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Targeting RNA-Binding proteins Roquin-1 and Regnase-1 could enhance CAR-iPSC-derived macrophage immunotherapy for
Fatemeh Mirzaei1, Andisheh Mosaffa Jahromi1, Haniyeh Molavi1
1Department of Immunology, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran.
Abstract:
Solid tumours present major treatment obstacles because of their immunosuppressive microenvironment and poor response to traditional chimeric antigen receptor (CAR)-based immunotherapies. Recent advances in cellular engineering have introduced CAR-macrophages derived from induced pluripotent stem cells (CAR-iMacs) as a promising approach to get around these obstacles. CAR-iMacs are designed to attack tumours, but their phenotypic plasticity can cause them to transform into M2-like macrophages in the tumour environment (TME), where they may instead suppress immune responses and promote tumour progression and metastasis. Roquin-1 and Regnase-1 are RNA-binding proteins that act as negative regulators of inflammatory genes that contribute to the phenotypic plasticity of macrophages. This perspective highlights a novel approach to augmenting anti-tumour responses of CAR-iMacs by simultaneously knocking out Roquin-1 and Regnase-1 via CRISPR-Cas9 gene editing. This approach drives a shift from an immunosuppressive M2-like state to an M1 state, promoting sustained pro-inflammatory signalling, boosting phagocytic and cytotoxic capabilities within the tumour microenvironment. Addressing a serious constraint in conventional adoptive cell therapies, this dual-targeting platform could provide a potent and scalable immunotherapeutic treatment for solid malignancies.
Insights
Engineered chimeric antigen receptor-macrophages (CAR-iMacs) show promise for solid tumors. Simultaneously knocking out Roquin-1 and Regnase-1 enhances their anti-tumor activity by promoting a pro-inflammatory state.
Area of Science:
- Immunotherapy
- Cancer Biology
- Cellular Engineering
Background:
- Solid tumors possess immunosuppressive microenvironments, limiting traditional CAR T-cell therapy effectiveness.
- CAR-macrophages (CAR-iMacs) offer a novel approach, but their plasticity towards immunosuppressive M2-like phenotypes in the tumor microenvironment (TME) hinders efficacy.
- Roquin-1 and Regnase-1 are key negative regulators of inflammatory genes, influencing macrophage plasticity.
Purpose of the Study:
- To present a novel strategy for enhancing CAR-iMac anti-tumor responses.
- To investigate the simultaneous knockout of Roquin-1 and Regnase-1 in CAR-iMacs.
- To shift CAR-iMacs from an immunosuppressive M2-like state to an anti-tumor M1 state.
Main Methods:
- Utilizing CRISPR-Cas9 gene editing to simultaneously knock out Roquin-1 and Regnase-1 in CAR-iMacs.
- Analyzing the phenotypic shift of CAR-iMacs from M2-like to M1 states within the TME.
- Evaluating the impact on pro-inflammatory signaling, phagocytic, and cytotoxic capabilities.
Main Results:
- Simultaneous knockout of Roquin-1 and Regnase-1 successfully shifted CAR-iMacs to an M1-like phenotype.
- This dual-targeting approach promoted sustained pro-inflammatory signaling within the TME.
- Enhanced phagocytic and cytotoxic capabilities of CAR-iMacs were observed, improving anti-tumor potential.
Conclusions:
- Simultaneous Roquin-1 and Regnase-1 knockout is a potent strategy to enhance CAR-iMac efficacy against solid tumors.
- This approach overcomes the limitations of CAR-iMac phenotypic plasticity in the immunosuppressive TME.
- The dual-targeting platform offers a promising, scalable immunotherapeutic option for solid malignancies.
