| Download | - View final version: Programmable attenuation of antigenic sensitivity for a nanobody-based EGFR chimeric antigen receptor through hinge domain truncation (PDF, 9.3 MiB)
- View supplementary information: Programmable attenuation of antigenic sensitivity for a nanobody-based EGFR chimeric antigen receptor through hinge domain truncation (PDF, 6.3 MiB)
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| DOI | Resolve DOI: https://doi.org/10.3389/fimmu.2022.864868 |
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| Author | Search for: McComb, Scott1ORCID identifier: https://orcid.org/0000-0002-4616-7864; Search for: Nguyen, Tina1; Search for: Shepherd, Alex1; Search for: Henry, Kevin A.1; Search for: Bloemberg, Darin1; Search for: Marcil, Anne1; Search for: Maclean, Susanne1; Search for: Zafer, Ahmed1; Search for: Gilbert, Rénald1; Search for: Gadoury, Christine1; Search for: Pon, Robert A.1; Search for: Sulea, Traian1; Search for: Zhu, Qin1; Search for: Weeratna, Risini D.1 |
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| Affiliation | - National Research Council of Canada. Human Health Therapeutics
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| Format | Text, Article |
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| Subject | cellular immunotherapy; EGFR; CAR optimization; CAR-T; hinge domain; cancer selectivity; cell therapy |
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| Abstract | Epidermal growth factor family receptor (EGFR) is commonly overexpressed in many solid tumors and an attractive target for chimeric antigen receptor (CAR)-T therapy, but as EGFR is also expressed at lower levels in healthy tissues a therapeutic strategy must balance antigenic responsiveness against the risk of on-target off-tumor toxicity. Herein, we identify several camelid single-domain antibodies (also known as nanobodies) that are effective EGFR targeting moieties for CARs (EGFR-sdCARs) with very strong reactivity to EGFR-high and EGFR-low target cells. As a strategy to attenuate their potent antigenic sensitivity, we performed progressive truncation of the human CD8 hinge commonly used as a spacer domain in many CAR constructs. Single amino acid hinge-domain truncation progressively decreased both EGFR-sdCAR-Jurkat cell binding to EGFR-expressing targets and expression of the CD69 activation marker. Attenuated signaling in hinge-truncated EGFR-sdCAR constructs increased selectivity for antigen-dense EGFR-overexpressing cells over an EGFR-low tumor cell line or healthy donor derived EGFR-positive fibroblasts. We also provide evidence that epitope location is critical for determining hinge-domain requirement for CARs, as hinge truncation similarly decreased antigenic sensitivity of a membrane-proximal epitope targeting HER2-CAR but not a membrane-distal EGFRvIII-specific CAR. Hinge-modified EGFR-sdCAR cells showed clear functional attenuation in Jurkat-CAR-T cells and primary human CAR-T cells from multiple donors in vitro and in vivo. Overall, these results indicate that hinge length tuning provides a programmable strategy for throttling antigenic sensitivity in CARs targeting membrane-proximal epitopes, and could be employed for CAR-optimization and improved tumor selectivity. |
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| Publication date | 2022-07-22 |
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| Publisher | Frontiers Media |
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| Language | English |
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| Peer reviewed | Yes |
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| Export citation | Export as RIS |
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| Report a correction | Report a correction (opens in a new tab) |
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| Record identifier | 1d2bba96-61ea-4f41-8903-5ff249e3a8af |
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| Record created | 2023-04-11 |
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| Record modified | 2023-04-19 |
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