancer patient carrying KRAS G12V mutation. Tumor-reactive TCR was obtained by single-cell RT-5' rapid-amplification of cDNA ends PCR analysis and introduced into peripheral blood lymphocytes to generate TCR-Ts.
RESULTS:We obtained a high-affinity TCR sequence that specifically recognized the HLA-A*11:01-restricted KRAS G12V8-16 epitope: KVA11-01. KVA11-01 TCR-T could significantly kill various tumor cells such as PANC-1, SW480 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V), and secreting high levels of interferon-γ (IFN-γ). Non-specific killing experiments suggested KVA11-01 specifically recognized tumor cells expressing both mutant KRAS G12V and HLA-A*11:01. In vivo assay, tumor inhibition experiments demonstrated that infusion of approximately 1E7 KVA11-01 TCR-T could significantly inhibit the growth of subcuta-neously transplanted tumors of PANC-1 and HeLa (overexpressing HLA-A*11:01 and KRAS G12V) cells in nude mice. No destruction of the morphologies of the liver, spleen and brain were observed. We also found that KVA11-01 TCR-T could significantly infiltrate into tumor tissue and had a better homing ability.
CONCLUSION:KVA11-01 TCR-T cells can effectively target a variety of malignant tumor cells carrying KRAS G12V mutation through in vitro and in vivo assay. KVA11-01 TCR-T cells have excellent biological activity, high specificity of target antigen and homing ability into solid tumor tissue. KVA11-01 TCR-T is expected to be an effective treatment for patients with KRAS G12V mutant solid malignancies.
