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The development of novel chimeric antigen receptor gamma-delta T cells against multiple myeloma and single-cell RNA
Yang Hong1, Cuiling Zhang1, Jue Wang2
1Department of Hematology, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China.
Abstract:
Chimeric antigen receptor (CAR) gamma-delta (γδ) T cells offer a major histocompatibility complex-independent, off-the-shelf alternative to conventional CAR alpha-beta (αβ) Tcells, but their efficacy in multiple myeloma (MM) remains underexplored. In this work, we developed a series of novel CAR γδ T cells redirected to B-cell maturation antigen (BCMA) and performed functional assays compared to CAR αβ T cells. As a result, novel CAR γδ T cells demonstrated similar activation upon antigen stimulation compared with CAR αβ T cells but showed lower proliferative capacity and less durable cytotoxicity. In vivo, higher doses of CAR γδ T cells were required to achieve tumour control comparable to CAR αβ T cells. Among CAR constructs, those with cluster of differentiation 3 (CD3ε or CD3ζ) intracellular domain exhibited superior cytotoxicity and in vivo efficacy. Single-cell transcriptomic profiling revealed the low ratio of cycling CAR γδ T cells after antigen stimulation limited the persistence of CAR γδ T cells. In conclusion, our developed novel CAR γδ T cells effectively eliminated MM cells in vitro, but exhibited limited in vivo persistence compared to traditional CAR αβT cells. These findings establish a baseline for utilizing γδ T cells as an off-the-shelf platform and highlight that addressing the proliferation bottleneck through further engineering is essential for future clinical translation.
Insights
Novel chimeric antigen receptor (CAR) gamma-delta T cells targeting BCMA show promise for multiple myeloma but have limited persistence. Further engineering is needed to overcome proliferation challenges for clinical use.
Area of Science:
- Immunology
- Cell Therapy
- Oncology
Background:
- Chimeric antigen receptor (CAR) gamma-delta (γδ) T cells are an MHC-independent, off-the-shelf alternative to CAR alpha-beta (αβ) T cells.
- Their efficacy in multiple myeloma (MM) is not well understood.
Purpose of the Study:
- To develop and evaluate novel CAR γδ T cells targeting B-cell maturation antigen (BCMA) for multiple myeloma.
- To compare their efficacy against conventional CAR αβ T cells.
Main Methods:
- Development of novel BCMA-targeted CAR γδ T cells.
- In vitro functional assays (activation, cytotoxicity) and in vivo studies.
- Single-cell transcriptomic profiling to analyze cell behavior.
Main Results:
- CAR γδ T cells showed antigen-specific activation comparable to CAR αβ T cells.
- CAR γδ T cells exhibited lower proliferation and less durable cytotoxicity in vitro.
- Higher doses of CAR γδ T cells were needed for comparable in vivo tumor control.
- CAR constructs with CD3ε or CD3ζ domains showed better cytotoxicity and in vivo efficacy.
- Single-cell analysis revealed limited cycling CAR γδ T cells, impacting persistence.
Conclusions:
- Novel CAR γδ T cells effectively eliminated MM cells in vitro but had limited in vivo persistence compared to CAR αβ T cells.
- These findings provide a baseline for using γδ T cells as an off-the-shelf therapy.
- Addressing the proliferation bottleneck through further engineering is crucial for clinical translation.
