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Updated: May 10, 2025
High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model
Published on: August 16, 2019
CAR T cells re-directed by a rationally designed human peptide tag demonstrate efficacy in preclinical models
Nele Knelangen1, Ulrika Bader2, Evangelia Maniaki2
1Research and Development, Miltenyi Biotec, Bergisch Gladbach, Germany; Division of Functional Immune Cell Modulation, Leibniz Institute for Immunotherapy, Regensburg, Germany.
Abstract:
Currently, only a few chimeric antigen receptor (CAR) T cell therapies have been approved by the Food and Drug Administration and European Medicines Agency for the treatment of B-cell malignancies. To enable broader application of the CAR T cell technology in other indications, improved control and flexible targeting of multiple tumor antigens are required. Here, we developed a novel adapter CAR (AdCAR) T cell platform for flexible targeting of multiple tumor antigens. This platform is based on a short peptide tag derived from an interdomain region of fibroblast growth factor receptor 2 (FGFR2), commonly mutated in cancer. To select AdCARs specific for mutated FGFR2-derived peptide tags, a multistep pooled screening approach in primary T cells was employed, incorporating MACS separation and next-generation sequencing. The resulting AdCAR was highly specific for the FGFR2-derived peptide tag. Using different in vitro and in vivo model systems, the activity of AdCAR T cells was shown to be strictly dependent on the presence of the adapter and corresponding target antigen. Moreover, AdCAR T cells could be redirected to different target antigens by the addition of respective adapter molecules (AM). Finally, in situ expression of functional AM in primary T cells under control of a drug-inducible promoter system was demonstrated, highlighting the potential for controlling the activity of AdCAR T cells by cellular micropharmacies.
Insights
A new adapter CAR (AdCAR) T cell platform offers flexible targeting of multiple tumor antigens. This innovation allows precise control over CAR T cell activity for broader cancer treatment applications.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Limited approved chimeric antigen receptor (CAR) T cell therapies exist, primarily for B-cell malignancies.
- Broader application of CAR T cell technology requires enhanced control and flexible targeting of multiple tumor antigens.
Purpose of the Study:
- To develop a novel adapter CAR (AdCAR) T cell platform for versatile targeting of multiple tumor antigens.
- To engineer AdCAR T cells with specificity for a peptide tag derived from mutated fibroblast growth factor receptor 2 (FGFR2).
Main Methods:
- A multistep pooled screening approach using MACS separation and next-generation sequencing was employed to select AdCARs.
- In vitro and in vivo model systems were utilized to evaluate AdCAR T cell activity.
- Functional adapter molecules (AM) were expressed in primary T cells under a drug-inducible promoter.
Main Results:
- A highly specific AdCAR for the FGFR2-derived peptide tag was successfully developed.
- AdCAR T cell activity was demonstrated to be dependent on the presence of the adapter and target antigen.
- AdCAR T cells could be redirected to different antigens using adapter molecules, and in situ expression of functional AM was achieved.
Conclusions:
- The novel AdCAR T cell platform enables flexible and controlled targeting of multiple tumor antigens.
- This platform holds potential for expanding CAR T cell therapy applications beyond current indications.
- Controlling AdCAR T cell activity via cellular micropharmacies presents a promising therapeutic strategy.
