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Updated: Jan 9, 2026
A Real-time Potency Assay for Chimeric Antigen Receptor T Cells Targeting Solid and Hematological Cancer Cells
Published on: November 12, 2019
Preclinical development and selection of nanobody-based CAR-T cells targeting HER2-positive solid tumors
Fien Meeus1,2, Arne Van der Vreken3, Dorien Autaers1
1Vrije Universiteit Brussel (VUB), Translational Oncology Research Center, Department of Biomedical Sciences, Molecular and Cellular Therapy Research Group, Brussels, Belgium.
Abstract:
Chimeric antigen receptor (CAR)-T cell therapy revolutionized treatment of hematological malignancies, but translation to solid tumors remains difficult. Therefore, different avenues are explored to develop novel CAR-T cell products. Nanobodies have advantageous features for CAR design, such as ease of engineering and low immunogenicity, but predictive values on nanobody characteristics for optimal CAR-T cell function remain unclear. We performed a side-by-side evaluation of 12 nanobody-based (nano)CARs against human epidermal growth factor receptor 2 (HER2). All nanoCARs showed the ability to activate a reporter T cell line upon recognition of HER2pos cells. When evaluated in primary T cells, 6/12 nanoCARs showed the strongest reactivity and cytotoxicity against HER2pos glioblastoma and breast cancer cells. Moreover, only 1/12 was able to confer potent cytotoxicity against HER2pos melanoma cells. These data add to the current view that nanobody selection for CAR design remains subject to extensive side-by-side screening. Still, superior cytotoxicity across multiple solid tumor cell lines-determined using in vitro assays evaluating activation, cytokine secretion, and target cell-specific killing-led to selection and further characterization of a lead nanoCAR 1R59b, showing tumor control in an in vivo xenograft model, providing a promising HER2 nanoCAR for further (pre)clinical investigation.
Insights
This study evaluated 12 nanobody-based chimeric antigen receptor (CAR)-T cell therapies targeting HER2. A lead candidate demonstrated potent anti-tumor activity in solid tumors, offering promise for clinical development.
Area of Science:
- Immunotherapy
- Oncology
- Biotechnology
Background:
- Chimeric antigen receptor (CAR)-T cell therapy shows success in blood cancers but faces challenges in solid tumors.
- Nanobodies offer advantages for CAR design, yet their impact on CAR-T cell function requires further investigation.
Purpose of the Study:
- To evaluate and compare the efficacy of 12 distinct nanobody-based CAR-T cell constructs targeting HER2 in solid tumors.
- To identify optimal nanobody CAR designs for enhanced anti-tumor activity and to select a lead candidate for further development.
Main Methods:
- Side-by-side screening of 12 nanobody-based CARs (nanoCARs) against HER2-expressing cells.
- In vitro assessment of T cell activation, cytokine secretion, and cytotoxicity against glioblastoma, breast cancer, and melanoma cell lines.
- In vivo evaluation of a lead nanoCAR in a xenograft tumor model.
Main Results:
- All nanoCARs recognized HER2-positive cells, with 6/12 showing significant reactivity and cytotoxicity against HER2-positive glioblastoma and breast cancer cells.
- Only one nanoCAR demonstrated potent cytotoxicity against HER2-positive melanoma cells.
- A lead nanoCAR (1R59b) exhibited superior cytotoxicity across multiple solid tumor cell lines and demonstrated tumor control in vivo.
Conclusions:
- Nanobody selection for CAR-T cell therapy necessitates extensive screening, as efficacy varies significantly.
- The identified lead nanoCAR (1R59b) shows promise as a potential therapeutic agent for HER2-positive solid tumors, warranting further preclinical investigation.
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