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Engineering a single-chain variable fragment of cetuximab for CAR T-cell therapy against head and neck squamous cell
Corinna Haist1, Zoe Poschinski2, Arthur Bister1
1Department of Otorhinolaryngology, Head & Neck Surgery, Heinrich Heine University, Düsseldorf, Germany; Department of Pediatrics III, University Children's Hospital, University of Duisburg-Essen, Essen, Germany.
Abstract:
The monoclonal antibody cetuximab recognizes domain III of the epithelial growth factor receptor (EGFR) with high-affinity and is an important element in the treatment of several malignancies that overexpress non-mutated wild-type EGFR. In order to create an EGFR recognizing chimeric antigen receptor (CAR) for cellular immunotherapy of head and neck squamous cell carcinoma (HNSCC), we rationally designed single chain fragments of different lengths based on the cetuximab variable heavy and light chains. We then cloned the different cetuximab fragments into our second generation CAR construct, expressed CARs on primary human T-cells from healthy donors using mono- or biscistronic lentiviral vectors and tested the stability, functionality and specificity of the CARs. Our smallest CAR construct was most efficient with greatly improved vector production and T-cell transduction efficacy. Finally, we demonstrated that the new cetuximab CAR construct expressed on T-cells is highly reactive against EGFR-positive HNSCCs and also malignant cells from other solid cancer entities. In conclusion, we generated an optimized high-affinity EGFR CAR construct for the next steps in cancer immunotherapy, which need to focus on the development of armored CAR T-cells that will be more resistant and effective in the hostile microenvironment present in solid cancers.
Insights
Researchers developed a novel EGFR-targeting chimeric antigen receptor (CAR) T-cell therapy for head and neck cancer. This optimized CAR T-cell therapy shows high reactivity against EGFR-positive solid tumors, paving the way for advanced cancer immunotherapy.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Cetuximab, a monoclonal antibody, targets epithelial growth factor receptor (EGFR) domain III, crucial for treating EGFR-overexpressing cancers.
- Head and neck squamous cell carcinoma (HNSCC) often overexpresses wild-type EGFR, making it a target for novel therapies.
Purpose of the Study:
- To engineer an optimized EGFR-targeting chimeric antigen receptor (CAR) construct for cellular immunotherapy of HNSCC.
- To evaluate the stability, functionality, and specificity of novel CAR constructs derived from cetuximab.
Main Methods:
- Rational design of single-chain fragments based on cetuximab variable chains.
- Cloning fragments into a second-generation CAR construct and expression on primary human T-cells via lentiviral vectors.
- Assessment of CAR stability, functionality, and specificity against EGFR-positive cancer cells.
Main Results:
- The smallest CAR construct demonstrated superior efficiency, with enhanced vector production and T-cell transduction rates.
- Engineered T-cells expressing the cetuximab CAR construct exhibited high reactivity against EGFR-positive HNSCC and other solid tumor cells.
- The optimized CAR construct shows promise for targeting various EGFR-positive malignancies.
Conclusions:
- An optimized, high-affinity EGFR CAR construct was successfully generated for cancer immunotherapy.
- This CAR T-cell therapy demonstrates significant potential for treating EGFR-positive solid tumors, including HNSCC.
- Future research should focus on developing armored CAR T-cells for enhanced efficacy in the tumor microenvironment.
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