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Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors
Received date: 2022-06-27
Online published: 2022-10-14
Supported by
the Cultivation of Pharmaceutical Innovation Varieties and Platforms of Beijing Municipal Science and Technology Commission and Zhongguancun Administrative Committee(Z211100002521027);the Peking University Cancer Hospital Science Foundation(2021-8);the Pilot Project (3rd Round) to Reform Public Development of Beijing Municipal Medical Research Institute(京医研2019-01)
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.
Xiao-jing CHENG , Dong JIANG , Lian-hai ZHANG , Jiang-hua WANG , Ya-zhen LI , Jia-hui ZHAI , Bao-qi YAN , Lu-lu ZHANG , Xing-wang XIE , Zi-yu LI , Jia-fu JI . Preclinical study of T cell receptor specifically reactive with KRAS G12V mutation in the treatment of malignant tumors[J]. Journal of Peking University(Health Sciences), 2022 , 54(5) : 884 -895 . DOI: 10.19723/j.issn.1671-167X.2022.05.016
| 1 | Bos JL . Ras oncogenes in human cancer: A review[J]. Cancer Res, 1989, 49 (17): 4682- 4689. |
| 2 | Singh H , Longo DL , Chabner BA . Improving prospects for targeting RAS[J]. J Clin Oncol, 2015, 33 (31): 3650- 3659. |
| 3 | Hobbs GA , Der CJ , Rossman KL . RAS isoforms and mutations in cancer at a glance[J]. J Cell Sci, 2016, 129 (7): 1287- 1292. |
| 4 | Windon AL , Loaiza-Bonilla A , Jensen CE , et al. A KRAS wild type mutational status confers a survival advantage in pancreatic ductal adenocarcinoma[J]. J Gastrointest Oncol, 2018, 9 (1): 1- 10. |
| 5 | Shin SH , Kim SC , Hong SM , et al. Genetic alterations of K-ras, p53, c-erbB-2, and DPC4 in pancreatic ductal adenocarcinoma and their correlation with patient survival[J]. Pancreas, 2013, 42 (2): 216- 222. |
| 6 | Bournet B , Muscari F , Buscail C , et al. KRAS G12D mutation subtype is a prognostic factor for advanced pancreatic adenocarcinoma[J]. Clin Transl Gastroenterol, 2016, 7 (3): e157. |
| 7 | Haas M , Ormanns S , Baechmann S , et al. Extended RAS analysis and correlation with overall survival in advanced pancreatic cancer[J]. Br J Cancer, 2017, 116 (11): 1462- 1469. |
| 8 | Marabese M , Ganzinelli M , Garassino MC , et al. KRAS mutations affect prognosis of non-small-cell lung cancer patients treated with first-line platinum containing chemotherapy[J]. Oncotarget, 2015, 6 (32): 34014- 34022. |
| 9 | Laghi L , Orbetegli O , Bianchi P , et al. Common occurrence of multiple KRAS mutations in pancreatic cancers with associated precursor lesions and in biliary cancers[J]. Oncogene, 2002, 21 (27): 4301- 4306. |
| 10 | Russo AL , Borger DR , Szymonifka J , et al. Mutational analysis and clinical correlation of metastatic colorectal cancer[J]. Can-cer, 2014, 120 (10): 1482- 1490. |
| 11 | Tran E , Ahmadzadeh M , Lu YC , et al. Immunogenicity of soma-tic mutations in human gastrointestinal cancers[J]. Science, 2015, 350 (6266): 1387- 1390. |
| 12 | Chatani PD , Yang JC . Mutated RAS. Targeting the "untargetable" with T cells[J]. Clin Cancer Res, 2020, 26 (3): 537- 544. |
| 13 | Tran E , Robbins PF , Lu YC , et al. T-cell transfer therapy targeting mutant KRAS in cancer[J]. N Engl J Med, 2016, 375 (23): 2255- 2262. |
| 14 | Leidner R , Sanjuan Silva N , Huang H , et al. Neoantigen T-cell receptor gene therapy in pancreatic cancer[J]. N Engl J Med, 2022, 386 (22): 2112- 2119. |
| 15 | Wang QJ , Yu Z , Griffith K , et al. Identification of T-cell receptors targeting KRAS-mutated human tumors[J]. Cancer Immunol Res, 2015, 4 (3): 204- 214. |
| 16 | Sim MJW , Lu J , Spencer M , et al. High-affinity oligoclonal TCRs define effective adoptive T cell therapy targeting mutant KRAS-G12D[J]. Proc Natl Acad Sci USA, 2020, 117 (23): 12826- 12835. |
| 17 | Cohen CJ , Zhao Y , Zheng Z , et al. Enhanced antitumor activity of murine-human hybrid T-cell receptor (TCR) in human lymphocytes is associated with improved pairing and TCR/CD3 stability[J]. Cancer Res, 2006, 66 (17): 8878- 8886. |
| 18 | Morgan RA , Chinnasamy N , Abate-Daga D , et al. Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy[J]. J Immunother, 2013, 36 (2): 133- 151. |
| 19 | Linette GP , Stadtmauer EA , Maus MV , et al. Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma[J]. Blood, 2013, 122 (6): 863- 871. |
| 20 | Cameron BJ , Gerry AB , Dukes J , et al. Identification of a Titin-derived HLA-A1-presented peptide as a cross-reactive target for engineered MAGE A3-directed T cells[J]. Sci Transl Med, 2013, 5 (197): 103- 126. |
| 21 | Suchin EJ , Langmuir PB , Palmer E , et al. Quantifying the frequency of alloreactive T cells in vivo: New answers to an old question[J]. J Immunol, 2001, 166 (2): 973- 981. |
| 22 | Li J , Li W , Huang K , et al. Chimeric antigen receptor T cell (CAR-T) immunotherapy for solid tumors: lessons learned and strategies for moving forward[J]. J Hematol Oncol, 2018, 11 (1): 22. |
| 23 | Idorn M , Skadborg SK , Kellermann L , et al. Chemokine receptor engineering of T cells with CXCR2 improves homing towards subcutaneous human melanomas in xenograft mouse model[J]. Oncoimmunology, 2018, 7 (8): e1450715. |
| 24 | Draper LM , Kwong ML , Gros A , et al. Targeting of HPV-16+ epithelial cancer cells by TCR gene engineered T cells directed against E6[J]. Clin Cancer Res, 2015, 21 (19): 4431- 4439. |
| 25 | Karapetis CS , Khambata-Ford S , Jonker DJ , et al. K-ras mutations and benefit from cetuximab in advanced colorectal cancer[J]. N Engl J Med, 2008, 359 (17): 1757- 1765. |
| 26 | Moore AR , Rosenberg SC , McCormick F , et al. RAS-targeted therapies: Is the undruggable drugged?[J]. Nat Rev Drug Discov, 2020, 19 (8): 533- 552. |
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