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Targeting of low ALK antigen density neuroblastoma using AND logic-gate engineered CAR-T cells
Emma Halliwell1, Alice Vitali1, Henrike Muller1
1Cancer Section, UCL Great Ormond Street Institute of Child Health, London, UK.
Background Aims:
The targeting of solid cancers with chimeric antigen receptor (CAR) T cells faces many technological hurdles, including selection of optimal target antigens. Promising pre-clinical and clinical data of CAR T-cell activity have emerged from targeting surface antigens such as GD2 and B7H3 in childhood cancer neuroblastoma. Anaplastic lymphoma kinase (ALK) is expressed in a majority of neuroblastomas at low antigen density but is largely absent from healthy tissues.
Methods:
To explore an alternate target antigen for neuroblastoma CAR T-cell therapy, the authors generated and screened a single-chain variable fragment library targeting ALK extracellular domain to make a panel of new anti-ALK CAR T-cell constructs.
Results:
A lead novel CAR T-cell construct was capable of specific cytotoxicity against neuroblastoma cells expressing low levels of ALK, but with only weak cytokine and proliferative T-cell responses. To explore strategies for amplifying ALK CAR T cells, the authors generated a co-CAR approach in which T cells received signal 1 from a first-generation ALK construct and signal 2 from anti-B7H3 or GD2 chimeric co-stimulatory receptors. The co-CAR approach successfully demonstrated the ability to avoid targeting single-antigen-positive targets as a strategy for mitigating on-target off-tumor toxicity.
Conclusions:
These data provide further proof of concept for ALK as a neuroblastoma CAR T-cell target.
Insights
Researchers explored targeting anaplastic lymphoma kinase (ALK) for neuroblastoma treatment using chimeric antigen receptor (CAR) T cells. A novel co-CAR approach enhanced T-cell responses and mitigated toxicity, showing ALK is a promising target.
Area of Science:
- Oncology
- Immunotherapy
- Cancer Research
Background:
- Chimeric antigen receptor (CAR) T-cell therapy faces challenges in solid tumor targeting, particularly antigen selection.
- Neuroblastoma, a childhood cancer, has shown promise with CAR T-cells targeting GD2 and B7H3.
- Anaplastic lymphoma kinase (ALK) is a potential target for neuroblastoma due to its expression in tumors and limited presence in healthy tissues.
Purpose of the Study:
- To develop novel CAR T-cell constructs targeting anaplastic lymphoma kinase (ALK) for neuroblastoma treatment.
- To investigate strategies for enhancing the efficacy and safety of ALK-targeted CAR T-cells.
Main Methods:
- Generated and screened a single-chain variable fragment library to create anti-ALK CAR T-cell constructs.
- Developed a co-CAR approach combining an ALK-targeting CAR with co-stimulatory receptors (anti-B7H3 or anti-GD2).
- Assessed cytotoxicity, cytokine production, and proliferative responses of engineered T-cells against neuroblastoma cells.
Main Results:
- A novel CAR T-cell construct demonstrated specific cytotoxicity against low-ALK-expressing neuroblastoma cells.
- The co-CAR approach improved T-cell responses and showed potential for mitigating on-target, off-tumor toxicity by avoiding single-antigen targeting.
- Engineered T-cells showed enhanced responses when receiving dual signals from the co-CAR system.
Conclusions:
- Anaplastic lymphoma kinase (ALK) is validated as a viable target antigen for neuroblastoma CAR T-cell therapy.
- The developed co-CAR strategy offers a promising approach to enhance CAR T-cell efficacy and safety in solid tumors.
- Further development of ALK-targeted CAR T-cells, particularly with co-stimulatory elements, holds potential for improved neuroblastoma treatment outcomes.
