Related Experiment Video
Updated: Sep 27, 2025
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
A TCR mimic CAR T cell specific for NDC80 is broadly reactive with solid tumors and hematologic malignancies
Martin G Klatt1, Tao Dao1, Zhiyuan Yang2
1Molecular Pharmacology Program, Sloan Kettering Institute, New York, NY.
Abstract:
Target identification for chimeric antigen receptor (CAR) T-cell therapies remains challenging due to the limited repertoire of tumor-specific surface proteins. Intracellular proteins presented in the context of cell surface HLA provide a wide pool of potential antigens targetable through T-cell receptor mimic antibodies. Mass spectrometry (MS) of HLA ligands from 8 hematologic and nonhematologic cancer cell lines identified a shared, non-immunogenic, HLA-A*02-restricted ligand (ALNEQIARL) derived from the kinetochore-associated NDC80 gene. CAR T cells directed against the ALNEQIARL:HLA-A*02 complex exhibited high sensitivity and specificity for recognition and killing of multiple cancer types, especially those of hematologic origin, and were efficacious in mouse models against a human leukemia and a solid tumor. In contrast, no toxicities toward resting or activated healthy leukocytes as well as hematopoietic stem cells were observed. This shows how MS can inform the design of broadly reactive therapeutic T-cell receptor mimic CAR T-cell therapies that can target multiple cancer types currently not druggable by small molecules, conventional CAR T cells, T cells, or antibodies.
Insights
Researchers identified a novel cancer target using mass spectrometry, enabling the development of T-cell receptor mimic chimeric antigen receptor (CAR) T-cell therapies. These therapies show promise for treating various cancers without harming healthy cells.
Area of Science:
- Oncology
- Immunotherapy
- Molecular Biology
Background:
- Chimeric antigen receptor (CAR) T-cell therapy faces challenges in identifying suitable tumor-specific targets.
- Intracellular proteins presented via cell surface HLA molecules offer a broader antigenic landscape for T-cell receptor mimic (TCRM) therapies.
Purpose of the Study:
- To identify novel, broadly applicable cancer targets for TCRM CAR T-cell therapy.
- To develop and evaluate the efficacy and safety of TCRM CAR T-cells targeting a specific HLA-presented intracellular antigen.
Main Methods:
- Mass spectrometry (MS) was used to analyze HLA ligands from diverse cancer cell lines.
- T-cell receptor mimic (TCRM) CAR T-cells were engineered to target the identified HLA-ligand complex.
- In vitro and in vivo (mouse models) assays were conducted to assess CAR T-cell activity and toxicity.
Main Results:
- A shared, non-immunogenic HLA-A*02-restricted ligand (ALNEQIARL) from the NDC80 gene was identified.
- Engineered TCRM CAR T-cells demonstrated high sensitivity and specificity against multiple cancer types, including leukemia and solid tumors.
- No observed toxicity in healthy leukocytes or hematopoietic stem cells.
Conclusions:
- Mass spectrometry is a valuable tool for discovering novel targets for TCRM CAR T-cell therapy.
- TCRM CAR T-cells targeting the ALNEQIARL:HLA-A*02 complex offer a potential therapeutic strategy for cancers currently lacking effective treatments.
- This approach broadens the applicability of CAR T-cell therapy to intracellular antigens and multiple cancer types.
Related Concept Videos
01:27Cytotoxic T Cells-mediated Immune Response
Immunological surveillance is the ability of immune cells to monitor and eliminate infected cells with intracellular pathogens, neoplastically transformed cells, and cells with non-self antigens. Cytotoxic T cells and NK...
01:22T Cell Activation and Clonal Selection
Naive T cells that have not yet encountered an antigen express two primary CD...
01:27Tumor Immunotherapy