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Patient-derived 3D organoid platform for functional assessment of GPC3-targeted CAR T cell cytotoxic activity in head
Han-Na Yoon1, Jin-Ha Kim2, Doeon Gu1
1Rare and Pediatric Cancer Branch, Research Institute, National Cancer Center, Goyang 10408, the Republic of Korea.
Background:
Head and neck squamous cell carcinoma (HNSCC) remains a therapeutic challenge owing to its marked heterogeneity and limited immunotherapy efficacy, underscoring the need for improved therapeutic strategies and preclinical systems supporting clinical translation.
Method:
We established a simplified and optimized 3D head and neck cancer organoid (HNCO)-chimeric antigen receptor (CAR) T cell co-culture platform that maintains uniform spheroid architecture (>500 μm) to recapitulate physiological hypoxia while preserving the tumor secretome. CAR T cell cytotoxic activity was assessed through multimodal readouts-structural disruption, ATP-based viability, and granzyme B secretion. Following initial organoid seeding, the workflow proceeded without physical manipulation, enabling concurrent multimodal assessment of CAR T cell activity from a single co-culture.
Results:
Analysis of The Cancer Genome Atlas (TCGA) data revealed that high glypican-3 (GPC3) expression was associated with poor survival in patients with HNSCC. We generated GPC3-targeted CAR T (GPC3-CAR T) cells. Using our co-culture platform, we evaluated the cytotoxic activity of GPC3-CAR T cells against five HNCOs harboring diverse genetic alterations and variable GPC3 expression. Organoids with high or moderate GPC3 expression consistently exhibited structural disintegration, reduced viability, and increased granzyme B secretion, whereas GPC3-low organoids showed heterogeneous responses.
Conclusions:
This proof-of-concept study introduces a patient-derived 3D organoid platform for functional assessment of CAR T cell cytotoxic activity in HNSCC. Our findings suggest that GPC3-CAR T therapy may be clinically applicable to subsets of patients with HNSCC, while emphasizing the need for functional validation to account for interpatient heterogeneity in clinical translation.
Insights
A new 3D organoid platform effectively assesses chimeric antigen receptor (CAR) T cell therapy for head and neck squamous cell carcinoma (HNSCC). Glypican-3 (GPC3)-targeted CAR T cells show promise against GPC3-expressing HNSCC organoids.
Area of Science:
- Oncology
- Immunotherapy
- Biotechnology
Background:
- Head and neck squamous cell carcinoma (HNSCC) presents significant therapeutic challenges due to heterogeneity and limited immunotherapy response.
- Improved preclinical models are crucial for advancing HNSCC treatment strategies and clinical translation.
Purpose of the Study:
- To develop and validate a simplified 3D head and neck cancer organoid (HNCO)-chimeric antigen receptor (CAR) T cell co-culture platform.
- To assess the efficacy of glypican-3 (GPC3)-targeted CAR T cells in a preclinical HNSCC model.
Main Methods:
- Established a 3D HNCO platform maintaining physiological hypoxia and tumor secretome.
- Utilized multimodal readouts (structural disruption, ATP viability, granzyme B secretion) to assess CAR T cell activity.
- Co-cultured GPC3-targeted CAR T cells with diverse HNCOs expressing variable GPC3 levels.
Main Results:
- High GPC3 expression in HNSCC correlates with poor patient survival (TCGA data).
- GPC3-CAR T cells induced significant structural disintegration, reduced viability, and increased granzyme B secretion in GPC3-high/moderate HNCOs.
- GPC3-low HNCOs exhibited heterogeneous responses to GPC3-CAR T cell therapy.
Conclusions:
- Introduced a patient-derived 3D organoid platform for functional CAR T cell assessment in HNSCC.
- GPC3-CAR T cell therapy shows potential for specific HNSCC patient subsets.
- Functional validation is essential to address interpatient heterogeneity for clinical translation.
