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Published on: June 14, 2024
ZFP36 disruption is insufficient to enhance the function of mesothelin-targeting human CAR-T cells
David Mai1,2, Tifara Boyce3, Aakash Mehta4,5
1Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, USA. damai@seas.upenn.edu.
Abstract:
Loss of inflammatory effector function, such as cytokine production and proliferation, is a fundamental driver of failure in T cell therapies against solid tumors. Here, we used CRISPR/Cas9 to genetically disrupt ZFP36, an RNA binding protein that regulates the stability of mRNAs involved in T cell inflammatory function, such as the cytokines IL2 and IFNγ, in human T cells engineered with a clinical-stage mesothelin-targeting CAR to determine whether its disruption could enhance antitumor responses. ZFP36 disruption slightly increased antigen-independent activation and cytokine responses but did not enhance overall performance in vitro or in vivo in a xenograft tumor model with NSG mice. While ZFP36 disruption does not reduce the function of CAR-T cells, these results suggest that singular disruption of ZFP36 is not sufficient to improve their function and may benefit from a multiplexed approach.
Insights
Disrupting ZFP36 in CAR T-cells did not improve their ability to fight solid tumors. Further research may require combining ZFP36 disruption with other genetic modifications for enhanced antitumor responses.
Area of Science:
- Immunology
- Cancer Biology
- Genetic Engineering
Background:
- T cell therapies, including CAR T-cells, often fail against solid tumors due to loss of inflammatory effector function.
- ZFP36 is an RNA-binding protein that regulates mRNA stability, impacting T cell functions like cytokine production (IL2, IFNγ).
Purpose of the Study:
- To investigate if genetically disrupting ZFP36 in human T cells engineered with a mesothelin-targeting CAR could enhance antitumor responses.
- To assess the impact of ZFP36 disruption on CAR T-cell function in vitro and in vivo.
Main Methods:
- CRISPR/Cas9 gene editing was used to disrupt ZFP36 in human T cells.
- CAR T-cells targeting mesothelin were engineered and ZFP36 disruption was performed.
- Antitumor responses were evaluated in vitro and in a xenograft tumor model using NSG mice.
Main Results:
- ZFP36 disruption led to a slight increase in antigen-independent T cell activation and cytokine responses.
- No significant enhancement in overall CAR T-cell performance was observed in vitro or in vivo.
- ZFP36 disruption did not impair the fundamental function of CAR T-cells.
Conclusions:
- Singular disruption of ZFP36 is insufficient to improve CAR T-cell efficacy against solid tumors.
- A multiplexed genetic engineering approach, potentially combining ZFP36 disruption with other modifications, may be necessary to enhance CAR T-cell antitumor activity.
