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Updated: Jul 5, 2026
Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody
Published on: May 16, 2020
A novel CLDN6 nanobody-based bispecific T-cell engager delivered by self-amplifying RNA platform exhibits potent
Zhixiong Zhu1, Jia Li1, Qizhong Lu1
1State Key Laboratory of Biotherapy and Cancer Center, Collaborative Innovation Center of Biotherapy, West China Hospital, Sichuan University, Chengdu, 610041, China.
Abstract:
Ovarian cancer remains a highly lethal gynecologic malignancy with limited early detection and frequent relapse. Claudin-6 (CLDN6), minimally expressed in normal tissues but frequently upregulated in ovarian cancer, represents a promising tumor-specific target. Bispecific T-cell engagers (BiTEs) effectively redirect cytotoxic T cells toward tumor cells. However, previous studies have reported that protein-based BiTEs, particularly scFv-based constructs, may encounter challenges related to protein folding, aggregation, and in vivo stability. To address these limitations, CLDN6-nanobodies were developed through our screening platform, which display cross-species reactivity to human, murine and non-human primate CLDN6. Using this binder, we constructed CLDN6 nanobody-based BiTE (CLDN6-Nb-BiTE) encoded by self-amplifying mRNA (sa-mRNA) and delivered by lipid nanoparticles (LNPs), providing a functional alternative to protein administration. The LNP-sa-mRNA formulation was stable and uniform, enabling robust CLDN6-Nb-BiTE expression, antigen-dependent cytotoxicity, and cytokine release in vitro, while sustained CLDN6-Nb-BiTE production in vivo promoted T-cell recruitment and durable tumor control without systemic toxicity. Overall, our work introduces the first CLDN6-Nb-BiTE delivered by sa-mRNA, establishing a foundation for next-generation ovarian cancer immunotherapy.
Insights
This study introduces a novel ovarian cancer immunotherapy using Claudin-6 (CLDN6) nanobody-based bispecific T-cell engagers (BiTEs) delivered via self-amplifying mRNA (sa-mRNA) and lipid nanoparticles (LNPs). This approach demonstrates effective tumor control and T-cell engagement without systemic toxicity.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Ovarian cancer is a lethal malignancy with poor early detection and high relapse rates.
- Claudin-6 (CLDN6) is a promising tumor-specific target due to its upregulation in ovarian cancer.
- Protein-based bispecific T-cell engagers (BiTEs) face challenges with stability and delivery.
Purpose of the Study:
- To develop a novel CLDN6-targeted immunotherapy overcoming limitations of protein-based BiTEs.
- To create a CLDN6 nanobody-based BiTE (CLDN6-Nb-BiTE) using self-amplifying mRNA (sa-mRNA) and lipid nanoparticles (LNPs).
Main Methods:
- Developed CLDN6-specific nanobodies with cross-species reactivity.
- Constructed CLDN6-Nb-BiTE encoded by sa-mRNA and formulated with LNPs.
- Evaluated in vitro cytotoxicity, cytokine release, and in vivo tumor control and toxicity.
Main Results:
- The LNP-sa-mRNA formulation was stable and facilitated robust CLDN6-Nb-BiTE expression.
- Demonstrated antigen-dependent cytotoxicity and cytokine release in vitro.
- Achieved sustained in vivo BiTE production, T-cell recruitment, and durable tumor control with no systemic toxicity.
Conclusions:
- Introduced the first CLDN6-Nb-BiTE delivered via sa-mRNA for ovarian cancer immunotherapy.
- Established a foundation for next-generation immunotherapies targeting CLDN6.
- Validated sa-mRNA/LNP delivery as a viable alternative to protein administration for BiTEs.
