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Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity
Published on: December 7, 2019
Engineered TCR T-cell therapy targeting mass spectrometry-identified natural epitope in PDAC
Jianxin Wang1, Tengyi Zhang2, Pan Li1
1Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA; The Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA; The Skip Viragh Pancreatic Cancer Center, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA; The Pancreatic Cancer Precision Medicine Center of Excellence Program, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA; The Bloomberg Kimmel Institute for Cancer Immunotherapy, Johns Hopkins University School of Medicine, Baltimore, MD, 21287, USA; Zhejiang Provincial Key Laboratory of Pancreatic Disease, The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Abstract:
Tumor antigens are crucial targets for T-cell-based therapy to induce tumor-specific rejection. However, identifying pancreatic ductal adenocarcinoma (PDAC)-specific T-cell epitopes has been challenging. Using advanced mass spectrometry (MS) analysis, we previously identified cancer-associated, class I MHC-bound epitopes shared by multiple PDAC patients with different HLA-A types. Here, we investigated one of these epitopes, LAMC2203-211, a naturally occurring nonmutated epitope on the LAMC2 protein. Following stimulation with the LAMC2203-211 peptide, we cloned T-cell receptors (TCRs) and transduced them into the Jurkat human T-cell line using a lentiviral vector. We found that Jurkat cells expressing LAMC2203-211-specific TCRs resulted in potent, LAMC2 specific, in vitro cytotoxic effects on PDAC cells. Furthermore, in mice that harbored either subcutaneously or orthotopically implanted tumors originating from both HLA-A allele-matched and unmatched PDAC patients, tumor growth was suppressed in a LAMC2-dependent manner following the infusion of LAMC2-targeting T cells. We have therefore developed a LAMC2-specific TCR-based T-cell therapy strategy likely suitable for many PDAC patients. This is the first study to adopt MS analysis to identify natural CD8+ T-cell epitopes in PDAC that could potentially serve as targets for PDAC immunotherapy.
Insights
Researchers identified a specific T-cell target, LAMC2203-211, for pancreatic ductal adenocarcinoma (PDAC) immunotherapy. This LAMC2-targeting T-cell therapy shows potent anti-PDAC effects in vitro and in vivo, offering a promising strategy for many patients.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Tumor antigens are vital for T-cell therapies to eliminate cancer.
- Identifying specific T-cell epitopes in pancreatic ductal adenocarcinoma (PDAC) has been difficult.
- Previous mass spectrometry (MS) analysis identified shared, cancer-associated epitopes in PDAC patients.
Purpose of the Study:
- To investigate the LAMC2203-211 epitope as a potential target for PDAC immunotherapy.
- To develop and evaluate a T-cell receptor (TCR)-based therapy targeting LAMC2 in PDAC.
Main Methods:
- Mass spectrometry (MS) was used to identify naturally occurring T-cell epitopes.
- T-cell receptors (TCRs) specific to LAMC2203-211 were cloned and engineered into T-cells.
- In vitro cytotoxicity assays and in vivo mouse models (subcutaneous and orthotopic) were used to assess therapeutic efficacy.
Main Results:
- Engineered T-cells expressing LAMC2203-211-specific TCRs demonstrated potent, specific cytotoxic effects against PDAC cells in vitro.
- Infusion of LAMC2-targeting T-cells suppressed tumor growth in mouse models, irrespective of HLA-A matching.
- Tumor suppression was dependent on the LAMC2 target.
Conclusions:
- A LAMC2-specific TCR-based T-cell therapy strategy has been developed for PDAC.
- This approach is potentially applicable to a broad range of PDAC patients.
- This study pioneers the use of MS for identifying natural CD8+ T-cell epitopes for PDAC immunotherapy.
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